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Sample records for clonorchis sinensis adult

  1. Adult Opisthorchis felineus major protein fractions deduced from transcripts: comparison with liver flukes Opisthorchis viverrini and Clonorchis sinensis.

    PubMed

    Pomaznoy, Mikhail; Tatkov, Sergey; Katokhin, Alexey; Afonnikov, Dmitry; Babenko, Vladimir; Furman, Dagmara; Brusentsov, Ilya; Belavin, Pavel; Najakshin, Alexandr; Guselnikov, Sergey; Vasiliev, Gennady; Sivkov, Anton; Prokhortchouk, Egor; Skryabin, Konstantin; Mordvinov, Viatcheslav

    2013-10-01

    The epidemiologically important liver flukes Opisthorchis felineus, Opisthorchis viverrini, and Clonorchis sinensis are of interest to health professionals, epidemiologists, pharmacologists, and molecular biologists. Recently the transcriptomes of the latter two species were intensively investigated. However our knowledge on molecular biology of O. felineus is scarce. We report the first results of the O. felineus transcriptome analysis. We isolated and annotated a total of 2560 expressed sequence tag (EST) sequences from adult O. felineus (deposited within the database of expressed sequence tags (dbEST), under accession numbers GenBank: JK624271-JK626790, JK006511-JK006547, JK649790-JK649792). Clustering and analysis resulted in the detection of 267 contigs. Of the protein sequences deduced from these, 82% had homologs in the NCBI (nr) protein database and 63% contained conserved domains, allowing the functions to be interpreted using the Gene Ontology terms. Comprehensive analysis of Opisthorchiidae- and Trematoda-specific substitutions within amino acid sequences deduced for the proteins myoglobin, vitelline precursor protein, cathepsin F, and 28kDa glutathione transferase was carried out. The gene set of the 32 ribosomal proteins for the three Opisthorchiidae species with the addition of available Schistosoma and Fasciola orthologs was created and is provided in the supplementary. The orthologous gene set created was used for inferring phylogeny within the Trematoda with special attention to interrelations within the Opisthorchiidae. The phylogenetic analysis revealed a closer relationship between C. sinensis and O. viverrini and some divergence of O. felineus from either O. viverrini or C. sinensis.

  2. Developmental Transcriptomic Features of the Carcinogenic Liver Fluke, Clonorchis sinensis

    PubMed Central

    Cho, Pyo Yun; Kim, Tae Im; Cho, Shin-Hyeong; Choi, Sang-Haeng; Park, Hong-Seog; Kim, Tong-Soo; Hong, Sung-Jong

    2011-01-01

    Clonorchis sinensis is the causative agent of the life-threatening disease endemic to China, Korea, and Vietnam. It is estimated that about 15 million people are infected with this fluke. C. sinensis provokes inflammation, epithelial hyperplasia, and periductal fibrosis in bile ducts, and may cause cholangiocarcinoma in chronically infected individuals. Accumulation of a large amount of biological information about the adult stage of this liver fluke in recent years has advanced our understanding of the pathological interplay between this parasite and its hosts. However, no developmental gene expression profiles of C. sinensis have been published. In this study, we generated gene expression profiles of three developmental stages of C. sinensis by analyzing expressed sequence tags (ESTs). Complementary DNA libraries were constructed from the adult, metacercaria, and egg developmental stages of C. sinensis. A total of 52,745 ESTs were generated and assembled into 12,830 C. sinensis assembled EST sequences, and then these assemblies were further categorized into groups according to biological functions and developmental stages. Most of the genes that were differentially expressed in the different stages were consistent with the biological and physical features of the particular developmental stage; high energy metabolism, motility and reproduction genes were differentially expressed in adults, minimal metabolism and final host adaptation genes were differentially expressed in metacercariae, and embryonic genes were differentially expressed in eggs. The higher expression of glucose transporters, proteases, and antioxidant enzymes in the adults accounts for active uptake of nutrients and defense against host immune attacks. The types of ion channels present in C. sinensis are consistent with its parasitic nature and phylogenetic placement in the tree of life. We anticipate that the transcriptomic information on essential regulators of development, bile chemotaxis, and

  3. [Studies on the immunodiagnosis of rabbit clonorchiasis II. Immunoaffinity purification of whole worm antigen and characterization of egg, metacercaria and adult antigens of Clonorchis sinensis

    PubMed

    Lee, Ok Ran; Chung, Pyung Rim; Nam, Hae Seon

    1988-06-01

    The sensitivity and specificity of crude and affinity-purified antigens of Clonorchis sinensis obtained from the infected rabbits were studied. Stage-specific antigenic proteins from the eggs, metacercariae and adult worms were characterized by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and enzyme-linked immunosorbent assay (ELISA) The results were as follows: The antibody-binding antigen (ABA) purified from whole worm crude antigen (WWA) by CNBr-activated Sepharose 4B affinity chromatography made 4 specific bands against rabbit anti-sera on Ouchterlony gel diffusion plate, while WWA made 7 bands. Major WWA protein bands by SDS-PAGE were found at 16,300-18,500 and 28,000-29,000 daltons, while major ABA protein bands were at 18,000-21,000 and 29,000-31,000 daltons. The reactivity of ABA with rabbit anti-sera in ELISA was remarkably less sensitive than that of WWA. Molecular weights of egg antigen (EGA), metacercarial antigen (MEA) and adult worm antigen (WWA) of C. sinensis ranged from 15,000-200,000 daltons, 15,000-100,000 daltons and 11,000-80,000 daltons, respectively. Major WWA proteins consisted mainly of polypeptide bands of low molecular weight, less than 31,000 daltons, while those of EGA and MEA consisted of higher molecular weights than 30,000 daltons. The ELISA reactivities of WWA to rabbit anti-sera were remarkably greater than those of MEA. EGA showed negative reaction throughout the experiments. WWA showed higher optical density (O.D.) than 1.0, when reacted with rabbit anti-sera obtained at 4-6 weeks after the infection. In the rabbit anti-sera later than 12 weeks after the infection, the O.D. reacting with WWA showed a plateau without variation. MEA showed relatively low O.D. values (<0.6), when reacted with anti-sera from lightly infected groups throughout the experiments, although there were some weak positive cases (O.D.>0.6) in heavily infected groups. MEA reacted with rabbit anti-sera showed negative results on Ouchterlony gel diffusion

  4. Clonorchis sinensis and Opisthorchis spp. in Vietnam: current status and prospects.

    PubMed

    Doanh, Pham N; Nawa, Yukifumi

    2016-01-01

    Clonorchis sinensis and Opisthorchis viverrini are clinically important small liver flukes because of their known association with development of cholangiocarcinoma. In Vietnam, high prevalence of C. sinensis infection in humans was previously reported in northern provinces, and O. viverrini infection has been detected in several central provinces. However, diagnosis of C. sinensis and O. viverrini infections in the past was merely based on faecal egg examination. This method alone can lead to misidentification at the species level because of morphological similarity between the eggs of these liver flukes and minute intestinal trematodes of the family Heterophyidae. In fact, recent surveys in Vietnam revealed that infection with several minute intestinal flukes, such as Haplorchis pumilio and H. taichui, are much more common than infection with C. sinensis or O. viverrini, and they often co-infect humans. Thus, previously reported prevalence of small liver fluke infection in Vietnam was likely over-estimated due to mis identification of parasites in copro-parasitological examinations. In addition, there is some confusion about identification of cercariae, metacercariae and also adults of C. sinensis and O. viverrini in intermediate and definitive hosts. The aim of this review is, therefore, to draw realistic pictures of the past and present scientific reports on the epidemiology and biology of C. sinensis and Opisthorchis spp. infection in Vietnam.

  5. Molecular Cloning and Characterization of Taurocyamine Kinase from Clonorchis sinensis: A Candidate Chemotherapeutic Target

    PubMed Central

    Tokuhiro, Shinji; Nagataki, Mitsuru; Jarilla, Blanca R.; Nomura, Haruka; Kim, Tae Im; Hong, Sung-Jong; Agatsuma, Takeshi

    2013-01-01

    Background Adult Clonorchis sinensis lives in the bile duct and causes endemic clonorchiasis in East Asian countries. Phosphagen kinases (PK) constitute a highly conserved family of enzymes, which play a role in ATP buffering in cells, and are potential targets for chemotherapeutic agents, since variants of PK are found only in invertebrate animals, including helminthic parasites. This work is conducted to characterize a PK from C. sinensis and to address further investigation for future drug development. Methology/Principal findings A cDNA clone encoding a putative polypeptide of 717 amino acids was retrieved from a C. sinensis transcriptome. This polypeptide was homologous to taurocyamine kinase (TK) of the invertebrate animals and consisted of two contiguous domains. C. sinensis TK (CsTK) gene was reported and found consist of 13 exons intercalated with 12 introns. This suggested an evolutionary pathway originating from an arginine kinase gene group, and distinguished annelid TK from the general CK phylogenetic group. CsTK was found not to have a homologous counterpart in sequences analysis of its mammalian hosts from public databases. Individual domains of CsTK, as well as the whole two-domain enzyme, showed enzymatic activity and specificity toward taurocyamine substrate. Of the CsTK residues, R58, I60 and Y84 of domain 1, and H60, I63 and Y87 of domain 2 were found to participate in binding taurocyamine. CsTK expression was distributed in locomotive and reproductive organs of adult C. sinensis. Developmentally, CsTK was stably expressed in both the adult and metacercariae stages. Recombinant CsTK protein was found to have low sensitivity and specificity toward C. sinensis and platyhelminth-infected human sera on ELISA. Conclusion CsTK is a promising anti-C. sinensis drug target since the enzyme is found only in the C. sinensis and has a substrate specificity for taurocyamine, which is different from its mammalian counterpart, creatine. PMID:24278491

  6. Worm recovery and precipitin antibody response in guinea pigs and rats infected with Clonorchis sinensis.

    PubMed

    Su, K E; Wang, F Y; Chi, P Y

    1998-12-01

    Guinea pigs (Hartley strain) and rats (Wistar strain) were each fed 200 and 100 Clonorchis sinensis metacercariae, respectively. Five animals from each species were sacrificed weekly between 1-8 weeks postinfection (WPI) and then at 12, 16, 20 and 30 WPI for collection of worms, bile and sera. The overall worm recovery rates for guinea pigs and rats were 18.7% and 12.4%, respectively. Only one of the five rats examined at 20 WPI still harbored one worm, while all were worm-free at 30 WPI. By a double diffusion test, no antibodies were detected against C. sinensis adult antigens in the bile juice. Serum antibodies were detected in at least 95% of the infected guinea pigs between 4-30 WPI and rats between 3-16 WPI. Precipitin antibodies seemed to be correlated with the presence of live worms in rats that had been infected for more than 12 weeks.

  7. Painless Jaundice Caused by Clonorchis sinensis Infection: A Case Report.

    PubMed

    Hao, Yuhua; Bao, Wanguo; Jin, Meishan; Li, Yuxiang; Wang, Feng

    2016-06-01

    A man with only yellowing of the skin and eye sclera was diagnosed with clonorchiasis, which rarely manifested jaundice as the initial symptom. However, because of a lack of evidence for a diagnostic gold standard, the time until definitive diagnosis was more than a week. The diagnostic process relied on inquiring about the patient's history, including the place of residence, dietary habits, and symptoms, as well as on serological findings, an imaging examination, and pathological findings. MRCP and CT results showed mild dilatation of intrahepatic ducts and increased periductal echogenicity. The eggs were ultimately found in stool by water sedimentation method after the negative report through direct smear. DNA sequencing of PCR production of the eggs demonstrated 98-100% homology with ITS2 of Clonorchis sinensis. After anti-parasite medical treatment, the patient's symptoms were gradually relieved. Throughout the diagnostic procedure, besides routine examinations, the sedimentation method or concentration method could be used as a sensitive way for both light and heavy C. sinensis infection in the definite diagnosis.

  8. Proteomic analysis of different period excretory secretory products from Clonorchis sinensis adult worms: molecular characterization, immunolocalization, and serological reactivity of two excretory secretory antigens-methionine aminopeptidase 2 and acid phosphatase.

    PubMed

    Zheng, Minghui; Hu, Kunhua; Liu, Wei; Li, Hongyu; Chen, Jingfang; Yu, Xinbing

    2013-03-01

    The excretory secretory products (ESP) of Clonorchis sinensis are the causative agents of clonorchiasis and biliary diseases. The parasites' ESP play important roles in host-parasite interactions. The protein compositions of ESP at different secretory times are different and have not been systemically investigated so far. In this study, we collected ESP from six different periods (0-3 h, 3-6 h, 6-12 h, 12-24 h, 24-36 h, and 36-48 h) from C. sinensis adults. Using a shotgun LC-MS/MS analysis, we found 187, 80, 103, 58, 248, and 383 proteins, respectively. Among these proteins, we selected methionine aminopeptidase 2 (MAP-2, presented in 24-36 h and 36-48 h ESP) and acid phosphatase (AP, presented in 3-6 h, 12-24 h, 24-36 h, and 36-48 h ESP) for further study. Bioinformatics analysis showed that CsMAP-2 has metallopeptidase family M24, unique lysine residue-rich and acidic residue-rich domain, SGTS motif, and auto-cleavage point; and that CsAP has possible signal sequence cleavage site, acid phosphate domain, and two histidine acid phosphatases active regions. CsMAP-2 and CsAP's cDNA have 1,425 bp and1,410 bp ORF, encoding 475 and 470 amino acid proteins and weighing 55.3840 kDa and 55.2875 kDa, respectively. MAP-2 and AP were identified as antigens present in the ESP and circulating antigens by immunoblot analysis, which were also found expressing in the eggs, metacercaria, and adult stages of C. sinensis. Immunofluorescence analysis showed that they were located in tegument and intestinal cecum of adult. MTT assay showed that they could inhibit hepatic stellate cell line (LX-2) proliferation. These findings presented the compositions of different period excretory secretary products from C. sinensis adults.

  9. Identification and characterization of microRNAs in Clonorchis sinensis of human health significance

    PubMed Central

    2010-01-01

    Background Clonorchis sinensis is a zoonotic parasite causing clonorchiasis-associated human disease such as biliary calculi, cholecystitis, liver cirrhosis, and it is currently classified as carcinogenic to humans for cholangiocarcinoma. MicroRNAs (miRNAs) are non-coding, regulating small RNA molecules which are essential for the complex life cycles of parasites and are involved in parasitic infections. To identify and characterize miRNAs expressed in adult C. sinensis residing chronically in the biliary tract, we developed an integrative approach combining deep sequencing and bioinformatic predictions with stem-loop real-time PCR analysis. Results Here we report the use of this approach to identify and clone 6 new and 62,512 conserved C. sinensis miRNAs which belonged to 284 families. There was strong bias on families, family members and sequence nucleotides in C. sinensis. Uracil was the dominant nucleotide, particularly at positions 1, 14 and 22, which were located approximately at the beginning, middle and end of conserved miRNAs. There was no significant "seed region" at the first and ninth positions which were commonly found in human, animals and plants. Categorization of conserved miRNAs indicated that miRNAs of C. sinensis were still innovated and concentrated along three branches of the phylogenetic tree leading to bilaterians, insects and coelomates. There were two miRNA strategies in C. sinensis for its parasitic life: keeping a large category of miRNA families of different animals and keeping stringent conserved seed regions with high active innovation in other places of miRNAs mainly in the middle and the end, which were perfect for the parasite to perform its complex life style and for host changes. Conclusions The present study represented the first large scale characterization of C. sinensis miRNAs, which have implications for understanding the complex biology of this zoonotic parasite, as well as miRNA studies of other related species such as

  10. Current status and perspectives of Clonorchis sinensis and clonorchiasis: epidemiology, pathogenesis, omics, prevention and control.

    PubMed

    Tang, Ze-Li; Huang, Yan; Yu, Xin-Bing

    2016-07-06

    Clonorchiasis, caused by Clonorchis sinensis (C. sinensis), is an important food-borne parasitic disease and one of the most common zoonoses. Currently, it is estimated that more than 200 million people are at risk of C. sinensis infection, and over 15 million are infected worldwide. C. sinensis infection is closely related to cholangiocarcinoma (CCA), fibrosis and other human hepatobiliary diseases; thus, clonorchiasis is a serious public health problem in endemic areas. This article reviews the current knowledge regarding the epidemiology, disease burden and treatment of clonorchiasis as well as summarizes the techniques for detecting C. sinensis infection in humans and intermediate hosts and vaccine development against clonorchiasis. Newer data regarding the pathogenesis of clonorchiasis and the genome, transcriptome and secretome of C. sinensis are collected, thus providing perspectives for future studies. These advances in research will aid the development of innovative strategies for the prevention and control of clonorchiasis.

  11. Clonorchis sinensis Infestation Promotes Three-Dimensional Aggregation and Invasion of Cholangiocarcinoma Cells

    PubMed Central

    Won, Jihee; Ju, Jung-Won; Kim, Sun Min; Shin, Yoojin; Chung, Seok; Pak, Jhang Ho

    2014-01-01

    Numerous experimental and epidemiological studies have demonstrated a correlation between Clonorchis sinensis (C. sinensis) infestation and cholangiocarcinoma (CCA). However, the role of C. sinensis in the increased invasiveness and proliferation involved in the malignancy of CCA has not been addressed yet. Here, we investigated the possibility that C. sinensis infestation promotes expression of focal and cell-cell adhesion proteins in CCA cells and secretion of matrix metalloproteinases (MMPs). Adhesion proteins help maintain cell aggregates, and MMPs promote the three-dimensional invasion of cells into the neighboring extracellular matrix (ECM). Using a novel microfluidic assay, we quantitatively addressed the role of excretory-secretory products (ESPs) gradients from C. sinensis in promoting the invasion of cells into the neighboring ECM. PMID:25340585

  12. Molecular identification and characterization of leucine aminopeptidase 2, an excretory-secretory product of Clonorchis sinensis.

    PubMed

    Deng, Chuanhuan; Sun, Jiufeng; Li, Xuerong; Wang, Lexun; Hu, Xuchu; Wang, Xiaoyun; Chen, Wenjun; Lv, Xiaoli; Liang, Chi; Li, Wenfang; Huang, Yan; Li, Ran; Wu, Zhongdao; Yu, Xinbing; Xu, Jin

    2012-10-01

    Aminopeptidases serve vital roles in metabolism of hormones, neurotransmission, turnover of proteins and immunological regulations. Leucine aminopeptidases catalyze the hydrolysis of amino-acid residues from the N-terminus of proteins and peptides. In the present study, leucine aminopeptidase 2 (LAP2) gene of Clonorchis sinensis (C. sinensis) was isolated and identified from an adult cDNA library of C. sinensis. Recombinant CsLAP2 was expressed and purified in Escherichia coli BL21. The open reading frame of LAP2 contains 1,560 bp equivalent to 519 amino acids, a similarity analysis showed a relatively low homology with Homo sapiens (19.0 %), Trypanosoma cruzi (18.0 %), Mus musculus (19.3 %), and relatively high homology with Schistosoma mansoni (65.6 %). The optimum condition of rCsLAP2 enzyme activity was investigated using a fluorescent substrate of Leu-MCA at 37 °C and pH 7.5. The K (m) and V (max) values of rCsLAP2 were 18.2 μM and 10.7 μM/min, respectively. CsLAP2 gene expression can be detected at the stages of the adult worm, metacercaria, excysted metacercaria and egg of C. sinensis using real-time PCR, no difference was observed at the stages of the adult worm, metacercaria and egg. However, CsLAP2 showed a higher expression level at the stage of excysted metacercaria than the adult worm (3.90-fold), metacercaria (4.60-fold) and egg (4.59-fold). Histochemistry analysis showed that CsLAP2 was located at the tegument and excretory vesicle of metacercaria, and the tegument and intestine of adult worm. The immune response specific to rCsLAP2 was characterized by a mixed response patterns of Th1 and Th2, indicating a compounded humoral and cellular immune response. The combined results from the present study indicate that CsLAP2 was an important antigen exposed to host immune system, and probably implicated as potential role in interaction with host cells in clonorchiasis.

  13. Interleukin-13 is involved in the formation of liver fibrosis in Clonorchis sinensis-infected mice.

    PubMed

    Xu, Yanquan; Liang, Pei; Bian, Meng; Chen, Wenjun; Wang, Xiaoyun; Lin, Jinsi; Shang, Mei; Qu, Hongling; Wu, Zhongdao; Huang, Yan; Yu, Xinbing

    2016-07-01

    Clonorchiasis is a chronic infection disease often accompanied by formation of liver fibrosis. Previous study has identified that Clonorchis sinensis (C. sinensis, Cs) infection and CsRNASET2 (a member of CsESPs) immunization can drive Th2 immune response. IL-13, a multifunctional Th2 cytokine, has been widely confirmed to be profibrotic mediator. We want to determine whether IL-13 is involved in the generation of liver fibrosis during C. sinensis infection. A part of mice were infected with C. sinensis or immunized with CsRNASET2, respectively. Another part of mice were intravenously injected with rIL-13. Liver tissues of C. sinensis-infected mice were stained with hematoxylin-eosin and Masson's trichrome, respectively. The transcriptional levels of collagen-I, collagen-III, α-SMA, and TIMP-1 in the livers of infected mice and rIL-13-treated mice were measured by quantitative RT-PCR. Besides, splenocytes of C. sinensis-infected and CsRNASET2-immunized mice were isolated, respectively. The levels of IL-13 in splenocytes were detected by ELISA. Our results displayed that the livers of C. sinensis-infected mice had serious chronic inflammation and collagen deposition. The transcriptional levels of collagen-I, collagen-III, α-SMA, and TIMP-1 in the livers of C. sinensis-infected mice were obviously increased. Splenocytes from both C. sinensis-infected and CsRNASET2-immunized mice expressed high levels of IL-13. Moreover, rIL-13 treatment markedly promoted the transcriptional levels of collagen-I, collagen-III, α-SMA, and TIMP-1. These data implied that hepatic fibrosis was formed in the livers of C. sinensis-infected mice, and IL-13 induced by C. sinensis infection and CsRNASET2 immunization might favor this progression.

  14. Clonorchis sinensis Co-infection Could Affect the Disease State and Treatment Response of HBV Patients

    PubMed Central

    Huang, Yan; Chen, Tingjin; Kong, Xiangzhan; Sun, Hengchang; Yu, Xinbing; Xu, Jin

    2016-01-01

    Background Clonorchis sinensis (C. sinensis) is considered to be an important parasitic zoonosis because it infects approximately 35 million people, while approximately 15 million were distributed in China. Hepatitis B virus (HBV) infection is a major public health issue. Two types of pathogens have the potential to cause human liver disease and eventually hepatocellular carcinoma. Concurrent infection with HBV and C. sinensis is often observed in some areas where C. sinensis is endemic. However, whether C. sinensis could impact HBV infection or vice versa remains unknown. Principal Findings Co-infection with C. sinensis and HBV develops predominantly in males. Co-infected C. sinensis and HBV patients presented weaker liver function and higher HBV DNA titers. Combination treatment with antiviral and anti-C. sinensis drugs in co-infected patients could contribute to a reduction in viral load and help with liver function recovery. Excretory-secretory products (ESPs) may, in some ways, increase HBV viral replication in vitro. A mixture of ESP and HBV positive sera could induce peripheral blood mononuclear cells (PBMCs) to produce higher level of Th2 cytokines including IL-4, IL-6 and IL-10 compared to HBV alone, it seems that due to presence of ESP, the cytokine production shift towards Th2. C. sinensis/HBV co-infected patients showed higher serum IL-6 and IL-10 levels and lower serum IFN-γ levels. Conclusions/Significance Patients with concomitant C. sinensis and HBV infection presented weaker liver function and higher HBV DNA copies. In co-infected patients, the efficacy of anti-viral treatment was better in patients who were prescribed with entecavir and praziquantel than entecavir alone. One possible reason for the weaker response to antiviral therapies in co-infected patients was the shift in cytokine production from Th1 to Th2 that may inhibit viral clearance. C. sinensis/HBV co-infection could exacerbate the imbalance of Th1/Th2 cytokine. PMID:27348302

  15. Clonorchis sinensis, an oriental liver fluke, as a human biological agent of cholangiocarcinoma: a brief review

    PubMed Central

    Kim, Tong-Soo; Pak, Jhang Ho; Kim, Jong-Bo; Bahk, Young Yil

    2016-01-01

    Parasitic diseases remain an unarguable public health problem worldwide. Liver fluke Clonorchis sinensis is a high risk pathogenic parasitic helminth which is endemic predominantly in Asian countries, including Korea, China, Taiwan, Vietnam, and the far eastern parts of Russia, and is still actively transmitted. According to the earlier 8th National Survey on the Prevalence of Intestinal Parasitic Infections in 2012, C. sinensis was revealed as the parasite with highest prevalence of 1.86% in general population among all parasite species surveyed in Korea. This fluke is now classified under one of the definite Group 1 human biological agents (carcinogens) by International Agency of Research on Cancer (IARC) along with two other parasites, Opisthorchis viverrini and Schistosoma haematobium. C. sinensis infestation is mainly linked to liver and biliary disorders, especially cholangiocarcinoma (CCA). For the purposes of this mini-review, we will only focus on C. sinensis and review pathogenesis and carcinogenesis of clonorchiasis, disease condition by C. sinensis infestation, and association between C. sinensis infestation and CCA. In this presentation, we briefly consider the current scientific status for progression of CCA by heavy C. sinensis infestation from the food-borne trematode and development of CCA. PMID:27418285

  16. Microscopic examination of gallbladder stones improves rate of detection of Clonorchis sinensis infection.

    PubMed

    Qiao, Tie; Ma, Rui-hong; Luo, Xiao-bing; Zheng, Pei-ming; Luo, Zhen-liang; Yang, Liu-qing

    2013-08-01

    To improve the rate of detection of Clonorchis sinensis infection, we compared different specimens from patients with cholecystolithiasis. Feces, gallbladder bile, and gallbladder stones collected from 179 consecutive patients with cholecystolithiasis underwent microscopic examination, and according to the results, 30 egg-positive and 30 egg-negative fecal, gallbladder bile, and gallbladder stone specimens, respectively, underwent real-time fluorescent PCR. The detection rates of eggs in feces, bile, and gallbladder stones were 30.7%, 44.7%, and 69.8%, respectively, and the differences were statistically significant (P<0.01). The PCR results confirmed that the eggs in the specimens were C. sinensis eggs. Eggs in the feces were "fresh" and in the gallbladder stones were "old." Microscopic examination of gallbladder stones may improve the detection rates of C. sinensis infection, which is important for developing individualized treatments to prevent the recurrence of gallbladder stones and to prevent the occurrence of severe liver damage and cholangiocarcinoma.

  17. The zoonotic, fish-borne liver flukes Clonorchis sinensis, Opisthorchis felineus and Opisthorchis viverrini.

    PubMed

    Petney, Trevor N; Andrews, Ross H; Saijuntha, Weerachai; Wenz-Mücke, Alexandra; Sithithaworn, Paiboon

    2013-11-01

    Clonorchis sinensis, Opisthorchis felineus and Opisthorchis viverrini are the three most important liver flukes involved in human health, infecting more than 45 million people worldwide. Both C. sinensis and O. viverrini, and possibly O. felineus, can induce human cholangiocarcinoma as well as inducing other hepatobiliary pathology. Although the life cycles of all three species are similar, only that of O. felineus in Europe remains predominantly zoonotic, while O. felineus in Asia and C. sinensis have a stronger mixture of zoonotic and anthroponotic components in their life cycles. Opisthorchis viverrini from the Mekong area of southeastern Asia is predominantly anthroponotic. Here we discuss the comparative epidemiology of these three taxa comparing in detail the use of first, second and final animal hosts, and consider the potential role of humans in spreading these pathogens. In addition we discuss the genetic structure of all three species in relation to potentially cryptic species complexes.

  18. Mitochondrial genomic comparison of Clonorchis sinensis from South Korea with other isolates of this species.

    PubMed

    Wang, Daxi; Young, Neil D; Koehler, Anson V; Tan, Patrick; Sohn, Woon-Mok; Korhonen, Pasi K; Gasser, Robin B

    2017-02-22

    Clonorchiasis is a neglected tropical disease that affects >35 million people mainly in China, Vietnam, South Korea and some parts of Russia. The disease-causing agent, Clonorchis sinensis, is a liver fluke of humans and other piscivorous animals, and has a complex aquatic life cycle involving snails and fish intermediate hosts. Chronic infection in humans causes liver disease and associated complications including malignant bile duct cancer. Central to control and to understanding the epidemiology of this disease is knowledge of the specific identity of the causative agent as well as genetic variation within and among populations of this parasite. Although most published molecular studies seem to suggest that C. sinensis represents a single species and that genetic variation within the species is limited, karyotypic variation within C. sinensis among China, Korea (2n=56) and Russian Far East (2n=14) suggests that this taxon might contain sibling species. Here, we assessed and applied a deep sequencing-bioinformatic approach to sequence and define a reference mitochondrial (mt) genome for a particular isolate of C. sinensis from Korea (Cs-k2), to confirm its specific identity, and compared this mt genome with homologous data sets available for this species. Comparative analyses revealed consistency in the number and structure of genes as well as in the lengths of protein-coding genes, and limited genetic variation among isolates of C. sinensis. Phylogenetic analyses of amino acid sequences predicted from mt genes showed that representatives of C. sinensis clustered together, with absolute nodal support, to the exclusion of other liver fluke representatives, but sub-structuring within C. sinensis was not well supported. The plan now is to proceed with the sequencing, assembly and annotation of a high quality draft nuclear genome of this defined isolate (Cs-k2) as a basis for a detailed investigation of molecular variation within C. sinensis from disparate

  19. [Detection of Clonorchis sinensis eggs in the ground gallbladder stones by microscopy].

    PubMed

    Ma, Rui-Hong; Qiao, Tie; Luo, Xiao-Bing

    2012-08-30

    Sera, feces, bile and gallbladder stones were collected from 179 patients who accepted gallbladder-preserving cholelithotomy during the period of January to June 2010 at the general surgery department in the Second People's Hospital of Panyu District in Guangzhou. Rapid colloidal gold immunochromatography was used to detect IgG against Clonorchis sinensis. C. sinensis eggs were examined by fecal direct smear, and in bile sediments and ground gallbladder stones. The results showed that the positive rate of rapid colloidal gold immunochromatographic assay for IgG was 51.4%, and the egg positive rate in feces, bile sediments and gallbladder stones was 30.7%, 44.7% and 69.8%, respectively. The detection rate of fecal direct smear was the lowest, while that of the gallbladder stone examination was the highest (P < 0.05). Those patients with egg-positive feces and/or bile sediments were all with egg-positive gallbladder stones.

  20. Investigation on oxidative stress of nitric oxide synthase interacting protein from Clonorchis sinensis.

    PubMed

    Bian, Meng; Xu, Qingxia; Xu, Yanquan; Li, Shan; Wang, Xiaoyun; Sheng, Jiahe; Wu, Zhongdao; Huang, Yan; Yu, Xinbing

    2016-01-01

    Numerous evidences indicate that excretory-secretory products (ESPs) from liver flukes trigger the generation of free radicals that are associated with the initial pathophysiological responses in host cells. In this study, we first constructed a Clonorchis sinensis (C. sinensis, Cs)-infected BALB/c mouse model and examined relative results respectively at 3, 5, 7, and 9 weeks postinfection (p.i.). Quantitative reverse transcription (RT)-PCR indicated that the transcriptional level of both endothelial nitric oxide synthase (eNOS) and superoxide dismutase (SOD) gradually decreased with lastingness of infection, while the transcriptional level of inducible NOS (iNOS) significantly increased. The level of malondialdehyde (MDA) in sera of infected mouse significantly increased versus the healthy control group. These results showed that the liver of C. sinensis-infected mouse was in a state with elevated levels of oxidation stress. Previously, C. sinensis NOS interacting protein coding gene (named CsNOSIP) has been isolated and recombinant CsNOSIP (rCsNOSIP) has been expressed in Escherichia coli, which has been confirmed to be a component present in CsESPs and confirmed to play important roles in immune regulation of the host. In the present paper, we investigated the effects of rCsNOSIP on the lipopolysaccharide (LPS)-induced activated RAW264.7, a murine macrophage cell line. We found that endotoxin-free rCsNOSIP significantly promoted the levels of nitric oxide (NO) and reactive oxygen species (ROS) after pretreated with rCsNOSIP, while the level of SOD decreased. Furthermore, rCsNOSIP could also increase the level of lipid peroxidation MDA. Taken together, these results suggested that CsNOSIP was a key molecule which was involved in the production of nitric oxide (NO) and its reactive intermediates, and played an important role in oxidative stress during C. sinensis infection.

  1. Comparison of ancient and modern Clonorchis sinensis based on ITS1 and ITS2 sequences.

    PubMed

    Liu, Wen-Qi; Liu, Juan; Zhang, Jun-Hua; Long, Xiao-Chun; Lei, Jia-Hui; Li, Yong-Long

    2007-02-01

    In 1975, an ancient corpse buried in 167 BC was found at Jiangling County, Hubei Province of China. The eggs of Clonorchis sinensis found in the gall bladder of the corpse were preserved well. In the present paper, we extracted the genomic DNA from the ancient eggs and modern eggs, respectively, and the internal transcribed spacer 1 and 2 (ITS1 and ITS2) at ribosomal RNA genes were studied. The results show that ITS2 sequences from the ancient sample were identical with those from modern samples, but in ITS1 differences in 15 nucleotide positions were found between the ancient and modern samples. The results demonstrated that it is possible to extract and sequence DNA from ancient parasite eggs. The ITS1 sequence obtained differed from all modern ones available to date. This might indicate sequence divergence through time, or might reflect a sequence polymorphism that may eventually be found also in modern samples.

  2. Epidemiological study on Clonorchis sinensis infection in Shenzhen area of Zhujiang delta in China.

    PubMed

    Zhang, Renli; Gao, Shitong; Geng, Yijie; Huang, Dana; Yu, Lei; Zhang, Shunxiang; Cheng, Jingquan; Fu, Yucai

    2007-06-01

    To study the transmission route and epidemiological features of Clonorchis sinensis infection in Shenzhen area, which is the biggest immigration city in the south of China, we examined 1,473 individuals (710 males and 763 females) to assess the current status of C. sinensis infection among the people in a village of Shenzhen in Zhujiang delta of Guangdong province, China. Freshwater snails, 630, of different species known as the first intermediate host of C. sinensis were collected and examined for cercaria infection, and 430 freshwater fishes of different species as the second intermediate host were examined for metacercaria infection. Among 1,473 people examined, 70 (4.75%) were found infected with C. sinensis. By counting eggs per gram feces (EPG), it was found that the intensity of infection in males was stronger than that of females, and the average EPG was 41.87 in all population. Snails, 1.15%, were infected with cercariae of C. sinensis. The average infection rate of freshwater fishes of 15 species with metacercariae of C. sinensis was 16.97%, and the carps reached the highest infection rate (40.74%). A questionnaire was designed with 12 questions covering socioeconomic conditions and human behavior, contamination of the environment, and fishponds. Of 1,473 interviewees, 54% did not know about fluke disease or its transmission route, 12% of those who knew about the fluke believed that the infection causes no harm or only slight harm to their health. Of the interviewees, 27%, ate raw fish at least one to two times per month. Of families, 5% used the same utensils for both raw fish and cooked food. Of the fishpond owners, 40% fed their fishes with feces of domestic animals and humans. All these factors of unhealthy behaviors, poor knowledge, inappropriate farming/fishery practices, and eating raw fish have made the prevalence of clonorchiasis increase in humans in the Shenzhen area. It is urgent to perform a control program, including health education

  3. Development of a real-time PCR assay for the detection of Clonorchis sinensis DNA in gallbladder bile and stone samples from patients with cholecystolithiasis.

    PubMed

    Qiao, Tie; Zheng, Pei-Ming; Ma, Rui-Hong; Luo, Xiao-Bing; Luo, Zhen-Liang

    2012-10-01

    High prevalence of cholecystolithiasis in parts of East Asia has been postulated to be associated with Clonorchis sinensis infection. This study describes the development of a TaqMan-based real-time PCR assay for the detection of C. sinensis DNA in gallbladder bile and stone samples from patients with cholecystolithiasis. Primers and probe targeting the cytochrome c oxidase subunit 1 gene of mitochondrial DNA proved to be highly specific for C. sinensis and did not amplify other related heterogeneous DNA samples. The detection limit of this assay was 0.1 pg of adult C. sinensis genomic DNA. All of the egg-positive samples determined by microscopy yielded positive results by real-time PCR assay and that genetic testing of gallbladder stones using real-time PCR was considered as the most effective means for assessing C. sinensis infection status. This assay not only contributes to a greater understanding of stone pathogenesis but also benefits patients with cholecystolithiasis by facilitating effective diagnosis, treatment, and relapse prevention.

  4. Comparative biochemical and functional properties of two leucine aminopeptidases of Clonorchis sinensis.

    PubMed

    Kang, Jung-Mi; Ju, Hye-Lim; Ju, Jung-Won; Sohn, Woon-Mok; Kim, Tong-Soo; Bahk, Young-Yil; Hong, Sung-Jong; Na, Byoung-Kuk

    2012-01-01

    Leucine aminopeptidases (LAP; EC 3.4.11.1) are a group of metalloexopeptidases, which catalyze the sequential removal of leucine amino acids from the N-termini of the polypeptides or proteins. In this study, we identified two novel genes that encode LAPs of Clonorchis sinensis (CsLAP1 and CsLAP2) and characterized their biochemical and functional properties. Multiple sequence alignment of the deduced amino acid sequences of CsLAP1 and CsLAP2 with those of other organisms revealed that typical metal-binding coordinating and active site residues for LAPs were well conserved in CsLAP1 and CsLAP2. Recombinant CsLAP1 and CsLAP2 showed similar biochemical properties such as pH optima at pH 8.0 and stability at neutral pHs. Both enzymes were specifically inhibited by bestatin and showed preferential substrate specificity for Leu-MCA. However, the enzymes differed in that they required different metal ions for maximum activity. Expressions of CsLAP1 and CsLAP2 were detected throughout the various developmental stages of C. sinensis, and their transcription levels increased gradually in accordance with the maturation of the parasite. Both enzymes were identified in soluble worm extract of C. sinensis, but not in excretory and secretory products. Immunolocalization studies showed that both enzymes were co-localized to the intestinal epithelial cells and gastrodermis of the parasite. These results collectively suggest that CsLAP1 and CsLAP2 are synthesized in the intestinal epithelial and gastrodermal cells of C. sinensis and may be involved in the final digestion of peptides that hydrolyzed within intestinal lumen followed by absorbed into gastrodermal cells of the parasite.

  5. Genetic diversity of the Chinese liver fluke Clonorchis sinensis from Russia and Vietnam.

    PubMed

    Chelomina, Galina N; Tatonova, Yulia V; Hung, Nguyen Manh; Ngo, Ha Duy

    2014-10-01

    Clonorchiasis is a parasitic disease of high public health importance in many countries in southeastern Asia and is caused by the Chinese liver fluke Clonorchis sinensis. However, the genetic structure and demographic history of its populations has not been sufficiently studied throughout the geographic range of the species and available data are based mainly on partial gene sequencing. In this study, we explored the genetic diversity of the complete 1560 bp cytochrome c oxidase subunit 1 (cox1) gene sequence for geographically isolated C. sinensis populations in Russia and Vietnam, to our knowledge for the first time. The results demonstrated low nucleotide and high haplotype differentiation within and between the two compared regions and a clear geographical vector for the distribution of genetic diversity patterns among the studied populations. These results suggest a deep local adaptation of the parasite to its environment including intermediate hosts and the existence of gene flow across the species' range. Additionally, we have predicted an amino acid substitution in the functional site of the COX1 protein among the Vietnamese populations, which were reported to be difficult to treat with praziquantel. The haplotype networks consisted of several region-specific phylogenetic lineages, the formation of which could have occurred during the most extensive penultimate glaciations in the Pleistocene Epoch. The patterns of genetic diversity and demographics are consistent with population growth of the liver fluke in the late Pleistocene following the Last Glacial Maximum, indicating the lack of a population bottleneck during the recent past in the species' history. The data obtained have important implications for understanding the phylogeography of C. sinensis, its host-parasite interactions, the ability of this parasite to evolve drug resistance, and the epidemiology of clonorchiasis under global climate change.

  6. Epidemiological investigation of Clonorchis sinensis infection in freshwater fishes in the Pearl River Delta.

    PubMed

    Chen, Daixiong; Chen, Jieyun; Huang, Ji; Chen, Xueying; Feng, Dana; Liang, Baofang; Che, Yuchuan; Liu, Xiaodan; Zhu, Cuihua; Li, Xiaomin; Shen, Haoxian

    2010-09-01

    Pearl River Delta region is a high clonorchiasis-endemic area in China. However, no complete epidemiological data exist regarding its infection in freshwater fishes, an important epidemic factor for Clonorchis sinensis. The present study collected freshwater fishes and shrimps from 32 sites of nine cities in the Pearl River Delta, and the encysted metacercariae of C. sinensis were detected by digesting these specimens with artificial gastric juice. The mean infection rate of freshwater fishes was 37.09% (2,160/5,824) with a mean number of 14.269 encysted metacercariae in every infected fish and 0.460 encysted metacercariae in every gram of fish meat. Of these freshwater fishes, 5,219 were domesticated, and the infection rate was 36.69% with a mean number of 10.743 encysted metacercariae in every infected fish and 0.312 encysted metacercariae in every gram of fish meat; the other 605 were wild, and the infection rate was 40.50% with a mean number of 41.829 encysted metacercariae in every infected fish and 8.812 encysted metacercariae in every gram of fish meat. A total of 228 shrimps were examined, and 3.07% of them were infected with a mean number of 1.00 encysted metacercariae in every infected shrimp. Pseudorasbora parva and Ctenopharyngodon idellus had the highest infection rate and degree of infection in the fishes studied. The results demonstrated a high incidence of C. sinensis infection in freshwater fishes and shrimps within Pearl River Delta region and a great difference in the infection rate among different collection sites and different fish species.

  7. Molecular differentiation of Opisthorchis viverrini and Clonorchis sinensis eggs by multiplex real-time PCR with high resolution melting analysis.

    PubMed

    Kaewkong, Worasak; Intapan, Pewpan M; Sanpool, Oranuch; Janwan, Penchom; Thanchomnang, Tongjit; Laummaunwai, Porntip; Lulitanond, Viraphong; Doanh, Pham Ngoc; Maleewong, Wanchai

    2013-12-01

    Opisthorchis viverrini and Clonorchis sinensis are parasites known to be carcinogenic and causative agents of cholangiocarcinoma in Asia. The standard method for diagnosis for those parasite infections is stool examination to detect parasite eggs. However, the method has low sensitivity, and eggs of O. viverrini and C. sinensis are difficult to distinguish from each other and from those of some other trematodes. Here, we report a multiplex real-time PCR coupled with high resolution melting (HRM) analysis for the differentiation of O. viverrini and C. sinensis eggs in fecal samples. Using 2 pairs of species-specific primers, DNA sequences from a portion of the mitochondrial NADH dehydrogenase subunit 2 (nad 2) gene, were amplified to generate 209 and 165 bp products for O. viverrini and C. sinensis, respectively. The distinct characteristics of HRM patterns were analyzed, and the melting temperatures peaked at 82.4±0.09℃ and 85.9±0.08℃ for O. viverrini and C. sinensis, respectively. This technique was able to detect as few as 1 egg of O. viverrini and 2 eggs of C. sinensis in a 150 mg fecal sample, which is equivalent to 7 and 14 eggs per gram of feces, respectively. The method is species-specific, rapid, simple, and does not require fluorescent probes or post-PCR processing for discrimination of eggs of the 2 species. It offers a new tool for differentiation and detection of Asian liver fluke infections in stool specimens.

  8. Effectiveness of mefloquine against Clonorchis sinensis in rats and Paragonimus westermani in dogs.

    PubMed

    Xiao, Shu-hua; Xue, Jian; Li-li, Xu; Zhang, Yong-nian; Qiang, Hui-qing

    2010-11-01

    The aim of the study is to explore the effect of mefloquine against Clonorchis sinensis and Paragonimus westermani. For anti-C. sinensis study, a total of 71 rats were divided into four batches for oral infection of each rat with 50 C. sinensis metacercariae. Five to 7 weeks post-infection, groups of rats were treated orally with mefloquine at single doses or multiple daily doses while infected, but untreated rats served as control. All treated rats were euthanized 2 weeks post-treatment for assessment of efficacy. For anti-P. westermani study, two batches of eight and ten dogs were each infected intraperitoneally with 100 P. westermani metacercariae. Eighty-five to 96 days post-infection, groups of two or three dogs were treated orally with mefloquine and groups of two dogs were treated with praziquantel at a single dose or multiple doses. In each batch of test, three untreated but infected dogs served as control. All treated dogs were euthanized 26-30 days post-treatment for evaluation of efficacy. In rats infected with C. sinensis and treated orally with mefloquine at a single dose of 75 and 150 mg/kg, no effect against C. sinensis was observed. When the dose of mefloquine was increased to 250 mg/kg, one third (five out of 15) rats died 3-5 days post-treatment. Although the mean worm burden was lower than that of the control, the difference between the treated and control groups was not statistically significant (P>0.05) with worm burden reduction of 22.4%. Whereas, the group of infected rats received mefloquine at a daily dose of 100 mg/kg for 3 days, one out of five rats died after the last administration. The mean worm burden was significantly lower than that of the control with worm burden reduction of 67.6% (P<0.01). In the first test of mefloquine against P. westermani, three infected dogs received two oral doses of the drug, 50 mg/kg, given at a 4-h interval, the mean worm burden were similar to that of the control. While other two dogs were treated with

  9. Sequences and gene organization of the mitochondrial genomes of the liver flukes Opisthorchis viverrini and Clonorchis sinensis (Trematoda).

    PubMed

    Cai, X Q; Liu, G H; Song, H Q; Wu, C Y; Zou, F C; Yan, H K; Yuan, Z G; Lin, R Q; Zhu, X Q

    2012-01-01

    Opisthorchis viverrini and Clonorchis sinensis are important trematodes infecting humans and animals, belonging to the family Opisthorchiidae. In the present study, we sequenced the nearly complete mitochondrial (mt) DNA (mtDNA) sequences of O. viverrini from Laos, obtained the complete mtDNA sequences of C. sinensis from China and Korea, and revealed their gene annotations and genome organizations. The mtDNA sequences of O. viverrini, C. sinensis (China isolate), C. sinensis (Korea isolate) were 13,510, 13,879, and 13,877 bp in size, respectively. Each of the three mt genomes comprises 36 genes, consisting of 12 genes coding for proteins, two genes for rRNA, and 20 genes (O. viverrini) or 22 genes (C. sinensis) for tRNA. The gene content and arrangement are identical to that of Fasciola hepatica, and Paragonimus westermani, but distinct from Schistosoma spp. All genes are transcribed in the same direction and have a nucleotide composition high in T. The contents of A + T of the mt genomes were 59.39% for O. viverrini, 60.03% for C. sinensis (China isolate), and 59.99% for C. sinensis (Korea isolate). Phylogenetic analyses using concatenated amino acid sequences of the 12 protein-coding genes, with three different computational algorithms [maximum parsimony, maximum likelihood, and Bayesian analysis], all revealed distinct groups with high statistical support, indicating that O. viverrini and C. sinensis represent sister taxa. These data provide additional novel mtDNA markers for studying the molecular epidemiology and population genetics of the two liver flukes and should have implications for the molecular diagnosis, prevention, and control of opisthorchiasis and clonorchiasis in humans and animals.

  10. Rapid detection and differentiation of Clonorchis sinensis and Opisthorchis viverrini using real-time PCR and high resolution melting analysis.

    PubMed

    Cai, Xian-Quan; Yu, Hai-Qiong; Li, Rong; Yue, Qiao-Yun; Liu, Guo-Hua; Bai, Jian-Shan; Deng, Yan; Qiu, De-Yi; Zhu, Xing-Quan

    2014-01-01

    Clonorchis sinensis and Opisthorchis viverrini are both important fish-borne pathogens, causing serious public health problem in Asia. The present study developed an assay integrating real-time PCR and high resolution melting (HRM) analysis for the specific detection and rapid identification of C. sinensis and O. viverrini. Primers targeting COX1 gene were highly specific for these liver flukes, as evidenced by the negative amplification of closely related trematodes. Assays using genomic DNA extracted from the two flukes yielded specific amplification and their identity was confirmed by sequencing, having the accuracy of 100% in reference to conventional methods. The assay was proved to be highly sensitive with a detection limit below 1 pg of purified genomic DNA, 5 EPG, or 1 metacercaria of C. sinensis. Moreover, C. sinensis and O. viverrini were able to be differentiated by their HRM profiles. The method can reduce labor of microscopic examination and the contamination of agarose electrophoresis. Moreover, it can differentiate these two flukes which are difficult to be distinguished using other methods. The established method provides an alternative tool for rapid, simple, and duplex detection of C. sinensis and O. viverrini.

  11. Molecular and Structural Characterization of the Tegumental 20.6-kDa Protein in Clonorchis sinensis as a Potential Druggable Target

    PubMed Central

    Kim, Yu-Jung; Yoo, Won Gi; Lee, Myoung-Ro; Kang, Jung-Mi; Na, Byoung-Kuk; Cho, Shin-Hyeong; Park, Mi-Yeoun; Ju, Jung-Won

    2017-01-01

    The tegument, representing the membrane-bound outer surface of platyhelminth parasites, plays an important role for the regulation of the host immune response and parasite survival. A comprehensive understanding of tegumental proteins can provide drug candidates for use against helminth-associated diseases, such as clonorchiasis caused by the liver fluke Clonorchis sinensis. However, little is known regarding the physicochemical properties of C. sinensis teguments. In this study, a novel 20.6-kDa tegumental protein of the C. sinensis adult worm (CsTegu20.6) was identified and characterized by molecular and in silico methods. The complete coding sequence of 525 bp was derived from cDNA clones and encodes a protein of 175 amino acids. Homology search using BLASTX showed CsTegu20.6 identity ranging from 29% to 39% with previously-known tegumental proteins in C. sinensis. Domain analysis indicated the presence of a calcium-binding EF-hand domain containing a basic helix-loop-helix structure and a dynein light chain domain exhibiting a ferredoxin fold. We used a modified method to obtain the accurate tertiary structure of the CsTegu20.6 protein because of the unavailability of appropriate templates. The CsTegu20.6 protein sequence was split into two domains based on the disordered region, and then, the structure of each domain was modeled using I-TASSER. A final full-length structure was obtained by combining two structures and refining the whole structure. A refined CsTegu20.6 structure was used to identify a potential CsTegu20.6 inhibitor based on protein structure-compound interaction analysis. The recombinant proteins were expressed in Escherichia coli and purified by nickel-nitrilotriacetic acid affinity chromatography. In C. sinensis, CsTegu20.6 mRNAs were abundant in adult and metacercariae, but not in the egg. Immunohistochemistry revealed that CsTegu20.6 localized to the surface of the tegument in the adult fluke. Collectively, our results contribute to a

  12. Molecular and Structural Characterization of the Tegumental 20.6-kDa Protein in Clonorchis sinensis as a Potential Druggable Target.

    PubMed

    Kim, Yu-Jung; Yoo, Won Gi; Lee, Myoung-Ro; Kang, Jung-Mi; Na, Byoung-Kuk; Cho, Shin-Hyeong; Park, Mi-Yeoun; Ju, Jung-Won

    2017-03-04

    The tegument, representing the membrane-bound outer surface of platyhelminth parasites, plays an important role for the regulation of the host immune response and parasite survival. A comprehensive understanding of tegumental proteins can provide drug candidates for use against helminth-associated diseases, such as clonorchiasis caused by the liver fluke Clonorchis sinensis. However, little is known regarding the physicochemical properties of C. sinensis teguments. In this study, a novel 20.6-kDa tegumental protein of the C. sinensis adult worm (CsTegu20.6) was identified and characterized by molecular and in silico methods. The complete coding sequence of 525 bp was derived from cDNA clones and encodes a protein of 175 amino acids. Homology search using BLASTX showed CsTegu20.6 identity ranging from 29% to 39% with previously-known tegumental proteins in C. sinensis. Domain analysis indicated the presence of a calcium-binding EF-hand domain containing a basic helix-loop-helix structure and a dynein light chain domain exhibiting a ferredoxin fold. We used a modified method to obtain the accurate tertiary structure of the CsTegu20.6 protein because of the unavailability of appropriate templates. The CsTegu20.6 protein sequence was split into two domains based on the disordered region, and then, the structure of each domain was modeled using I-TASSER. A final full-length structure was obtained by combining two structures and refining the whole structure. A refined CsTegu20.6 structure was used to identify a potential CsTegu20.6 inhibitor based on protein structure-compound interaction analysis. The recombinant proteins were expressed in Escherichia coli and purified by nickel-nitrilotriacetic acid affinity chromatography. In C. sinensis, CsTegu20.6 mRNAs were abundant in adult and metacercariae, but not in the egg. Immunohistochemistry revealed that CsTegu20.6 localized to the surface of the tegument in the adult fluke. Collectively, our results contribute to a

  13. Clonorchis sinensis ferritin heavy chain triggers free radicals and mediates inflammation signaling in human hepatic stellate cells.

    PubMed

    Mao, Qiang; Xie, Zhizhi; Wang, Xiaoyun; Chen, Wenjun; Ren, Mengyu; Shang, Mei; Lei, Huali; Tian, Yanli; Li, Shan; Liang, Pei; Chen, Tingjin; Liang, Chi; Xu, Jin; Li, Xuerong; Huang, Yan; Yu, Xinbing

    2015-02-01

    Clonorchiasis, caused by direct and continuous contact with Clonorchis sinensis, is associated with hepatobiliary damage, inflammation, periductal fibrosis, and the development of cholangiocarcinoma. Hepatic stellate cells respond to liver injury through production of proinflammatory mediators which drive fibrogenesis; however, their endogenous sources and pathophysiological roles in host cells were not determined. C. sinensis ferritin heavy chain (CsFHC) was previously confirmed as a component of excretory/secretory products and exhibited a number of extrahepatic immunomodulatory properties in various diseases. In this study, we investigated the expression pattern and biological role of CsFHC in C. sinensis. CsFHC was expressed throughout life stages of C. sinensis. More importantly, we found that treatment of human hepatic stellate cell line LX-2 with CsFHC triggered the production of free radicals via time-dependent activation of NADPH oxidase, xanthine oxidase, and inducible nitric oxide synthase. The increase in free radicals substantially promoted the degradation of cytosolic IκBα and nuclear translocation of NF-κB subunits (p65 and p50). CsFHC-induced NF-κB activation was markedly attenuated by preincubation with specific inhibitors of corresponding free radical-producing enzyme or the antioxidant. In addition, CsFHC induced an increased expression level of proinflammatory cytokines, IL-1β and IL-6, in NF-κB-dependent manner. Our results indicate that CsFHC-triggered free radical-mediated NF-κB signaling is an important factor in the chronic inflammation caused by C. sinensis infection.

  14. Toll-Like Receptor-Mediated Free Radical Generation in Clonorchis sinensis Excretory-Secretory Product-Treated Cholangiocarcinoma Cells

    PubMed Central

    Bahk, Young Yil; Pak, Jhang Ho

    2016-01-01

    Clonorchiasis, caused by direct contact with Clonorchis sinensis worms and their excretory-secretory products (ESPs), is associated with chronic inflammation, malignant changes in bile ducts, and even cholangiocarcinogenesis. Our previous report revealed that intracellular free radicals enzymatically generated by C. sinensis ESPs cause NF-κB-mediated inflammation in human cholangiocarcinoma cells (HuCCT1). Therefore, the present study was conducted to examine the role of upstream Toll-like receptors (TLRs) on the initial host innate immune responses to infection. We found that treatment of HuCCT1 cells with native ESPs induced changes in TLR mRNA levels in a time-dependent manner, concomitant with the generation of free radicals. ESP-mediated free radical generation was markedly attenuated by preincubation of the cells with TLR1-4-neutralizing antibodies, indicating that at least TLR1 through 4 participate in stimulation of the host innate immune responses. These findings indicate that free radicals triggered by ESPs are critically involved in TLR signal transduction. Continuous signaling by this pathway may function in initiating C. sinensis infection-associated inflammation cascades, a detrimental event leading to progression to more severe hepatobiliary diseases. PMID:27853127

  15. Toll-Like Receptor-Mediated Free Radical Generation in Clonorchis sinensis Excretory-Secretory Product-Treated Cholangiocarcinoma Cells.

    PubMed

    Bahk, Young Yil; Pak, Jhang Ho

    2016-10-01

    Clonorchiasis, caused by direct contact with Clonorchis sinensis worms and their excretory-secretory products (ESPs), is associated with chronic inflammation, malignant changes in bile ducts, and even cholangiocarcinogenesis. Our previous report revealed that intracellular free radicals enzymatically generated by C. sinensis ESPs cause NF-κB-mediated inflammation in human cholangiocarcinoma cells (HuCCT1). Therefore, the present study was conducted to examine the role of upstream Toll-like receptors (TLRs) on the initial host innate immune responses to infection. We found that treatment of HuCCT1 cells with native ESPs induced changes in TLR mRNA levels in a time-dependent manner, concomitant with the generation of free radicals. ESP-mediated free radical generation was markedly attenuated by preincubation of the cells with TLR1-4-neutralizing antibodies, indicating that at least TLR1 through 4 participate in stimulation of the host innate immune responses. These findings indicate that free radicals triggered by ESPs are critically involved in TLR signal transduction. Continuous signaling by this pathway may function in initiating C. sinensis infection-associated inflammation cascades, a detrimental event leading to progression to more severe hepatobiliary diseases.

  16. C3H/He Mice as an Incompatible Cholangiocarcinoma Model by Clonorchis sinensis, Dicyclanil and N-Nitrosodimethylamine

    PubMed Central

    Uddin, Md. Hafiz; Li, Shunyu; Jin, Yan; Choi, Min-Ho; Jang, Ja June; Hong, Sung-Tae

    2016-01-01

    Clonorchis sinensis is a Group-I bio-carcinogen, associated with cholangiocarcinoma (CCA). The hamster is the only experimental model of C. sinensis-mediated CCA, but we oblige another animal model. The present study intended to develop a C. sinensis (Cs) mediated CCA model using C3H/He mice, co-stimulated with N-nitrosodimethyl-amine (NDMA) and dicyclanil (DC). The mice were divided into 8 groups with different combinations of Cs, NDMA, and DC. Six months later the mice were sacrificed and subjected to gross and histopathological examination. The body weights were significantly reduced among the groups treated with 2 or more agents (eg. Cs+NDMA, Cs+DC, NDMA+DC, and Cs+NDMA+DC). In contrast, liver weight percentages to body weight were increased in above groups by 4.1% to 4.7%. A Change of the spleen weight was observed only in Cs+NDMA group. Though C. sinensis infection is evident from hyperplastic changes, only 1 worm was recovered. T wo mice, 1 from Cs and the other from Cs+DC group, showed mass forming lesions; 1 (281.2 mm3) from the Cs group was a hepatocellular adenoma and the other (280.6 mm3) from the Cs+DC group was a cystic mass (peliosis). Higher prevalence of gray-white nodules was observed in Cs group (42.9%) followed by Cs+NDMA+DC group (21.4%). The mice of the Cs+NDMA+DC group showed hyper-proliferation of the bile duct with fibrotic changes. No characteristic change for CCA was recognized in any of the groups. In conclusion, C3H/He mice produce no CCA but extensive fibrosis when they are challenged by Cs, NDMA, and DC together. PMID:27417082

  17. Inducible nitric oxide synthase response and associated cytokine gene expression in the spleen of mice infected with Clonorchis sinensis.

    PubMed

    Shen, Ji-Qing; Yang, Qing-Li; Xue, Yan; Cheng, Xiao-Bing; Jiang, Zhi-Hua; Yang, Yi-Chao; Chen, Ying-Dan; Zhou, Xiao-Nong

    2015-05-01

    Clonorchis sinensis is a food-borne parasite that induces a permanent increase of nitrosation in the body upon infection. The spleen is an important secondary lymphoid organ for the regulation of immune responses locally and in the whole body. However, the functions and mechanisms of the spleen in nitric oxide (NO) responses after C. sinensis infection remain unknown. In this study, BALB/c mice were infected with 20, 40, and 80 C. sinensis metacercariae to simulate mild, moderate, and severe infections, respectively. We examined the expression of inducible nitric oxide synthase (iNOS) in the spleen and the relevant cytokine transcription in splenocytes from the mice infected with different amounts of metacercariae. The iNOS of the mice infected with 80 metacercariae was expressed in the spleen as early as 10 days post-infection (dpi) and gradually increased until 90 dpi. The iNOS expression in the mice infected with 40 metacercariae was detected only at 45 and 90 dpi, but not in the mice infected with 20 metacercariae. The level of interferon (IFN)-γ messenger RNA (mRNA) transcription in splenocytes significantly increased at 10 and 20 dpi (P < 0.05) in response to mild/moderate infection but gradually decreased to normal levels after 45 dpi. The level of IL-12p35 mRNA transcription did not change at 10 and 20 dpi but significantly decreased after 45 dpi under moderate/severe infection (P < 0.05/0.01/0.001). The level of IL-18 mRNA transcription significantly increased at 10 dpi (P < 0.05/0.01) but significantly decreased after 20 dpi (P < 0.05/0.01/0.001). These results suggest that spleen is an important organ for iNOS/NO responses, which correspond to the severity of C. sinensis infection, but cannot be attributed to the expression of the Th1 cytokines.

  18. Immune response induced by oral delivery of Bacillus subtilis spores expressing enolase of Clonorchis sinensis in grass carps (Ctenopharyngodon idellus).

    PubMed

    Jiang, Hongye; Chen, Tingjin; Sun, Hengchang; Tang, Zeli; Yu, Jinyun; Lin, Zhipeng; Ren, Pengli; Zhou, Xinyi; Huang, Yan; Li, Xuerong; Yu, Xinbing

    2017-01-01

    Clonorchiasis, caused by the consumption of raw or undercooked freshwater fish containing infective metacercariae of Clonorchis sinensisis (C.sinensis), remains a common public health problem. New effective prevention strategies are still urgent to control this food-borne infectious disease. The previous studies suggested Bacillus subtilis (B. subtilis) spores was an ideal vaccines delivery system, and the C.sinensis enolase (CsENO) was a potential vaccine candidate against clonorchiasis. In the current study, we detected CsENO-specific IgM levels by ELISA in sera, intestinal mucus and skin mucus in grass carps (Ctenopharyngodon idella) through oral administration with B. subtilis spores surface expressing CsENO. In addition, immune-related genes expression was also measured by qRT-PCR. Grass carps orally treated with B. subtilis spores or normal forages were used as controls. The results of ELISA manifested that specific IgM levels of grass carps in CsENO group in sera, intestine mucus and skin mucus almost significantly increased from week 4 post the first oral administration when compared to the two control groups. The levels of specific IgM reached its peak in intestine mucus firstly, then in sera, and last in skin mucus. qRT-PCR results showed that 5 immune-related genes expression had different degree of rising trend in CsENO group when compared to the two control groups. Our study demonstrated that orally administrated with B. subtilis spores expressing CsENO induced innate and adaptive immunity, systemic and local mucosal immunity, and humoral and cellular immunity. Our work may pave the way to clarify the exact mechanisms of protective efficacy elicited by B. subtilis spores expressing CsENO and provide new ideas for vaccine development against C. sinensis infection.

  19. [A Study On The Fine Structure Of Clonorchis Sinensis, A Liver Fluke: II. The Alimentary Tract And The Excretory System

    PubMed

    Jeong, Kye Heon; Rim, Han Jong; Kim, Woo Kap; Kim, Chang Whan; Yang, He Young

    1980-06-01

    A morphological study on the ultrastructures of the alimentary tract and the excretory system of Clonorchis sinensis was conducted. The liver flukes were collected from rabbit liver six months after the experimental infection The worms were washed with 0.85% saline solution and immediately moved to cold 2.5% glutaraldehyde in 0.1 M phosphate buffer pH 7.4. The materials were dissected and fixed for two hours. The blocks were post-fixed in 1% osmium tetroxide. The blocks were embedded in Epon 812. Ultra thin sections were cut with Sovall MT-2 ultramicrotome and stained with uranyl acetate and lead citrate. Sections were then observed with Hitachi HS-7S electron microscope. The following results were obtained in a series of observations. The walls of oral cavity and esophagus comprised tegumental syncytium, basement membrane, loose connective tissue, muscular layer and parenchymal cells. The apical surface and the base of the syncytium were covered with a protoplasmic membrane for each forming numerous invaginations. Granular endoplasmic reticulum was developed in the epithelium of the oesophagus. The gastrodermis of Clonorchis sinensis comprised two types of cells in general. The first cell type was numerous one forming a single continuous layer of epithelial cells. Each of the cells had outfolded cytoplasm into the caecal lumen and lamellae along the cell surface. Among the above epithelial cells, no considerable differences in structure reflecting their functional states were identified. The second cell type was less differentiated in nature and lay within the gastrodermis above the basement membrane but not in contact with the caecal lumen, being overlapped by neighboring gastrodermal cells of the type described above. At this portion the gastrodermis seemed to be a pseudostratified epithelium. There were well-developed lamellae along the surface of epithelia of all canals or duct concerning evacuation. The excretory pore was 7.5 micrometer in diameter and dorso

  20. Systemic and local mucosal immune responses induced by orally delivered Bacillus subtilis spore expressing leucine aminopeptidase 2 of Clonorchis sinensis.

    PubMed

    Qu, Hongling; Xu, Yanquan; Sun, Hengchang; Lin, Jinsi; Yu, Jinyun; Tang, Zeli; Shen, Jiqing; Liang, Chi; Li, Shan; Chen, Wenjun; Li, Xuerong; Wu, Zhongdao; Huang, Yan; Yu, Xinbing

    2014-08-01

    Human clonorchiasis caused by Clonorchis sinensis (C. sinensis) has been increasingly prevalent in recent years so that an effective measure is essential and urgent to control the infectious disease. Oral delivery of antigens from C. sinensis may be an important approach to effectively induce both systemic and local immune responses to anti-infection of the parasite. In the current study, we used Bacillus subtilis (B. subtilis) spores as a delivery vehicle to introduce leucine aminopeptidase 2 of C. sinensis (CsLAP2), an excretory/secretory antigen with high immunogenicity, expressing on their surface. SDS-PAGE, western blotting, and flow cytometry indicated that CsLAP2 was successfully expressed on the surface of B. subtilis spores (CotC-CsLAP2 spores). BALB/c mice were treated with spores intragastrically. On day 31 after the treatment, we found that mice intragastrically treated with CotC-CsLAP2 spores exhibited higher IgG, IgG1, IgG2a, and IgA level in sera as well as higher sIgA level in bile and intestinal lavage fluid compared to mice orally administrated with spores not expressing CsLAP2 (CotC spores) and naïve mice. The peak titer of IgG/IgA presented on day 31/49 after oral administration. IgG1 level was lower than IgG2a in group administrated with CotC-CsLAP2 spores. sIgA-secreting cells were obviously observed in intestinal epithelium of mice orally treated with CotC-CsLAP2 spores. After incubated with CotC-CsLAP2, the levels of IFN-γ, IL-6, IL-10, IL-17A, and TNF significantly increased in the supernatant of splenocytes isolated from mice orally treated with CotC-CsLAP2 spores, while there was no statistically significant difference of IL-4 level representing Th2 response among the groups. Our study demonstrated that oral administration of CsLAP2 delivered by B. subtilis spore elicited obvious systemic and local mucosal immunity. Secretory IgA and Th1-Th17 cellular immunity might involved in mechanisms of the immune response.

  1. Cell-free translational screening of an expression sequence tag library of Clonorchis sinensis for novel antigen discovery.

    PubMed

    Kasi, Devi; Catherine, Christy; Lee, Seung-Won; Lee, Kyung-Ho; Kim, Yu Jung; Ro Lee, Myeong; Ju, Jung Won; Kim, Dong-Myung

    2017-01-27

    The rapidly evolving cloning and sequencing technologies have enabled understanding of genomic structure of parasite genomes, opening up new ways of combatting parasite-related diseases. To make the most of the exponentially accumulating genomic data, however, it is crucial to analyze the proteins encoded by these genomic sequences. In this study, we adopted an engineered cell-free protein synthesis system for large-scale expression screening of an expression sequence tag (EST) library of Clonorchis sinensis to identify potential antigens that can be used for diagnosis and treatment of clonorchiasis. To allow high-throughput expression and identification of individual genes comprising the library, a cell-free synthesis reaction was designed such that both the template DNA and the expressed proteins were co-immobilized on the same microbeads, leading to microbead-based linkage of the genotype and phenotype. This reaction configuration allowed streamlined expression, recovery, and analysis of proteins. This approach enabled us to identify 21 antigenic proteins. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 2017.

  2. Effects of excretory/secretory products from Clonorchis sinensis and the carcinogen dimethylnitrosamine on the proliferation and cell cycle modulation of human epithelial HEK293T cells.

    PubMed

    Kim, Eun-Min; Kim, June-Sung; Choi, Min-Ho; Hong, Sung-Tae; Bae, Young Mee

    2008-09-01

    Clonorchis sinensis is one of the most prevalent parasitic helminths in Korea. Although cholangiocarcinoma can be induced by C. sinensis infection, the underlying mechanism is not clearly understood. To assess the role of C. sinensis infection in carcinogenesis, an in vitro system was established using the human epithelial cell line HEK293T. In cells exposed to the excretory/secretory products (ESP) of C. sinensis and the carcinogen dimethylnitrosamine (DMN), cellular proliferation and the proportion of cells in the G2/M phase increased. Moreover, the expression of the cell cycle proteins E2F1, p-pRb, and cyclin B was dramatically increased when ESP and DMN were added together. Similarly, the transcription factor E2F1 showed its highest level of activity when ESP and DMN were added simultaneously. These findings indicate that DMN and ESP synergistically affect the regulation of cell cycle-related proteins. Our results suggest that exposure to C. sinensis and a small amount of a carcinogen such as DMN can promote carcinogenesis in the bile duct epithelium via uncontrolled cellular proliferation and the upregulation of cell cycle-related proteins.

  3. Clonorchis sinensis-derived total protein attenuates airway inflammation in murine asthma model by inducing regulatory T cells and modulating dendritic cell functions

    SciTech Connect

    Jeong, Young-Il; Kim, Seung Hyun; Ju, Jung Won; Cho, Shin Hyeong; Lee, Won Ja; Park, Jin Wook; Park, Yeong-Min; Lee, Sang Eun

    2011-04-22

    Highlights: {yields} Treatment with Clonorchis sinensis-derived total protein attenuates OVA-induced airway inflammation and AHR to methacholine. {yields} Induction of CD4{sup +}CD25{sup +}Foxp3{sup +} T cells and IL-10 along with suppression of splenocyte proliferation by C. sinensis-derived total protein. {yields} C. sinensis-derived total protein interferes with the expression of co-stimulatory molecules in DCs. -- Abstract: Asthma is characterized by Th2-mediated inflammation, resulting in airway hyperresponsiveness (AHR) through airway remodeling. Recent epidemiological and experimental reports have suggested an inverse relationship between the development of allergy and helminth infections. Infection by Clonorchis sinensis, a liver fluke that resides in the bile duct of humans, is endemic predominantly in Asia including Korea and China. Using a murine model for asthma, we investigated the effects of C. sinensis-derived total protein (Cs-TP) on allergen-induced airway inflammation and the mechanism underlying the protective effects of Cs-TP administration on asthma. Treatment with Cs-TP attenuated OVA-induced airway inflammation and methacholine-induced AHR, as well as eosinophilia development, lymphocyte infiltration into the lung, and goblet cell metaplasia. This protective effect of Cs-TP is associated with markedly reduced OVA-specific IgE and Th1/Th2 cytokine production. Moreover, Cs-TP increased the number of CD4{sup +}CD25{sup +}Foxp3{sup +} regulatory T (Treg) cells as well as their suppressive activity. In fact, proliferation of OVA-restimulated splenocytes was suppressed significantly. Cs-TP also inhibited the expression of such co-stimulatory molecules as CD80, CD86, and CD40 in LPS- or OVA-stimulated dendritic cells (DCs), suggesting that Cs-TP could interfere with the capacity of airway DCs to prime naive T cells. These data demonstrate the capacity of C. sinensis to ameliorate allergic asthma and broaden our understanding of the paradoxical

  4. The Dynamics of Treg/Th17 and the Imbalance of Treg/Th17 in Clonorchis sinensis-Infected Mice.

    PubMed

    Yan, Chao; Zhang, Bei-Bei; Hua, Hui; Li, Bo; Zhang, Bo; Yu, Qian; Li, Xiang-Yang; Liu, Ying; Pan, Wei; Liu, Xiang-Ye; Tang, Ren-Xian; Zheng, Kui-Yang

    2015-01-01

    Clonorchiasis, caused by the liver fluke Clonorchis sinensis, is a chronic parasitic infection regulated by T cell subsets. An imbalance of CD4+CD25+ Foxp3+regulatory T (Treg) and interleukin (IL)-17-secreting T cells (Th17) may control inflammation and play an important role in the pathogenesis of immune evasion. In the present study, we assessed the dynamics of Treg/Th17 and determined whether the Treg/Th17 ratio is altered in C. sinensis-infected mice. The results showed that the percentages of splenic Treg cells in CD4+ T cells were suppressed on day 14 post-infection (PI) but increased on day 56 PI, while Th17 cells were increased on day 56 PI compared with normal control (NC) mice. The Treg/Th17 ratio steadily increased from day 28 to day 56 PI. The hepatic levels of their specific transcription factors (Foxp3 for Treg and RORγt for Th17) were increased in C. sinensis-infected mice from day 14 to 56 PI, and significantly higher than those in NC mice. Meanwhile, serum levels of IL-2 and IL-17 were profoundly increased in C. sinensis-infected mice throughout the experiment; while the concentrations of IL-6 and transforming growth factor β1 (TGF-β1) peaked on day 14 PI, but then decreased on day 28 and 56 PI. Our results provide the first evidence of an increased Treg/Th17 ratio in C. sinensis-infected mice, suggesting that a Treg/Th17 imbalance may play a role in disease outcomes of clonorchiasis.

  5. The Dynamics of Treg/Th17 and the Imbalance of Treg/Th17 in Clonorchis sinensis-Infected Mice

    PubMed Central

    Hua, Hui; Li, Bo; Zhang, Bo; Yu, Qian; Li, Xiang-Yang; Liu, Ying; Pan, Wei; Liu, Xiang-Ye; Tang, Ren-Xian; Zheng, Kui-Yang

    2015-01-01

    Clonorchiasis, caused by the liver fluke Clonorchis sinensis, is a chronic parasitic infection regulated by T cell subsets. An imbalance of CD4+CD25+ Foxp3+regulatory T (Treg) and interleukin (IL)-17-secreting T cells (Th17) may control inflammation and play an important role in the pathogenesis of immune evasion. In the present study, we assessed the dynamics of Treg/Th17 and determined whether the Treg/Th17 ratio is altered in C. sinensis-infected mice. The results showed that the percentages of splenic Treg cells in CD4+ T cells were suppressed on day 14 post-infection (PI) but increased on day 56 PI, while Th17 cells were increased on day 56 PI compared with normal control (NC) mice. The Treg/Th17 ratio steadily increased from day 28 to day 56 PI. The hepatic levels of their specific transcription factors (Foxp3 for Treg and RORγt for Th17) were increased in C. sinensis-infected mice from day 14 to 56 PI, and significantly higher than those in NC mice. Meanwhile, serum levels of IL-2 and IL-17 were profoundly increased in C. sinensis-infected mice throughout the experiment; while the concentrations of IL-6 and transforming growth factor β1 (TGF-β1) peaked on day 14 PI, but then decreased on day 28 and 56 PI. Our results provide the first evidence of an increased Treg/Th17 ratio in C. sinensis-infected mice, suggesting that a Treg/Th17 imbalance may play a role in disease outcomes of clonorchiasis. PMID:26599407

  6. [The reagents kit to detect Metyorchis biis, Opisthorchis viverrini and Clonorchis sinensis, Opisthorchis felineus--agents of opisthorchiasis using technique of polymerase chain reaction in real-time].

    PubMed

    Seredina, T A; Petrenko, V A; Tronin, A V; Sazonov, A Iu; Sapugol'tseva, O B; Katokhin, A V; Odintsova, E S

    2014-08-01

    The helminths Opisthorchis felineus, Opisthorchis viverrini, Clonorchis sinensis, Metorchis bilis are the agents of opisthorchiasis. The actual diagnostic of parasitic diseases based on microscope analysis of samples of human feces to detect presence of ova of parasites suffers of many shortcomings, in particular low sensitivity especially at earlier stages. The purpose of this study was to compare results of detection of parasites using both classical technique and technique of specific differentiation based on extraction of nucleic acids from samples of human feces and implementation of reaction of amplification of the chosen fragment of DNA with detection of products of polymerase chain reaction in the real time. The study detected 150 out of 165 positive samples and also 6 out of 37 negative samples both validated by coproovoscopy.

  7. Clonorchis sinensis excretory-secretory products promote the migration and invasion of cholangiocarcinoma cells by activating the integrin β4-FAK/Src signaling pathway.

    PubMed

    Pak, Jhang Ho; Bashir, Qudsia; Kim, In Ki; Hong, Sung-Jong; Maeng, Sejung; Bahk, Young Yil; Kim, Tong-Soo

    2017-03-08

    Cholangiocarcinoma (CCA) is a slow-growing but highly metastatic cancer. Its metastatic potential largely explains its high mortality rate. A recognized risk factor for CCA development is infection with the liver flukes Opisthorchis viverrini and Clonorchis sinensis. We previously reported that the excretory-secretory products (ESPs) of C. sinensis promoted the three-dimensional aggregation and invasion of CCA cells. In the present study, a quantitative real-time PCR array of extracellular matrix (ECM) and adhesion molecules was used to examine the regulatory mechanism of ESP-mediated CCA cell migration and invasion. In particular, the expression levels of integrin α isoforms and β4 were upregulated in response to ESPs. Increased expression of integrin β4 was probably correlated with activation of focal adhesion kinase (FAK) and the steroid receptor coactivator (Src) family kinase and the subsequent activation of two downstream focal adhesion molecules, paxillin and vinculin. Moreover, inhibition of FAK/Src activation reduced paxillin and vinculin phosphorylation and attenuated ESP-induced CCA cell migration and invasion. These findings suggest that the integrin β4-FAK/Src signaling axis may play a crucial role in clonorchiasis-associated CCA metastasis during tumor progression.

  8. Free radicals enzymatically triggered by Clonorchis sinensis excretory-secretory products cause NF-κB-mediated inflammation in human cholangiocarcinoma cells.

    PubMed

    Nam, Joo-Hyun; Moon, Ju Hyun; Kim, In Ki; Lee, Myoung-Ro; Hong, Sung-Jong; Ahn, Joong Ho; Chung, Jong Woo; Pak, Jhang Ho

    2012-01-01

    Chronic clonorchiasis, caused by direct and continuous contact with Clonorchis sinensis worms and their excretory-secretory products, is associated with hepatobiliary damage, inflammation, periductal fibrosis and even development of cholangiocarcinoma. Our previous report revealed that intracellular reactive oxygen species were generated in C. sinensis excretory-secretory product-treated human cholangiocarcinoma cells; however, their endogenous sources and pathophysiological roles in host cells were not determined. In the present study, we found that treatment of human cholangiocarcinoma cells with excretory-secretory products triggered increases in free radicals via a time-dependent activation of NADPH oxidase, xanthine oxidase and inducible nitric oxide synthase. This increase in free radicals substantially promoted the degradation of cytosolic IκB-α, nuclear translocation of nuclear factor-κB subunits (RelA and p50), and increased κB consensus DNA-binding activity. Excretory-secretory product-induced nuclear factor-κB activation was markedly attenuated by preincubation with specific inhibitors of each free radical-producing enzyme or the antioxidant, N-acetylcysteine. Moreover, excretory-secretory products induced an increase in the mRNA and protein expression of the proinflammatory cytokines, IL-1β and IL-6, in an nuclear factor-κB-dependent manner, indicating that enzymatic production of free radicals in ESP-treated cells participates in nuclear factor-κB-mediated inflammation. These findings provide new insights into the pathophysiological role of C. sinensis excretory-secretory products in host chronic inflammatory processes, which are initial events in hepatobiliary diseases.

  9. Increased hepatic Th2 and Treg subsets are associated with biliary fibrosis in different strains of mice caused by Clonorchis sinensis

    PubMed Central

    Fang, Fan; Du, Ying; Ma, Rui; Li, Xiang-Yang; Yu, Qian; Meng, Di; Tang, Ren-Xian; Zheng, Kui-Yang

    2017-01-01

    Previous studies showed that CD4+T cells responses might be involved in the process of biliary fibrosis. However, the underlying mechanism resulting in biliary fibrosis caused by Clonorchis sinensis remains not yet fully elucidated. The objectives of the present study were to investigate the different profiles of hepatic CD4+T cell subsets (Th1, Th2, Th17 and Treg cells) and their possible roles in the biliary fibrosis of different strains of mice (C57BL/6, BALB/c and FVB mice) induced by C. sinensis infection. C57BL/6, BALB/c and FVB mice were orally gavaged with 45 metacercariae. All mice were sacrificed on 28 days post infection in deep anesthesia conditions. The leukocytes in the liver were separated to examine CD4+T cell subsets by flow cytometry and the left lobe of liver was used to observe pathological changes, collagen depositions and the concentrations of hydroxyproline. The most serious cystic and fibrotic changes appeared in FVB infected mice indicated by gross observation, Masson’s trichrome staining and hydroxyproline content detection. In contrast to C57BL/6 infected mice, diffuse nodules and more intensive fibrosis were observed in the BALB/c infected mice. No differences of the hepatic Th1 subset and Th17 subset were found among the three strains, but the hepatic Th2 and Treg cells and their relative cytokines were dramatically increased in the BALB/c and FVB infected groups compared with the C57BL/6 infected group (P<0.01). Importantly, increased Th2 subset and Treg subset all positively correlated with hydroxyproline contents (P<0.01). This result for the first time implied that the increased hepatic Th2 and Treg cell subsets were likely to play potential roles in the formation of biliary fibrosis in C. sinensis-infected mice. PMID:28151995

  10. The characteristics of the expression of heat shock proteins and COX-2 in the liver of hamsters infected with Clonorchis sinensis, and the change of endocrine hormones and cytokines.

    PubMed

    Choi, WonHyung; Chu, JongPhil

    2012-12-01

    The liver fluke Clonorchis sinensis (Digenea) is a high-risk parasite that causes serious diseases such as cirrhosis, carcinogenic liver damage and clonorchiasis in East Asia. This study was conducted to evaluate the relationship between stress/endocrine hormones and inflammation induced by infection as well as the expression of heat shock proteins (hsp-27, hsp-90), cox-2 and cytokines in the livers of hamsters infected with C. sinensis. The average body weight of infected hamsters decreased up to 25% compared with that of the control group, and bile duct hyperplasia with inflammation, liver fibrosis and hepatic necrosis were observed in C. sinensis-infected livers. The expression of hsp-27, hsp-90, and cox-2 was significantly increased in the livers of C. sinensis-infected hamsters compared with the control group. Moreover, the expression levels of inflammatory cytokines (IL-1beta, IL-2, TGF-beta2 and IFN-alpha1) were markedly increased in the livers of the infected group compared with those of the control group. Consistently, plasma IL-3 and IL-6 levels gradually increased during the infection period, and the concentration levels of testosterone, luteinizing hormone (LH), follicle-stimulating hormone (FSH), corticosterone, and adrenocorticotropic hormone (ACTH) in C. sinensis-infected hamsters increased over 25%, compared with those of the uninfected normal group. These results demonstrate that C. sinensis infection may increase the expression of hsp27, hsp90 and cox-2 as well as it may cause periductal fibrosis, chronic inflammation and hepatic necrosis in the liver. Furthermore, the results indicate that C. sinensis infection induces not only stress-induced hormone imbalance but also the sustained secretion of inflammatory cytokines through chronic stress/stimuli.

  11. Rapid detection and differentiation of Clonorchis sinensis and Opisthorchis viverrini eggs in human fecal samples using a duplex real-time fluorescence resonance energy transfer PCR and melting curve analysis.

    PubMed

    Sanpool, Oranuch; Intapan, Pewpan M; Thanchomnang, Tongjit; Janwan, Penchom; Lulitanond, Viraphong; Doanh, Pham Ngoc; Van Hien, Hoang; Dung, Do Trung; Maleewong, Wanchai; Nawa, Yukifumi

    2012-07-01

    We developed a single step duplex real-time fluorescence resonance energy transfer (FRET) PCR merged with melting curve analysis for the fast detection and differentiation of Clonorchis sinensis and Opisthorchis viverrini eggs in human fecal samples. Two species of mitochondrial NADH dehydrogenase subunit 2 (nad2) DNA elements, the 165-bp nad2 product of C. sinensis and the 209-bp nad2 product of O. viverrini, were amplified by species-specific primers, and the fluorescence melting curve analyses were generated from hybrid of amplicons and two pairs of species-specific fluorophore-labeled probes. By their different fluorescence channels and melting temperatures, both C. sinensis and O. viverrini eggs in infected human fecal samples were detected and differentiated with high (100%) sensitivity and specificity. Detection limit was as little as a single C. sinensis egg and two O. viverrini eggs in 100 mg of fecal sample. The assay could distinguish the DNA of both parasites from the DNA of negative fecal samples and fecal samples with other parasitosis, as well as from the well-defined genomic DNA of human leukocytes and other parasites. It can reduce labor time of microscopic examination and is not prone to carry over contamination of agarose electrophoresis. Our duplex real-time FRET PCR method would be useful to determine the accurate range of endemic areas and/or to discover the co-endemic areas of two liver flukes, C. sinensis and O. viverrini, in Asia. This method also would be helpful for the differential diagnosis of the suspected cases of liver fluke infections among travelers who had visited the endemic countries of those parasites.

  12. Clonorchis sinensis excretory-secretory products regulate migration and invasion in cholangiocarcinoma cells via extracellular signal-regulated kinase 1/2/nuclear factor-κB-dependent matrix metalloproteinase-9 expression.

    PubMed

    Pak, Jhang Ho; Shin, Jimin; Song, In-Sung; Shim, Sungbo; Jang, Sung-Wuk

    2017-01-01

    Matrix metalloproteinase-9 plays an important role in the invasion and metastasis of various types of cancer cells. We have previously reported that excretory-secretory products from Clonorchis sinensis increases matrix metalloproteinase-9 expression. However, the regulatory mechanisms through which matrix metalloproteinase-9 expression affects cholangiocarcinoma development remain unclear. In the current study, we examined the potential role of excretory-secretory products in regulating the migration and invasion of various cholangiocarcinoma cell lines. We demonstrated that excretory-secretory products significantly induced matrix metalloproteinase-9 expression and activity in a concentration-dependent manner. Reporter gene and chromatin immunoprecipitation assays showed that excretory-secretory products induced matrix metalloproteinase-9 expression by enhancing the activity of nuclear factor-kappa B. Moreover, excretory-secretory products induced the degradation and phosphorylation of IκBα and stimulated nuclear factor-kappa B p65 nuclear translocation, which was regulated by extracellular signal-regulated kinase 1/2. Taken together, our findings indicated that the excretory-secretory product-dependent enhancement of matrix metalloproteinase-9 activity and subsequent induction of IκBα and nuclear factor-kappa B activities may contribute to the progression of cholangiocarcinoma.

  13. Genetic diversity of nuclear ITS1-5.8S-ITS2 rDNA sequence in Clonorchis sinensis Cobbold, 1875 (Trematoda: Opisthorchidae) from the Russian Far East.

    PubMed

    Tatonova, Yulia V; Chelomina, Galina N; Besprosvannykh, Vladimir V

    2012-12-01

    The present study examined the molecular organisation and sequence variation in the nuclear ribosomal DNA (rDNA) region, including the two internal transcribed spacers (ITS1 and ITS2) and the 5.8S gene of the Clonorchis sinensis from the Russian Far East. The relevant sequences from other parts of this species' area were downloaded from GenBank. The results showed 100% identity for all investigated 5.8S-ITS2 rDNA sequences. In contrast, two levels of intraspecific variations were revealed in the complete ITS1 sequences. The intra-genomic variation resulted from a C/T polymorphism in a single position. The inter-individual differences between the ITS1 sequences were both due to nucleotide and size polymorphisms resulting from a varying number of five-nucleotide repeats and followed by two ITS1 length variants. These variant frequencies correlate with the clonorchiasis level in some geographical localities. ITS1 differences, both in the mutation profile and mutation localisation, were revealed between northern and southern geographical samples. The presence of GC boxes that are identical to known regulatory motifs in eukaryotes was detected within the ITS1 sub-repeats. The predicted secondary structures for ITS1 consist of two large branches, one of which was invariable, while another depended on ITS1 length. The predicted secondary structure for ITS2 includes four helices around the core. The main differences between C. sinensis and other opisthorchids were localised on the tops of helices 2, 3, and 4. A phylogenetic MST reconstruction subdivided all ITS1 sequences into two well differentiated clusters, each with the major widespread ribotype, and showed that ribotype diversity in both Russia and Korea is much lower than in China. The results obtained demonstrate the feasibility of complete ITS1 sequences in C. sinensis population genetics and can be considered as a basis for further studies of the parasite infection because they may help to elucidate the molecular

  14. Evolutionary course of CsRn1 long-terminal-repeat retrotransposon and its heterogeneous integrations into the genome of the liver fluke, Clonorchis sinensis

    PubMed Central

    Bae, Young-An

    2003-01-01

    The evolutionary course of the CsRn1 long-terminal-repeat (LTR) retrotransposon was predicted by conducting a phylogenetic analysis with its paralog LTR sequences. Based on the clustering patterns in the phylogenetic tree, multiple CsRn1 copies could be grouped into four subsets, which were shown to have different integration times. Their differential sequence divergences and heterogeneous integration patterns strongly suggested that these subsets appeared sequentially in the genome of C. sinensis. Members of recently expanding subset showed the lowest level of divergence in their LTR and reverse transcriptase gene sequences. They were also shown to be highly polymorphic among individual genomes of the trematode. The CsRn1 element exhibited a preference for repetitive, agenic chromosomal regions in terms of selecting integration targets. Our results suggested that CsRn1 might induce a considerable degree of intergenomic variation and, thereby, have influenced the evolution of the C. sinensis genome. PMID:14699262

  15. Clonorcis sinensis eggs are associated with calcium carbonate gallbladder stones.

    PubMed

    Qiao, Tie; Ma, Rui-hong; Luo, Zhen-liang; Yang, Liu-qing; Luo, Xiao-bing; Zheng, Pei-ming

    2014-10-01

    Calcium carbonate gallbladder stones were easily neglected because they were previously reported as a rare stone type in adults. The aim of this study was to investigate the relationship between calcium carbonate stones and Clonorchis sinensis infection. A total of 598 gallbladder stones were studied. The stone types were identified by FTIR spectroscopy. The C. sinensis eggs and DNA were detected by microscopic examination and real-time fluorescent PCR respectively. And then, some egg-positive stones were randomly selected for further SEM examination. Corresponding clinical characteristics of patients with different types of stones were also statistically analyzed. The detection rate of C. sinensis eggs in calcium carbonate stone, pigment stone, mixed stone and cholesterol stone types, as well as other stone types was 60%, 44%, 36%, 6% and 30%, respectively, which was highest in calcium carbonate stone yet lowest in cholesterol stone. A total of 182 stones were egg-positive, 67 (37%) of which were calcium carbonate stones. The C. sinensis eggs were found adherent to calcium carbonate crystals by both light microscopy and scanning electron microscopy. Patients with calcium carbonate stones were mainly male between the ages of 30 and 60, the CO2 combining power of patients with calcium carbonate stones were higher than those with cholesterol stones. Calcium carbonate gallbladder stones are not rare, the formation of which may be associated with C. sinensis infection.

  16. Clonorchis sinensis lysophospholipase inhibits TGF-β1-induced expression of pro-fibrogenic genes through attenuating the activations of Smad3, JNK2, and ERK1/2 in hepatic stellate cell line LX-2.

    PubMed

    Zhou, Lina; Shang, Mei; Shi, Mengchen; Zhao, Lu; Lin, Zhipeng; Chen, Tingjin; Wu, Yinjuan; Tang, Zeli; Sun, Hengchang; Yu, Jinyun; Huang, Yan; Yu, Xinbing

    2016-02-01

    Liver fibrosis is a wound healing response associated with chronic liver injury. Hepatic stellate cells (HSCs) activation is a key event in the development of liver fibrosis. Since helminths have the ability to live for decades in the host by establishing an adaptive relationship in the interplay with its hosts, we hypothesize that whether Clonochis sinensis LysophospholipaseA (CsLysoPLA), a component of excretory/secretory proteins, can attenuate the fibrogenic response by inhibiting activation of LX-2 cells, thereby balancing the pro-fibrotic and anti-fibrotic response during the Clonochis sinensis (C. sinensis) infection. In the present study, LX-2 cells were stimulated with CsLysoPLA in the presence of TGF-β1, and the expressions of collagen type I (COL1A1), α-smooth muscle actin (α-SMA), and matrix metalloproteinase 2 (MMP2) were decreased. In addition, CsLysoPLA significantly inhibited the proliferation and migration of LX-2 cells stimulated by TGF-β1. Pretreatment of LX-2 cells with CsLysoPLA attenuated the phosphorylation of Smad3 as well as JNK2 and ERK1/2 in response to the stimulation of TGF-β1. For the first time, our results showed an anti-fibrogenic effect of CsLysoPLA by attenuating the response of LX-2 cells to TGF-β1 through inhibiting the activations of Smad3, ERK1/2, and JNK2.

  17. STUDIES ON CHEMOTHERAPY AND SERODIAGNOSIS FOR CLONORCHIS SINENSIS INFECTION.

    DTIC Science & Technology

    CHEMOTHERAPEUTIC AGENTS, *SERODIAGNOSIS, PARASITIC DISEASES , CHEMOTHERAPY, PARASITIC DISEASES , DOSAGE, TOXICITY, BODY WEIGHT, PATHOLOGY, MORTALITY RATE, HEMATOLOGY, SODIUM COMPOUNDS, BIOASSAY, JAPAN.

  18. Effect of green tea (Camellia sinensis L.) extract on morphological and functional changes in adult male gonads of albino rats.

    PubMed

    Chandra, Amar K; Choudhury, Shyamosree Roy; De, Neela; Sarkar, Mahitosh

    2011-09-01

    Green tea, prepared from the steamed and dried leaves of the shrub Camellia sinensis, is known for its antioxidant and anti-carcinogenic effects. However, its effects on male gonadal functions have not been explored adequately and the present investigation has been undertaken to evaluate the effect of green tea extract on gonads of adult male albino rats. Results of in vivo studies showed that green tea extract (GTE) at mild (1.25 g%, identical to 5 cups of tea/day), moderate (2.5 g%, identical to 10 cups of tea/day) and high (5.0 g%, identical to 20 cups of tea/day) doses, for a period of 26 days, altered morphology and histology of testis and accessory sex organs. A significant dose-dependent decrease in the sperm counts, inhibited activities of testicular delta(5)3beta-and 17beta-hydroxysteroid dehydrogenase (delta5-3beta3-HSD and 17beta3-HSD respectively) and decreased serum testosterone level were noticed. Significant increase in serum LH level was observed after moderate and high doses; serum FSH level also increased but not significantly. Histopathological examination showed inhibition of spermatogenesis evidenced by preferential loss of matured and elongated spermatids. Results of this study showed that GTE at relatively high dose may cause impairment of both the morphological and normal functional status of testis in rodents and thus its consumption at relatively high doses raises concern on male reproductive function in spite of its other beneficial effects.

  19. Cell-Free Expression and In Situ Immobilization of Parasite Proteins from Clonorchis sinensis for Rapid Identification of Antigenic Candidates

    PubMed Central

    Ju, Jung Won; Kim, Ho-Cheol; Shin, Hyun-Il; Kim, Yu Jung; Kim, Dong-Myung

    2015-01-01

    Progress towards genetic sequencing of human parasites has provided the groundwork for a post-genomic approach to develop novel antigens for the diagnosis and treatment of parasite infections. To fully utilize the genomic data, however, high-throughput methodologies are required for functional analysis of the proteins encoded in the genomic sequences. In this study, we investigated cell-free expression and in situ immobilization of parasite proteins as a novel platform for the discovery of antigenic proteins. PCR-amplified parasite DNA was immobilized on microbeads that were also functionalized to capture synthesized proteins. When the microbeads were incubated in a reaction mixture for cell-free synthesis, proteins expressed from the microbead-immobilized DNA were instantly immobilized on the same microbeads, providing a physical linkage between the genetic information and encoded proteins. This approach of in situ expression and isolation enables streamlined recovery and analysis of cell-free synthesized proteins and also allows facile identification of the genes coding antigenic proteins through direct PCR of the microbead-bound DNA. PMID:26599101

  20. A review of the control of clonorchiasis sinensis and Taenia solium taeniasis/cysticercosis in China.

    PubMed

    Wu, Wei; Qian, Xiaohua; Huang, Yixin; Hong, Qingbiao

    2012-11-01

    Clonorchiasis sinensis and Taenia solium taeniasis/cysticercosis are major foodborne parasitoses. Clonorchiasis sinensis is actively transmitted in some areas of China, Korea, Russia, Vietnam, etc. Currently, it is estimated that more than 200 million people are at risk of infection, 15-20 million people are infected, and 1.5-2 million show symptoms or complications. In China, it is relatively heavily transmitted in Zhujiang River Delta, including Hong Kong and Macao, and Northeast China, where many Korean people live. The transmission is related to the unhealthy habits of residents who like to have raw fish or half-raw fish. The infection of Clonorchis sinensis could result in serious liver and biliary system damages, and chronic cases may induce liver and bile duct cancers. T. solium taeniasis/cysticercosis is distributed around the world except the areas where the residents have a taboo against pork for religious reasons. Recent years, the urban inhabitants infected with T. solium/Cysticercus are increasing in China. T. solium results in intestinal diseases, and cysticercosis is a very serious disease, especially nervous system cysticercosis. Its symptoms include headache, epilepsy, sudden death, etc. Health education and health promotion, environmental reconstruction, and chemotherapy are the main control measures for these diseases. Through several decades of efforts in China, the achievements of control of clonorchiasis and T. solium taeniasis/cysticercosis are great. For example, in one of the main clonorchiasis-endemic provinces, Shandong Province, clonorchiasis has been controlled. In 31 T. solium taeniasis/cysticercosis-endemic counties of Henan Province, through a 6-year control program, the decline rates of T. solium taeniasis and cysticercosis were 90.8 and 96.8 %, respectively. This paper reviews the researches on the control of clonorchiasis and T. solium taeniasis/cysticercosis in China past decades so as to provide references for other countries

  1. [A study on the fine structure of Clonorchus sinensis, a liver Fluke V. The mature spermatozoa

    PubMed

    Jeong, Kye Heon; Rim, Han Jong

    1984-06-01

    An ultrastructural study on the mature spermatozoa of Clonorchis sinensis was carried out. For this study, the liver flukes were collected from the livers of rabbits and rats artificially infected with the metacercariae obtained from the fresh water fish, Pseudorasbora parva. Six-month old worms were used. The collected liver flukes were washed with 0.85 percent saline solution and then immediately moved to cold 2 percent glutaraldehyde buffered with 0.l M Millonig's phosphate buffer (pH 7.4). The materials were dissected into appropriate pieces in the fixative about 30 minutes after beginning of the fixation. Two hours later the materials containing the seminal receptacle were rinsed several times with the buffer and were secondarily fixed with cold, buffered 1 percent osmium tetroxide for 2 hours. The fully fixed tissue blocks were dehydrated in a series of graded concentrations of acetone and were embedded in Epon 812 mixture. Thin sections obtained from LKB-5 ultramicrotome were stained with uranyl acetate and Reynold's lead citrate. Observations of the sections were carried out with JEM-100CX II electron microscope. In general, the mature sperm was long thread-like form with a sickle-shaped head. According to the longitudinal sectioned view of the sperm tail, the nucleus seemed to be spirally coiled and run a little far along the tail. The acrosome was not observed. The cytoplasm of the tail was biflagellated as usual in trematodes. Unlike other platyhelminth spermatozoa, the sperm tail of Clonorchis sinensis showed the [9+2] pattern in the microtubular arrangement. The mitochondria with poorly developed cristae were observed throughout the middle piece. The middle piece of the tail showed dull ladder or triangular shapes with the two flagella at the bottom. But, the principal piece of the tail was slightly flattened cylindrical shape with two flagella within the cytoplasm. The end piece was uniflagellated. It was not clearly identified whether the end piece

  2. Digestive enzyme activities in mudskipper Boleophthalmus pectinirostris and Chinese black sleeper Bostrichthys sinensis

    NASA Astrophysics Data System (ADS)

    Wu, Renxie; Hong, Wanshu; Zhang, Qiyong

    2010-07-01

    The mudskipper Boleophthalmus pectinirostris and Chinese black sleeper Bostrichthys sinensis occupy the intertidal zone. However, both species have their own unique diet. The former is an herbivore and the latter is a carnivore. In order to reveal the relationship between digestive enzyme activities and diets in the two species, the activities of protease (P), non-specific bile salt-activated lipase (BAL) and α-amylase (A) were determined in the stomach and intestine of adult mudskipper B. pectinirostris and Chinese black sleeper B. sinensis. The results showed that the activities of protease, BAL and α-amylase in the intestine of B. pectinirostris were significantly ( P<0.05) higher than those in the stomach. In B. sinensis, gastric protease activity was not different from the intestinal protease ( P>0.05), while BAL and α-amylase activities of the intestine were significantly ( P<0.05) higher than those of the stomach. The activity of gastric protease in B. sinensis was significantly ( P<0.05) higher than that in B. pectinirostris, while the activities of intestinal protease were not different between the two fish species ( P>0.05). BAL activities of the stomach and intestine in B. sinensis were significantly ( P<0.05) higher than those in B. pectinirostris, while α-amylase activities of the stomach and intestine in B. pectinirostris were significantly ( P<0.05) higher than those in B. sinensis. The ratios of P/BAL, A/P and A/BAL of the digestive tract in B. pectinirostris were 1.5, 107.3 and 158.6, respectively; and those in B. sinensis were 0.2, 1.6 and 0.2, respectively. It can be concluded that food digestion in the adult B. pectinirostris is mainly carried out in the intestine, whereas in the adult B. sinensis it is initiated in the stomach and finishes in the intestine. The activities of BAL and α-amylase in B. pectinirostris and B. sinensis are well correlated with their diets. However, a clear-cut correlation between protease activity and diets is

  3. HISTOPATHOLOGY OF GASTRIC WALL IN CHINESE ALLIGATOR ALLIGATOR SINENSIS INFECTED WITH ORTLEPPASCARIS SINENSIS (NEMATODA: ASCARIDOIDEA).

    PubMed

    Zhao, Jinhong; Wang, Shaosheng; Tu, Genjun; Zhou, Yongkang; Wu, Xiaobing; Li, Chaopin

    2015-09-01

    Crocodiles are susceptible to infection with a wide array of external and internal gastrointestinal helminths, yet little is known on the histopathology following infection or the effects of these parasites. The present study was aimed at evaluating the impact of infection by Ortleppascaris sinensis (Nematoda: Ascaridoidea) on the stomach of captive Alligator sinensis. The histological examination of the stomach revealed presence of superficial ulcer in mucous layer and granulomatous inflammation in submucous layer at entire gastric walls of the Alligator sinensis. Our findings also confirm that development of Ortleppascaris sinensis is in close association with the wall of the stomach.

  4. The Chemical Constituents and Pharmacological Actions of Cordyceps sinensis

    PubMed Central

    Liu, Yi; Wang, Jihui; Wang, Wei; Zhang, Hanyue; Zhang, Xuelan; Han, Chunchao

    2015-01-01

    Cordyceps sinensis, also called DongChongXiaCao (winter worm, summer grass) in Chinese, is becoming increasingly popular and important in the public and scientific communities. This study summarizes the chemical constituents and their corresponding pharmacological actions of Cordyceps sinensis. Many bioactive components of Cordyceps sinensis have been extracted including nucleoside, polysaccharide, sterol, protein, amino acid, and polypeptide. In addition, these constituents' corresponding pharmacological actions were also shown in the study such as anti-inflammatory, antioxidant, antitumour, antiapoptosis, and immunomodulatory actions. Therefore can use different effects of C. sinensis against different diseases and provide reference for the study of Cordyceps sinensis in the future. PMID:25960753

  5. Chemistry and Pharmacology of Citrus sinensis.

    PubMed

    Favela-Hernández, Juan Manuel J; González-Santiago, Omar; Ramírez-Cabrera, Mónica A; Esquivel-Ferriño, Patricia C; Camacho-Corona, María del Rayo

    2016-02-22

    Presently the search for new drugs from natural resources is of growing interest to the pharmaceutical industry. Natural products have been the source of new drugs since ancient times. Plants are a good source of secondary metabolites which have been found to have beneficial properties. The present study is a review of the chemistry and pharmacology of Citrus sinensis. This review reveals the therapeutic potential of C. sinensis as a source of natural compounds with important activities that are beneficial for human health that could be used to develop new drugs.

  6. First insight into CD59-like molecules of adult Fasciola hepatica.

    PubMed

    Shi, Yunliang; Toet, Hayley; Rathinasamy, Vignesh; Young, Neil D; Gasser, Robin B; Beddoe, Travis; Huang, Weiyi; Spithill, Terry W

    2014-09-01

    The present study focussed on investigating CD59-like molecules of Fasciola hepatica. A cDNA encoding a CD59-like protein (termed FhCD59-1) identified previously in the membrane fraction of the F. hepatica tegument was isolated. This homologue was shown to encode a predicted open reading frame (ORF) of 122 amino acids (aa) orthologous to human CD59 with a 25 aa signal peptide, a mature protein containing 10 cysteines and a conserved CD59/Ly-6 family motif "CCXXXXCN". An analysis of cDNAs from two different adult specimens of F. hepatica revealed seven variable types of FhCD59-1 sequences, designated FhCD59-1.1 to FhCD59-1.7, which had 94.3-99.7% amino acid sequence identity upon pairwise comparison. Molecular modeling of FhCD59-1.1 with human CD59 confirmed the presence of the three-finger protein domain found in the CD59 family and predicted three disulphide bonds in the F. hepatica sequence. The interrogation of F. hepatica databases identified two additional sequences, designated FhCD59-2 and FhCD59-3, which had only 23.4-29.5% amino acid identity to FhCD59-1.1. Orthologues of the inferred CD59 protein sequences of F. hepatica were also identified in other flatworms, including Fasciola gigantica, Fascioloides magna, Schistosoma haematobium, Schistosoma japonicum, Schistosoma mansoni, Clonorchis sinensis, Opisthorchis viverrini, Taenia solium, Echinococcus granulosus and the free living Schmidtea mediterannea. The results revealed a considerable degree of sequence complexity in the CD59-like sequence families in F. hepatica and flatworms. Phylogenetic analysis of CD59-like aa sequences from F. hepatica and flatworms showed that FhCD59-2 clustered with the known surface-associated protein SmCD59-2 of S. mansoni. Relatively well-supported clades specific to schistosomes, fasciolids and opisthorchiids were identified. The qPCR analysis of gene transcription showed that the relative expression of these 3 FhCD59-like sequences varied by 11-47-fold during fluke

  7. De novo assembly, characterization and annotation for the transcriptome of Sarcocheilichthys sinensis

    PubMed Central

    Zhu, Chuankun; Pan, Zhengjun; Wang, Hui; Chang, Guoliang; Wu, Nan; Ding, Huaiyu

    2017-01-01

    The Chinese lake gudgeon Sarcocheilichthys sinensis is a small cyprinid fish with great aquaculture potential both for its edible and ornamental values. Nevertheless, available genomic and transcriptomic information for this fish is extremely deficient. In this study, a normalized cDNA library was constructed using 13 mixed tissues of an adult male S. sinensis, and was sequenced by the Illumina HiSeq2500 platform. De novo assembly was performed using 38,911,511 obtained clean reads, and a total of 147,282 unigenes with an average length of 900 bp were finally achieved. 96.2% of these unigenes were annotated in 9 public databases, and 16 segments of growth-related genes were identified for future studies. In addition, 28,493 unigenes were assigned to 61 subcategories of Gene Ontology (GO), and 10,483 unigenes were assigned to 25 categories of Cluster of Orthologous Group (COG). Moreover, 14,943 unigenes were classified into 225 pathways of the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. A total of 30,666 microsatellites were detected from 17,627 unigenes with an average distribution density of 1:2405 bp. This transcriptome data set will be valuable for researches on discovery, expression and evolution on genes of interest. Meanwhile, the identified microsatellites would be useful tools for genetic and genomic studies in S. sinensis. PMID:28196101

  8. Random repeated cross sectional study on breeding site characterization of Anopheles sinensis larvae in distinct villages of Yongcheng City, People's Republic of China

    PubMed Central

    2012-01-01

    Background Characterizing the breeding site of Anopheles sinensis is of major importance for the transition from malaria control to elimination in China. However, little information is available especially regarding the characteristics and influencing factors of breeding sites of An. sinensis in Yongcheng City, a representative region of unstable malaria transmission in the Huang-Huai River region of central China. The aims of this study were to determine the breeding site characteristics of An. sinensis and related environmental and physicochemical parameters, to find out which breeding site characteristics could best explain the presence of An. sinensis larvae, and to determine whether the breeding habit of An. sinensis has changed or not. Methods Random repeated cross sectional study was undertaken in six villages of the Yongcheng city characterized by different levels of the historical incidence of P. vivax malaria. The potential breeding sites of An. sinensis larvae in each village were examined twice per month both in the household courtyards and the village surroundings. The larval sampling was done by the standard dipping method. Some important breeding site characterizations were recorded and characterized. The anopheline mosquito larvae and emerged adults were identified to the species level morphologically and to sub-species by the ribosomal DNA PCR technique. Chi-square analysis and logistic regression analysis were applied to determine the importance of factors for explaining the presence or absence of An. sinensis larvae. Results According to the ribosomal DNA PCR assay, all sampled anopheline mosquito larvae and emerged adults belonged to An. sinensis. Only 3 containers that were sampled from the household courtyards were found to contain An. sinensis larvae. There were no differences in the species composition of mosquito larvae among containers that contained water in the household courtyards (P > 0.05). An. sinensis larvae were shown to be present

  9. An Experimental Novel Study: Angelica sinensis Prevents Epidural Fibrosis in Laminectomy Rats via Downregulation of Hydroxyproline, IL-6, and TGF-β1

    PubMed Central

    Kong, Xiaohong; Zhao, Xuechao; Zhou, Xianhu; Su, Yanhua; Sharma, Hari S.; Feng, Shiqing

    2013-01-01

    With laminectomy being widely accepted as the treatment for lumbar disorders, epidural fibrosis (EF) is a common complication for both the patients and the surgeons alike. Currently, EF is thought to cause recurrent postoperative pain after laminectomy or after discectomy. Angelica sinensis is a traditional Chinese medicine which has shown anti-inflammatory, antifibrotic, and antiproliferative properties. The object of this study was to investigate the effects of Angelica sinensis on the prevention of post-laminectomy EF formation in a rat model. A controlled double-blinded study was conducted in sixty healthy adult Wistar rats that underwent laminectomy at the L1-L2 levels. They were divided randomly into 3 groups according to the treatment method, with 20 in each group: (1) Angelica sinensis treatment group, (2) saline treatment group, and (3) sham group (laminectomy without treatment). All rats were euthanized humanely 4 weeks after laminectomy. The hydroxyproline content, Rydell score, vimentin cells density, fibroblasts density, inflammatory cells density, and inflammatory factors expressions all suggested better results in Angelica sinensis group than the other two groups. Topical application of Angelica sinensis could inhibit fibroblasts proliferation and TGF-β1 and IL-6 expressions and prevent epidural scar adhesion in postlaminectomy rat model. PMID:24069047

  10. Draft Genome Sequence of Paecilomyces hepiali, Isolated from Cordyceps sinensis.

    PubMed

    Yu, Yi; Wang, Wenting; Wang, Linping; Pang, Fang; Guo, Lanping; Song, Lai; Liu, Guiming; Feng, Chengqiang

    2016-07-07

    Paecilomyces hepiali is an endoparasitic fungus that commonly exists in the natural Cordyceps sinensis Here, we report the draft genome sequence of P. hepiali, which will facilitate the exploitation of medicinal compounds produced by the fungus.

  11. Draft Genome Sequence of Paecilomyces hepiali, Isolated from Cordyceps sinensis

    PubMed Central

    Yu, Yi; Wang, Wenting; Wang, Linping; Pang, Fang; Guo, Lanping; Song, Lai

    2016-01-01

    Paecilomyces hepiali is an endoparasitic fungus that commonly exists in the natural Cordyceps sinensis. Here, we report the draft genome sequence of P. hepiali, which will facilitate the exploitation of medicinal compounds produced by the fungus. PMID:27389266

  12. Cordyceps collected from Bhutan, an appropriate alternative of Cordyceps sinensis.

    PubMed

    Wu, Ding-Tao; Lv, Guang-Ping; Zheng, Jian; Li, Qian; Ma, Shuang-Cheng; Li, Shao-Ping; Zhao, Jing

    2016-11-22

    Natural Cordyceps collected in Bhutan has been widely used as natural Cordyceps sinensis, an official species of Cordyceps used as Chinese medicines, around the world in recent years. However, whether Cordyceps from Bhutan could be really used as natural C. sinensis remains unknown. Therefore, DNA sequence, bioactive components including nucleosides and polysaccharides in twelve batches of Cordyceps from Bhutan were firstly investigated, and compared with natural C. sinensis. Results showed that the fungus of Cordyceps from Bhutan was C. sinensis and the host insect belonged to Hepialidae sp. In addition, nucleosides and their bases such as guanine, guanosine, hypoxanthine, uridine, inosine, thymidine, adenine, and adenosine, as well as compositional monosaccharides, partial acid or enzymatic hydrolysates, molecular weights and contents of polysaccharides in Cordyceps from Bhutan were all similar to those of natural C. sinensis. All data suggest that Cordyceps from Bhutan is a rational alternative of natural C. sinensis, which is beneficial for the improvement of their performance in health and medicinal food areas.

  13. Cordyceps collected from Bhutan, an appropriate alternative of Cordyceps sinensis

    PubMed Central

    Wu, Ding-Tao; Lv, Guang-Ping; Zheng, Jian; Li, Qian; Ma, Shuang-Cheng; Li, Shao-Ping; Zhao, Jing

    2016-01-01

    Natural Cordyceps collected in Bhutan has been widely used as natural Cordyceps sinensis, an official species of Cordyceps used as Chinese medicines, around the world in recent years. However, whether Cordyceps from Bhutan could be really used as natural C. sinensis remains unknown. Therefore, DNA sequence, bioactive components including nucleosides and polysaccharides in twelve batches of Cordyceps from Bhutan were firstly investigated, and compared with natural C. sinensis. Results showed that the fungus of Cordyceps from Bhutan was C. sinensis and the host insect belonged to Hepialidae sp. In addition, nucleosides and their bases such as guanine, guanosine, hypoxanthine, uridine, inosine, thymidine, adenine, and adenosine, as well as compositional monosaccharides, partial acid or enzymatic hydrolysates, molecular weights and contents of polysaccharides in Cordyceps from Bhutan were all similar to those of natural C. sinensis. All data suggest that Cordyceps from Bhutan is a rational alternative of natural C. sinensis, which is beneficial for the improvement of their performance in health and medicinal food areas. PMID:27874103

  14. Morphological and molecular characterization of Ortleppascaris sinensis sp. nov. (Nematoda: Ascaridoidea) from the Chinese alligator Alligator sinensis.

    PubMed

    Zhao, J H; Wang, S S; Tu, G J; Zhou, Y K; Wu, X B

    2016-05-01

    A new nematode species, Ortleppascaris sinensis sp. nov. (Ascaridoidea), is described from specimens found in the stomach and intestine of the Chinese alligator Alligator sinensis Fauvel, 1879 (Crocodilian: Alligatoridae) in the National Nature Reserve of Chinese Alligator (Chinese Crocodile Lake) in Anhui Province, China. This is the first description of O. sinensis sp. nov. in both China and this crocodile host, increasing its distribution in South Asia as well as expanding the number of helminths known to infect this crocodile. The detailed description of O. sinensis sp. nov., based on light and scanning electron microscopic examination, provides new taxonomic data for this species, and we also report sequences of the internal transcribed spacers (ITS), small subunit DNA segments (18S) and the cytochrome oxidase I (COI) gene.

  15. Antihyperlipidemic effects of Citrus sinensis, Citrus paradisi, and their combinations

    PubMed Central

    Mallick, Neelam; Khan, Rafeeq Alam

    2016-01-01

    Objective: Hyperlipidemia, extensively contributes in the progression of coronary heart diseases and atherosclerosis, but may be managed through alterations in the nutritional pattern. Several studies show that diet rich in polyphenols and antioxidants have antiatherogenic effects. Citrus sinensis and Citrus paradisi are widely known for health benefits and have found to produce antioxidant, anti-inflammatory, and hypolipidemic effects, hence current research was planned to determine the hypolipidemic effects of C. sinensis and C. paradisi in rats receiving diet rich in cholesterol. Materials and Methods: All rats were divided into 11 groups each comprising 10 animals: Normal control group and hyperlipidemic control. C. sinensis treated three groups, C. paradisi treated three groups, C. sinensis and C. paradisi combination treated two groups, and group treated atorvastatin. All rats in the respective groups were treated orally with sterile water, juices, and standard drug for 8 weeks and lipid profile was estimated at the end of dosing. Results: Cholesterol, triglycerides (TGs), and low-density lipoprotein (LDL) were decreased at all the three doses of C. sinensis and C. paradisi but rise in high-density lipoprotein (HDL) was only significant at 8 ml/kg, and 0.3 ml/kg, respectively. Animals received the combination doses of C. sinensis and C. paradisi also showed a highly significant fall in cholesterol, LDL, and TGs, however HDL level was significantly elevated by SPJ-2 combination. Conclusion: Results suggest that C. sinensis and C. paradisi possess antihyperlipidemic activity due to phytochemicals and other essential nutrients, hence may serve as cardioprotective by preventing thrombosis. PMID:27134462

  16. Larvicidal, pupicidal, repellent and adulticidal activity of Citrus sinensis orange peel extract against Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus (Diptera: Culicidae).

    PubMed

    Murugan, Kadarkarai; Mahesh Kumar, Palanisamy; Kovendan, Kalimuthu; Amerasan, Duraisamy; Subrmaniam, Jayapal; Hwang, Jiang-Shiou

    2012-10-01

    Mosquitoes are the carriers of severe and well-known illnesses such as malaria, arboviral encephalitis, dengue fever, chikunguniya fever, West Nile virus and yellow fever. These diseases produce significant morbidity and mortality in humans and livestock around the world. The present study explored the effects of orange peel ethanol extract of Citrus sinensis on larvicidal, pupicidal, repellent and adulticidal activity against Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus. The orange peel material was shade dried at room temperature and powdered coarsely. From orange peel, 300 g powdered was macerated with 1 L of ethanol sequentially for a period of 72 h each and filtered. The yields of the orange peel ethanol crude extract of C. sinensis 13.86 g, respectively. The extracts were concentrated at reduced temperature on a rotary vacuum evaporator and stored at a temperature of 4 °C. The larvicidal, pupicidal and adult mortality was observed after 24 h of exposure; no mortality was observed in the control group. For C. sinensis, the median lethal concentration values (LC(50)) observed for the larvicidal and pupicidal activities against mosquito vector species A. stephensi first to fourth larval instars and pupae were 182.24, 227.93, 291.69, 398.00 and 490.84 ppm; A. aegypti values were 92.27, 106.60, 204.87, 264.26, 342.45, 436.93 and 497.41 ppm; and C. quinquefasciatus values were 244.70, 324.04, 385.32, 452.78 and 530.97 ppm, respectively. The results of maximum repellent activity were observed at 450 ppm in ethanol extracts of C. sinensis and the mean complete protection time ranged from 150 to 180 min was tested. The ethanol extract of C. sinensis showed 100% repellency in 150 min and showed complete protection in 90 min at 350 ppm against A. stephensi, A. aegypti and C. quinquefasciatus, respectively. The adult mortality was found in ethanol extract of C. sinensis with the LC(50) and LC(90) values of 272.19 and 457.14 ppm, A. stephensi; 289.62 and

  17. Larvicidal and irritant activities of hexane leaf extracts of Citrus sinensis against dengue vector Aedes aegypti L.

    PubMed Central

    Warikoo, Radhika; Ray, Ankita; Sandhu, Jasdeep Kaur; Samal, Roopa; Wahab, Naim; Kumar, Sarita

    2012-01-01

    Objective To assess the larvicidal and irritant activities of the hexane extracts of leaves of Citrus sinensis (C. sinensis) against the early fourth instars and female adults of Aedes aegypti (Ae. aegypti). Methods The larvicidal potential of the prepared leaf extract was evaluated against early fourth instar larvae of Ae. aegypti using WHO protocol. The mortality counts were made after 24 h and LC50 and LC90 values were calculated. The efficacy of extract as mosquito irritant was assessed by contact irritancy assays. Extract-impregnated paper was placed on a glass plate over which a perspex funnel with a hole on the top was kept inverted. Single female adult, 3-day old unfed/blood-fed, was released inside the funnel. After 3 min of acclimatization time, the time taken for the first take-off and total number of flights undertaken during 15 min were scored. Results The citrus leaf extracts from hexane possessed moderate larvicidal efficiency against dengue vector. The bioassays resulted in an LC50 and LC90 value of 446.84 and 1 370.96 ppm, respectively after 24 h of exposure. However, the extracts were proved to be remarkable irritant against adults Ae. aegypti, more pronounced effects being observed on blood-fed females than unfed females. The extract-impregnated paper was thus proved to be 7–11 times more irritable as compared with the control paper. Conclusions The hexane extracts from C. sinensis leaves are proved to be reasonably larvicidal but remarkably irritant against dengue vector. Further studies are needed to identify the possible role of extract as adulticide, oviposition deterrent and ovicidal agent. The isolation of active ingredient from the extract could help in formulating strategies for mosquito control. PMID:23569887

  18. Effects of temperature and salinity on the development of the amphipod crustacean Eogammarus sinensis

    NASA Astrophysics Data System (ADS)

    Xue, Suyan; Fang, Jianguang; Zhang, Jihong; Jiang, Zengjie; Mao, Yuze; Zhao, Fazhen

    2013-09-01

    The amphipod crustacean Eogammarus sinensis has useful features that make it suitable for use in the aquaculture of fish and large decapod crustaceans. In this study, we investigated the effects of temperature and salinity on the development, fecundity, survival, and growth rate of E. sinensis. The results show that temperature significantly affected E. sinensis development, but salinity. As temperature increased, the duration of E. sinensis embryonic development decreased. Fecundity was affected significantly by temperature and the combination of temperature and salinity, but by salinity alone. In addition, high temperatures accelerated E. sinensis juvenile growth rates, whereas high salinity reduced it. Therefore, our data suggest that E. sinensis tolerates a wide range of salinities and that temperature has more significant effects than salinity on the embryonic development, fecundity, and growth of E. sinensis. Our results shall be useful for mass production of this species for use in aquaculture.

  19. Identification of Chinese medicinal fungus Cordyceps sinensis by depth-profiling mid-infrared photoacoustic spectroscopy

    NASA Astrophysics Data System (ADS)

    Du, Changwen; Zhou, Jianmin; Liu, Jianfeng

    2017-02-01

    With increased demand for Cordyceps sinensis it needs rapid methods to meet the challenge of identification raised in quality control. In this study Cordyceps sinensis from four typical natural habitats in China was characterized by depth-profiling Fourier transform infrared photoacoustic spectroscopy. Results demonstrated that Cordyceps sinensis samples resulted in typical photoacoustic spectral appearance, but heterogeneity was sensed in the whole sample; due to the heterogeneity Cordyceps sinensis was represented by spectra of four groups including head, body, tail and leaf under a moving mirror velocity of 0.30 cm s- 1. The spectra of the four groups were used as input of a probabilistic neural network (PNN) to identify the source of Cordyceps sinensis, and all the samples were correctly identified by the PNN model. Therefore, depth-profiling Fourier transform infrared photoacoustic spectroscopy provides novel and unique technique to identify Cordyceps sinensis, which shows great potential in quality control of Cordyceps sinensis.

  20. Identification of Chinese medicinal fungus Cordyceps sinensis by depth-profiling mid-infrared photoacoustic spectroscopy.

    PubMed

    Du, Changwen; Zhou, Jianmin; Liu, Jianfeng

    2017-02-15

    With increased demand for Cordyceps sinensis it needs rapid methods to meet the challenge of identification raised in quality control. In this study Cordyceps sinensis from four typical natural habitats in China was characterized by depth-profiling Fourier transform infrared photoacoustic spectroscopy. Results demonstrated that Cordyceps sinensis samples resulted in typical photoacoustic spectral appearance, but heterogeneity was sensed in the whole sample; due to the heterogeneity Cordyceps sinensis was represented by spectra of four groups including head, body, tail and leaf under a moving mirror velocity of 0.30cms(-1). The spectra of the four groups were used as input of a probabilistic neural network (PNN) to identify the source of Cordyceps sinensis, and all the samples were correctly identified by the PNN model. Therefore, depth-profiling Fourier transform infrared photoacoustic spectroscopy provides novel and unique technique to identify Cordyceps sinensis, which shows great potential in quality control of Cordyceps sinensis.

  1. Pharmacological effects of Radix Angelica Sinensis (Danggui) on cerebral infarction

    PubMed Central

    2011-01-01

    Radix Angelica Sinensis, the dried root of Angelica sinensis (Danggui), is a herb used in Chinese medicine to enrich blood, promote blood circulation and modulate the immune system. It is also used to treat chronic constipation of the elderly and debilitated as well as menstrual disorders. Research has demonstrated that Danggui and its active ingredients, as anti-arthrosclerotic, anti-hypertensive, antioxidant anti-inflammatory agents which would limit platelet aggregation, are effective in reducing the size of cerebral infarction and improving neurological deficit scores. PMID:21867503

  2. The occurrence and distribution of Tuckerella japonica (Acari: Tuckerellidae) on tea bushes, Camellia sinensis and C. assamica, in Alabama, Georgia and South Carolina, USA.

    PubMed

    Childers, Carl C; Ebert, Timothy A; Rogers, Michael E; Shepard, Merle

    2016-08-01

    Adults, immatures and eggs of Tuckerella japonica (Ehara) were collected from unknown clones or varieties of Camellia sinensis (L.) O. Kuntze tea bushes in the Clemson University Farm, Coastal Research and Education Center, Charleston, South Carolina; from Assam hybrids in The Caw Caw Nature Preserve in Ravenel, SC; from C. sinensis and C. assamica (Masters) in the Charleston Tea Plantation on Wadmalaw Island, SC; C. sinensis in the Fairhope Tea Plantation in Fairhope, Alabama; and from C. sinensis 'Rosea' and a C. sinensis and C. assamica hybrid in Savannah and Ellabell, Georgia, between 1994 and 2015. This mite was consistently collected from 1-, 2- and 3+-year-old wood of tea plants with significantly greater numbers collected from 2-year-old wood. All stages of the mite were found within longitudinally split areas of the wood where underlying green bark tissues were exposed. As 1-year-old wood matured, there was increased splitting of the bark with increased mite presence. Mature green fruit (= developing seed pods) of tea were also frequented by T. japonica between June-July and October and their numbers were no greater than those on 1- or 3+-year wood. When the fruit were small (March-May) or as they hardened in late fall, they were not suitable feeding sites for this mite. Very few T. japonica were collected from 50 mature, inner or outer leaf samples with none usually found. Tuckerella japonica has multiple, overlapping generations and occurs on tea throughout the year in Alabama, Georgia and South Carolina, USA.

  3. Detection of drug-resistant Klebsiella pneumoniae in Chinese hares (Lepus sinensis).

    PubMed

    Du, Yingchun; Luo, Jing; Wang, Chengmin; Wen, Qingna; Duan, Mingxing; Zhang, Hong; He, Hongxuan

    2014-01-01

    We investigated an outbreak of acute pneumonia among adult Chinese hares (Lepus sinensis) and diarrhea among juvenile hares in Hebei Province, China, in 2012. Diagnosis was based on necropsy examination, microbial characteristics, biochemical identification, and nucleotide sequence analysis. The isolated bacteria from tissue samples of dead hares were identified as Klebsiella pneumoniae ssp. pneumoniae (K. pneumoniae). This K. pneumoniae was resistant to the antimicrobials imipenem, meropenem, penicillin, and vancomycin, but was highly sensitive to cefepime, cotrimoxazole, and enrofloxacin. Klebsiella pneumoniae is an important opportunistic pathogen, which often causes nosocomial infections in immunocompromised patients. However, the emergence of drug-resistant K. pneumoniae in hares indicates the existence of increasing risk of pathogen transmission between humans and wildlife. Given the close association between wildlife, livestock, and humans, it is important to identify epidemiologic factors associated with infection in these hares to minimize the risk of K. pneumoniae transmission.

  4. [Cordyceps sinensis, a fungi used in the Chinese traditional medicine].

    PubMed

    Illana Esteban, Carlos

    2007-12-31

    Cordyceps sinensis (Berk.) Sacc. is an ascomycete fungus known in China since antiquity, which is still being used today. A summary, showing relevant papers about this fungus, regarding habitat, history, marketing, consumption, nomenclature, pharmacological composition, culture and medical use, is presented.

  5. Investigation and analysis of microbiological communities in natural Ophiocordyceps sinensis.

    PubMed

    Xia, Fei; Liu, Yan; Shen, Guang-Rong; Guo, Lian-Xian; Zhou, Xuan-Wei

    2015-02-01

    Ophiocordyceps sinensis is a fungus that parasitizes caterpillars, and more than 30 species of filamentous fungi have been isolated from its fruiting body. However, its microbiological diversity remains unclear. Based on the clone library and quantitative PCR techniques, the bacterial flora and mycobiota of 3 different samples (larva, stromata/sclerotia, and surface soil) from natural O. sinensis specimens were investigated using primer sets that targeted the 16S rRNA gene and internal transcribed spacer region of ribosomal DNA. The results showed that the abundance of bacterial and fungal communities in the soil attached to the surface of O. sinensis was (6.4 ± 1.4) × 10(6) and (6.0 ± 0.3) × 10(7) copies/g dry matter, respectively, which was the highest compared with that in the larva and stromal samples. The main groups of bacteria in the O. sinensis samples were Proteobacteria and Actinobacteria, while Ascomycota was the most dominant fungal group in the 3 samples. At the genus level, Geomyces, Phoma, and Trichocladium were the dominant genera in the larval sample, while Geomyces and Cladosporium were the dominant genera in the stromal sample. In conclusion, a great number of bacterial and fungal species were present in naturally occurring O. sinensis specimens, and there was a high diversity of bacterial and fungal communities. These findings contribute to the understanding of the bacterial and fungal community structure of this valuable medicinal fungus and lay the foundation for the future discovery of new medicinal microorganism resources.

  6. Habitats and Distribution of Anopheles Sinensis and Associated Anopheles Hyrcanus Group in Japan

    DTIC Science & Technology

    2005-01-01

    SCIENTIFIC NOTE HABITATS AND DISTRffiUTION OF ANOPHELES SINENSIS AND ASSOCIATED ANOPHELES HYRCANUS GROUP IN JAPAN LEOPOLDO M. RUEDA.’ MASASHIRO IWAKAMI.2.b...collections were carried out in August 2002 and July 2003 in Japan . Anopheles sinensis of the Hyrcanus Group. Myzomyia Series of Anopheles. was the...Anopheles sinensis. Hyrcanus Group, Aedes, Clllex, Uranotaenia. Japan . Culicidae The Anopheles Hyrcanus Group consists of sev- eral species that are vectors

  7. Isolation, Culture and Characterization of Hirsutella sinensis Mycelium from Caterpillar Fungus Fruiting Body

    PubMed Central

    Ko, Yun-Fei; Liau, Jian-Ching; Lee, Chien-Sheng; Chiu, Chen-Yaw; Martel, Jan; Lin, Chuan-Sheng; Tseng, Shun-Fu; Ojcius, David M.; Lu, Chia-Chen; Lai, Hsin-Chih; Young, John D.

    2017-01-01

    The caterpillar fungus Ophiocordyceps sinensis (previously called Cordyceps sinensis) has been used for centuries in Asia as a tonic to improve health and longevity. Recent studies show that O. sinensis produces a wide range of biological effects on cells, laboratory animals and humans, including anti-fatigue, anti-infection, anti-inflammatory, antioxidant, and anti-tumor activities. In view of the rarity of O. sinensis fruiting bodies in nature, cultivation of its anamorph mycelium represents a useful alternative for large-scale production. However, O. sinensis fruiting bodies harvested in nature harbor several fungal contaminants, a phenomenon that led to the isolation and characterization of a large number of incorrect mycelium strains. We report here the isolation of a mycelium from a fruiting body of O. sinensis and we identify the isolate as O. sinensis’ anamorph (also called Hirsutella sinensis) based on multi-locus sequence typing of several fungal genes (ITS, nrSSU, nrLSU, RPB1, RPB2, MCM7, β-tubulin, TEF-1α, and ATP6). The main characteristics of the isolated mycelium, including its optimal growth at low temperature (16°C) and its biochemical composition, are similar to that of O. sinensis fruiting bodies, indicating that the mycelium strain characterized here may be used as a substitute for the rare and expensive O. sinensis fruiting bodies found in nature. PMID:28046129

  8. Quality control of Cordyceps sinensis, a valued traditional Chinese medicine.

    PubMed

    Li, S P; Yang, F Q; Tsim, Karl W K

    2006-08-28

    Cordyceps sinensis, a well-known and valued traditional Chinese medicine, is also called DongChongXiaCao (winter worm summer grass) in Chinese. It is commonly used to replenish the kidney and soothe the lung for the treatment of fatigue, night sweating, hyposexualities, hyperglycemia, hyperlipidemia, asthemia after severe illness, respiratory disease, renal dysfunction and renal failure, arrhythmias and other heart disease, and liver disease. As the rarity and upstanding curative effects of natural Cordyceps, several mycelial strains have been isolated from natural Cordyceps and manufactured in large quantities by fermentation technology, and they are commonly sold as health food products in Asia. In addition, some substitutes such as Cordyceps militaris also have been used and adulterants also confused the market. Therefore, quality control of C. sinensis and its products is very important to ensure their safety and efficacy. Herein, markers and analytical methods for quality control of Cordyceps were reviewed and discussed.

  9. Traditional uses and medicinal potential of Cordyceps sinensis of Sikkim

    PubMed Central

    Panda, Ashok Kumar; Swain, Kailash Chandra

    2011-01-01

    Cordyceps sinensis has been described as a medicine in old Chinese medical books and Tibetan medicine. It is a rare combination of a caterpillar and a fungus and found at altitudes above 4500m in Sikkim. Traditional healers and local people of North Sikkim recommend the mushroom, i.e., Yarsa gumba, Keera jhar (C. sinensis) for all diseases either as a single drug or combined with other herbs. The present study was undertaken to collect information regarding the traditional uses of cordyceps in Sikkim. It was found that most local folk healers/traditional healers use cordyceps for the treatment of 21 ailments. A modern literature search was carried out to assess whether the curative effects are valid or just blind faith of local people. Chemical constituents of cordyceps are given and pharmacological and biological studies reviewed. More mechanism-based and disease-oriented clinical studies are recommended. PMID:21731381

  10. Chemical Examination of Citrus sinensis Flavedo Variety Pineapple

    PubMed Central

    Rani, Geeta; Yadav, Lalita; Kalidhar, S. B.

    2009-01-01

    Phytochemical examination of Citrus sinensis flavedo var. Pineapple resulted in the isolation of six compounds characterized as tetracosane, ethyl pentacosanoate, tetratriacontanoic acid, tangertin, β-sitosteryl-β-D-glucoside and 3,5,4'-trihydroxy-7,3'-dimethoxy flavanone 3-O-β-glucoside. Of these 3,5,4'-trihydroxy-7,3'-dimethoxy flavanone 3-O-β-glucoside is a hitherto unreported compound. PMID:20376223

  11. Thesis Abstract Fermented milk elaborated with Camellia sinensis.

    PubMed

    Ribeiro, O A S; Silva, M I A; Boari, C A

    2016-05-13

    This study aimed to develop and to characterize fermented dairy beverage formulated with Camellia sinensis. The infusion was elaborated with the addiction of dehydrated leaves of C. sinensis in whey (1g/100g) which added in sweetened milk (10% sucrose w/w) coagulated by Streptococcus salivarius subspecies thermophilus and Lactobacillus delbrueckii subspecies bulgaricus in proportions of 10, 20, 30 and 40% (v/w). The control treatment consisted of yogurt added with sucrose (10% w/w). Analysis were performed to quantify dry mass, moisture, ash, protein, fat, sodium, acidity, total quantification of lactic acid bacteria, total antioxidant activity and viscosity at the initial time of production and at 15 and 30 days of storage. Chromatographic determination of volatile compounds and sensory tests of acceptance and consumption intention were conducted at the initial time of production. Dry matter content, moisture, ash and total count of lactic acid bacteria from fermented milk drink formulations were not significantly affected by the amount of infusion of C. sinensis. However, the content of protein, fat and sodium were significantly lower with the increase of the proportion of infusion incorporated into the product. Significant reduction in apparent viscosity occurs with the increase in the amount of infusion added. The total antioxidant activity of the formulations was significantly higher as higher were the amount of added infusion. The addition of infusion contributed to the diversification of volatile aroma and taste makers in the product. The formulation of fermented dairy drink with addition of 30% infusion C. sinensis was better evaluated in sensory tests, with greater acceptance and greater consumer intent of consumption.

  12. Anthocyanins from red flower tea (Benibana-cha), Camellia sinensis.

    PubMed

    Terahara, N; Takeda, Y; Nesumi, A; Honda, T

    2001-02-01

    Three anthocyanins were isolated from the leaves of red flower tea (Benibana-cha), Camellia sinensis, and their structures were determined by means of chemical and spectroscopic analyses. Two are the anthocyanins, delphinidin and cyanidin 3-O-beta-D-galactosides, respectively. Whereas the third, delphinidin 3-0-beta-D-(6-(E)-p-coumaryl)galactopyranoside. The anthocyanins were also contained in the flowers of Benibana-cha in different compositions.

  13. Complete mitochondrial genome of the medicinal fungus Ophiocordyceps sinensis

    PubMed Central

    Li, Yi; Hu, Xiao-Di; Yang, Rui-Heng; Hsiang, Tom; Wang, Ke; Liang, De-Quan; Liang, Fan; Cao, De-Ming; Zhou, Fan; Wen, Ge; Yao, Yi-Jian

    2015-01-01

    As part of a genome sequencing project for Ophiocordyceps sinensis, strain 1229, a complete mitochondrial (mt) genome was assembled as a single circular dsDNA of 157,510 bp, one of the largest reported for fungi. Conserved genes including the large and small rRNA subunits, 27 tRNA and 15 protein-coding genes, were identified. In addition, 58 non-conserved open reading frames (ncORFs) in the intergenic and intronic regions were also identified. Transcription analyses using RNA-Seq validated the expression of most conserved genes and ncORFs. Fifty-two introns (groups I and II) were found within conserved genes, accounting for 68.5% of the genome. Thirty-two homing endonucleases (HEs) with motif patterns LAGLIDADG (21) and GIY-YIG (11) were identified in group I introns. The ncORFs found in group II introns mostly encoded reverse transcriptases (RTs). As in other hypocrealean fungi, gene contents and order were found to be conserved in the mt genome of O. sinensis, but the genome size was enlarged by longer intergenic regions and numerous introns. Intergenic and intronic regions were composed of abundant repetitive sequences usually associated with mobile elements. It is likely that intronic ncORFs, which encode RTs and HEs, may have contributed to the enlarged mt genome of O. sinensis. PMID:26370521

  14. How cell wall complexity influences saccharification efficiency in Miscanthus sinensis

    PubMed Central

    De Souza, Amanda P.; Kamei, Claire L. Alvim; Torres, Andres F.; Pattathil, Sivakumar; Hahn, Michael G.; Trindade, Luisa M.; Buckeridge, Marcos S.

    2015-01-01

    The production of bioenergy from grasses has been developing quickly during the last decade, with Miscanthus being among the most important choices for production of bioethanol. However, one of the key barriers to producing bioethanol is the lack of information about cell wall structure. Cell walls are thought to display compositional differences that lead to emergence of a very high level of complexity, resulting in great diversity in cell wall architectures. In this work, a set of different techniques was used to access the complexity of cell walls of different genotypes of Miscanthus sinensis in order to understand how they interfere with saccharification efficiency. Three genotypes of M. sinensis displaying different patterns of correlation between lignin content and saccharification efficiency were subjected to cell wall analysis by quantitative/qualitative analytical techniques such as monosaccharide composition, oligosaccharide profiling, and glycome profiling. When saccharification efficiency was correlated negatively with lignin, the structural features of arabinoxylan and xyloglucan were found to contribute positively to hydrolysis. In the absence of such correlation, different types of pectins, and some mannans contributed to saccharification efficiency. Different genotypes of M. sinensis were shown to display distinct interactions among their cell wall components, which seem to influence cell wall hydrolysis. PMID:25908240

  15. How cell wall complexity influences saccharification efficiency in Miscanthus sinensis

    DOE PAGES

    De Souza, Amanda P.; Kamei, Claire L. Alvim; Torres, Andres F.; ...

    2015-04-23

    The production of bioenergy from grasses has been developing quickly during the last decade, with Miscanthus being among the most important choices for production of bioethanol. However, one of the key barriers to producing bioethanol is the lack of information about cell wall structure. Cell walls are thought to display compositional differences that lead to emergence of a very high level of complexity, resulting in great diversity in cell wall architectures. In this work, a set of different techniques was used to access the complexity of cell walls of different genotypes of Miscanthus sinensis in order to understand how theymore » interfere with saccharification efficiency. Three genotypes of M. sinensis displaying different patterns of correlation between lignin content and saccharification efficiency were subjected to cell wall analysis by quantitative/qualitative analytical techniques such as monosaccharide composition, oligosaccharide profiling, and glycome profiling. When saccharification efficiency was correlated negatively with lignin, the structural features of arabinoxylan and xyloglucan were found to contribute positively to hydrolysis. In the absence of such correlation, different types of pectins, and some mannans contributed to saccharification efficiency. In conclusion, different genotypes of M. sinensis were shown to display distinct interactions among their cell wall components, which seem to influence cell wall hydrolysis.« less

  16. Temperature-dependent sex determination ruled out in the Chinese soft-shelled turtle (Pelodiscus sinensis) via molecular cytogenetics and incubation experiments across populations.

    PubMed

    Mu, Yi; Zhao, Bo; Tang, Wen-Qi; Sun, Bao-Jun; Zeng, Zhi-Gao; Valenzuela, Nicole; Du, Wei-Guo

    2015-01-01

    The sex determination mechanism for the Chinese soft-shelled turtle (Pelodiscus sinensis) is subject to controversy. Some populations have been shown to possess sex chromosomes and thus genotypic sex determination (GSD), while others were reported to exhibit temperature-dependent sex determination (TSD). To test whether TSD and GSD coexist in this species or whether populations differ in their sex-determining system, we conducted egg incubation experiments to investigate how temperature influences hatchling sex in a wide range of populations of this species in China. In parallel, we used comparative genome hybridization (CGH) to study the micro-sex chromosomes of adult P. sinensis in the 2 populations that were previously identified to be TSD. The incubation experiments showed that temperature did not affect hatchling sex in any of the studied populations. CGH indicated that turtles have micro-sex chromosomes of the female heterogametic (ZZ/ZW) system in the 2 disputed populations. These results indicate that P. sinensis is a GSD rather than a TSD species. Thus, the apparent coexistence of TSD and GSD in this species is the result of previous misdiagnosis in purportedly TSD populations.

  17. Hypoglycemic activity of the fungi Cordyceps militaris, Cordyceps sinensis, Tricholoma mongolicum, and Omphalia lapidescens in streptozotocin-induced diabetic rats.

    PubMed

    Zhang, Guoqing; Huang, Yuedong; Bian, Yong; Wong, Jack H; Ng, T B; Wang, Hexiang

    2006-10-01

    Crude extracts were prepared from fruiting bodies and mycelia of the medicinal fungus Cordyceps militaris, and a polysaccharide-enriched fraction was obtained after extraction with hot water and ethanol precipitation. Polysaccharide-enriched fractions were similarly prepared from Cordyceps sinensis, Omphalia lapidescens, and Tricholoma mongolicum. The various aforementioned preparations were orally administered into different groups of adult rats 24 h before an intraperitoneal injection of streptozotocin (40 mg/kg body weight), and subsequently daily for another 4 days. The dosage used was 10 mg/kg body weight for polysaccharide-enriched preparations and 100 mg/kg body weight for crude extracts. Control rats received distilled water instead of crude extract or polysaccharide-enriched preparation. It was found in the control rats that plasma glucose level rose from about 90 mg/dl before streptozotocin injection to levels that were maintained at about 300 mg/dl postinjection. All preparations produced hypoglycemic effects. C. militaris polysaccharide-enriched fraction displayed a more prominent effect than that of C. sinensis polysaccharide-enriched fraction which in turn was more potent than that of O. lapidescens and T. mongolicum polysaccharide-enriched fractions. The hypoglycemic effect of C. militaris polysaccharide-enriched fraction was dose-dependent.

  18. Bioactivity of six plant extracts on adults of Demotispa neivai (Coleoptera: Chrysomelidae).

    PubMed

    Martínez, Luis C; Plata-Rueda, Angelica; Zanuncio, José C; Serrão, José E

    2015-01-01

    Demotispa neivai Bondar (Coleoptera: Chrysomelidae) damage oil palm fruits, which makes it necessary to develop products to control this insect. The mortality, repellency, and antifeeding effects on adults of D. neivai of six plant extracts of Azadirachta indica A. Juss. (Sapindales: Meliaceae), Ricinus communis (L.) (Malpighiaes: Euphorbiaceae), Citrus sinensis Oesbek (Sapindales: Rutaceae), Nicotiana tabacum (L.) (Slanales: Solanaceae), Capsicum annuum (L.) (Solanales: Solanaceae), and Artemisia absinthium (L.) (Asterales: Asteraceae) were determined: 1) the lethal concentration LC50-90, lethal time of D. neivai was evaluated after spraying the fruits of oil palm; 2) repellent effects of each ingredient were evaluated by calculating the index of repellency; 3) antifeeding effects with the rate of inhibition calculated between doses of 20 and 24 g/liter. The mortality of D. neivai was higher with the extracts Ci. sinensis, R. communis, N. tabacum, and Ca. annuum. The mortality of D. neivai increased in the first 72 hr in all treatments. The extracts of N. tabacum, Ca. annuum, and A. indica were more repellent to D. neivai that those of Ci. sinensis, Ar. Absinthium, and R. communis. Antifeeding effect was higher with Ci. sinensis and R. communis. The increased mortality of D. neivai by Ci. sinensis can be explained by the effect of this compound on the respiratory system of insects. Extracts of Ci. sinensis, R. communis, N. tabacum, and Ca. annuum repelled and caused mortality of D. neivai and, thus, can be used in integrate pest management programs of this pest in oil palm plantations.

  19. Antimycotic Activity and Genotoxic Evaluation of Citrus sinensis and Citrus latifolia Essential Oils.

    PubMed

    Ruiz-Pérez, Nancy J; González-Ávila, Marisela; Sánchez-Navarrete, Jaime; Toscano-Garibay, Julia D; Moreno-Eutimio, Mario A; Sandoval-Hernández, Teresa; Arriaga-Alba, Myriam

    2016-05-03

    The aim of this study was to evaluate the antifungal activity of essential oils (EOs) of Citrus sinensis (C. sinensis) and Citrus latifolia (C. latifolia) against five Candida species: Candida albicans, Candida tropicalis, Candida glabrata, Candida lusitaniae and Candida guilliermondii; and perform its genotoxic evaluation. The EOs of C. sinensis and C. latifolia were obtained from the peel by hydro-distillation. The major components determined by GC-MS were in C. sinensis, d-limonene (96%) and α-myrcene (2.79%); and in C. latifolia, d-limonene (51.64%), β-thujene (14.85%), β-pinene (12.79%) and γ-terpinene (12.8%). Antifungal properties were studied by agar diffusion method, where C. sinensis presented low activity and C. latifolia essential oil was effective to inhibit growing of C. lusitaniae and C. guilliermondii with IC50 of 6.90 and 2.92 μg respectively. The minimum inhibitory concentrations (MIC) for C. sinensis were in a range of 0.42-3.71 μg and for C. latifolia of 0.22-1.30 μg. Genotoxic evaluation was done by Ames test where none of the oils induced point mutations. Flow cytometry was used to measure toxicity in human oral epithelial cells, C. sinensis was not cytotoxic and C. latifolia was toxic at 21.8 μg. These properties might bestow different odontological applications to each essential oil.

  20. Profiling metals in Cordyceps sinensis by using inductively coupled plasma mass spectrometry.

    PubMed

    Wei, Xin; Hu, Hankun; Zheng, Baogeng; Arslan, Zikri; Huang, Hung-Chung; Mao, Weidong; Liu, Yi-Ming

    2017-01-28

    Cordyceps sinensis (C. sinensis) is a natural product that has diverse nutritional and medicinal values. Since the availability of natural C. sinensis becomes limited its authentication and quality control is of high significance. Herein we report on profiling of metals in C. sinensis by using inductively coupled plasma mass spectrometry (ICP-MS). The analysis reveals that C. sinensis contains a wide array of essential elements, including P, Mg, Zn, Cu, Fe, etc. Toxic metals detected are Cd, Pb, and As. In all five samples analyzed Pb contents are below 2.0 ppm. Arsenic level in C. sinensis caterpillar is significantly higher than that in its mycelium and varies from 3.0 to 32 ppm likely due to soil contamination. It's for the first time demonstrated in this work that clustering analysis on the proposed metal profiles consisting of 24 elements is very useful to identify "abnormal" C. sinensis samples, thus adding another dimension to the effective means for authentication and quality assessment of this highly demanded previous natural product.

  1. Tyrosine Hydroxylase is crucial for maintaining pupal tanning and immunity in Anopheles sinensis

    PubMed Central

    Qiao, Liang; Du, Minghui; Liang, Xin; Hao, Youjin; He, Xiu; Si, Fengling; Mei, Ting; Chen, Bin

    2016-01-01

    Tyrosine hydroxylase (TH), the initial enzyme in the melanin pathway, catalyzes tyrosine conversion into Dopa. Although expression and regulation of TH have been shown to affect cuticle pigmentation in insects, no direct functional studies to date have focused on the specific physiological processes involving the enzyme during mosquito development. In the current study, silencing of AsTH during the time period of continuous high expression in Anopheles sinensis pupae led to significant impairment of cuticle tanning and thickness, imposing a severe obstacle to eclosion in adults. Meanwhile, deficiency of melanin in interference individuals led to suppression of melanization, compared to control individuals. Consequently, the ability to defend exogenous microorganisms declined sharply. Accompanying down-regulation of the basal expression of five antimicrobial peptide genes resulted in further significant weakening of immunity. TH homologs as well as the composition of upstream transcription factor binding sites at the pupal stage are highly conserved in the Anopheles genus, implying that the TH-mediated functions are crucial in Anopheles. The collective evidence strongly suggests that TH is essential for Anopheles pupae tanning and immunity and provides a reference for further studies to validate the utility of the key genes involved in the melanization pathway in controlling mosquito development. PMID:27416870

  2. Conspecific reproductive success affects age of recruitment in a great cormorant, Phalacrocorax carbo sinensis, colony.

    PubMed

    Frederiksen, M; Bregnballe, T

    2001-07-22

    Few studies have addressed the proximate factors affecting the age at which individuals of long-lived bird species are recruited into the breeding population. We use capture-recapture analysis of resightings of 16 birth cohorts of colour-ringed great cormorants, Phalacrocorax carbo sinensis, in a Danish colony to assess the evidence for two hypotheses: conspecific attraction (earlier recruitment when the colony is large) and conspecific reproductive success (earlier recruitment following years of high breeding success). For both males and females, conspecific reproductive success was the most important covariate explaining the interannual variation in age of recruitment; colony size was also important for females. These covariates explained nearly 60% of the year-to-year variation for both sexes. The age of recruitment increased for cohorts born after 1990, and this increase was correlated with a decline in breeding success in the colony; we interpret this as an indirect and delayed density-dependent effect. Females were recruited earlier than males (mean age of recruitment for cohorts born before 1990: 2.98 years versus 3.53 years); the most plausible reason for this is a skewed sex ratio in favour of males in the adult population. Recruitment of males may thus, to some extent, be constrained by the availability of females. This study provides the first evidence that conspecific reproductive success can affect the age at which individual birds start to breed.

  3. [Discrimination of polysaccharides from Angelica sinensis and its different processed products based on Fourier transform infrared spectroscopy].

    PubMed

    Ji, Peng; Wei, Yan-Ming; Hua, Yong-Li; Zhang, Wen-quan

    2014-05-01

    A new rapid and nondestructive method for identifying polysaccharides from Angelica sinensis and its different processed products was developed, and this method was based on Fourier transform infrared spectroscopy (FTIR). In the clinic of traditional Chinese medicine, unprocessed Angelica sinensis(UAS) is of ten used after processed, the common processed products are Angelica sinensis parched with wine(WAS), Angelica sinensis parched with soil(SAS), Angelica sinensis parched with oil(OAS) and Charred Angelica sinensis(CAS). In order to use polysaccharides from Angelica sinensis and its processed products effectively and reasonably in clinic, it is very necessary to identify them. FTIR of polysaccharides from Angelica sinensis and its different processed products was determined, and then it was decomposed by discrete wavelet transform (DWT). The high frequency information in scale 2, 3 and 4 was selected as feature information, from which the each wavelet entropy was extracted as characteristic value. BP neural network was trained with these characteristic values. The trained BP neural network was used to identify polysaccharides from Angelica sinensis and its different processed products. According to 30 prediction samples, the correct rate for recognition was 93. 3%, which indicates that: it has better feasibility to identify polysaccharides from Angelica sinensis and its different processed products by this method, which is based on FTIR, discrete wavelet transform and BP neural network

  4. A newly found cadmium accumulator--Malva sinensis Cavan.

    PubMed

    Zhang, Shirong; Chen, Mingying; Li, Ting; Xu, Xiaoxun; Deng, Liangji

    2010-01-15

    Screening hyperaccumulators and accumulators is a key step in the phytoremediation of soils contaminated by heavy metals. A pot experiment was conducted involving a soil Cd concentration gradient (0, 50, 75, 100, 125, 150, 175, and 200 mg kg(-1)) to determine if Malva sinensis Cavan. from two lead-zinc mines in Kangding and Yajiang in western Sichuan, China, is a Cd-hyperaccumulator. The highest Cd concentrations in plant shoots from Kangding and Yajiang were 154.30 and 122.77 mg kg(-1), respectively, at a soil Cd concentration of 200 mg kg(-1). The largest amounts of accumulation in plant shoots from Kangding and Yajiang were 700.5 and 1403.2 microg pot(-1), respectively. The bioconcentration factors in shoots were 0.53-1.03 for Kangding and 0.69-1.25 for Yajiang. Moreover, all translocation factors of plants from the two sites were over 1.0. Therefore, M. sinensis can be classified as a Cd-accumulator or non-standard Cd-hyperaccumulator.

  5. Effect of lipopolysaccharide on the hemocyte apoptosis of Eriocheir sinensis *

    PubMed Central

    Xu, Hai-sheng; Lyu, Sun-jian; Xu, Jie-hao; Lu, Bin-jie; Zhao, Jing; Li, Song; Li, Yi-qun; Chen, Yu-yin

    2015-01-01

    In the present study, we investigated the possible toxicity mechanism of lipopolysaccharide (LPS) extracted from Gram-negative bacteria in Eriocheir sinensis hemocytes. Apoptotic hemocytes and reactive oxygen species (ROS) production induced by the LPS were monitored by the combination of flow cytometry and microscope observation. It was shown that LPS induced serious damage on the DNA and morphological changes in hemocytes, including cell shrinkage, fracture of nucleus membrane, margination, condensation and fragmentation of chromatin, and formation of apoptotic bodies indicating obvious hemocyte apoptosis. As compared with the control group, the apoptotic cell ratio increased to 30.61% and 39.01% after 1-h exposure and 57.72% and 75.01% after 2-h exposure to 1 and 10 μg/ml LPS, respectively (P<0.05). Significant outburst of ROS production was observed in LPS-treated hemocytes with approximately 176.6% of relative dichlorofluorescein mean fluorescence at 1-h exposure, followed by a drastic decline (P<0.05). These results indicated that LPS would induce oxidative stress on hemocytes from E. sinensis and cause ROS burst, DNA damage, and subsequently apoptosis. The process of ROS-mediated apoptosis might be one of the potential toxicity mechanisms of LPS on crustacean hemocytes. PMID:26642180

  6. A transcriptome analysis of mitten crab testes (Eriocheir sinensis)

    PubMed Central

    Zhang, Wei; Wan, Haolei; Jiang, Hui; Zhao, Yunlong; Zhang, Xiaowei; Hu, Songnian; Wang, Qun

    2011-01-01

    The identification of expressed genes involved in sexual precocity of the mitten crab (Eriocheir sinensis) is critical for a better understanding of its reproductive development. To this end, we constructed a cDNA library from the rapid developmental stage of testis of E. sinensis and sequenced 3,388 randomly picked clones. After processing, 2,990 high-quality expressed sequence tags (ESTs) were clustered into 2,415 unigenes including 307 contigs and 2,108 singlets, which were then compared to the NCBI non-redundant (nr) protein and nucleotide (nt) database for annotation with Blastx and Blastn, respectively. After further analysis, 922 unigenes were obtained with concrete annotations and 30 unigenes were found to have functions possibly related to the process of reproduction in male crabs – six transcripts relevant to spermatogenesis (especially Cyclin K and RecA homolog DMC1), two transcripts involved in nuclear protein transformation, two heat-shock protein genes, eleven transcription factor genes (a series of zinc-finger proteins), and nine cytoskeleton protein-related genes. Our results, besides providing valuable information related to crustacean reproduction, can also serve as a base for future studies of reproductive and developmental biology. PMID:21637557

  7. Enhancement of Neuromuscular Activity by Natural Specimens and Cultured Mycelia of Cordyceps sinensis in Mice

    PubMed Central

    Singh, K. P.; Meena, H. S.; Negi, P. S.

    2014-01-01

    The present study was aimed to evaluate the effect of natural specimen and laboratory cultured mycelia of Cordyceps sinensis on neuromuscular activity in mice. The powder of natural specimen and laboratory cultured Cordyceps sinensis was orally administered at the dose rate of 100, 300 and 500 mg/kg for 30 days. Natural specimen and in vitro propagated Cordyceps sinensis showed significant (P<0.05) enhancement in neuromuscular endurance and antidepressant activity at 300 and 500 mg/kg as compared to the control group. However, the fungus did not proved to be as effective as fluoxetine in exhibiting antidepressant action. Muscular endurance was determined on a Rota rod apparatus while antidepressant (mood elevating) activity was measured on a photoactometer in Swiss albino mice. The effects produced by both natural specimens and laboratory cultured Cordyceps sinensis were comparable and showed almost equal potency. PMID:25425763

  8. Complete Chloroplast Genome Sequence of Aquilaria sinensis (Lour.) Gilg and Evolution Analysis within the Malvales Order

    PubMed Central

    Wang, Ying; Zhan, Di-Feng; Jia, Xian; Mei, Wen-Li; Dai, Hao-Fu; Chen, Xiong-Ting; Peng, Shi-Qing

    2016-01-01

    Aquilaria sinensis (Lour.) Gilg is an important medicinal woody plant producing agarwood, which is widely used in traditional Chinese medicine. High-throughput sequencing of chloroplast (cp) genomes enhanced the understanding about evolutionary relationships within plant families. In this study, we determined the complete cp genome sequences for A. sinensis. The size of the A. sinensis cp genome was 159,565 bp. This genome included a large single-copy region of 87,482 bp, a small single-copy region of 19,857 bp, and a pair of inverted repeats (IRa and IRb) of 26,113 bp each. The GC content of the genome was 37.11%. The A. sinensis cp genome encoded 113 functional genes, including 82 protein-coding genes, 27 tRNA genes, and 4 rRNA genes. Seven genes were duplicated in the protein-coding genes, whereas 11 genes were duplicated in the RNA genes. A total of 45 polymorphic simple-sequence repeat loci and 60 pairs of large repeats were identified. Most simple-sequence repeats were located in the noncoding sections of the large single-copy/small single-copy region and exhibited high A/T content. Moreover, 33 pairs of large repeat sequences were located in the protein-coding genes, whereas 27 pairs were located in the intergenic regions. Aquilaria sinensis cp genome bias ended with A/T on the basis of codon usage. The distribution of codon usage in A. sinensis cp genome was most similar to that in the Gonystylus bancanus cp genome. Comparative results of 82 protein-coding genes from 29 species of cp genomes demonstrated that A. sinensis was a sister species to G. bancanus within the Malvales order. Aquilaria sinensis cp genome presented the highest sequence similarity of >90% with the G. bancanus cp genome by using CGView Comparison Tool. This finding strongly supports the placement of A. sinensis as a sister to G. bancanus within the Malvales order. The complete A. sinensis cp genome information will be highly beneficial for further studies on this traditional medicinal

  9. Complete Chloroplast Genome Sequence of Aquilaria sinensis (Lour.) Gilg and Evolution Analysis within the Malvales Order.

    PubMed

    Wang, Ying; Zhan, Di-Feng; Jia, Xian; Mei, Wen-Li; Dai, Hao-Fu; Chen, Xiong-Ting; Peng, Shi-Qing

    2016-01-01

    Aquilaria sinensis (Lour.) Gilg is an important medicinal woody plant producing agarwood, which is widely used in traditional Chinese medicine. High-throughput sequencing of chloroplast (cp) genomes enhanced the understanding about evolutionary relationships within plant families. In this study, we determined the complete cp genome sequences for A. sinensis. The size of the A. sinensis cp genome was 159,565 bp. This genome included a large single-copy region of 87,482 bp, a small single-copy region of 19,857 bp, and a pair of inverted repeats (IRa and IRb) of 26,113 bp each. The GC content of the genome was 37.11%. The A. sinensis cp genome encoded 113 functional genes, including 82 protein-coding genes, 27 tRNA genes, and 4 rRNA genes. Seven genes were duplicated in the protein-coding genes, whereas 11 genes were duplicated in the RNA genes. A total of 45 polymorphic simple-sequence repeat loci and 60 pairs of large repeats were identified. Most simple-sequence repeats were located in the noncoding sections of the large single-copy/small single-copy region and exhibited high A/T content. Moreover, 33 pairs of large repeat sequences were located in the protein-coding genes, whereas 27 pairs were located in the intergenic regions. Aquilaria sinensis cp genome bias ended with A/T on the basis of codon usage. The distribution of codon usage in A. sinensis cp genome was most similar to that in the Gonystylus bancanus cp genome. Comparative results of 82 protein-coding genes from 29 species of cp genomes demonstrated that A. sinensis was a sister species to G. bancanus within the Malvales order. Aquilaria sinensis cp genome presented the highest sequence similarity of >90% with the G. bancanus cp genome by using CGView Comparison Tool. This finding strongly supports the placement of A. sinensis as a sister to G. bancanus within the Malvales order. The complete A. sinensis cp genome information will be highly beneficial for further studies on this traditional medicinal

  10. Variation of theanine, phenolic, and methylxanthine compounds in 21 cultivars of Camellia sinensis harvested in different seasons.

    PubMed

    Fang, Rui; Redfern, Sally P; Kirkup, Don; Porter, Elaine A; Kite, Geoffrey C; Terry, Leon A; Berry, Mark J; Simmonds, Monique S J

    2017-04-01

    This is the first study to use chemometric methods to differentiate among 21 cultivars of Camellia sinensis from China and between leaves harvested at different times of the year using 30 compounds implicated in the taste and quality of tea. Unique patterns of catechin derivatives were observed among cultivars and across harvest seasons. C. sinensis var. pubilimba (You 510) differed from the cultivars of C. sinensis var. sinensis, with higher levels of theobromine, (+)-catechin, gallocatechin, gallocatechin gallate and theasinensin B, and lower levels of (-)-epicatechin, (-)-epigallocatechin (EGC) and (-)-epigallocatechin gallate (EGCG), respectively. Three cultivars of C. sinensis var. sinensis, Fuyun 7, Qiancha 7 and Zijuan contained significantly more caffeoylquinic acids than others cultivars. A Linear Discriminant Analysis model based on the abundance of 12 compounds was able to discriminate amongst all 21 tea cultivars. Harvest time impacted the abundance of EGC, theanine and afzelechin gallate.

  11. Studies on Anopheles sinensis, the vector species of vivax malaria in Korea

    PubMed Central

    2005-01-01

    Extensive previous studies on taxonomy, behavior/bionomics and control of Anopheles sinensis are reviewed and summarized. Recent molecular identification revealed that the population of An. sinensis complex includes An. sinensis, An. pullus, An. lesteri and at least two new species, and An. yatsushiroensis is synonmy of An. pullus. An. sinensis is the main vector specie of vivax malaria in Korea. Larvae of An. sinensis breed in wide range of habitats which are naturally-made clean water, stagnant or flowing; main habitats include rice fields, ditches, streams, irrigation cannals, marshes, ponds, ground pools, etc. Their host preferences are highly zoophilic. Human blood rate is very low (0.7-1.7%); nevertheless An. sinensis readily feeds on man when domestic animals are not found near by. They feed on hosts throughout the night from dusk to dawn with a peak period of 02:00-04:00 hours; they are slightly more exophagic (biting outdoors); much larger numbers come into the room when light is on. Main resting places are outdoors such as grasses, vegetable fields and rice fields. A mark-release-recapture study resulted that 37.1% was recaptured within 1 km, 29.4% at 1-3 km, 21.1% at 3-6 km, 10.3% at 6-9 km and 2.1% at 9-12 km distance. An. sinensis hibernate outdoors (mostly under part of dense grasses) during October-March. At the end of the hibernation period (March-April) they feed on cows at daytime. Until today any single measure to effectively control An. sinensis population has not been found. Indoor residual spray with a long-lasting insecticide can not reduce vector population densities, but shorten their life spans in some degree, so contributes to malaria control. PMID:16192749

  12. Cordyceps sinensis attenuates renal fibrosis and suppresses BAG3 induction in obstructed rat kidney

    PubMed Central

    Du, Feng; Li, Si; Wang, Tian; Zhang, Hai-Yan; Zong, Zhi-Hong; Du, Zhen-Xian; Li, De-Tian; Wang, Hua-Qin; Liu, Bo; Miao, Jia-Ning; Bian, Xiao-Hui

    2015-01-01

    BAG3 regulates a number of cellular processes, including cell proliferation, apoptosis, adhesion and migration, and epithelial-mesenchymal transition (EMT). However, the role of BAG3 in renal tubular EMT and renal interstitial fibrosis remains elusive. This study aimed to examine the dynamic expression of BAG3 during renal fibrosis, and to investigate the efficacy of Cordyceps sinensis (C. sinensis) on renal fibrosis. A rat model of unilateral ureteral obstruction (UUO) was established, and the expression of BAG3 and α-SMA, and the efficacy of C. sinensis on renal fibrosis induced by UUO were examined. The results showed that UUO led to collagen accumulation, which was significantly suppressed by C. sinensis. UUO increased the expression of BAG3 and α-SMA, a mesenchymal marker, while UUO induced BAG3 and α-SMA expression was significantly inhibited by C. sinensis. In addition, immunohistochemical staining demonstrated that BAG3 immunoreactivity was restricted to tubular epithelium. In conclusion, BAG3 is a potential target for the prevention and/or treatment of renal fibrosis, and C. Sinensis is a promising agent for renal fibrosis. PMID:26175854

  13. Systematic revelation of the protective effect and mechanism of Cordycep sinensis on diethylnitrosamine-induced rat hepatocellular carcinoma with proteomics

    PubMed Central

    Wang, Pei-Wen; Hung, Yu-Chiang; Li, Wen-Tai; Yeh, Chau-Ting; Pan, Tai-Long

    2016-01-01

    Cordyceps sinensis (C. sinensis) has been reported to treat liver diseases. Here, we investigated the inhibitory effect of C. sinensis on hepatocarcinoma in a diethylnitrosamine (DEN)-induced rat model with functional proteome tools. In the DEN-exposed group, levels of serum alanine aminotransferase and aspartate aminotransferase were increased while C. sinensis application remarkably inhibited the activities of these enzymes. Histopathological analysis also indicated that C. sinensis could substantially restore hypertrophic hepatocytes caused by DEN, suggesting that C. sinensis is effective in preventing DEN-induced hepatocarcinogenesis. We therefore comprehensively delineated the global protein alterations using a proteome platform. The most meaningful changes were found among proteins involved in oxidative stress and detoxification. Meanwhile, C. sinensis application could attenuate the carbonylation level of several enzymes as well as chaperone proteins. Network analysis implied that C. sinensis could obviously alleviate hepatocarcinoma via modulating redox imbalance, protein ubiquitination and tumor growth–associated transcription factors. Our findings provide new insight into the potential effects of C. sinensis in preventing carcinogenesis and might help in developing novel therapeutic strategies against chemical-induced hepatocarcinoma. PMID:27531890

  14. The complete mitochondrial genome of Chinese sturgeon (Acipenser sinensis).

    PubMed

    Liao, Xiaolin; Tian, Hua; Zhu, Bin; Chang, Jianbo

    2016-01-01

    The complete mitochondrial genome of Chinese sturgeon (Acipenser sinensis) was determined by direct sequencing of PCR products. The Chinese sturgeon mitochondrial DNA is a circular molecule (16,688 bp in length) with the typical gene arrangement of vertebrate mtDNA, containing 13 protein-coding genes, two ribosomal RNA and 22 transfer RNA genes, and a non-coding control region. Its control region contains 4.5 copies of unit with 82 bp long at 5' end, which has been reported before for this species. Phylogenetic tree based on 13 protein-coding genes confirmed that the complete mtDNA sequence of Chinese sturgeon was reported here for the first time.

  15. Acoustic signals of Chinese alligators (Alligator sinensis): social communication.

    PubMed

    Wang, Xianyan; Wang, Ding; Wu, Xiaobing; Wang, Renping; Wang, Chaolin

    2007-05-01

    This paper reports the first systematic study of acoustic signals during social interactions of the Chinese alligator (Alligator sinensis). Sound pressure level (SPL) measurements revealed that Chinese alligators have an elaborate acoustic communication system with both long-distance signal-bellowing-and short-distance signals that include tooting, bubble blowing, hissing, mooing, head slapping and whining. Bellows have high SPL and appear to play an important role in the alligator's long range intercommunion. Sounds characterized by low SPL are short-distance signals used when alligators are in close spatial proximity to one another. The signal spectrographic analysis showed that the acoustic signals of Chinese alligators have a very low dominant frequency, less than 500 Hz. These frequencies are consistent with adaptation to a habitat with high density vegetation. Low dominant frequency sound attenuates less and could therefore cover a larger spatial range by diffraction in a densely vegetated environment relative to a higher dominant frequency sound.

  16. The draft genome of sweet orange (Citrus sinensis).

    PubMed

    Xu, Qiang; Chen, Ling-Ling; Ruan, Xiaoan; Chen, Dijun; Zhu, Andan; Chen, Chunli; Bertrand, Denis; Jiao, Wen-Biao; Hao, Bao-Hai; Lyon, Matthew P; Chen, Jiongjiong; Gao, Song; Xing, Feng; Lan, Hong; Chang, Ji-Wei; Ge, Xianhong; Lei, Yang; Hu, Qun; Miao, Yin; Wang, Lun; Xiao, Shixin; Biswas, Manosh Kumar; Zeng, Wenfang; Guo, Fei; Cao, Hongbo; Yang, Xiaoming; Xu, Xi-Wen; Cheng, Yun-Jiang; Xu, Juan; Liu, Ji-Hong; Luo, Oscar Junhong; Tang, Zhonghui; Guo, Wen-Wu; Kuang, Hanhui; Zhang, Hong-Yu; Roose, Mikeal L; Nagarajan, Niranjan; Deng, Xiu-Xin; Ruan, Yijun

    2013-01-01

    Oranges are an important nutritional source for human health and have immense economic value. Here we present a comprehensive analysis of the draft genome of sweet orange (Citrus sinensis). The assembled sequence covers 87.3% of the estimated orange genome, which is relatively compact, as 20% is composed of repetitive elements. We predicted 29,445 protein-coding genes, half of which are in the heterozygous state. With additional sequencing of two more citrus species and comparative analyses of seven citrus genomes, we present evidence to suggest that sweet orange originated from a backcross hybrid between pummelo and mandarin. Focused analysis on genes involved in vitamin C metabolism showed that GalUR, encoding the rate-limiting enzyme of the galacturonate pathway, is significantly upregulated in orange fruit, and the recent expansion of this gene family may provide a genomic basis. This draft genome represents a valuable resource for understanding and improving many important citrus traits in the future.

  17. Anti-TB Polyynes from the roots of Angelica sinensis

    PubMed Central

    Deng, Shixin; Wang, Yuehong; Inui, Taichi; Chen, Shao-Nong; Farnsworth, Norman R.; Cho, Sanghyun; Franzblau, Scott G.; Pauli, Guido F.

    2008-01-01

    Following chemotaxonomic evidence, the PE and CHCl3 extracts of the roots of the botanical Angelica sinensis (Oliv.) Diels (Dang Gui) were investigated for in vitro anti-TB activity, in parallel to studying their serotonergic and GABAergic potential. The activities were confirmed to chemically overlap with the neurotropic active principles present in medium lipophilic fractions. Phytochemical investigations led to the isolation of five polyynes: falcarindiol (1), 9Z,17-octadecadiene-12,14-diyne-1,11,16-triol,1-acetate (2), oplopandiol (3), heptadeca-1-ene-9,10-epoxy-4,6-diyne-3,8-diol (4), and the new polyyne 8-hydroxy-1-methoxy-(Z)-9-heptadecene-4,6-diyn-3-one (5), as characterized by spectroscopic techniques including 1D, 2D NMR and HR-MS. All compounds were tested against two pathogenic strains of Mycobacterium tuberculosis (H37Rv and Erdman) in vitro in a microplate Alamar Blue assay (MABA). The most potent anti-TB constituents were 1 and 2, exhibiting MIC values of 1.4-26.7 μg/ml; 3 showed moderate MICs (49.5 and 50.2 μg/ml, respectively) while 4 and 5 were weakly active (MIC> 60 μg/ml). Notably, none of the five compounds exhibited significant cytotoxicity against VERO cells. These findings not only reveal a new potential area of therapeutic value for A. sinensis, but also underline the role of polyynes as anti-TB active principles in ethnobotanical preparations, and as lead compounds. PMID:18567055

  18. GC-FID/MS Profiling of Supercritical CO2 Extracts of Peels from Citrus aurantium, C. sinensis cv. Washington navel, C. sinensis cv. Tarocco and C. sinensis cv. Doppio Sanguigno from Dubrovnik Area (Croatia).

    PubMed

    Jerković, Igor; Drulžić, Jasmina; Marijanović, Zvonimir; Gugić, Mirko; Jokić, Stela; Roje, Marin

    2015-07-01

    The peels of Citrus aurantium L. and Citrus sinensis Osbeck cultivars from the Dubrovnik region (south Croatia) were extracted by supercritical CO2 at 40 degrees C and 10 MPa at 1.76 kg/h to obtain enriched extracts in comparison with simple pressing of the peels. The extracts were analyzed in detail by gas chromatography and mass spectrometry (GC-FID/MS). Relevant similarities among the peel oil compositions of C. aurantium and C. sinensis cultivars were found with limonene predominance (up to 54.3%). The principal oxygenated monoterpenes were linalool (3.0%-5.9%), α-terpineol (0.7%-2.4%), linalyl acetate (0.0%-5.0%), geranyl acetate (0.0%-0.4%), (Z)-citral (0.0%-1.8%) and (E)-citral (0.0%-1.9%). Several sesquiterpenes were found with minor percentages. Coumarin derivatives were identified in all the samples among the relevant compounds. Isogeijerin dominated in the peels of C. sinensis cv. Tarocco (15.3%) and C. aurantium (11.2%). Scoparone ranged from 0.1% to 0.5% in all the samples. Bergapten (up to 1.4%), osthole (up to 1.1%) and 7-methoxy-8-(2-formylpropyl)coumarin (up to 1.1%) were found mostly in C. sinensis cv. Doppio Sanguigno. It was possible to indicate a few other differences among the extracts such as higher percentage of linalool, linalyl and geranyl acetates, as well as the abundance of sabinene and isogeijerin in C. aurantium or the occurrence of β-sinensal in C. sinensis cultivars.

  19. Domestic cats (Felis catus) are definitive hosts for Sarcocystis sinensis from water buffaloes (Bubalus bubalis)

    PubMed Central

    GJERDE, Bjørn; HILALI, Mosaad

    2016-01-01

    The definitive hosts of Sarcocystis sinensis in water buffaloes have hitherto been unknown, but the close similarity of this species to the cat-transmitted Sarcocystis bovifelis in cattle suggested they were felids. In a previous study, two domestic cats were fed macroscopic sarcocysts of Sarcocystis fusiformis contained within or dissected from the esophageal muscles of water buffaloes, while no microscopic sarcocysts of S. sinensis were noticed. Both cats started shedding small numbers of sporocysts 8–10 days post infection (dpi) and were euthanized 15 dpi. Using a PCR-based molecular assay targeting the mitochondrial cox1 gene of S. fusiformis, both cats were shown to act as definitive hosts for this species. In the present study, DNA samples derived from oocysts/sporocysts in the intestinal mucosa of both cats were further examined by PCR for the presence of S. sinensis using 2 newly designed primers selectively targeting the cox1 gene of this species. All 6 DNA samples examined from each cat tested positive for S. sinensis. A 1,038-bp-long portion of cox1 was amplified and sequenced as 2 overlapping fragments from 5 of these DNA samples. The 5 sequences shared 99.3–100% identity with 7 previous cox1 sequences of S. sinensis obtained from sarcocysts in water buffaloes. Additionally, amplification of the ITS1 region with primers targeting various Sarcocystis spp., yielded amplicons of 2 different lengths, corresponding to those obtained from sarcocyst isolates of S. sinensis and S. fusiformis, respectively. This is the first study to show that cats act as definitive hosts for S. sinensis. PMID:27075117

  20. Antimycotic Activity and Genotoxic Evaluation of Citrus sinensis and Citrus latifolia Essential Oils

    PubMed Central

    Ruiz-Pérez, Nancy J.; González-Ávila, Marisela; Sánchez-Navarrete, Jaime; Toscano-Garibay, Julia D.; Moreno-Eutimio, Mario A.; Sandoval-Hernández, Teresa; Arriaga-Alba, Myriam

    2016-01-01

    The aim of this study was to evaluate the antifungal activity of essential oils (EOs) of Citrus sinensis (C. sinensis) and Citrus latifolia (C. latifolia) against five Candida species: Candida albicans, Candida tropicalis, Candida glabrata, Candida lusitaniae and Candida guilliermondii; and perform its genotoxic evaluation. The EOs of C. sinensis and C. latifolia were obtained from the peel by hydro-distillation. The major components determined by GC-MS were in C. sinensis, d-limonene (96%) and α-myrcene (2.79%); and in C. latifolia, d-limonene (51.64%), β-thujene (14.85%), β-pinene (12.79%) and γ-terpinene (12.8%). Antifungal properties were studied by agar diffusion method, where C. sinensis presented low activity and C. latifolia essential oil was effective to inhibit growing of C. lusitaniae and C. guilliermondii with IC50 of 6.90 and 2.92 μg respectively. The minimum inhibitory concentrations (MIC) for C. sinensis were in a range of 0.42–3.71 μg and for C. latifolia of 0.22–1.30 μg. Genotoxic evaluation was done by Ames test where none of the oils induced point mutations. Flow cytometry was used to measure toxicity in human oral epithelial cells, C. sinensis was not cytotoxic and C. latifolia was toxic at 21.8 μg. These properties might bestow different odontological applications to each essential oil. PMID:27137128

  1. [Use of cowpea (Vigna sinensis) as a chicken complement in an infant formula].

    PubMed

    Modernell, Marisa Guerra; Granito, Marisela; Paolini, Mariangel; Olaizola, Cristina

    2008-09-01

    Legumes represent an important protein source worldwide. In Venezuela, they are generally prepared at home and are consumed by adults, as soup or stew, while children eat them in very small quantities. In order to include legumes in the children's diet, the following work was done using cowpea (Vigna sinensis) as an complement of chicken in the preparation of a nutritionally balanced formula, adapted to the requirements of children. Several formulas were developed and three of them were selected based on their acceptability. In the first formula, the protein source was only of chicken. In the second formula, the chicken was partially substituted by cowpea, and in the third formula, the protein source was only made of cowpea. Other formula ingredients included rice, pumpkin (Curcubita maxima), carrot and some seasonings. Proximal analysis, protein quality (as protein efficiency ratio and protein digestibility) and sensory evaluation (7-point hedonic scale) were performed on the formulas. The proximal composition was similar in the three formulas: protein (3.5%), fat (1.3%) and carbohydrates (19.7%), with a good distribution of the energy contribution (98.9 kcal/100 g or 413.8 kJ/100 g). The protein quality and protein digestibility were higher for the chicken-cowpea formula than for the cowpea one. The acceptability with the mothers was higher for the chicken-cowpea formula than for the cowpea one. The acceptability of the chicken-cowpea formula with children was 77% (7-point hedonic facial scale) and 92% (measuring consumption). Due to the high acceptability and good protein quality, the chicken-cowpea formula could be included in the lunch meal of the children in daycare homes.

  2. Importance of wine-treated Angelica Sinensis Radix in Si Wu Tang, a traditional herbal formula for treating women's ailments.

    PubMed

    Zhan, Janis Y X; Zheng, Ken Y Z; Zhu, Kevin Y; Zhang, Wendy L; Bi, Cathy W C; Chen, J P; Du, Crystal Y Q; Dong, Tina T X; Lau, David T W; Tsim, Karl W K

    2013-05-01

    Si Wu Tang (Four Agents Decoction), a traditional Chinese decoction composed of Angelica Sinensis Radix, Chuanxiong Rhizoma, Paeoniae Radix Alba, and Rehmanniae Radix Praeparata in a ratio of 1 : 1 : 1 : 1, has been used to treat women's diseases for more than a thousand years. According to the original description of Si Wu Tang, Angelica Sinensis Radix should be treated with wine. However, the importance of this wine-treated Angelica Sinensis Radix in Si Wu Tang's function has not been identified. In this article, the chemical and biological properties of two decoctions processed in different ways (Si Wu Tang with crude Angelica Sinensis Radix and Si Wu Tang with wine-treated Angelica Sinensis Radix) were compared for examination. The herbal decoction Si Wu Tang prepared from wine-treated Angelica Sinensis Radix contained much different amounts of its active compounds. Compared with Si Wu Tang using crude Angelica Sinensis Radix, Si Wu Tang prepared from wine-treated Angelica Sinensis Radix had better biological responses. Therefore, these findings accentuate the functional importance of herbs treated with wine in the Chinese decoction.

  3. GENETIC ANALYSIS OF THE CHINESE MITTEN CRAB (ERIOCHEIR SINENSIS) INTRODUCED TO THE NORTH AMERICAN GREAT LAKES AND ST. LAWRENCE SEAWAY

    EPA Science Inventory

    The Chinese mitten crab (Eriocheir sinensis) is a globally invasive organism, with established non-native populations in Europe and California, USA. Since 1965, there have been sixteen confirmed catches of E. sinensis in the North American Great Lakes and their associated waterw...

  4. Genetic structure of Miscanthus sinensis and Miscanthus sacchariflorus in Japan indicates a gradient of bidirectional but asymmetric introgression.

    PubMed

    Clark, Lindsay V; Stewart, J Ryan; Nishiwaki, Aya; Toma, Yo; Kjeldsen, Jens Bonderup; Jørgensen, Uffe; Zhao, Hua; Peng, Junhua; Yoo, Ji Hye; Heo, Kweon; Yu, Chang Yeon; Yamada, Toshihiko; Sacks, Erik J

    2015-07-01

    Unilateral introgression from diploids to tetraploids has been hypothesized to be an important evolutionary mechanism in plants. However, few examples have been definitively identified, perhaps because data of sufficient depth and breadth were difficult to obtain before the advent of affordable high-density genotyping. Throughout Japan, tetraploid Miscanthus sacchariflorus and diploid Miscanthus sinensis are common, and occasionally hybridize. In this study, 667 M. sinensis and 78 M. sacchariflorus genotypes from Japan were characterized using 20 704 SNPs and ten plastid microsatellites. Similarity of SNP genotypes between diploid and tetraploid M. sacchariflorus indicated that the tetraploids originated through autopolyploidy. Structure analysis indicated a gradient of introgression from diploid M. sinensis into tetraploid M. sacchariflorus throughout Japan; most tetraploids had some M. sinensis DNA. Among phenotypically M. sacchariflorus tetraploids, M. sinensis ancestry averaged 7% and ranged from 1-39%, with introgression greatest in southern Japan. Unexpectedly, rare (~1%) diploid M. sinensis individuals from northern Japan were found with 6-27% M. sacchariflorus ancestry. Population structure of M. sinensis in Japan included three groups, and was driven primarily by distance, and secondarily by geographic barriers such as mountains and straits. Miscanthus speciation is a complex and dynamic process. In contrast to limited introgression between diploid M. sacchariflorus and M. sinensis in northern China, selection for adaptation to a moderate maritime climate probably favoured cross-ploidy introgressants in southern Japan. These results will help guide the selection of Miscanthus accessions for the breeding of biomass cultivars.

  5. Genetic structure of Miscanthus sinensis and Miscanthus sacchariflorus in Japan indicates a gradient of bidirectional but asymmetric introgression

    DOE PAGES

    Clark, Lindsay V.; Stewart, J. Ryan; Nishiwaki, Aya; ...

    2015-01-24

    Unilateral introgression from diploids to tetraploids has been hypothesized to be an important evolutionary mechanism in plants. However, few examples have been definitively identified, perhaps because data of sufficient depth and breadth were difficult to obtain before the advent of affordable high-density genotyping. Throughout Japan, tetraploid Miscanthus sacchariflorus and diploid Miscanthus sinensis are common, and occasionally hybridize. In this study, 667 M. sinensis and 78 M. sacchariflorus genotypes from Japan were characterized using 20 704 SNPs and ten plastid microsatellites. Similarity of SNP genotypes between diploid and tetraploid M. sacchariflorus indicated that the tetraploids originated through autopolyploidy. Structure analysis indicatedmore » a gradient of introgression from diploid M. sinensis into tetraploid M. sacchariflorus throughout Japan; most tetraploids had some M. sinensis DNA. Among phenotypically M. sacchariflorus tetraploids, M. sinensis ancestry averaged 7% and ranged from 1-39%, with introgression greatest in southern Japan. Unexpectedly, rare (~1%) diploid M. sinensis individuals from northern Japan were found with 6-27% M. sacchariflorus ancestry. Population structure of M. sinensis in Japan included three groups, and was driven primarily by distance, and secondarily by geographic barriers such as mountains and straits. Miscanthus speciation is a complex and dynamic process. In contrast to limited introgression between diploid M. sacchariflorus and M. sinensis in northern China, selection for adaptation to a moderate maritime climate probably favoured cross-ploidy introgressants in southern Japan. Ultimately, these results will help guide the selection of Miscanthus accessions for the breeding of biomass cultivars.« less

  6. Angelica Sinensis attenuates inflammatory reaction in experimental rat models having spinal cord injury.

    PubMed

    Xu, Jun; E, Xiao-Qiang; Liu, Hui-Yong; Tian, Jun; Yan, Jing-Long

    2015-01-01

    This study was aimed to evaluate the effect of Angelica Sinensis on experimental rat models in which spinal cord injury was induced by studying different factors. Different factors causing inflammation play a key role in pathophysiology of SCI. Here three groups of rats (n=15, each was used). These included a sham control group where only laminectomy was performed, SCI group where SCI was induced and AS/SCI group where although SCI was induced but Angelica Sinensis was also administered to study its effect and draw a comparison with control. The expression of I-kBα and NF-kB p65 was also studied using western blotting and after recording optical density (OD) values of western blots. MPO activity was used to measure the effect of 20 mg/kg Angelica Sinensis. The levels of proinflammatory cytokines TNF-α, IL-1β and IL-6 were also studied. As compared with SCI group and sham control it was observed that Angelica Sinensis significantly reduced the expression of I-kBα and NF-kB p65, (P<0.05), while MPO activity was also significantly reduced. Proinflammatory cytokine level was also reduced in treated group as compared to both other groups. On the basis of this study we concluded that the use of 20 mg/kg Angelica Sinensis in rat models can attenuate the secondary damage caused by SCI and thus help in controlling the pathology of SCI in rats.

  7. Genomic Analysis of Detoxification Supergene Families in the Mosquito Anopheles sinensis.

    PubMed

    Zhou, Dan; Liu, Xianmiao; Sun, Yan; Ma, Lei; Shen, Bo; Zhu, Changliang

    2015-01-01

    Anopheles sinensis is an important malaria vector in China and other Southeast Asian countries, and the emergence of insecticide resistance in this mosquito poses a serious threat to the efficacy of malaria control programs. The recently published An. sinensis genome and transcriptome provide an opportunity to understand the molecular mechanisms of insecticide resistance. Analysis of the An. sinensis genome revealed 174 detoxification genes, including 93 cytochrome P450s (P450s), 31 glutathione-S-transferases (GSTs), and 50 choline/carboxylesterases (CCEs). The gene number was similar to that in An. gambiae, but represented a decrease of 29% and 42% compared with Aedes aegypti and Culex quinquefasciatus, respectively. The considerable contraction in gene number in Anopheles mosquitoes mainly occurred in two detoxification supergene families, P450s and CCEs. The available An. sinensis transcriptome was also re-analyzed to further identify key resistance-associated detoxification genes. Among 174 detoxification genes, 124 (71%) were detected. Several candidate genes overexpressed in a deltamethrin-resistant strain (DR-strain) were identified as belonging to the CYP4 or CYP6 family of P450s and the Delta GST class. These generated data provide a basis for identifying the resistance-associated genes of An. sinensis at the molecular level.

  8. [The mechanical wound transcriptome of three-year-old Aquilaria sinensis].

    PubMed

    Zhang, Zheng; Gao, Zhi-hui; Wei, Jian-he; Xu, Yan-hong; Li, Ying; Yang, Yun; Meng, Hui; Sui, Chun; Wang, Meng-xi

    2012-08-01

    Chinese agarwood is formed in the aromatic resinous wood formed in Aquilaria sinensis (Lour.) Gilg (botanical family: Thymelaeaceae). Only when suffering stress of wound, etc, can A. sinensis produce sesquiterpenes etc. compounds of agarwood around wounds. However, little is known about how wound induced the biosynthesis pathway of sesquiterpenes. To reveal the molecular mechanism of wound-induced agarwood formation, RNA sequencing (RNA-seq) technology was used to investigate the profile of gene expression in A. sinensis treated by mechanical wounding and elucidate its functional gene. A total of 40,295 ESTs with an average read length of 305 bp were generated and 22 095 unigenes were formed by initial gene splicing. 61.6% of these unigenes (13 611) were annotated using BLAST searches against the SwissProt, KEGG, Nr and Nt databases. Twenty-six unigenes (encoding 7 enzymes) were found to be involved in sesquiterpene of agarwood biosynthesis by bioinformatic tools of Gene Ontology and KEGG. Novel genes that are potentially involved in sesquiterpenes biosynthesis were identified in A. sinensis, providing data for further sesquiterpenes biosynthesis pathway by molecular methods and the EST data establish a foundation for future studies in the molecular mechanisms of wound-induce agarwood formation in A. sinensis.

  9. Angelica Sinensis attenuates inflammatory reaction in experimental rat models having spinal cord injury

    PubMed Central

    Xu, Jun; E, Xiao-Qiang; Liu, Hui-Yong; Tian, Jun; Yan, Jing-Long

    2015-01-01

    This study was aimed to evaluate the effect of Angelica Sinensis on experimental rat models in which spinal cord injury was induced by studying different factors. Different factors causing inflammation play a key role in pathophysiology of SCI. Here three groups of rats (n=15, each was used). These included a sham control group where only laminectomy was performed, SCI group where SCI was induced and AS/SCI group where although SCI was induced but Angelica Sinensis was also administered to study its effect and draw a comparison with control. The expression of I-kBα and NF-kB p65 was also studied using western blotting and after recording optical density (OD) values of western blots. MPO activity was used to measure the effect of 20 mg/kg Angelica Sinensis. The levels of proinflammatory cytokines TNF-α, IL-1β and IL-6 were also studied. As compared with SCI group and sham control it was observed that Angelica Sinensis significantly reduced the expression of I-kBα and NF-kB p65, (P<0.05), while MPO activity was also significantly reduced. Proinflammatory cytokine level was also reduced in treated group as compared to both other groups. On the basis of this study we concluded that the use of 20 mg/kg Angelica Sinensis in rat models can attenuate the secondary damage caused by SCI and thus help in controlling the pathology of SCI in rats. PMID:26261562

  10. Genomic Analysis of Detoxification Supergene Families in the Mosquito Anopheles sinensis

    PubMed Central

    Zhou, Dan; Liu, Xianmiao; Sun, Yan; Ma, Lei; Shen, Bo; Zhu, Changliang

    2015-01-01

    Anopheles sinensis is an important malaria vector in China and other Southeast Asian countries, and the emergence of insecticide resistance in this mosquito poses a serious threat to the efficacy of malaria control programs. The recently published An. sinensis genome and transcriptome provide an opportunity to understand the molecular mechanisms of insecticide resistance. Analysis of the An. sinensis genome revealed 174 detoxification genes, including 93 cytochrome P450s (P450s), 31 glutathione-S-transferases (GSTs), and 50 choline/carboxylesterases (CCEs). The gene number was similar to that in An. gambiae, but represented a decrease of 29% and 42% compared with Aedes aegypti and Culex quinquefasciatus, respectively. The considerable contraction in gene number in Anopheles mosquitoes mainly occurred in two detoxification supergene families, P450s and CCEs. The available An. sinensis transcriptome was also re-analyzed to further identify key resistance-associated detoxification genes. Among 174 detoxification genes, 124 (71%) were detected. Several candidate genes overexpressed in a deltamethrin-resistant strain (DR-strain) were identified as belonging to the CYP4 or CYP6 family of P450s and the Delta GST class. These generated data provide a basis for identifying the resistance-associated genes of An. sinensis at the molecular level. PMID:26588704

  11. Limnoithona sinensis as refuge for bacteria: protection from UV radiation and chlorine disinfection in drinking water treatment.

    PubMed

    Lin, Tao; Cai, Bo; Chen, Wei

    2014-11-01

    In this study, we tested the potential of Limnoithona sinensis to provide its attached bacteria refuge against disinfection. The experimental results indicated that in water devoid of zooplankton, both UV radiation and chlorine disinfection significantly decreased the viability of free-living bacteria. In the presence of L. sinensis, however, the attached bacteria could survive and rapidly recover from disinfection. This demonstrated that L. sinensis provided protection from external damage to various aquatic bacteria that were attached to its body. The surviving bacteria remained on L. sinensis after disinfection exposure, which enabled a rapid increase in the bacterial population followed by their subsequent release into the surrounding water. Compared with UV radiation, chlorine disinfection was more effective in terms of inactivating attached bacteria. Both UV radiation and chlorine disinfection had little effect in terms of preventing the spread of undesirable bacteria, due to the incomplete inactivation of the bacteria associated with L. sinensis.

  12. Extraction of Angelica sinensis polysaccharides using ultrasound-assisted way and its bioactivity.

    PubMed

    Zhao, Ying; Shi, Yongyong; Yang, Huixin; Mao, Lijuan

    2016-07-01

    In the present article, ultrasound-assisted extraction (UAE) of polysaccharides from Angelica sinensis were investigated. The aim of the study is to examine the extraction parameters such as ultrasound power (140-180W), the ratio of liquid to solid (5-7), extraction time (40-50min) and extraction temperature (80-100°C) and to obtain the best possible combinations of these parameters through response surface methodology (RSM). Based on contour plots and variance analysis, optimum operational conditions for maximizing polysaccharides yield were found to be 180w, 7, 45min and 90°C. Under the optimum operating conditions determined, 6.96% polysaccharides were achieved. In addition, the results showed that A. sinensis polysaccharides (ASP) could increase antioxidant enzymes activities and decrease the MDA levels in the skeletal muscle of exhaustive exercise rats. This study provides strong evidence that A. sinensis polysaccharides supplementation possessed protective effects against exhaustive exercise-induced oxidative stress.

  13. Metabolic fingerprinting of Angelica sinensis during growth using UPLC-TOFMS and chemometrics data analysis

    PubMed Central

    2013-01-01

    Background The radix of Angelica sinensis is widely used as a medicinal herbal and metabolomics research of this plant during growth is necessary. Results Principal component analysis of the UPLC-QTOFMS data showed that these 27 samples could be separated into 4 different groups. The chemical markers accounting for these separations were identified from the PCA loadings plot. These markers were further verified by accurate mass tandem mass and retention times of available reference standards. The study has shown that accumulation of secondary metabolites of Angelica sinensis is closely related to the growth periods. Conclusions The UPLC-QTOFMS based metabolomics approach has great potential for analysis of the alterations of secondary metabolites of Angelica sinensis during growth. PMID:23453085

  14. The extract of Cordyceps sinensis inhibited airway inflammation by blocking NF-κB activity.

    PubMed

    Chiou, Ya-Ling; Lin, Ching-Yuang

    2012-06-01

    Aiming the extract of Cordyceps sinensis significantly inhibits airway inflammation, airway hyperresponsiveness, and the infiltration of eosinophils in the airway of rats and may be related to the modulation of T helper (Th)1 and Th2 cells functions. The mechanisms of C. sinensis involved in modulation of suppression inflammation are not yet determined. In this study, the mechanism involved in the extract of C. sinensis-C.S.3-modulated suppression of inflammation was investigated in vivo and in vitro systems. The results showed that C.S.3 reduced airway inflammation in ovalbumin-induced allergic mice. Furthermore, we found C.S.3 could decrease extracellular signal-regulated kinase 1/2 signaling pathway to suppress activity of nuclear factor-κB in lung cells and cultured airway smooth muscle cells. Conclusion C.S.3 may provide clinical applications for asthma in the future.

  15. Review of Naturopathy of Medical Mushroom, Ophiocordyceps Sinensis, in Sexual Dysfunction

    PubMed Central

    Jiraungkoorskul, Kanitta; Jiraungkoorskul, Wannee

    2016-01-01

    Sexual dysfunctions including desire, arousal, orgasm, and pain disorders are increasing worldwide due to etiological factors and aging. Several types of treatment are claimed in modern medicine, but they have serious side effects and higher costs. In fact, alternative approaches, such as the intake of plants, fungi, and insects, or their extracts, have also been practiced to enhance sexuality and ameliorate illness with notable successes. However, the scientific evidence related to the mechanisms and efficacy of these alternative medicines is both scarce and all too often unconvincing. Ophiocordyceps sinensis is an Ascomycetes fungus parasitic to Lepidoptera larvae, and has long been used as medicine to treat many illnesses and promote longevity in Chinese society. Previous investigations have shown that O. sinensis has many pharmacological activities. This review has focused on illustrating that O. sinensis can enhance libido and sexual performance, and can restore impaired reproductive functions, such as impotency or infertility, in both sexes. PMID:27041868

  16. Green tea (Camellia sinensis) catechins and vascular function.

    PubMed

    Moore, Rosalind J; Jackson, Kim G; Minihane, Anne M

    2009-12-01

    The health benefits of green tea (Camellia sinensis) catechins are becoming increasingly recognised. Amongst the proposed benefits are the maintenance of endothelial function and vascular homeostasis and an associated reduction in atherogenesis and CVD risk. The mounting evidence for the influential effect of green tea catechins on vascular function from epidemiological, human intervention and animal studies is subject to review together with exploration of the potential mechanistic pathways involved. Epigallocatechin-3-gallate, one of the most abundant and widely studied catechin found in green tea, will be prominent in the present review. Since there is a substantial inconsistency in the published data with regards to the impact of green tea catechins on vascular function, evaluation and interpretation of the inter- and intra-study variability is included. In conclusion, a positive effect of green tea catechins on vascular function is becoming apparent. Further studies in animal and cell models using physiological concentrations of catechins and their metabolites are warranted in order to gain some insight into the physiology and molecular basis of the observed beneficial effects.

  17. Functional Characterization of a New Tea (Camellia sinensis) Flavonoid Glycosyltransferase.

    PubMed

    Zhao, Xianqian; Wang, Peiqiang; Li, Mingzhuo; Wang, Yeru; Jiang, Xiaolan; Cui, Lilan; Qian, Yumei; Zhuang, Juhua; Gao, Liping; Xia, Tao

    2017-03-15

    Tea (Camellia sinensis) is an important commercial crop, in which the high content of flavonoids provides health benefits. A flavonoid glycosyltransferase (CsUGT73A20), belonging to cluster IIIa, was isolated from tea plant. The recombinant CsUGT73A20 in Escherichia coli exhibited a broad substrate tolerance toward multiple flavonoids. Among them, kaempferol was the optimal substrate compared to quercetin, myricetin, naringenin, apigenin, and kaempferide. However, no product was detected when UDP-galactose was used as the sugar donor. The reaction assay indicated that rCsUGT73A20 performed multisite glycosidation toward flavonol compounds, mainly forming 3-O-glucoside and 7-O-glucoside in vitro. The biochemical characterization analysis of CsUGT73A20 showed more K7G product accumulated at pH 8.0, but K3G was the main product at pH 9.0. Kinetic analysis demonstrated that high pH repressed the glycosylation reaction at the 7-OH site in vitro. Besides, the content of five flavonol-glucosides was increased in CsUGT73A20-overexpressing tobaccos (Nicotiana tabacum).

  18. Spermiogenesis in soft-shelled turtle, Pelodiscus sinensis.

    PubMed

    Zhang, Li; Han, Xiang-Kun; Li, Mei-Ying; Bao, Hui-Jun; Chen, Qiu-Sheng

    2007-10-01

    Spermiogenesis in the soft-shelled turtle, Pelodiscus sinensis, was examined by transmission electron microscopy. The process includes nuclear elongation, chromatin condensation, acrosomal and flagellar development, and elimination of excess cytoplasm. In stage I, the proacrosomal vesicle occurs next to a shallow fossa of the nucleus, and a dense acrosomal granule forms beneath it. A smaller subacrosomal granule in the middle of the fibrous layer is related to the development of intranuclear tubules. The nucleus begins to move eccentrically. In stage II, the round proacrosomal vesicle is flattened by protrusion of the nuclear fossa, and the dense acrosomal granule diffuses into the vesicle, as the fibrous layer forms the subacrosomal cone. Circular manchettes develop around the nucleus, and the chromatin coagulates into small granules. The movement of the nucleus causes rearrangement of the cytoplasm. In stage III, the front of the elongating nucleus protrudes out of the spermatid and is covered by the flat acrosome; coarse granules replace the small ones within the nucleus. At the posterior pole of the head, mitochondria move backward. Numerous microtubules begin to assemble the axoneme of flagellum. In stage IV, the chromatin concentrates to dense homogeneous phase. The circular manchette is reorganized longitudinally. The Sertoli process covers the acrosome and the residues of the cytoplasmic lobes are eliminated. In stage V, the sperm head matures. After dissolution of the longitudinal manchette, the mitochondria arrange themselves around the proximal and distal centrioles. Caudal to the mitochondrial mass, a fibrous sheath surrounds the proximal portion of the flagellum.

  19. Fullerene-Based Symmetry in Hibiscus rosa-sinensis Pollen

    PubMed Central

    Andrade, Kleber; Guerra, Sara; Debut, Alexis

    2014-01-01

    The fullerene molecule belongs to the so-called super materials. The compound is interesting due to its spherical configuration where atoms occupy positions forming a mechanically stable structure. We first demonstrate that pollen of Hibiscus rosa-sinensis has a strong symmetry regarding the distribution of its spines over the spherical grain. These spines form spherical hexagons and pentagons. The distance between atoms in fullerene is explained applying principles of flat, spherical, and spatial geometry, based on Euclid’s “Elements” book, as well as logic algorithms. Measurements of the pollen grain take into account that the true spine lengths, and consequently the real distances between them, are measured to the periphery of each grain. Algorithms are developed to recover the spatial effects lost in 2D photos. There is a clear correspondence between the position of atoms in the fullerene molecule and the position of spines in the pollen grain. In the fullerene the separation gives the idea of equal length bonds which implies perfectly distributed electron clouds while in the pollen grain we suggest that the spines being equally spaced carry an electrical charge originating in forces involved in the pollination process. PMID:25003375

  20. Antioxidant activity of oils extracted from orange (Citrus sinensis) seeds.

    PubMed

    Jorge, Neuza; Silva, Ana Carolina da; Aranha, Caroline P M

    2016-05-31

    Due to the increasing production of food in the world with consequent increase of the production of waste, the importance of developing researches for its use is noticed. Thus, the interest in vegetable oils with bioactive compounds, such as the ones extracted from fruit seeds, is growing. Therefore, the present study aims to characterize the oils extracted from seeds of Hamlin, Natal, Pera-rio and Valencia orange varieties (Citrus sinensis), as to the levels of total carotenoids, total phenolic compounds, tocopherols and phytosterols, as well as to determine their antioxidant activity. The orange seed oils presented important content of total carotenoids (19.01 mg/kg), total phenolic compounds (4.43 g/kg), α-tocopherol (135.65 mg/kg) and phytosterols (1304.2 mg/kg). The antioxidant activity ranged from 56.0% (Natal) to 70.2% (Pera-rio). According to the results it is possible to conclude that the orange seed oils can be used as specialty oils in diet, since they contain considerable amounts of bioactive compounds and antioxidants.

  1. Identification of chemical markers in Cordyceps sinensis by HPLC-MS/MS.

    PubMed

    Hu, Hankun; Xiao, Ling; Zheng, Baogen; Wei, Xin; Ellis, Alexis; Liu, Yi-Ming

    2015-10-01

    Authentication and quality assessment of Cordyceps sinensis, a precious and pricey natural product that offers a variety of health benefits, is highly significant. To identify effective chemical markers, authentic C. sinensis was thoroughly screened by using HPLC-MS/MS. In addition to many previously reported ingredients, two glycosides, i.e., cyclo-Ala-Leu-rhamnose and Phe-o-glucose, were detected for the first time in this material. Six ingredients detected, including cordycepin, D-mannitol, Phe, Phe-o-glucose, cyclo-Gly-Pro, and cyclo-Ala-Leu-rhamnose, were selected as a collection of chemical markers. An HPLC-MS/MS method was developed to simultaneously quantify them with sensitivity and specificity. The method had limits of detection ranging from 0.008 μg mL(-1) for cordycepin to 0.75 μg mL(-1) for cyclo-Gly-Pro. Recovery was found between 96 and 103 % in all tests. To evaluate the effectiveness of the marker collection proposed, five authentic C. sinensis samples and five samples of its substitutes were analyzed. Cordycepin, D-mannitol, and Phe were found present in all samples. The contents ranged from 0.0076 to 0.029 % (w/w) for cordycepin, 0.33 to 18.9 % for mannitol, and 0.0013 to 0.642 % for Phe. Interestingly, the two glycosides, Phe-o-glucose and cyclo-Ala-Leu-rhamnose, were detected only in authentic C. sinensis samples. These results indicated that the proposed protocol based on HPLC-MS/MS quantification of the markers might have a great potential in authentication and quality assessment of C. sinensis. Graphical abstract Chemical markers of C. sinensis identified in this work.

  2. Dujardinascaris gigantea sp. n. (Nematoda: Ascaridida) from the critically endangered crocodile Alligator sinensis Fauvel (Reptilia: Crocodylia).

    PubMed

    Zhao, Jin-Hong; Li, Liang; Guo, Yan-Ning; Zhang, Lu-Ping

    2015-03-01

    The Chinese alligator Alligator sinensis Fauvel (Reptilia: Crocodylia) is considered as one of the most critically endangered species of the 23 extant crocodiles. However, our knowledge of the helminth parasites of this rare animal is completely lacking. During a helminthological survey of reptiles in China, we found a new ascaridoid nematode, Dujardinascaris gigantea sp. n. from A. sinensis. The morphology of D. gigantea sp. n. was studied using light and scanning electron microscopy. The new species was also characterised using molecular methods by sequencing and analysing the small ribosomal DNA (18S) and the second internal transcribed spacer (ITS-2).

  3. Transcriptome Sequencing of Chemically Induced Aquilaria sinensis to Identify Genes Related to Agarwood Formation

    PubMed Central

    Ye, Wei; Wu, Hongqing; He, Xin; Wang, Lei; Zhang, Weimin; Li, Haohua; Fan, Yunfei; Tan, Guohui; Liu, Taomei; Gao, Xiaoxia

    2016-01-01

    Background Agarwood is a traditional Chinese medicine used as a clinical sedative, carminative, and antiemetic drug. Agarwood is formed in Aquilaria sinensis when A. sinensis trees are threatened by external physical, chemical injury or endophytic fungal irritation. However, the mechanism of agarwood formation via chemical induction remains unclear. In this study, we characterized the transcriptome of different parts of a chemically induced A. sinensis trunk sample with agarwood. The Illumina sequencing platform was used to identify the genes involved in agarwood formation. Methodology/Principal Findings A five-year-old Aquilaria sinensis treated by formic acid was selected. The white wood part (B1 sample), the transition part between agarwood and white wood (W2 sample), the agarwood part (J3 sample), and the rotten wood part (F5 sample) were collected for transcriptome sequencing. Accordingly, 54,685,634 clean reads, which were assembled into 83,467 unigenes, were obtained with a Q20 value of 97.5%. A total of 50,565 unigenes were annotated using the Nr, Nt, SWISS-PROT, KEGG, COG, and GO databases. In particular, 171,331,352 unigenes were annotated by various pathways, including the sesquiterpenoid (ko00909) and plant–pathogen interaction (ko03040) pathways. These pathways were related to sesquiterpenoid biosynthesis and defensive responses to chemical stimulation. Conclusions/Significance The transcriptome data of the different parts of the chemically induced A. sinensis trunk provide a rich source of materials for discovering and identifying the genes involved in sesquiterpenoid production and in defensive responses to chemical stimulation. This study is the first to use de novo sequencing and transcriptome assembly for different parts of chemically induced A. sinensis. Results demonstrate that the sesquiterpenoid biosynthesis pathway and WRKY transcription factor play important roles in agarwood formation via chemical induction. The comparative analysis of

  4. Isosmotic points and their ecological significance for juvenile Chinese sturgeon Acipenser sinensis.

    PubMed

    Zhao, F; Zhuang, P; Zhang, T; Wang, Y; Hou, J; Liu, J; Zhang, L

    2015-04-01

    Serum osmolality and ion concentrations were measured in juvenile Chinese sturgeon Acipenser sinensis at different salinities to determine the isosmotic point. Isosmotic and isoionic concentrations were calculated from the regressions for serum and ambient osmolality, with Na(+) , Cl(-) and K(+) as salinities 9·19, 8·17, 7·89 and 9·70, respectively. These values were consistent with the salinity of the habitat where juvenile A. sinensis occur in the Yangtze Estuary, suggesting that an isosmotic salinity is an important factor driving their habitat choice.

  5. Chemical fingerprinting and quantitative analysis of two common Gleditsia sinensis fruits using HPLC-DAD.

    PubMed

    Chen, Jianping; Li, Zhonggui; Zheng, Ken Y Z; Guo, Ava J Y; Zhu, Kevin Y; Zhang, Wendy L; Zhan, Janis Y X; Dong, Tina T X; Su, Ziren; Tsim, Karl W K

    2013-12-01

    Gleditsiae Fructus Abnormalis and Gleditsiae Sinensis Fructus are obtained from different developmental stages of fruits from Gleditsia sinensis Lam. (Leguminosae). The possible interchangeable usage of the two fruits, however, has long been very controversial. Here, high performance liquid chromatography coupled with diode array detection was developed to explore their chemical fingerprinting profiles. Besides, the amounts of aglycones of saponin compounds, echinocystic acid and oleanolic acid in both fruits were quantified. The results indicated that there was no significant difference in the content of aglycones from the two types of fruits. However, their chromatographic fingerprints showed distinct characteristics. Therefore, the interchangeable application of these fruits has to be taken with a specific precaution.

  6. Citrus essential oil of Nigeria. Part V: Volatile constituents of sweet orange leaf oil (Citrus sinensis).

    PubMed

    Kasali, Adeleke A; Lawal, Oladipupo A; Eshilokun, Adeolu O; Olaniyan, Abayomi A; Opoku, Andy R; Setzer, William N

    2011-06-01

    The volatile oils extracted from leaves of eight cultivars of Citrus sinensis (L) Osbeck were comprehensively analysed by a combination of GC and GC-MS. Fifty four constituents accounting for 82.3-98.2% were identified. Sabinene (20.9-49.1%), delta-3-carene (0.3-14.3%), (E)-beta-ocimene (4.4-12.6%), linalool (3.7-11.1%) and terpinen-4-ol (1.7-12.5%) were the major constituents that are common to all the volatile oils. In addition, a cluster analysis was carried out and indicated at least four different chemotypes for the C. sinensis cultivars.

  7. Effect of Medium Supplements on Agrobacterium rhizogenes Mediated Hairy Root Induction from the Callus Tissues of Camellia sinensis var. sinensis

    PubMed Central

    Rana, Mohammad M.; Han, Zhuo-Xiao; Song, Da-Peng; Liu, Guo-Feng; Li, Da-Xiang; Wan, Xiao-Chun; Karthikeyan, Alagarsamy; Wei, Shu

    2016-01-01

    Tea (Camellia sinensis L.) is recalcitrant to Agrobacterium-mediated genetic transformation largely due to the bactericidal effects of tea polyphenols and phenolics oxidation induced by necrosis of explant tissue over the process of transformation. In this study, different antioxidants/adsorbents were added as supplements to the co-cultivation and post co-cultivation media to overcome these problems for the transformation improvement. Tea-cotyledon-derived calli were used as explants and Agrobacterium rhizognes strain ATCC 15834 was used as a mediator. Results showed that Agrobacterium growth, virulence (vir) gene expression and browning of explant tissue were greatly influenced by different supplements. Murashige and Skoog (MS) basal salts medium supplemented with 30 g·L−1 sucrose, 0.1 g·L−1 l-glutamine and 5 g·L−1 polyvinylpolypyrrolidone (PVPP) as co-cultivation and post co-cultivation media could maintain these parameters better that ultimately led to significant improvement of hairy root generation efficiency compared to that in the control (MS + 30 g·L−1 sucrose). Additionally, the reporter genes β-glucuronidase (gusA) and cyan fluorescent protein (cfp) were also stably expressed in the transgenic hairy roots. Our study would be helpful in establishing a feasible approach for tea biological studies and genetic improvement of tea varieties. PMID:27428960

  8. Bioactivity of Six Plant Extracts on Adults of Demotispa neivai (Coleoptera: Chrysomelidae)

    PubMed Central

    Martínez, Luis C.; Plata-Rueda, Angelica; Zanuncio, José C.; Serrão, José E.

    2015-01-01

    Demotispa neivai Bondar (Coleoptera: Chrysomelidae) damage oil palm fruits, which makes it necessary to develop products to control this insect. The mortality, repellency, and antifeeding effects on adults of D. neivai of six plant extracts of Azadirachta indica A. Juss. (Sapindales: Meliaceae), Ricinus communis (L.) (Malpighiaes: Euphorbiaceae), Citrus sinensis Oesbek (Sapindales: Rutaceae), Nicotiana tabacum (L.) (Slanales: Solanaceae), Capsicum annuum (L.) (Solanales: Solanaceae), and Artemisia absinthium (L.) (Asterales: Asteraceae) were determined: 1) the lethal concentration LC50-90, lethal time of D. neivai was evaluated after spraying the fruits of oil palm; 2) repellent effects of each ingredient were evaluated by calculating the index of repellency; 3) antifeeding effects with the rate of inhibition calculated between doses of 20 and 24 g/liter. The mortality of D. neivai was higher with the extracts Ci. sinensis, R. communis, N. tabacum, and Ca. annuum. The mortality of D. neivai increased in the first 72 hr in all treatments. The extracts of N. tabacum, Ca. annuum, and A. indica were more repellent to D. neivai that those of Ci. sinensis, Ar. Absinthium, and R. communis. Antifeeding effect was higher with Ci. sinensis and R. communis. The increased mortality of D. neivai by Ci. sinensis can be explained by the effect of this compound on the respiratory system of insects. Extracts of Ci. sinensis, R. communis, N. tabacum, and Ca. annuum repelled and caused mortality of D. neivai and, thus, can be used in integrate pest management programs of this pest in oil palm plantations. PMID:25843587

  9. Immunomodulation by Hibiscus rosa-sinensis: effect on the humoral and cellular immune response of Mus musculus.

    PubMed

    Mishra, Nidhi; Tandon, Vijay Lakshmi; Gupta, Rekha

    2012-03-15

    In West India, the Hibiscus rosa-sinensis L. (Malvaceae) is traditionally used as tea as a natural diuretic. Extract of this plant contains Vitamin C and is used traditionally as a mild medicine. In spite of a long history of traditional medicinal value of H. rosa-sinensis still no data are available for immunomodulatory activity. In present investigation, aqueous extract of H. rosa-sinensis (AEHrs) (500 mg kg(-1) BW) was intraperitoneally (IP) injected to the male Swiss albino mice (Mus musculus) to evaluate the immunomodulatory property of extract. In addition for evaluation of phytochemical constituents of flowers of H. rosa-sinensis HPTLC was performed. The crude extract of H. rosa-sinensis has immunomodulatory activity. After the 15 days treatment, the number of plaque forming cells increased by 0.6%, antibody titre was increased 38.15% and significant increase of 52% was observed in DTH response. At the same concentration of dose the level of serum IL-1alpha enhanced significantly (14.27%) whereas a considerable decrease (32.70%) in the concentration of IL-2 was observed among AEHrs treated mice in comparison to the control mice. HPTLC chromatogram revealed that H. rosa-sinensis posses alkaloid (Rf 0.93) and flavonoids (Rf 0.02, 0.06, 0.14) on the basis of Rf values. Results of investigation supports for the immunomodulatory activity of H. rosa-sinensis aqueous extract.

  10. rDNA-targeted PCR primers and FISH probe in the detection of Ophiocordyceps sinensis hyphae and conidia.

    PubMed

    Zhong, Xin; Peng, Qingyun; Qi, Lili; Lei, Wei; Liu, Xin

    2010-11-01

    Ophiocordyceps sinensis (Berk.) Sung, Sung, Hywel-Jones & Spatafora (syn. Cordyceps sinensis) one of the entomopathogenic fungi, is a rare Traditional Chinese Medicine (TCM) found in the Qinghai-Tibetan Plateau. Polymerase Chain Reaction (PCR) and Fluorescence in situ hybridization (FISH) methods are necessary to identify the mycelia or spores of O. sinensis from its habitat and to monitor its dispersal, colonization and infectivity. To develop both primers and probe specific to O. sinensis, ribosomal DNA (rDNA) amplified with universal primers from O. sinensis genomic DNA and seven closely related fungi were sequenced. According to these sequences, the upper and lower primers (OsT-F and OsT-R) were designed within internal transcribed spacer region 1 (ITS1) and ITS2 and flanked by universal primers ITS5 and ITS4, respectively. The designed primers were used for general PCR, touchdown PCR, or both together with the universal primers for nested-touchdown PCR. The results showed that only the extracted DNA of O. sinensis was specifically amplified. The sensitivity of nested-touchdown PCR with extracted DNA of O. sinensis is as low as 10(-14)g (10 fg) and at least 1000 times higher than the other PCR methods. In addition, Cy5-labeled probe (OsLSU) for cytoplasmic LSU rRNA was hybridized with the ascospores of O. sinensis. It showed a strong red fluorescence throughout the whole cell but did not cross-react with other entomopathogenic fungi. Taken together, these methods were useful for studying the biology and ecology of O. sinensis.

  11. [Sterol extracts from Begonia Sinensis Rhizome against respiratory inflammation].

    PubMed

    Yao, Yong; Jiang, Wei; Li, Yu-shan

    2015-08-01

    The acute and chronic respiratory tract inflammation models were made to investigate the effect and mechanism of sterol extracts from Begonia Sinensis Rhizome (BSR). The first model of acute lung injury was made with Kunming mice by inhaling cigarette smoke, then the mice were treated with different concentrations of BSR sterol extracts. Lung tissue morphology was detected by HE staining, TNF-alpha/MPO were detected by Elisa, and cPLA2 protein were, detected by Western blotting respectively. Results showed that in model group, lung sheet became real, alveolar space shrank or disappeared, alveolar septum was thickened, plenty of inflammatory cells were infiltrated, capillary blood vessels were congestive and the expression of TNF-α, MPO, cPLA2 increased; after administration, a small amount of inflammatory cells were infiltrated, alveolar septum became obvious, capillary congestion status was significantly relieved and the expression of TNF-α, MPO, cPLA2 decreased (P < 0.05). The second model of chronic respiratory tract inflammation in BALB/c mice with bronchial asthma was induced by OVA, then the mice were treated with different concentrations of BSR sterol extracts. Lung tissue morphology was detected by HE staining, indexes such as IL-4, IL-5, IL-13 were detected by Elisa, and the cPLA2 protein expression was detected by Western blotting respectively. Results showed that in model group, a lot of inflammatory cells around lung vessels and bronchi exuded, bronchial goblet cells proliferated and the expression of IL-4, IL-5, IL-13, cPLA2 increased; after administration, inflammatory and goblet cell hyperplasia reduced, the expression of IL-4, IL-5, IL-13, cPLA2 also decreased (P < 0.05). The above results showed BSR sterol extracts could resist against respiratory inflammation by inhibiting cPLA2 in a dose-dependent manner.

  12. Evaluation of Hirsutella sinensis mycelium for antifatigue effect.

    PubMed

    Wu, Lung-Yuan; Wu, Ming-Fang; Lu, Hsu-Feng; Liu, Chia-Hui; Lee, Ching-Hsiao; Chen, Yung-Liang; Hsueh, Shu-Ching; Yeh, Chun; Huang, Yi-Ping; Liu, Jia-You; Chung, Jing-Gung

    2015-01-01

    The aim of this study was to investigate whether Hirsutella sinensis mycelium (HSM) has any antifatigue effect, using a forced swimming model in rats. Forty rats were randomly divided into five groups, each containing eight animals. The control group received 2 ml/kg body weight of distilled water and a positive control group was administered 1.13 ml/kg Quaker Essence of Chicken. The treated swimming groups were administered HSM powder manufactured by Chang Gung Biotechnology Corporation, Ltd., at doses of 63 mg/kg, 189 mg/kg or 378 mg/kg body weight/day, respectively for a period of six weeks. The above experiment was repeated with another 40 rats but for a period of eight weeks. At the end of the experiments, rats were placed in a swimming apparatus and the total swimming time until exhaustion was recorded. Pre-/post-exercise concentrations of serum urea nitrogen (BUN) and lactic acid were also determined. There were no deaths during the study. Physical and behavioral examinations did not reveal any treatment-related adverse effects after dosing. Changes in lactate levels were dose-dependent for the 8- but not the 6-week treatment. BUN levels were more affected by the 8-week treatment of HSM but not significantly altered in the 6-week treatment groups. The 8-week treatment groups showed a significant increase in swimming time to exhaustion compared to the control groups, which was not dose-dependent. For the 6-week treatment, only the middle and high doses increased swimming time to exhaustion. Conjugated diene contents were significantly higher in rats treated at any HSM dose for 8-weeks than the control groups. Swimming did not alter levels of liver glycogen when compared to the control sub-groups. Results of this study demonstrate that HSM improves physical endurance, which may be beneficial in treating conditions where fatigue is a factor and other antifatigue treatments are contraindicated.

  13. Multilocus sequence typing analysis of Pseudomonas aeruginosa isolated from pet Chinese stripe-necked turtles (Ocadia sinensis)

    PubMed Central

    Wendt, Mitchell

    2016-01-01

    Our research sought to characterize the phylogeny of Pseudomonas aeruginosa isolated from pet Chinese stripe-necked turtles (Ocadia sinensis) to better understand its evolutionary relation to other isolates and increase understanding of a potential zoonotic pathogen transmitted through direct contact with pet turtles. Thirty-one Pseudomonas aeruginosa isolates were obtained from both immature and adult turtles sold in pet shops in Korea. To characterize the phylogenic position of Chinese stripe-necked turtle-borne P. aeruginosa relative to other strains, multilocus sequence typing (MLST) analysis was performed due to the accessibility and breadth of MLST databases. Seven housekeeping genes (acsA, aroE, guaA, mutL, nuoD, ppsA, and trpE) were sequenced and the results were compared with data from the MLST database. The genes were further used for phylogenetic analysis of P. aeruginosa using concatenated gene fragments. Both rooted and unrooted phylogenetic trees were generated. Eleven distinct sequence types were present within the isolates among which seven were new. Expanding an unrooted phylogenetic tree to include P. aeruginosa MLST sequences isolated from various other geographic locations and sources revealed a divergent cluster containing the majority of isolates obtained from turtles. This suggests that P. aeruginosa strains particularly well-adapted for inhabiting turtles occupy a distinct phylogenetic position. PMID:28053614

  14. Sarcocystis sinensis is an ultrastructurally distinct parasite of water buffalo that can cause foodborne illness but cannot complete its life-cycle in human beings.

    PubMed

    Chen, Xinwen; Zuo, Yangxian; Rosenthal, Benjamin M; He, Yongshu; Cui, Liwang; Yang, Zhaoqing

    2011-05-31

    In this study, we compared the morphology of Sarcocystis sinensis and Sarcocystis hominis, and assessed the infectiousness of S. sinensis for human volunteers. The cysts of S. sinensis were from water buffalo (Bubalus bubalis) and those of S. hominis were from cattle (Bos taurus). Transmission electron microscopy of S. sinensis cysts revealed that the cyst wall had leaning, finger-like protrusions measuring 1.44-5.08 μm in length and without invaginations on the tip surface of the protrusions. In contrast, the cyst wall of S. hominis had upright, finger-like protrusions measuring 9.43 μm×2.42 μm and with vesicle-like invaginations on the tip surface of the protrusions. Scanning electron microscopy revealed that surface of the protrusions was arranged as rectangles in S. sinensis, as compared to tongue-shaped in S. hominis. Other distinguishing features of S. sinensis include a thin ground substrate (GS) zone with microtubules and small, circle-like structures located at the base of the protrusions. Human volunteers, after consuming S. sinensis cysts, produced no sporocysts or oocysts in feces, suggesting that humans could not serve as definitive hosts for S. sinensis. By contrast, many sporocysts and oocysts were passed in feces of a human volunteer 11-29 days after ingestion of S. hominis cysts. These results showed that S. sinensis and S. hominis are separate species and S. sinensis cannot use human being as the definitive host.

  15. Chromatographic and mass spectrometric fingerprinting analyses of Angelica sinensis (Oliv.) Diels-derived dietary supplements.

    PubMed

    Zhao, Yang; Sun, Jianghao; Yu, Liangli Lucy; Chen, Pei

    2013-05-01

    Angelica sinensis (Oliv.) Diels ("Danggui" in Chinese) is one of the most commonly used traditional Chinese medicines. It has been used to invigorate blood circulation for the treatment of anemia, hypertension, chronic bronchitis, asthma, rheumatism, and cardiovascular diseases. There are a number of A. sinensis-derived dietary supplements in the US markets. However, no study have been conducted to investigate the quality of these dietary supplements. In this paper, high-performance liquid chromatographic and flow-injection mass spectrometric fingerprints were both evaluated to assess the consistency of A. sinensis-derived dietary supplements. Similarity analysis was carried out on the high-performance liquid chromatographic (HPLC) fingerprints. Meanwhile, principal component analysis (PCA) was performed on the data obtained from flow-injection mass spectrometric (FIMS) fingerprints, which can analyze each sample in 2 min, compared with 30 min required for the chromatographic fingerprint. Both methods show significant chemical differences between samples that may be due to differences in growing locations, growing conditions, harvesting times, and/or botanical processing. The loading plots obtained from PCA singled out the discriminatory ions that were responsible for chemical differences of A. sinensis-derived dietary supplements.

  16. Diverse Colletotrichum species cause anthracnose of tea plants (Camellia sinensis (L.) O. Kuntze) in China

    PubMed Central

    Wang, Yu-Chun; Hao, Xin-Yuan; Wang, Lu; Bin Xiao; Wang, Xin-Chao; Yang, Ya-Jun

    2016-01-01

    Anthracnose caused by Colletotrichum is one of the most severe diseases that can afflict Camellia sinensis. However, research on the diversity and geographical distribution of Colletotrichum in China remain limited. In this study, 106 Colletotrichum isolates were collected from diseased leaves of Ca. sinensis cultivated in the 15 main tea production provinces in China. Multi-locus phylogenetic analysis coupled with morphological identification showed that the collected isolates belonged to 11 species, including 6 known species (C. camelliae, C. cliviae, C. fioriniae, C. fructicola, C. karstii, and C. siamense), 3 new record species (C. aenigma, C. endophytica, and C. truncatum), 1 novel species (C. wuxiense), and 1 indistinguishable strain, herein described as Colletotrichum sp. Of these species, C. camelliae and C. fructicola were the dominant species causing anthracnose in Ca. sinensis. In addition, our study provided further evidence that phylogenetic analysis using a combination of ApMat and GS sequences can be used to effectively resolve the taxonomic relationships within the C. gloeosporioides species complex. Finally, pathogenicity tests suggested that C. camelliae, C. aenigma, and C. endophytica are more invasive than other species after the inoculation of the leaves of Ca. sinensis. PMID:27782129

  17. Draft Genome Sequence of Methylobacterium mesophilicum Strain SR1.6/6, Isolated from Citrus sinensis

    PubMed Central

    Marinho Almeida, Diogo; Dini-Andreote, Francisco; Camargo Neves, Aline Aparecida; Jucá Ramos, Rommel Thiago; Andreote, Fernando Dini; Carneiro, Adriana Ribeiro; Oliveira de Souza Lima, André; Caracciolo Gomes de Sá, Pablo Henrique; Ribeiro Barbosa, Maria Silvanira

    2013-01-01

    Methylobacterium mesophilicum strain SR1.6/6 is an endophytic bacterium isolated from a surface-sterilized Citrus sinensis branch. Ecological and biotechnological aspects of this bacterium, such as the genes involved in its association with the host plant and the primary oxidation of methanol, were annotated in the draft genome. PMID:23788544

  18. Volatile compounds of healthy and insect-damaged Hippophae rhamnoides sinensis in natural and planted forests.

    PubMed

    Zong, Shixiang; Luo, Youqing; Zhou, Jiao; Liu, Shujing

    2012-01-01

    Volatile compounds of healthy and insect-damaged stems of Hippophae rhamnoides sinensis were analysed using dynamic headspace and thermal-desorption cold-trap injector gas chromatography/mass spectroscopy (TCT-GC/MS). Sixteen compounds, belonging to alkanes, alcohols, aldehydes, esters, ketones, and ethers, were identified in the stems of healthy H. rhamnoides sinensis; the compounds in H. rhamnoides sinensis occurring naturally or cultivated in plantations were similar, but the relative contents were significantly different. In plants damaged by Holcocerus hippophaecolus, the nature and content of the volatile compounds were greatly changed. Butanedione and butyl glyoxylate were newly generated after damage by the pest, and the relative levels of pentanal, heptanal, eucalyptol, terpineol, and camphor were sharply increased in both naturally occurring and plantation-grown plants. n-Decane, trans-2-nonen-1-ol, and n-hexadecane levels increased in plants cultivated in the plantation and decreased in natural forests, whereas the levels of other types were reduced. Thus, both the nature and the content of volatile compounds of H. rhamnoides sinensis are affected by H. hippophaecolus damage, providing a theoretical basis to identify the mechanism of pest destruction.

  19. De Novo Assembly and Comparative Transcriptome Analysis Provide Insight into Lysine Biosynthesis in Toona sinensis Roem

    PubMed Central

    Zhang, Xia; Song, Zhenqiao; Liu, Tian; Guo, Linlin; Li, Xingfeng

    2016-01-01

    Toona sinensis Roem is a popular leafy vegetable in Chinese cuisine and is also used as a traditional Chinese medicine. In this study, leaf samples were collected from the same plant on two development stages and then used for high-throughput Illumina RNA-sequencing (RNA-Seq). 125,884 transcripts and 54,628 unigenes were obtained through de novo assembly. A total of 25,570 could be annotated with known biological functions, which indicated that the T. sinensis leaves and shoots were undergoing multiple developmental processes especially for active metabolic processes. Analysis of differentially expressed unigenes between the two libraries showed that the lysine biosynthesis was an enriched KEGG pathway, and candidate genes involved in the lysine biosynthesis pathway in T. sinensis leaves and shoots were identified. Our results provide a primary analysis of the gene expression files of T. sinensis leaf and shoot on different development stages and afford a valuable resource for genetic and genomic research on plant lysine biosynthesis. PMID:27376077

  20. Inflorescence architecture affects pollinator behaviour and mating success in Spiranthes sinensis (Orchidaceae).

    PubMed

    Iwata, Tatsunori; Nagasaki, Osamu; Ishii, Hiroshi S; Ushimaru, Atushi

    2012-01-01

    • Despite the wide inflorescence diversity among angiosperms, the effects of inflorescence architecture (three-dimensional flower arrangement) on pollinator behaviour and mating success have not been sufficiently studied in natural plant populations. • Here, we investigated how inflorescence architecture affected inter- and intra-plant pollinator movements and consequent mating success in a field population of Spiranthes sinensis var. amoena (S. sinensis). In this species, the flowers are helically arranged around the stem, and the degree of twisting varies greatly among individuals. The large variation in inflorescence architecture in S. sinensis results from variation in a single structural parameter, the helical angle (the angular distance between neighbour-flower directions). • The numbers of visits per inflorescence and successive probes per visit by leaf-cutting bees decreased with helical angle, indicating that individual flowers of tightly twisted inflorescences received less visitations. As expected from pollinator behaviour, pollinia removal and fruit set of individual flowers decreased with helical angle. Meanwhile, geitonogamy decreased in tightly twisted inflorescences. • Our novel findings demonstrate that natural variation in inflorescence architecture significantly affects pollinator behaviour and reproductive success, suggesting that inflorescence architecture can evolve under pollinator-mediated natural selection in plant populations. We also discuss how diverse inflorescence architectures may have been maintained in S. sinensis populations.

  1. Chromatographic and mass spectrometric fingerprinting analyses of angelica sinensis (Oliv.) diels dietary supplements

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Angelica sinensis (Oliv.) Diels (“Danggui” in Chinese) is one of the most commonly used Traditional Chinese Medicines (TCMs). It has been used to invigorate blood circulation for the treatment of anemia, hypertension, chronic bronchitis, asthma, rheumatism and cardiovascular diseases. There are a lo...

  2. Exogenous abscisic acid significantly affects proteome in tea plant (Camellia sinensis) exposed to drought stress

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Tea [Camellia sinensis (L.) O. Kuntze] is an important economic crop, and drought is the most important abiotic stress affecting yield and quality. Abscisic acid (ABA) is an important phytohormone responsible for activating drought resistance. Increased understanding of ABA effects on tea plant unde...

  3. Hematology and plasma biochemistry values of great cormorant (Phalacrocorax carbo sinensis) nestlings.

    PubMed

    Minias, Piotr; Kaczmarek, Krzysztof; Janiszewski, Tomasz; Markowski, Janusz

    2013-01-01

    Hematology and plasma biochemistry values were determined in 92 free-living Great Cormorant (Phalacrocorax carbo sinensis) chicks at Jeziorsko reservoir, central Poland. Percentage distribution of leukocytes, packed cell volume, plasma concentrations of hemoglobin and basic biochemical parameters were evaluated. These values may be treated as reference ranges for free-living Great Cormorant nestlings.

  4. Synergistic antimicrobial activity of Camellia sinensis and Juglans regia against multidrug-resistant bacteria.

    PubMed

    Farooqui, Amber; Khan, Adnan; Borghetto, Ilaria; Kazmi, Shahana U; Rubino, Salvatore; Paglietti, Bianca

    2015-01-01

    Synergistic combinations of antimicrobial agents with different mechanisms of action have been introduced as more successful strategies to combat infections involving multidrug resistant (MDR) bacteria. In this study, we investigated synergistic antimicrobial activity of Camellia sinensis and Juglans regia which are commonly used plants with different antimicrobial agents. Antimicrobial susceptibility of 350 Gram-positive and Gram-negative strains belonging to 10 different bacterial species, was tested against Camellia sinensis and Juglans regia extracts. Minimum inhibitory concentrations (MICs) were determined by agar dilution and microbroth dilution assays. Plant extracts were tested for synergistic antimicrobial activity with different antimicrobial agents by checkerboard titration, Etest/agar incorporation assays, and time kill kinetics. Extract treated and untreated bacteria were subjected to transmission electron microscopy to see the effect on bacterial cell morphology. Camellia sinensis extract showed higher antibacterial activity against MDR S. Typhi, alone and in combination with nalidixic acid, than to susceptible isolates." We further explore anti-staphylococcal activity of Juglans regia that lead to the changes in bacterial cell morphology indicating the cell wall of Gram-positive bacteria as possible target of action. The synergistic combination of Juglans regia and oxacillin reverted oxacillin resistance of methicillin resistant Staphylococcus aureus (MRSA) strains in vitro. This study provides novel information about antimicrobial and synergistic activity of Camellia sinensis and Juglans regia against MDR pathogens.

  5. A Nucleotide Signature for the Identification of Angelicae Sinensis Radix (Danggui) and Its Products

    PubMed Central

    Wang, Xiaoyue; Liu, Yang; Wang, Lili; Han, Jianping; Chen, Shilin

    2016-01-01

    It is very difficult to identify Angelicae sinensis radix (Danggui) when it is processed into Chinese patent medicines. The proposed internal transcribed spacer 2 (ITS2) is not sufficient to resolve heavily processed materials. Therefore, a short barcode for the identification of processed materials is urgently needed. In this study, 265 samples of Angelicae sinensis radix and adulterants were collected. The ITS2 region was sequenced, and based on one single nucleotide polymorphism(SNP) site unique to Angelica sinensis, a nucleotide signature consisting of 37-bp (5′-aatccgcgtc atcttagtga gctcaaggac ccttagg-3′) was developed. It is highly conserved and specific within Angelica sinensis while divergent among other species. Then, we designed primers (DG01F/DG01R) to amplify the nucleotide signature region from processed materials. 15 samples procured online were analysed. By seeking the signature, we found that 7 of them were counterfeits. 28 batches of Chinese patent medicines containing Danggui were amplified. 19 of them were found to contain the signature, and adulterants such as Ligusticum sinense, Notopterygium incisum, Angelica decursiva and Angelica gigas were detected in other batches. Thus, this nucleotide signature, with only 37-bp, will broaden the application of DNA barcoding to identify the components in decoctions, Chinese patent medicines and other products with degraded DNA. PMID:27713564

  6. Metal Analysis in Citrus Sinensis Fruit Peel and Psidium Guajava Leaf

    PubMed Central

    Dhiman, Anju; Nanda, Arun; Ahmad, Sayeed

    2011-01-01

    The determination of metal traces is very important because they are involved in biological cycles and indicate high toxicity. The objective of the present study is to measure the levels of heavy metals and mineral ions in medicinally important plant species, Citrus sinensis and Psidium guajava. This study investigates the accumulation of Copper (Cu), Zinc (Zn), Cadmium (Cd), Aluminum (Al), Mercury (Hg), Arsenic (As), Selenium (Se) and inorganic minerals like Calcium (Ca) and Magnesium (Mg) in C. sinensis (sweet orange) fruit peel and P. guajava (guava) leaf, to measure the levels of heavy metal contamination. Dried powdered samples of the plants were digested using wet digestion method and elemental determination was done by atomic absorption spectrophotometer. Results are expressed as mean ± standard deviation and analysed by student's ‘t’ test. Values are considered significant at P < 0.05. The results were compared with suitable safety standards and the levels of Cu, Zn, Cd, Mg and Ca in C. sinensis fruit peel and P. guajava leaves were within the acceptable limits for human consumption. The order of concentration of elements in both the samples showed the following trend: Mg > Ca > Al > Zn > Cu > Cd > Hg = As = Se. The content of Hg, As and Se in C. sinensis fruit peel and P. guajava leaves was significantly low and below detection limit. The content of toxic metals in tested plant samples was found to be low when compared with the limits prescribed by various authorities (World Health Organization, WHO; International Centre for Materials Research, ICMR; American Public Health Association, APHA). The content of Hg, As and Se in C. sinensis fruit peel and P. guajava leaves was not detectable and met the appropriate safety standards. In conclusion, the tested plant parts taken in the present study were found to be safe. PMID:21976824

  7. Sequencing and De Novo Assembly of the Gonadal Transcriptome of the Endangered Chinese Sturgeon (Acipenser sinensis)

    PubMed Central

    Du, Hao; Zhang, Shuhuan; Wei, Qiwei

    2015-01-01

    Background The Chinese sturgeon (Acipenser sinensis) is endangered through anthropogenic activities including over-fishing, damming, shipping, and pollution. Controlled reproduction has been adopted and successfully conducted for conservation. However, little information is available on the reproductive regulation of the species. In this study, we conducted de novo transcriptome assembly of the gonad tissue to create a comprehensive dataset for A. sinensis. Results The Illumina sequencing platform was adopted to obtain 47,333,701 and 47,229,705 high quality reads from testis and ovary cDNA libraries generated from three-year-old A. sinensis. We identified 86,027 unigenes of which 30,268 were annotated in the NCBI non-redundant protein database and 28,281 were annotated in the Swiss-prot database. Among the annotated unigenes, 26,152 and 7,734 unigenes, respectively, were assigned to gene ontology categories and clusters of orthologous groups. In addition, 12,557 unigenes were mapped to 231 pathways in the Kyoto Encyclopedia of Genes and Genomes Pathway database. A total of 1,896 unigenes, potentially differentially expressed between the two gonad types, were found, with 1,894 predicted to be up-regulated in ovary and only two in testis. Fifty-five potential gametogenesis-related genes were screened in the transcriptome and 34 genes with significant matches were found. Besides, more paralogs of 11 genes in three gene families (sox, apolipoprotein and cyclin) were found in A. sinensis compared to their orthologs in the diploid Danio rerio. In addition, 12,151 putative simple sequence repeats (SSRs) were detected. Conclusions This study provides the first de novo transcriptome analysis currently available for A. sinensis. The transcriptomic data represents the fundamental resource for future research on the mechanism of early gametogenesis in sturgeons. The SSRs identified in this work will be valuable for assessment of genetic diversity of wild fish and genealogy

  8. [Fast Detection of Camellia Sinensis Growth Process and Tea Quality Informations with Spectral Technology: A Review].

    PubMed

    Peng, Ji-yu; Song, Xing-lin; Liu, Fei; Bao, Yi-dan; He, Yong

    2016-03-01

    The research achievements and trends of spectral technology in fast detection of Camellia sinensis growth process information and tea quality information were being reviewed. Spectral technology is a kind of fast, nondestructive, efficient detection technology, which mainly contains infrared spectroscopy, fluorescence spectroscopy, Raman spectroscopy and mass spectroscopy. The rapid detection of Camellia sinensis growth process information and tea quality is helpful to realize the informatization and automation of tea production and ensure the tea quality and safety. This paper provides a review on its applications containing the detection of tea (Camellia sinensis) growing status(nitrogen, chlorophyll, diseases and insect pest), the discrimination of tea varieties, the grade discrimination of tea, the detection of tea internal quality (catechins, total polyphenols, caffeine, amino acid, pesticide residual and so on), the quality evaluation of tea beverage and tea by-product, the machinery of tea quality determination and discrimination. This paper briefly introduces the trends of the technology of the determination of tea growth process information, sensor and industrial application. In conclusion, spectral technology showed high potential to detect Camellia sinensis growth process information, to predict tea internal quality and to classify tea varieties and grades. Suitable chemometrics and preprocessing methods is helpful to improve the performance of the model and get rid of redundancy, which provides the possibility to develop the portable machinery. Future work is to develop the portable machinery and on-line detection system is recommended to improve the further application. The application and research achievement of spectral technology concerning about tea were outlined in this paper for the first time, which contained Camellia sinensis growth, tea production, the quality and safety of tea and by-produce and so on, as well as some problems to be solved

  9. The oldest record of Alligator sinensis from the Late Pliocene of Western Japan, and its biogeographic implication

    NASA Astrophysics Data System (ADS)

    Iijima, Masaya; Takahashi, Keiichi; Kobayashi, Yoshitsugu

    2016-07-01

    The late Cenozoic fossil record of alligators in East Asia is crucial in understanding the origin and past distribution of Asian alligators that are now represented by a single species, Alligator sinensis. This study reports a partial skeleton of A. sinensis from the Late Pliocene (approximately 3.0 Ma) of western Japan. This Japanese A. sinensis is large in size (>200 cm total length), comparable to the maximum size of extant individuals. It demonstrates the oldest record of A. sinensis and wider distribution of this species in the past. Tectonic and geographic history of East Asia suggests that alligators presumably dispersed into Japan before 25 Ma or after 10 Ma, yet finally were wiped out from Japan due to the semi-isolated condition of the Japanese island arc and the deteriorated climate during the Plio-Pleistocene.

  10. Anxiolytic activity of aqueous extract of Camellia sinensis in rats

    PubMed Central

    Shastry, Rajeshwari; Ullal, Sheetal Dinkar; Karkala, Shreyas; Rai, Seema; Gadgade, Akash

    2016-01-01

    Objectives: The present study was undertaken to evaluate anxiolytic effect of Camellia sinensis (CS) and possible mechanism on acute and chronic administration in rats. Materials and Methods: Eight groups of rats with six in each group were used. Group I served as control. Group II received diazepam (1 mg/kg). Groups III, IV, and V received CS in doses of 3.3, 16.5, and 33 mg/kg, respectively. Three pharmacologically validated experimental models – elevated plus maze (EPM), light and dark box (LDB), and open field tests (OFT) – were employed. Each animal was tested initially in the EPM and then in the LDB, followed by the OFT in a single setting. In EMP, number of entries into, time spent in, and number of rears in each arm in a 5-min period were noted. In LDB, number of entries and time spent in bright arena, number of rears, and duration of immobility were noted. In OFT, number of peripheral and central squares crossed, time spent, and number of rears in central squares were observed for a 5-min period. One-way ANOVA followed by post hoc least significant difference test was performed. Results: In EPM and LDB, CS at 3.3, 16.5, and 33 mg/kg (acute and chronic models) increased the number of entries and time spent and rearing in the open arms and bright arena, respectively, compared to control. In the OFT, CS at 16.5 and 33 mg/kg significantly increased the number of squares crossed, time spent, and the number of rears in the central squares compared to control. Anxiolytic effect was dose dependent in EPM and LDB and CS at 33 mg/kg showed better anxiolytic activity compared to diazepam (1 mg/kg) in all models. Flumazenil (0.5 mg/kg) and bicuculline (1 mg/kg) completely inhibited while picrotoxin (1 mg/kg) partially inhibited the anxiolytic effect of CS. Diazepam and CS at 33 mg/kg reduced the locomotor activity in rats. Conclusion: CS has dose-dependent anxiolytic activity which is comparable to diazepam. Anxiolytic action of CS is likely mediated through GABAA

  11. Protective effect of extract of Cordyceps sinensis in middle cerebral artery occlusion-induced focal cerebral ischemia in rats

    PubMed Central

    2010-01-01

    Background Ischemic hypoxic brain injury often causes irreversible brain damage. The lack of effective and widely applicable pharmacological treatments for ischemic stroke patients may explain a growing interest in traditional medicines. From the point of view of "self-medication" or "preventive medicine," Cordyceps sinensis was used in the prevention of cerebral ischemia in this paper. Methods The right middle cerebral artery occlusion model was used in the study. The effects of Cordyceps sinensis (Caterpillar fungus) extract on mortality rate, neurobehavior, grip strength, lactate dehydrogenase, glutathione content, Lipid Peroxidation, glutathione peroxidase activity, glutathione reductase activity, catalase activity, Na+K+ATPase activity and glutathione S transferase activity in a rat model were studied respectively. Results Cordyceps sinensis extract significantly improved the outcome in rats after cerebral ischemia and reperfusion in terms of neurobehavioral function. At the same time, supplementation of Cordyceps sinensis extract significantly boosted the defense mechanism against cerebral ischemia by increasing antioxidants activity related to lesion pathogenesis. Restoration of the antioxidant homeostasis in the brain after reperfusion may have helped the brain recover from ischemic injury. Conclusions These experimental results suggest that complement Cordyceps sinensis extract is protective after cerebral ischemia in specific way. The administration of Cordyceps sinensis extract significantly reduced focal cerebral ischemic/reperfusion injury. The defense mechanism against cerebral ischemia was by increasing antioxidants activity related to lesion pathogenesis. PMID:20955613

  12. Cordyceps sinensis preserves intestinal mucosal barrier and may be an adjunct therapy in endotoxin-induced sepsis rat model: a pilot study

    PubMed Central

    Gu, Guo-Sheng; Ren, Jian-An; Li, Guan-Wei; Yuan, Yu-Jie; Li, Ning; Li, Jie-Shou

    2015-01-01

    Background: Cordyceps sinensis (C. sinensis), a traditional Chinese medicine, exhibits various pharmacological activities such as reparative, antioxidant, and apoptosis inhibitory effects. Intestinal barrier dysfunction plays a vital role in the progression of sepsis. We aimed to explore the effect of C. sinensis on the gut barrier and evaluate its efficacy in sepsis. Methods: A murine model of gut barrier dysfunction was created by intraperitoneal injection of endotoxin. C. sinensis or saline was administered orally after the induction of sepsis. Alterations of intestinal barrier were evaluated and compared in terms of epithelial cell apoptosis, proliferation index (PI), intercellular tight junction (TJ) and proliferating cell nuclear antigen (PCNA). Results: C. sinensis significantly decreased the percentage of apoptotic cells and promoted mucosal cells proliferation indicated by enhanced PI and PCNA expression in the intestinal mucosa compared to control group. The TJs between epithelial cells which were disrupted in septic rats were also restored by treatment of C. sinensis. In survival studies, C. sinensis was demonstrated to confer a protection against the lethal effect of sepsis. Conclusion: These results suggest that C. sinensis has gut barrier-protection effect in endotoxin-induced sepsis by promoting the proliferation and inhibiting the apoptosis of intestinal mucosal cells, as well as restoring the TJs of intestinal mucosa. C. sinensis may have the potential to be a useful adjunct therapy for sepsis. PMID:26221273

  13. Morphological Observations and Fatty Acid Composition of Indoor-Cultivated Cordyceps sinensis at a High-Altitude Laboratory on Sejila Mountain, Tibet

    PubMed Central

    Guo, Lian-Xian; Xu, Xiao-Ming; Liang, Fu-Rui; Yuan, Jian-Ping; Peng, Juan; Wu, Chou-Fei; Wang, Jiang-Hai

    2015-01-01

    Cordyceps sinensis, a caterpillar entomopathogenic fungus-host larva complex, is a rare medicinal herb found in the Qinghai-Tibetan Plateau and its surrounding high-altitude areas. The alternation of generations in the life cycle, whatever the fungus or its host insect, requires special growth conditions. However, it is difficult to simulate the growth conditions of C. sinensis, which hinders its artificial cultivation. In this work, the life cycle from the host larva to C. sinensis was observed in an indoor-cultivation laboratory at 4,200 m a.s.l. on Sejila Mountain, Tibet. Comparative examinations between indoor-cultivated and wild C. sinensis demonstrated that the indoor-cultivated C. sinensis preferred to germinate multiple long, slim stromata at diverse positions on dead larvae, including but not limited to their heads. Their fatty acid composition shows a significant difference in the levels of polyunsaturated fatty acids (PUFAs). In indoor-cultivated C. sinensis, PUFAs constituted 24.59% and 49.43%, respectively, of neutral and polar lipids; meanwhile, in wild C. sinensis, PUFAs represented 34.34% and 61.25% of neutral and polar lipids, respectively. These observations and fatty acid data suggest that environmental factors, particularly temperature, soil pressure and light intensity, strongly affect the growth of C. sinensis. Our new findings may provide important information for improving techniques for the large-scale artificial cultivation of C. sinensis. PMID:25938484

  14. Isolation and characterization of polymorphic microsatellite markers in the endangered species Bretschneidera sinensis Hemsl.

    PubMed

    Li, M; Chen, H F; Wang, Z F; Zhang, S

    2016-08-19

    Bretschneidera sinensis is an endangered species that is mainly distributed in South China. As a tertiary relict and the single species in the Bretschneideraceae family, it has a high conservation value. To investigate the influence of human disturbance on its mating system, 63 new microsatellites were developed using restriction-site-associated DNA sequencing and their polymorphisms were tested on 30 samples from one population. Among the 63 microsatellites, the number of alleles per locus ranged from 2 to 16. The observed and expected heterozygosities ranged from 0.133 to 0.967 and from 0.127 to 0.912, respectively. These microsatellites may be used for studying the mating system of B. sinensis as well as the within-population hereditary structure.

  15. [Effect of wound to growth of larva of host to Ophiocordyceps sinensis during artificial breeding].

    PubMed

    Liu, Fei; Zhang, De-li; Zeng, Wei; Li, Li; Luo, Qing-ming; Tu, Yong-qin; Chen, Shi-jiang; Yin, Ding-hua

    2015-01-01

    To clear the effect of the wound to the growth of the larva of the host to the Ophiocordyceps sinensis, the wounds of same severity at the same position were made artificially to the larva and which were artificial fed at the same environment and condition. The results indicated that, over the winter, the survival rate of the wounded of the infection larva was lower than that of the healthy larva, but the weight had no significant difference between the wounded and the healthy larva. The survival rate of the wounded of the no infection larva was lower than that of the healthy larva, but except with black skin, the wounded larva with offwhite and dusty red had no influence on the variety of the weight. In summery, wound had no advantage to the survival rate, but had no influence to the weight. The result had provided theoretical basis to the reforming of the system of the artificial culture O. sinensis.

  16. Gliocyte and Synapse Analyses in Cerebral Ganglia of the Chinese Mitten Crab, Eriocheir Sinensis: Ultrastructural Study

    PubMed Central

    Zhang, H.; Yu, P.; Zhong, S.; Ge, T.; Peng, S.; Zhou, Z.; Guo, X.

    2016-01-01

    The Chinese mitten crab Eriocheir sinensis is an economically important aquatic species in China. Many studies on gene structure, breeding, and diseases of the crab have been reported. However, knowledge about the organization of the nerve system of the crab remains largely unknown. To study the ultrastructure of the cerebral ganglia of E. sinensis and to compare the histological findings regarding the nerve systems of crustaceans, the cerebral ganglia were observed by transmission electron microscopy. The results showed that four types of gliocytes, including type I, II, III, and IV gliocytes were located in the cerebral ganglia. In addition, three types of synapses were present in the cerebral ganglia, including unidirectional synapses, bidirectional synapses, and combined type synapses. PMID:27734995

  17. Citrus sinensis annotation project (CAP): a comprehensive database for sweet orange genome.

    PubMed

    Wang, Jia; Chen, Dijun; Lei, Yang; Chang, Ji-Wei; Hao, Bao-Hai; Xing, Feng; Li, Sen; Xu, Qiang; Deng, Xiu-Xin; Chen, Ling-Ling

    2014-01-01

    Citrus is one of the most important and widely grown fruit crop with global production ranking firstly among all the fruit crops in the world. Sweet orange accounts for more than half of the Citrus production both in fresh fruit and processed juice. We have sequenced the draft genome of a double-haploid sweet orange (C. sinensis cv. Valencia), and constructed the Citrus sinensis annotation project (CAP) to store and visualize the sequenced genomic and transcriptome data. CAP provides GBrowse-based organization of sweet orange genomic data, which integrates ab initio gene prediction, EST, RNA-seq and RNA-paired end tag (RNA-PET) evidence-based gene annotation. Furthermore, we provide a user-friendly web interface to show the predicted protein-protein interactions (PPIs) and metabolic pathways in sweet orange. CAP provides comprehensive information beneficial to the researchers of sweet orange and other woody plants, which is freely available at http://citrus.hzau.edu.cn/.

  18. A novel sesquiterpene alcohol from Fimetariella rabenhorstii, an endophytic fungus of Aquilaria sinensis.

    PubMed

    Tao, Mei-Hua; Yan, Jian; Wei, Xiao-Yi; Li, Dong-Li; Zhang, Wei-Min; Tan, Jian-Wen

    2011-06-01

    A novel sesquiterpene alcohol, named frabenol (1), was isolated from liquid cultures of Fimetariella rabenhorstii A20, an endophytic fungus of the agarwood-forming plant Aquilaria sinensis (Lour.) Gilg. The structure of the new compound was elucidated on the basis of spectroscopic methods. The finding of a sesquiterpenoid compound in F. rabenhorstii A20 implied that endophytic fungi of agarwood-producing plants could also contribute to the generation of fragrant chemicals during the agarwood formation processes.

  19. Four new 2-(2-phenylethyl)chromone derivatives from withered wood of Aquilaria sinensis.

    PubMed

    Yagura, Toru; Ito, Michiho; Kiuchi, Fumiyuki; Honda, Gisho; Shimada, Yasuo

    2003-05-01

    Four new chromone derivatives, 5-hydroxy-6-methoxy-2-(2-phenylethyl)chromone (1), 6-hydroxy-2-(2-hydroxy-2-phenylethyl)chromone (2), 8-chloro-2-(2-phenylethyl)-5,6,7-trihydroxy-5,6,7,8-tetrahydrochromone (3), 6,7-dihydroxy-2-(2-phenylethyl)-5,6,7,8-tetrahydrochromone (4) were isolated from the MeOH extract of withered wood of Aquilaria sinensis, together with seven known constituents of agarwood.

  20. Qinanmer, a new compound from Chinese agarwood 'Qi-Nan' originating from Aquilaria sinensis.

    PubMed

    Shao, Hang; Kong, Fan-Dong; Wang, Hao; Mei, Wen-Li; Dai, Hao-Fu

    2017-01-02

    Qinanmer (1), a new compound comprising 2-(2-phenylethyl)chromone and sesquiterpene moieties, together with two known 2-(2-phenylethyl)chromone derivatives (2-3), was isolated from the high-quality Chinese agarwood "Qi-Nan" originating from Aquilaria sinensis (Lour.) Glig. The structure of 1 was elucidated by spectroscopic techniques (UV, IR, 1D and 2D NMR), MS analyses, ECD spectra analyses, and quantum (13)C NMR calculations.

  1. Amelioration of Gamma-hexachlorocyclohexane (Lindane) induced renal toxicity by Camellia sinensis in Wistar rats

    PubMed Central

    Prasad, W. L. N. V. Vara; Srilatha, Ch.; Sailaja, N.; Raju, N. K. B.; Jayasree, N.

    2016-01-01

    Aim: A study to assess the toxic effects of gamma-hexachlorocyclohexane (γ-HCH) (lindane) and ameliorative effects of Camellia sinensis on renal system has been carried out in male Wistar rats. Materials and Methods: Four groups of rats with 18 each were maintained under standard laboratory hygienic conditions and provided feed and water ad libitum. γ-HCH was gavaged at 20 mg/kg b.wt. using olive oil as vehicle to Groups II. C. sinensis at 100 mg/kg b.wt. was administered orally in distilled water to Group IV in addition to γ-HCH 20 mg/kg b.wt. up to 45 days to study ameliorative effects. Groups I and III were treated with distilled water and C. sinensis (100 mg/kg b.wt.), respectively. Six rats from each group were sacrificed at fortnight intervals. Serum was collected for creatinine estimation. The kidney tissues were collected in chilled phosphate buffer saline for antioxidant profile and in also 10% buffered formalin for histopathological studies. Results: γ-HCH treatment significantly increased serum creatinine and significantly reduced the renal antioxidative enzymes catalase, superoxide dismutase, and glutathione peroxidase. Grossly, severe congestion was noticed in the kidneys. Microscopically, kidney revealed glomerular congestion, atrophy, intertubular hemorrhages, degenerative changes in tubular epithelium with vacuolated cytoplasm, desquamation of epithelium and urinary cast formation. A significant reduction in serum creatinine levels, significant improvement in renal antioxidant enzyme activities and near to normal histological appearance of kidneys in Group IV indicated that the green tea ameliorated the effects of γ-HCH, on renal toxicity. Conclusion: This study suggested that C. sinensis extract combined with γ-HCH could enhance antioxidant/detoxification system which consequently reduced the oxidative stress thus potentially reducing γ-HCH toxicity and tissue damage. PMID:27956790

  2. Antioxidant and antibacterial activities of Hibiscus Rosa-sinensis Linn flower extracts.

    PubMed

    Khan, Zulfiqar Ali; Naqvi, Syed Ali-Raza; Mukhtar, Ammara; Hussain, Zaib; Shahzad, Sohail Anjum; Mansha, Asim; Ahmad, Matloob; Zahoor, Ameer Fawad; Bukhari, Iftikhar Hussain; Ashraf-Janjua, Muhammad Ramazan-Saeed; Mahmood, Nasir; Yar, Muhammad

    2014-05-01

    Antioxidant and antibacterial potential of different solvent extracts of locally grown Hibiscus rosa-sinensis Linn was evaluated. The antioxidant activity was assessed by estimation of total flavonoids contents, total phenolic contents, DPPH free radical scavenging activity and percentage inhibition of linoleic acid oxidation capacity. Agar disc diffusion method was used to assess antibacterial potential of crude extract of H. rosa-sinensis. The yield of the crude extracts (23.21 ± 3.67 and 18.36 ± 2.98% in 80% methanol and ethanol solvents was calculated, respectively. Methanol and ethanol extract of H. rosa-sinensis showed total phenolics 61.45 ± 3.23 and 59.31 ± 4.31 mg/100g as gallic acid equivalent, total flavonoids 53.28 ± 1.93 and 32.25±1.21 mg/100g as catechine equivalent, DPPH free radical scavenging activity 75.46±4.67 and 64.98 ± 2.11% and inhibition of linoleic acid oxidation potential 75.8 ±3.22 and 61.6 ± 2.01% respectively, was measured. Antibacterial study against three human pathogens such as staphlococus sp. Bacillus sp. and Escherichia coli showed growth inhibitory effect in the range of 12.75 ± 1.17 to 16.75 ± 2.10 mm. These results showed H. rosa-sinensis indigenous to Kallar Kahar and its allied areas bear promising medicinal values and could be used for developing herbal medicines to target oxidative stress and infectious diseases.

  3. Transcriptome using Illumina sequencing reveals the traits of spermatogenesis and developing testes in Eriocheir sinensis

    PubMed Central

    Qian, Hui

    2017-01-01

    Chinese mitten crab (Eriocheir sinensis) has the spermatozoa with typical aflagellate, decondensed chromatin, cup-shaped nuclei, and radial arms. However, the mechanism of spermatogenesis during which the specific spermatozoa are generated in this species is yet unclear. Here, the transcriptome of developing testis in E. sinensis was analyzed using the ways of RNA-seq and bioinformatics analysis to identify candidate genes potentially involved in development of testis and spermatogenesis. The Illumina HiSeq2500 sequencing of three replicons of samples produced a total of 145.19 M clean reads representing with a total of 21.34 Gb bases and 45.48% GC content. 56.30% clean reads were mapped to the draft genome of E. sinensis. The assembly of the transcriptome yielded contigs of 5691802 sequences and unigenes of 406527 sequences. Total 24246 and 40793 transcripts were annotated using Swissprot and Nr database, respectively. There were 48213 (70.31%) and 7858 (46.25%) transcripts with identity of more than 99 matching to mature testis unigenes in the databases of Nr and EST, respectively. The analytic results of KOG, GO and KEGG showed wide potential molecular functions of transcripts in the developing testes. KEGG analysis of unigenes yielded total 9422 predicted genes. Those predicted genes were involved in total 216 KEGG pathways related to the physiological activities of developing testis. 1975 predicted genes were involved in cellular and subcellular structural alteration of male germ cells. There were important roles of some pathways in the processes of morphological and structural biogenesis pertaining to testis development and spermatogenesis. Other 583 unigenes encoding the genetic and epigenetic factors also be found, which might contribute to the decondensation and stability of decondensed nuclei in the spermatozoa. These predicted events provide a view of the potential molecular mechanisms of development of testis and spermatogenesis in E. sinensis. PMID

  4. Therapeutic Effect of Captopril, Pentoxifylline, and Cordyceps Sinensis in Pre-Hepatic Portal Hypertensive Rats

    PubMed Central

    Ahmed, Ahmed F.; El-Maraghy, Nabila N.; Ghaney, Rasha H. Abdel; Elshazly, Shimaa M.

    2012-01-01

    Background/Aim: Portal hypertension is an important and potentially fatal complication of liver disease whereby cellular and fibrotic alterations manifest to increase portal venous pressure. The aim of this study is to investigate the effect of captopril, pentoxifylline (PTX), and cordyceps sinensis in pre-hepatic portal hypertensive rats. Settings and Design: Wister male rats were divided at random into 3 main groups: the first group: control rats. The second group: sham-operated rats and the third group: prehepatic portal hypertensive rats (PHPHT) induced by regulated pre-hepatic portal vein ligation. After 14 days, Group 3 was subdivided into 5 subgroups. Subgroup (1): portal vein-ligated (PVL) was killed at once; Subgroup (2): received distilled water for 30 days (untreated PVL group); subgroups 3-5 were treated with captopril (60 mg/kg, orally); PTX (100 mg/kg, orally); and C. sinensis (200 mg/kg, orally), respectively, as a single daily dose for 30 days. Patients and Methods: Portal pressure, nitric oxide (NO), antioxidant enzymes, Liver enzymes, and creatinine levels were measured to evaluate the status of the liver state. Results: Portal vein ligation produced significant increments in liver enzymes, NO, creatinine and portal pressure concomitant with significant decrements in glutathione content and superoxide dismutase activity. Treatment with captopril, PTX, and C. sinensis resulted in a significant reduction in liver enzymes, NO, creatinine and portal pressure and observable increase in antioxidant enzymes. Conclusions: captopril, PTX, and C. sinensis have promising effect in controlling PHPHT and reducing hyperdynamic circulatory state through reduction of portal pressure and NO level. PMID:22626797

  5. Chasing ghosts: Allopolyploid origin of Oxyria sinensis (Polygonaceae) from its only diploid congener and an unknown ancestor.

    PubMed

    Luo, Xin; Hu, Quanjun; Zhou, Pingping; Zhang, Dan; Wang, Qian; Abbott, Richard J; Liu, Jianquan

    2017-03-12

    Reconstructing the origin of a polyploid species is particularly challenging when an ancestor has become extinct. Under such circumstances the extinct donor of a genome found in the polyploid may be treated as a 'ghost' species in that its prior existence is recognised through the presence of its genome in the polyploid. In this study, we aimed to determine the polyploid origin of Oxyria sinensis (2n=40) for which only one congeneric species is known, i.e. diploid O. digyna (2n=14). Genomic in situ hybridization (GISH), transcriptome, phylogenetic and demographic analyses, and ecological niche modeling were conducted for this purpose. GISH revealed that O. sinensis comprised 14 chromosomes from O. digyna and 26 chromosomes from an unknown ancestor. Transcriptome analysis indicated that following divergence from O. digyna, involving genome duplication around 12 million years ago (Ma), a second genome duplication occurred approximately 6 Ma to give rise to O. sinensis. Oxyria sinensis was shown to contain homologous gene sequences divergent from those present in O. digyna in addition to a set that clustered with those in O. digyna. Coalescent simulations indicated that O. sinensis expanded its distribution approximately 6-7 Ma, possibly following the second polyploidization event, whereas O. digyna expanded its range much later. It was also indicated that the distributions of both species contracted and re-expanded during the Pleistocene climatic oscillations. Ecological niche modeling similarly suggested that both species experienced changes in their distributional ranges in response to Quaternary climatic changes. The extinction of the unknown 'ghost' tetraploid species implicated in the origin of O. sinensis could have resulted from superior adaptation of O. sinensis to repeated climatic changes in the region where it now occurs. This article is protected by copyright. All rights reserved.

  6. The Chinese soft-shelled turtle, Pelodiscus sinensis, decreases nitrogenous excretion, reduces urea synthesis and suppresses ammonia production during emersion.

    PubMed

    Ip, Yuen K; Lee, Serene M L; Wong, Wai P; Chew, Shit F

    2013-05-01

    The objective of this study was to examine the effects of 6 days of emersion on nitrogen metabolism and excretion in the Chinese soft-shelled turtle, Pelodiscus sinensis. Despite having a soft shell with a cutaneous surface that is known to be water permeable, P. sinensis lost only ~2% of body mass and was able to maintain its hematocrit and plasma osmolality, [Na(+)] and [Cl(-)] during 6 days of emersion. During emersion, it ameliorated water loss by reducing urine output, which led to a reduction (by 29-76%) in ammonia excretion. In comparison, there was a more prominent reduction (by 82-99%) in urea excretion during emersion due to a lack of water to flush the buccopharyngeal epithelium, which is known to be the major route of urea excretion. Consequently, emersion resulted in an apparent shift from ureotely to ammonotely in P. sinensis. Although urea concentration increased in several tissues, the excess urea accumulated could only account for 13-22% of the deficit in urea excretion. Hence, it can be concluded that a decrease (~80%) in urea synthesis occurred in P. sinensis during the 6 days of emersion. Indeed, emersion led to significant decreases in the activity of some ornithine-urea cycle enzymes (argininosuccinate synthetase/argininosuccinate lyase and arginase) from the liver of P. sinensis. As a decrease in urea synthesis occurred without the accumulation of ammonia and total free amino acids, it can be deduced that ammonia production through amino acid catabolism was suppressed with a proportional reduction in proteolysis in P. sinensis during emersion. Indeed, calculated results revealed that there could be a prominent decrease (~88%) in ammonia production in turtles after 6 days of emersion. In summary, despite being ureogenic and ureotelic in water, P. sinensis adopted a reduction in ammonia production, instead of increased urea synthesis, as the major strategy to ameliorate ammonia toxicity and problems associated with dehydration during

  7. Oral Administration of a Hot Water Extract of the Softshell Turtle (Trionyx sinensis) Improves Exercise Performance

    PubMed Central

    Harwanto, Dicky; Lee, Gong-Hyeon; Park, Sun-Mee; Choi, Jae-Suk; Kim, Mi-Ryung; Hong, Yong-Ki

    2015-01-01

    Freshwater softshell turtle (Trionyx sinensis) extract has been used traditionally as a tonic soup, and to recover from physical fatigue. To support these claims, the forelimb grip strength of mice was measured after feeding a soft-shell turtle extract for 7 days. The T. sinensis extract significantly increased the grip strength to 1.25±0.07 N (P<0.01), which is 16.8% higher than the force on day 0. After exercising, the blood glucose levels in extract-fed mice were 202% higher and urea levels were 73% lower, which were both significantly different than the levels observed after control treatment. Lactate dehydrogenase was significantly higher by 314%, and glutathione peroxidase increased by 165%. In addition, the obesity markers, serum triglyceride and cholesterol, decreased to 62% and 49%, respectively, after mice were fed the extract. These data show that the T. sinensis extract provided more energy for forelimb exercise, prevented protein catabolism and muscle fatigue, and decreased the oxidative stress caused by an exhaustive workout. PMID:26176001

  8. Computational identification and analysis of MADS box genes in Camellia sinensis.

    PubMed

    Gogoi, Madhurjya; Borchetia, Sangeeta; Bandyopadhyay, Tanoy

    2015-01-01

    MADS (Minichromosome Maintenance1 Agamous Deficiens Serum response factor) box genes encode transcription factors and they play a key role in growth and development of flowering plants. There are two types of MADS box genes- Type I (serum response factor (SRF)-like) and Type II (myocyte enhancer factor 2 (MEF2)-like). Type II MADS box genes have a conserved MIKC domain (MADS DNA-binding domain, intervening domain, keratin-like domain, and c-terminal domain) and these were extensively studied in plants. Compared to other plants very little is known about MADS box genes in Camellia sinensis. The present study aims at identifying and analyzing the MADS-box genes present in Camellia sinensis. A comparative bioinformatics and phylogenetic analysis of the Camellia sinensis sequences along with Arabidopsis thaliana MADS box sequences available in the public domain databases led to the identification of 16 genes which were orthologous to Type II MADS box gene family members. The protein sequences were classified into distinct clades which are associated with the conserved function of flower and seed development. The identified genes may be used for gene expression and gene manipulation studies to elucidate their role in the development and flowering of tea which may pave the way to improve the crop productivity.

  9. Short communication: in vitro assessment of antioxidant, antibacterial and phytochemical analysis of peel of Citrus sinensis.

    PubMed

    Mehmood, Basharat; Dar, Kamran Khurshid; Ali, Shaukat; Awan, Uzma Azeem; Nayyer, Abdul Qayyum; Ghous, Tahseen; Andleeb, Saiqa

    2015-01-01

    Antibacterial effect of Citrus sinensis peel extracts was evaluated against several pathogenic bacteria associated with human and fish infections viz., Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumonia, Staphylococcus aureus, Streptococcus pyogenes, Staphylococcus epidermidis, Serratia marcesnces, Shigella flexneri, Enterobacter amnigenus, Salmonella Typhimurium and Serratia odorifera. Methanol, ethanol, chloroform and diethyl ether solvents were used for extraction. In vitro antibacterial activity was analyzed by agar well and agar disc diffusion methods. It was found that ethanol extract showed highly significant inhibition of E. coli and K. pneumonia (12.6±0.94 mm and 11.6±1.2 mm) whereas methanol extract of C. sinensis also showed high zone of inhibition of S. odorifera (10.0±2.16 mm). The potential activity of active extracts was assessed and also compared with standard antibiotics through activity index formulation. The order of antioxidant activity through ABTS·+ and DPPH free radical scavenging activity was ethanol>methanol>chloroform>diethyl ether. Phytochemical screening of all solvents had determined the presence of terpenoids, alkaloids, steroids, glycosides and flavonoids. It was also found that Chloroform/Methanol (5:5) and Butanol/Ethanol/Water (4:1:2.2) solvent systems showed significant separation of active phytochemical constituents. These findings reveal the potential use of C. sinensis peel to treat infectious diseases, which are being caused by microorganisms.

  10. A Contemporary Treatment Approach to Both Diabetes and Depression by Cordyceps sinensis, Rich in Vanadium

    PubMed Central

    Guo, Jian-You; Han, Chun-Chao

    2010-01-01

    Diabetes mellitus is accompanied by hormonal and neurochemical changes that can be associated with anxiety and depression. Both diabetes and depression negatively interact, in that depression leads to poor metabolic control and hyperglycemia exacerbates depression. We hypothesize one novel vanadium complex of vanadium-enriched Cordyceps sinensis (VECS), which is beneficial in preventing depression in diabetes, and influences the long-term course of glycemic control. Vanadium compounds have the ability to imitate the action of insulin, and this mimicry may have further favorable effects on the level of treatment satisfaction and mood. C. sinensis has an antidepressant-like activity, and attenuates the diabetes-induced increase in blood glucose concentrations. We suggest that the VECS may be a potential strategy for contemporary treatment of depression and diabetes through the co-effect of C. sinensis and vanadium. The validity of the hypothesis can most simply be tested by examining blood glucose levels, and swimming and climbing behavior in streptozotocin-induced hyperglycemic rats. PMID:19948751

  11. Relationship between knockdown resistance, metabolic detoxification and organismal resistance to pyrethroids in Anopheles sinensis.

    PubMed

    Zhong, Daibin; Chang, Xuelian; Zhou, Guofa; He, Zhengbo; Fu, Fengyang; Yan, Zhentian; Zhu, Guoding; Xu, Tielong; Bonizzoni, Mariangela; Wang, Mei-Hui; Cui, Liwang; Zheng, Bin; Chen, Bin; Yan, Guiyun

    2013-01-01

    Anopheles sinensis is the most important vector of malaria in Southeast Asia, including China. Currently, the most effective measure to prevent malaria transmission relies on vector control through the use of insecticides, primarily pyrethroids. Extensive use of insecticides poses strong selection pressure on mosquito populations for resistance. Resistance to insecticides can arise due to mutations in the insecticide target site (target site resistance), which in the case of pyrethroids is the para-type sodium channel gene, and/or the catabolism of the insecticide by detoxification enzymes before it reaches its target (metabolic detoxification resistance). In this study, we examined deltamethrin resistance in An. sinensis from China and investigated the relative importance of target site versus metabolic detoxification mechanisms in resistance. A high frequency (>85%) of nonsynonymous mutations in the para gene was found in populations from central China, but not in populations from southern China. Metabolic detoxification as measured by the activity of monooxygenases and glutathione S-transferases (GSTs) was detected in populations from both central and southern China. Monooxygenase activity levels were significantly higher in the resistant than the susceptible mosquitoes, independently of their geographic origin. Stepwise multiple regression analyses in mosquito populations from central China found that both knockdown resistance (kdr) mutations and monooxygenase activity were significantly associated with deltamethrin resistance, with monooxygenase activity playing a stronger role. These results demonstrate the importance of metabolic detoxification in pyrethroid resistance in An. sinensis, and suggest that different mechanisms of resistance could evolve in geographically different populations.

  12. How cell wall complexity influences saccharification efficiency in Miscanthus sinensis

    SciTech Connect

    De Souza, Amanda P.; Kamei, Claire L. Alvim; Torres, Andres F.; Pattathil, Sivakumar; Hahn, Michael G.; Trindade, Luisa M.; Buckeridge, Marcos S.

    2015-04-23

    The production of bioenergy from grasses has been developing quickly during the last decade, with Miscanthus being among the most important choices for production of bioethanol. However, one of the key barriers to producing bioethanol is the lack of information about cell wall structure. Cell walls are thought to display compositional differences that lead to emergence of a very high level of complexity, resulting in great diversity in cell wall architectures. In this work, a set of different techniques was used to access the complexity of cell walls of different genotypes of Miscanthus sinensis in order to understand how they interfere with saccharification efficiency. Three genotypes of M. sinensis displaying different patterns of correlation between lignin content and saccharification efficiency were subjected to cell wall analysis by quantitative/qualitative analytical techniques such as monosaccharide composition, oligosaccharide profiling, and glycome profiling. When saccharification efficiency was correlated negatively with lignin, the structural features of arabinoxylan and xyloglucan were found to contribute positively to hydrolysis. In the absence of such correlation, different types of pectins, and some mannans contributed to saccharification efficiency. In conclusion, different genotypes of M. sinensis were shown to display distinct interactions among their cell wall components, which seem to influence cell wall hydrolysis.

  13. [Cloning and expression analysis of farnesyl pyrophosphate synthase from Aquilaria sinensis].

    PubMed

    Yang, Xin; Wei, Jian-He; Liu, Juan; Xu, Yan-Hong

    2013-10-01

    Farnesyl diphosphate synthase (FPS) is one of the key rate-limiting enzymes in the sesquiterpene metabolic pathways. In this study, the open reading frame (ORF) of FPS was cloned by PCR based on the transcript sequence of AsFPS1 from the Aquilaria sinensis transcriptome database and sequenced. Total RNA was extracted from the root, stem and leaves of three-year-old A. sinensis, and from healthy and wounded A. sinensis calli, and then reverse-transcribed into single-stranded cDNA as a template for real-time PCR, to detect the expression specificity of AsFPSI in different tissues and its expression profile in responding to different treatments. The result showed that the full length of AsFPS1 was 1 342 bp with the ORF 1 029 bp, encoding 342 amino acids. Tissue expression analysis indicated that AsFPS1 was mainly expressed in root and stem, and was lower in leaves. Inducible-experiments showed that the genes was induced by mechanical wound as well as chemical liquid induction, and reached the highest expression level at 6 h and 12 h, respectively. The full-length cDNA clone of AsFPSI and its expression patterns analysis will provide a foundation for follow-up study on its biological function and agarwood sesquiterpene biosynthesis mechanism.

  14. Computational identification and analysis of MADS box genes in Camellia sinensis

    PubMed Central

    Gogoi, Madhurjya; Borchetia, Sangeeta; Bandyopadhyay, Tanoy

    2015-01-01

    MADS (Minichromosome Maintenance1 Agamous Deficiens Serum response factor) box genes encode transcription factors and they play a key role in growth and development of flowering plants. There are two types of MADS box genes- Type I (serum response factor (SRF)-like) and Type II (myocyte enhancer factor 2 (MEF2)-like). Type II MADS box genes have a conserved MIKC domain (MADS DNA-binding domain, intervening domain, keratin-like domain, and c-terminal domain) and these were extensively studied in plants. Compared to other plants very little is known about MADS box genes in Camellia sinensis. The present study aims at identifying and analyzing the MADS-box genes present in Camellia sinensis. A comparative bioinformatics and phylogenetic analysis of the Camellia sinensis sequences along with Arabidopsis thaliana MADS box sequences available in the public domain databases led to the identification of 16 genes which were orthologous to Type II MADS box gene family members. The protein sequences were classified into distinct clades which are associated with the conserved function of flower and seed development. The identified genes may be used for gene expression and gene manipulation studies to elucidate their role in the development and flowering of tea which may pave the way to improve the crop productivity. PMID:25914445

  15. Comparison of the functional features of the pump organs of Anopheles sinensis and Aedes togoi

    PubMed Central

    Ha, Young-Ran; Lee, Seung-Chul; Seo, Seung-Jun; Ryu, Jeongeun; Lee, Dong-Kyu; Lee, Sang-Joon

    2015-01-01

    Mosquitoes act as vectors for severe tropical diseases. Mosquito-borne diseases are affected by various factors such as environmental conditions, host body susceptibility, and mosquito feeding behavior. Among these factors, feeding behavior is affected by the feeding pump system located inside the mosquito head and also depends on the species of mosquito. Therefore, the 3D morphological structures of the feeding pumps of Aedes togoi and Anopheles sinensis were comparatively investigated using synchrotron X-ray microscopic computed tomography. In addition, the feeding behaviors of their pumping organs were also investigated using a 2D X-ray micro-imaging technique. An. sinensis, a malarial vector mosquito, had a larger feeding pump volume than Ae. togoi in the static or resting position. Interestingly, the two species of mosquitoes exhibited different feeding behaviors. Ae. togoi had a higher feeding frequency and expansion ratio than An. sinensis. Ae. togoi also exhibited F-actin localization more clearly. These distinctive variations in feeding volumes and behaviors provide essential insight into the blood-feeding mechanisms of female mosquitoes as vectors for tropical diseases. PMID:26464043

  16. Antimicrobial effects of Citrus sinensis peel extracts against dental caries bacteria: An in vitro study

    PubMed Central

    Shetty, Sapna B.; Mahin-Syed-Ismail, Prabu; Varghese, Shaji; Thomas-George, Bibin; Kandathil- Thajuraj, Pathinettam; Baby, Deepak; Haleem, Shaista; Sreedhar, Sreeja

    2016-01-01

    Background Ethnomedicine is gaining admiration since years but still there is abundant medicinal flora which is unrevealed through research. The study was conducted to assess the in vitro antimicrobial potential and also determine the minimum inhibitory concentration (MIC) of Citrus sinensis peel extracts with a view of searching a novel extract as a remedy for dental caries pathogens. Material and Methods Aqueous and ethanol (cold and hot) extracts prepared from peel of Citrus sinensis were screened for in vitro antimicrobial activity against Streptococcus mutans and Lactobacillus acidophilus, using agar well diffusion method. The lowest concentration of every extract considered as the minimal inhibitory concentration (MIC) values were determined for both test organisms. One way ANOVA with Post Hoc Bonferroni test was applied for statistical analysis. Confidence level and level of significance were set at 95% and 5% respectively. Results Dental caries pathogens were inhibited most by hot ethanolic extract of Citrus sinensispeel followed by cold ethanolic extract. Aqueous extracts were effective at very high concentrations. Minimum inhibitory concentration of hot and cold ethanolic extracts of Citrus sinensis peel ranged between 12-15 mg/ml against both the dental caries pathogens. Conclusions Citrus sinensispeels extract was found to be effective against dental caries pathogens and contain compounds with therapeutic potential. Nevertheless, clinical trials on the effect of these plants are essential before advocating large-scale therapy. Key words:Agar well diffusion, antimicrobial activity, dental caries, Streptococcus mutans, Lactobacillus acidophilus. PMID:26855710

  17. Fermentation optimization for the production of bioactive polysaccharides from Cordyceps sinensis fungus UM01.

    PubMed

    Wang, Lan-Ying; Cheong, Kit-Leong; Wu, Ding-Tao; Meng, Lan-Zhen; Zhao, Jing; Li, Shao-Ping

    2015-08-01

    The optimal fermentation conditions and medium for the production of bioactive polysaccharides from the mycelium of Cordyceps sinensis fungus UM01 were investigated by using orthogonal design and high performance size exclusion chromatography coupled with multi-angel laser light scattering and refractive index detector (HPSEC-MALLS-RID). Results showed that the optimal temperature, initial pH, rotation speed, medium capacity (ratio of medium volume to the volume of flask bottle) and inoculums volume for the mycelium growth were 15 °C, pH 6.0, 150 rpm, 2/5 (v/v), and 3% (v/v), respectively. Furthermore, bioactive polysaccharides from the mycelium of C. sinensis fungus UM01 were determined as polysaccharide fractions with the molecular weight above 10 kDa. The optimal fermentation medium was determined as a composition of glucose 30.0 g/L, sucrose 30.0 g/L, KH2PO4 1.0 g/L, CaCl2 0.5 g/L, yeast extract 3.0 g/L, and MgCl2 0.1g/L according to the maximum amount of the bioactive polysaccharides (486.16±19.60 mg/L) measured by HPSEC-MALLS/RID. Results are helpful to establish an efficient and controllable fermentation process for the industrial production of bioactive polysaccharides from C. sinensis UM01, and beneficial to develop a unique health and functional product in future.

  18. In vitro antibacterial activity of Hibiscus rosa-sinensis flower extract against human pathogens

    PubMed Central

    Ruban, P; Gajalakshmi, K

    2012-01-01

    Objective To access the in vitro antibacterial activity of Hibiscus rosa-sinensis (H. rosa- sinensis) flower extract against human pathogens. Methods Antibacterial activity was evaluated by using disc and agar diffusion methods. The protein was run through poly acrylmide gel electrophoresis to view their protein profile. Results The results showed that the cold extraction illustrates a maximum zone of inhibition against Bacillus subtillis (B. subtillis), Escherichia coli (E. coli) viz., (17.00 ± 2.91), (14.50 ± 1.71) mm, followed by hot extraction against, E. coli, Salmonella sp. as (11.66 ± 3.14), (10.60 ± 3.09) mm. In methanol extraction showed a highest zone of inhibition recorded against B. subtillis, E. coli as (18.86 ± 0.18), (18.00 ± 1.63) mm pursued by ethanol extraction showed utmost zone of inhibition recorded against Salmonella sp. at (20.40 ± 1.54) mm. The crude protein from flower showed a maximum inhibitory zone observed against Salmonella sp., E. coli viz., (16.55 ± 1.16), (14.30 ± 2.86) mm. The flower material can be taken as an alternative source of antibacterial agent against the human pathogens. Conclusions The extracts of the H. rosa-sinensis are proved to have potential antibacterial activity, further studies are highly need for the drug development. PMID:23569938

  19. Bioactive Peptides from Angelica sinensis Protein Hydrolyzate Delay Senescence in Caenorhabditis elegans through Antioxidant Activities

    PubMed Central

    Wang, Qiangqiang; Huang, Yunxuan; Qin, Chuixin; Liang, Ming; Mao, Xinliang; Li, Shuiming; Zou, Yongdong; Jia, Weizhang; Li, Haifeng; Ma, Chung Wah; Huang, Zebo

    2016-01-01

    Since excessive reactive oxygen species (ROS) is known to be associated with aging and age-related diseases, strategies modulating ROS level and antioxidant defense systems may contribute to the delay of senescence. Here we show that the protein hydrolyzate from Angelica sinensis was capable of increasing oxidative survival of the model animal Caenorhabditis elegans intoxicated by paraquat. The hydrolyzate was then fractionated by ultrafiltration, and the antioxidant fraction (<3 kDa) was purified by gel filtration to obtain the antioxidant A. sinensis peptides (AsiPeps), which were mostly composed of peptides with <20 amino acid residues. Further studies demonstrate that AsiPeps were able to reduce the endogenous ROS level, increase the activities of the antioxidant enzymes superoxide dismutase and catalase, and decrease the content of the lipid peroxidation product malondialdehyde in nematodes treated with paraquat or undergoing senescence. AsiPeps were also shown to reduce age pigments accumulation and extend lifespan but did not affect the food-intake behavior of the nematodes. Taken together, our results demonstrate that A. sinensis peptides (AsiPeps) are able to delay aging process in C. elegans through antioxidant activities independent of dietary restriction. PMID:26941890

  20. The Chinese soft-shelled turtle, Pelodiscus sinensis, excretes urea mainly through the mouth instead of the kidney.

    PubMed

    Ip, Yuen K; Loong, Ai M; Lee, Serene M L; Ong, Jasmine L Y; Wong, Wai P; Chew, Shit F

    2012-11-01

    The Chinese soft-shelled turtle, Pelodiscus sinensis, is well adapted to aquatic environments, including brackish swamps and marshes. It is ureotelic, and occasionally submerges its head into puddles of water during emersion, presumably for buccopharyngeal respiration. This study was undertaken to test the hypothesis that the buccophyaryngeal cavity constitutes an important excretory route for urea in P. sinensis. Results indicate that a major portion of urea was excreted through the mouth instead of the kidney during immersion. When restrained on land, P. sinensis occasionally submerged their head into water (20-100 min), during which urea excretion and oxygen extraction occurred simultaneously. These results indicate for the first time that buccopharyngeal villiform processes (BVP) and rhythmic pharyngeal movements were involved in urea excretion in P. sinensis. Urea excretion through the mouth was sensitive to phloretin inhibition, indicating the involvement of urea transporters (UTs). In addition, saliva samples collected from the buccopharyngeal surfaces of P. sinensis injected intraperitoneally with saline contained ~36 mmol N l(-1) urea, significantly higher than that (~2.4 mmol N l(-1)) in the plasma. After intraperitoneal injection with 20 μmol urea g(-1) turtle, the concentration of urea in the saliva collected from the BVP increased to an extraordinarily high level of ~614 μmol N ml(-1), but the urea concentration (~45 μmol N ml(-1)) in the plasma was much lower, indicating that the buccopharyngeal epithelium of P. sinensis was capable of active urea transport. Subsequently, we obtained from the buccopharyngeal epithelium of P. sinensis the full cDNA sequence of a putative UT, whose deduced amino acid sequence had ~70% similarity with human and mouse UT-A2. This UT was not expressed in the kidney, corroborating the proposition that the kidney had only a minor role in urea excretion in P. sinensis. As UT-A2 is known to be a facilitative urea

  1. Bioactivity-Guided Fractionation and GC/MS Fingerprinting of Angelica sinensis and Angelica archangelica Root Components for Antifungal and Mosquito Deterrent Activity

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Angelica sinensis and A. archangelica belong to the Umbelliferae and both are used in traditional Chinese medicine to treat gynecological and intestinal disorders. In this study, oils from three different A. sinensis collections and one A. archangelica root were analyzed by GC and GC/MS. The domin...

  2. Complete Chloroplast Genomes of Erianthus arundinaceus and Miscanthus sinensis: Comparative Genomics and Evolution of the Saccharum Complex

    PubMed Central

    Tsuruta, Shin-ichi; Ebina, Masumi; Kobayashi, Makoto; Takahashi, Wataru

    2017-01-01

    The genera Erianthus and Miscanthus, both members of the Saccharum complex, are of interest as potential resources for sugarcane improvement and as bioenergy crops. Recent studies have mainly focused on the conservation and use of wild accessions of these genera as breeding materials. However, the sequence data are limited, which hampers the studies of phylogenetic relationships, population structure, and evolution of these grasses. Here, we determined the complete chloroplast genome sequences of Erianthus arundinaceus and Miscanthus sinensis by using 454 GS FLX pyrosequencing and Sanger sequencing. Alignment of the E. arundinaceus and M. sinensis chloroplast genome sequences with the known sequence of Saccharum officinarum demonstrated a high degree of conservation in gene content and order. Using the data sets of 76 chloroplast protein-coding genes, we performed phylogenetic analysis in 40 taxa including E. arundinaceus and M. sinensis. Our results show that S. officinarum is more closely related to M. sinensis than to E. arundinaceus. We estimated that E. arundinaceus diverged from the subtribe Sorghinae before the divergence of Sorghum bicolor and the common ancestor of S. officinarum and M. sinensis. This is the first report of the phylogenetic and evolutionary relationships inferred from maternally inherited variation in the Saccharum complex. Our study provides an important framework for understanding the phylogenetic relatedness of the economically important genera Erianthus, Miscanthus, and Saccharum. PMID:28125648

  3. Effects of Hibiscus rosa sinensis L (Malvaceae) on wound healing activity: a preclinical study in a Sprague Dawley rat.

    PubMed

    Shivananda Nayak, B; Sivachandra Raju, S; Orette, F A; Chalapathi Rao, A V

    2007-06-01

    Hibiscus rosa sinensis (H rosa sinensis), a plant product, has been used for the treatment of a variety of diseases as well as to promote wound healing. The wound-healing activity of the ethanol extract of H rosa sinensis flower was determined in rats, using excision, incision, and dead space wound models and is presented in this report. The animals were randomly divided into 2 groups of 6 each in all the models. Test group animals in each model were treated with the ethanol extract of H rosa sinensis orally by mixing in drinking water (120 mg kg(-1) day(-1)), and the control group animals were maintained with plain drinking water. Healing was assessed by the rate of wound contraction, period of epithelialization, tensile strength (skin breaking strength), granulation tissue weight, and hydroxyproline content. The antimicrobial activity of the flower extract against selected microorganisms that infect the wounds was also assessed. Animals treated with the extract exhibited an 86% reduction in the wound area compared with controls, who exhibited a 75% reduction. The extract-treated animals were found to epithelize their wounds significantly faster than controls (P < .002) and have shown significantly higher skin-breaking strength than controls (P < .002). The dry and wet weight of granulation tissue and hydroxyproline content were also increased significantly when compared with controls. The reported observations suggest H rosa sinensis aids wound healing in the rat model.

  4. Authentication of Cordyceps sinensis by DNA Analyses: Comparison of ITS Sequence Analysis and RAPD-Derived Molecular Markers.

    PubMed

    Lam, Kelly Y C; Chan, Gallant K L; Xin, Gui-Zhong; Xu, Hong; Ku, Chuen-Fai; Chen, Jian-Ping; Yao, Ping; Lin, Huang-Quan; Dong, Tina T X; Tsim, Karl W K

    2015-12-15

    Cordyceps sinensis is an endoparasitic fungus widely used as a tonic and medicinal food in the practice of traditional Chinese medicine (TCM). In historical usage, Cordyceps specifically is referring to the species of C. sinensis. However, a number of closely related species are named themselves as Cordyceps, and they are sold commonly as C. sinensis. The substitutes and adulterants of C. sinensis are often introduced either intentionally or accidentally in the herbal market, which seriously affects the therapeutic effects or even leads to life-threatening poisoning. Here, we aim to identify Cordyceps by DNA sequencing technology. Two different DNA-based approaches were compared. The internal transcribed spacer (ITS) sequences and the random amplified polymorphic DNA (RAPD)-sequence characterized amplified region (SCAR) were developed here to authenticate different species of Cordyceps. Both approaches generally enabled discrimination of C. sinensis from others. The application of the two methods, supporting each other, increases the security of identification. For better reproducibility and faster analysis, the SCAR markers derived from the RAPD results provide a new method for quick authentication of Cordyceps.

  5. Tapeworm Khawia sinensis: review of the introduction and subsequent decline of a pathogen of carp, Cyprinus carpio.

    PubMed

    Oros, Mikulás; Hanzelová, Vladimíra; Scholz, Tomás

    2009-10-14

    The Asian tapeworm Khawia sinensis Hsü, 1935 (Cestoda: Caryophyllidea) is a large-sized (body length up to 11.5 cm) monozoic (unsegmented) parasite of common carp (Cyprinus carpio L.) that may cause mortality of young fish (fry). Since the 1960s, this cestode successfully colonized a large part of Europe, including the British Isles, North America and Japan. However, a review of published records provides evidence that the tapeworm K. sinensis, invasive parasite of carp, has become less common during the last two decades. Decline of K. sinensis may have been related to the recent introduction of another invasive tapeworm, the caryophyllidean Atractolytocestus huronensis Anthony, 1958 to Europe. Other factors that may have caused that K. sinensis is much less common than previously are also briefly discussed. A comparison of K. sinensis from feral and cultured carp, published to date, with those recently found for the first time in wild populations of carp in Slovakia did not reveal any marked differences in their morphology or measurements.

  6. Genetic variations of ND5 gene of mtDNA in populations of Anopheles sinensis (Diptera: Culicidae) malaria vector in China

    PubMed Central

    2013-01-01

    Background Anopheles sinensis is a principal vector for Plasmodium vivax malaria in most parts of China. Understanding of genetic structure and genetic differentiation of the mosquito should contribute to the vector control and malaria elimination in China. Methods The present study investigated the genetic structure of An. sinensis populations using a 729 bp fragment of mtDNA ND5 among 10 populations collected from seven provinces in China. Results ND5 was polymorphic by single mutations within three groups of An. sinensis that were collected from 10 different geographic populations in China. Out of 140 specimens collected from 10 representative sites, 84 haplotypes and 71 variable positions were determined. The overall level of genetic differentiation of An. sinensis varied from low to moderate across China and with a FST range of 0.00065 – 0.341. Genealogy analysis clustered the populations of An. sinensis into three main clusters. Each cluster shared one main haplotype. Pairwise variations within populations were higher (68.68%) than among populations (31.32%) and with high fixation index (FST = 0.313). The results of the present study support population growth and expansion in the An. sinensis populations from China. Three clusters of An. sinensis populations were detected in this study with each displaying different proportion patterns over seven Chinese provinces. No correlation between genetic and geographic distance was detected in overall populations of An. sinensis (R2 = 0.058; P = 0.301). Conclusions The results indicate that the ND5 gene of mtDNA is highly polymorphic in An. sinensis and has moderate genetic variability in the populations of this mosquito in China. Demographic and spatial results support evidence of expansion in An. sinensis populations. PMID:24192424

  7. Quantitative analysis of marker compounds in Angelica gigas, Angelica sinensis, and Angelica acutiloba by HPLC/DAD.

    PubMed

    Jeong, Su Yang; Kim, Hye Mi; Lee, Kyu Ha; Kim, Kyu Yeob; Huang, Dae Sun; Kim, Jong Hwan; Seong, Rack Seon

    2015-01-01

    Although Danggui is the root of Angelica gigas NAKAI in the Korean Pharmacopoeia, it is determined that Danggui is also the root of Angelica sinensis (OLIV.) DIELS in China and Hong Kong, as well as the root of Angelica acutiloba KITAGAWA in Japan. Accordingly, we tried to develop an identification method using the main compounds in A. gigas, A. sinensis, and A. acutiloba through HPLC/diode-array detector (DAD). This method was fully validated for linearity, accuracy, precision, recovery, and robustness. Multivariate analysis was also implemented after pattern analysis and monitoring. As a result, each compound pattern of A. gigas, A. sinensis, and A. acutiloba was identified, making it possible to distinguish them from each other.

  8. Genetic diversity and distribution patterns of host insects of Caterpillar Fungus Ophiocordyceps sinensis in the Qinghai-Tibet Plateau.

    PubMed

    Quan, Qing-Mei; Chen, Ling-Ling; Wang, Xi; Li, Shan; Yang, Xiao-Ling; Zhu, Yun-Guo; Wang, Mu; Cheng, Zhou

    2014-01-01

    The caterpillar fungus Ophiocordyceps sinensis is one of the most valuable medicinal fungi in the world, and it requires host insects in family Hepialidae (Lepidoptera) to complete its life cycle. However, the genetic diversity and phylogeographic structures of the host insects remain to be explored. We analyzed the genetic diversity and temporal and spatial distribution patterns of genetic variation of the host insects throughout the O. sinensis distribution. Abundant haplotype and nucleotide diversity mainly existed in the areas of Nyingchi, ShangriLa, and around the edge of the Qinghai-Tibet Plateau, where are considered as the diversity center or micro-refuges of the host insects of O. sinensis. However, there was little genetic variation among host insects from 72.1% of all populations, indicating that the host species composition might be relatively simple in large-scale O. sinensis populations. All host insects are monophyletic except for those from four O. sinensis populations around Qinghai Lake. Significant phylogeographic structure (NST>GST, P<0.05) was revealed for the monophyletic host insects, and the three major phylogenetic groups corresponded with specific geographical areas. The divergence of most host insects was estimated to have occurred at ca. 3.7 Ma, shortly before the rapid uplift of the QTP. The geographical distribution and star-like network of the haplotypes implied that most host insects were derived from the relicts of a once-widespread host that subsequently became fragmented. Neutrality tests, mismatch distribution analysis, and expansion time estimation confirmed that most host insects presented recent demographic expansions that began ca. 0.118 Ma in the late Pleistocene. Therefore, the genetic diversity and distribution of the present-day insects should be attributed to effects of the Qinghai-Tibet Plateau uplift and glacial advance/retreat cycles during the Quaternary ice age. These results provide valuable information to guide

  9. High resolution genetic mapping by genome sequencing reveals genome duplication and tetraploid genetic structure of the diploid Miscanthus sinensis.

    PubMed

    Ma, Xue-Feng; Jensen, Elaine; Alexandrov, Nickolai; Troukhan, Maxim; Zhang, Liping; Thomas-Jones, Sian; Farrar, Kerrie; Clifton-Brown, John; Donnison, Iain; Swaller, Timothy; Flavell, Richard

    2012-01-01

    We have created a high-resolution linkage map of Miscanthus sinensis, using genotyping-by-sequencing (GBS), identifying all 19 linkage groups for the first time. The result is technically significant since Miscanthus has a very large and highly heterozygous genome, but has no or limited genomics information to date. The composite linkage map containing markers from both parental linkage maps is composed of 3,745 SNP markers spanning 2,396 cM on 19 linkage groups with a 0.64 cM average resolution. Comparative genomics analyses of the M. sinensis composite linkage map to the genomes of sorghum, maize, rice, and Brachypodium distachyon indicate that sorghum has the closest syntenic relationship to Miscanthus compared to other species. The comparative results revealed that each pair of the 19 M. sinensis linkages aligned to one sorghum chromosome, except for LG8, which mapped to two sorghum chromosomes (4 and 7), presumably due to a chromosome fusion event after genome duplication. The data also revealed several other chromosome rearrangements relative to sorghum, including two telomere-centromere inversions of the sorghum syntenic chromosome 7 in LG8 of M. sinensis and two paracentric inversions of sorghum syntenic chromosome 4 in LG7 and LG8 of M. sinensis. The results clearly demonstrate, for the first time, that the diploid M. sinensis is tetraploid origin consisting of two sub-genomes. This complete and high resolution composite linkage map will not only serve as a useful resource for novel QTL discoveries, but also enable informed deployment of the wealth of existing genomics resources of other species to the improvement of Miscanthus as a high biomass energy crop. In addition, it has utility as a reference for genome sequence assembly for the forthcoming whole genome sequencing of the Miscanthus genus.

  10. Transcriptomic Analysis Reveals the Molecular Mechanisms of Drought-Stress-Induced Decreases in Camellia sinensis Leaf Quality

    PubMed Central

    Wang, Weidong; Xin, Huahong; Wang, Mingle; Ma, Qingping; Wang, Le; Kaleri, Najeeb A.; Wang, Yuhua; Li, Xinghui

    2016-01-01

    The tea plant [Camellia sinensis (L.) O. Kuntze] is an important commercial crop rich in bioactive ingredients, especially catechins, caffeine, theanine and other free amino acids, which the quality of tea leaves depends on. Drought is the most important environmental stress affecting the yield and quality of this plant. In this study, the effects of drought stress on the phenotype, physiological characteristics and major bioactive ingredients accumulation of C. sinensis leaves were examined, and the results indicated that drought stress resulted in dehydration and wilt of the leaves, and significant decrease in the total polyphenols and free amino acids and increase in the total flavonoids. In addition, HPLC analysis showed that the catechins, caffeine, theanine and some free amino acids in C. sinensis leaves were significantly reduced in response to drought stress, implying that drought stress severely decreased the quality of C. sinensis leaves. Furthermore, differentially expressed genes (DEGs) related to amino acid metabolism and secondary metabolism were identified and quantified in C. sinensis leaves under drought stress using high-throughput Illumina RNA-Seq technology, especially the key regulatory genes of the catechins, caffeine, and theanine biosynthesis pathways. The expression levels of key regulatory genes were consistent with the results from the HPLC analysis, which indicate a potential molecular mechanism for the above results. Taken together, these data provide further insights into the mechanisms underlying the change in the quality of C. sinensis leaves under environmental stress, which involve changes in the accumulation of major bioactive ingredients, especially catechins, caffeine, theanine and other free amino acids. PMID:27066035

  11. Identification and functional characterization of three type III polyketide synthases from Aquilaria sinensis calli.

    PubMed

    Wang, Xiaohui; Zhang, Zhongxiu; Dong, Xianjuan; Feng, Yingying; Liu, Xiao; Gao, Bowen; Wang, Jinling; Zhang, Le; Wang, Juan; Shi, Shepo; Tu, Pengfei

    2017-03-30

    Type III polyketide synthases (PKSs) play an important role in biosynthesis of various plant secondary metabolites and plant adaptation to environmental stresses. Aquilaria sinensis is the main plant species for production of agarwood, little is known about the PKS family. In this study, AsCHS1 and two new type III PKSs, AsPKS1 and AsPKS2, were isolated and characterized in Aquilaria sinensis calli. The comparative sequence and phylogenetic analysis indicated that AsPKS1 and AsPKS2 belong to non-CHS group different from AsCHS1. The recombinant AsPKS1 and AsPKS2 produced the lactone-type products, suggesting different enzyme activities with AsCHS1. Three PKS genes had a tissues-specific pattern in A. sinensis. Moreover, we examined the expression profiles of three PKS genes in calli under different abiotic stresses and hormone treatments. AsCHS1 transcript was significantly induced by salt stress, AsPKS1 abundance was most remarkably enhanced by CdCl2 treatment, while AsPKS2 expression was most significantly induced by mannitol treatment. Furthermore, AsCHS1, AsPKS1 and AsPKS2 transcript were enhanced upon gibberellins (GA3), methyl jasmonate (MeJA), salicylic acid (SA) treatments, while three PKS genes displayed low transcript levels at the early stage under abscisic acid (ABA) treatment. In addition, three GFP:PKSs fusion proteins were localized in the cytoplasm and cell wall in Nicotiana benthamiana cells. These results indicated the multifunctional role of three type III PKSs in polyketide biosynthesis, plant resistance in abiotic stresses and signal transduction.

  12. Cloning, expression, and characterization of Fe-SOD from Isöetes sinensis.

    PubMed

    Xu, Y; Dai, X L; Liu, B D; Wang, Q X

    2016-11-03

    Although the palynology and sporophyte stage of Isöetes sinensis have been well studied, the biology of its gametophyte and embryo is less well understood. To date, the functions of several genes of I. sinensis and the molecular mechanisms of enzymes encoded by them remain to be studied. In the present study, the Fe-SOD gene of I. sinensis was successfully cloned using RT-PCR and rapid amplification of cDNA ends (RACE), and termed IsFeSOD. IsFeSOD has certain reference value in the classification of system evolution. The study also accumulated data for further research on the SOD gene. Bioinformatic analysis was employed to compare IsFeSOD with gene sequences obtained from other plants present in the GenBank. Furthermore, the recombinant pET32-FeSOD plasmids were transformed into Escherichia coli BL21 for expression. IsFeSOD was observed to have 1469 nucleotides that were predicted to encode 247 amino acids. The bioinformatic analysis revealed that IsFeSOD contained conserved TGGGA sequences, similar to eight other species, in addition to five other conserved sequences. The recombinant protein was about 43 kDa. Recombinant FeSOD was expressed, purified, and confirmed by western blotting. Alignment of complete Fe-SOD mRNA sequences from 9 species revealed several conserved sequences. A phylogenetic tree was constructed using MEGA4.1 and ClustalX multiple-sequence alignment programs. This study could be helpful in further characterization of SOD genes and for classification of system evolution status.

  13. Calmodulin is a stress and immune response gene in Chinese mitten crab Eriocheir sinensis.

    PubMed

    Li, Shuo; Jia, Zirui; Li, Xuejing; Geng, Xuyun; Sun, Jinsheng

    2014-09-01

    Calmodulin (CaM) is a multifunctional calcium sensor protein that participates in various cellular processes under normal, stress and pathological conditions. In crabs, however, the involvement of CaM in response to environmental stress and immune challenges has not been revealed yet. In the present study, a CaM cDNA (EsCaM) was identified from Chinese mitten crab Eriocheir sinensis and its mRNA expression patterns in response to ambient (salinity and pH) stress and immune challenges was examined. EsCaM encodes a 149-amino-acid protein with a calculated molecular mass of 16.8 kDa and an isoelectric point of 4.09. In unstimulated healthy E. sinensis, EsCaM mRNA transcript was detected in all tested tissues with predominant expression in hepatopancreas and the lowest expression in haemocytes. Ambient salinity (15‰ and 30‰ salinities) and pH (pH 6 and 8.5) stress significantly altered EsCaM mRNA expression in gill, hepatopancreas, haemocytes, intestine and muscle in Chinese mitten crab. In addition, EsCaM gene expression was significantly and rapidly induced as early as 2 h after LPS and Poly(I:C) immune stimulations in haemocytes in vitro. Furthermore, EsCaM expression was significantly up-regulated in E. sinensis haemocytes, gill, hepatopancreas, intestine and muscle in response to Edwardsiella tarda and Vibrio anguillarum challenges. Collectively, our findings suggest that EsCaM is an important stress and immune response gene in Chinese mitten crab.

  14. Phylogeography and genetic structure of a Tertiary relict tree species, Tapiscia sinensis (Tapisciaceae): implications for conservation

    PubMed Central

    Zhang, Jinju; Li, Zuozhou; Fritsch, Peter W.; Tian, Hua; Yang, Aihong; Yao, Xiaohong

    2015-01-01

    Background and Aims The phylogeography of plant species in sub-tropical China remains largely unclear. This study used Tapiscia sinensis, an endemic and endangered tree species widely but disjunctly distributed in sub-tropical China, as a model to reveal the patterns of genetic diversity and phylogeographical history of Tertiary relict plant species in this region. The implications of the results are discussed in relation to its conservation management. Methods Samples were taken from 24 populations covering the natural geographical distribution of T. sinensis. Genetic structure was investigated by analysis of molecular variance (AMOVA) and spatial analysis of molecular variance (SAMOVA). Phylogenetic relationships among haplotypes were constructed with maximum parsimony and haplotype network methods. Historical population expansion events were tested with pairwise mismatch distribution analysis and neutrality tests. Species potential range was deduced by ecological niche modelling (ENM). Key Results A low level of genetic diversity was detected at the population level. A high level of genetic differentiation and a significant phylogeographical structure were revealed. The mean divergence time of the haplotypes was approx. 1·33 million years ago. Recent range expansion in this species is suggested by a star-like haplotype network and by the results from the mismatch distribution analysis and neutrality tests. Conclusions Climatic oscillations during the Pleistocene have had pronounced effects on the extant distribution of Tapiscia relative to the Last Glacial Maximum (LGM). Spatial patterns of molecular variation and ENM suggest that T. sinensis may have retreated in south-western and central China and colonized eastern China prior to the LGM. Multiple montane refugia for T. sinense existing during the LGM are inferred in central and western China. The populations adjacent to or within these refugia of T. sinense should be given high priority in the development of

  15. Comparative effect of Citrus sinensis and carbimazole on serum T4, T3 and TSH levels

    PubMed Central

    Uduak, Okon Akpan; Ani, Elemi John; Etoh, Emmauel Columba Inyang; Macstephen, Adienbo Ologbagno

    2014-01-01

    Background: There are previous independent reports on the anti-thyroid property of Citrus sinensis. This isoflavones and phenolic acid-rich natural agent is widely consumed as dietary supplement, thus the need to investigate its comparative effect with a standard anti-thyroid drug on T4, T3 and thyroid stimulating hormone (TSH) levels. Objective: To compare the effect of Citrus sinensis and carbimazole (CARB) on blood levels of thyroid hormones (T4 and T3) and TSH. Materials and Methods: Male wistar albino rats weighing 100-150 g were employed in this research. The rats were randomly assigned to four groups of seven rats per group. Group I served as control and were administered distilled water while groups II-IV were administered with 1500 mg/kg of Citrus sinensis (fresh orange juice; FOJ), 0.1 μg/g of levothyroxine (LVT) and 0.01 mg/g of CARB, respectively, per oral once daily for 28 days. The animals were sacrificed under chloroform anaesthesia and blood sample collected by cardiac puncture and processed by standard method to obtain serum. TSH, T4 and T3 were assayed with the serum using ARIA II automated radioimmunoassay instrument. Results: The results showed that TSH level was significantly (P < 0.05) decreased in LVT treated group compared with the FOJ group. T4 was significantly (P < 0.05) decreased in the FOJ and CARB groups compared with the control and LVT groups. LVT significantly increased T4 when compared with FOJ group. T3 was significantly (P < 0.05) decreased in the CARB group compared with the control. Conclusion: These findings suggest that FOJ alters thyroid hormones metabolism to reduce their serum levels with a compensatory elevations of TSH level in a direction similar to CARB. PMID:25013255

  16. Anti-inflammatory effects of Citrus sinensis L., Citrus paradisi L. and their combinations.

    PubMed

    Khan, Rafeeq Alam; Mallick, Neelam; Feroz, Zeeshan

    2016-05-01

    Citrus bioflavonoids embrace a wide group of phenolic compounds effecting the production and scavenging of reactive oxygen species and the processes relating free radical-mediated injury. Keeping in view of the antioxidant and anti-inflammatory properties of Citrus sinensis and Citrus paradisi, present study was undertaken to explore the effects of C. sinensis (orange juice) and C. paradisi (grapefruit juice) at three different doses alone and their two combinations with the objective to examine the effects of these compounds in an experimental model of rat colitis induced by trinitrobenzenesulphonic acid (TNBS). Hence biochemical parameters e.g. myeloperoxidase, alkaline phosphatase, C-reactive protein (CRP) and glutathione were assessed. Data entry and analysis was accomplished by Statistical Package for the Social Sciences version 17 and was presented as mean ± S.E.M with 95% confidence interval. Present result shows that these juices, mainly C. paradisi, may be efficacious for the management of inflammatory bowel disease. In acute colitis model, C. paradise encouraged a decrease in the extension of the lesion escorted by a decrease in the occurrence of diarrhea and reinstatement of the glutathione content. Related effects were produced by the administration of C. sinensis, which also prevented the myeloperoxidase and alkaline phosphatase actions in acute intestinal inflammatory process. The effect of the citrus juices on the inflammatory process may be associated to their antioxidant and anti-inflammatory properties, as revealed in present investigation. The favorable effects exerted were demonstrated both by histological and biochemical changes and were related with a progress in the colonic oxidative status.

  17. Biosynthesis of silver nanoparticles using citrus sinensis peel extract and its antibacterial activity

    NASA Astrophysics Data System (ADS)

    Kaviya, S.; Santhanalakshmi, J.; Viswanathan, B.; Muthumary, J.; Srinivasan, K.

    2011-08-01

    Biosynthesis of silver nanoparticles (AgNPs) was achieved by a novel, simple green chemistry procedure using citrus sinensis peel extract as a reducing and a capping agent. The effect of temperature on the synthesis of silver nanoparticles was carried out at room temperature (25 °C) and 60 °C. The successful formation of silver nanoparticles has been confirmed by UV-vis, FTIR, XRD, EDAX, FESEM and TEM analysis and their antibacterial activity against Escherichia coli, Pseudomonas aeruginosa (Gram-negative), and Staphylococcus aureus (Gram-positive) has been studied. The results suggest that the synthesized AgNPs act as an effective antibacterial agent.

  18. Genetic differentiation within Eriochoir sinensis (milne, edwards) revealed by allozyme and RAPD

    NASA Astrophysics Data System (ADS)

    Cui, Zhao-Xia; Xiang, Jian-Hai; Song, Lin-Sheng; Zhou, Ling-Hua; Shi, Wei-Liang

    2000-09-01

    We analyzed 17 allozymes, and 20 primers in order to detect the genetic differentiation between commercial populations (Changjiang River, Liaohe River) of Eriochoir sinensis. Ten allozymes (LDH, MDH, ME, IDH, EST, ALP, AAT, CTL, POD, SOD) showed 21 loci by vertically discontinuos buffer system polyacrylamide gel electrophoresis. RAPD profiles generated by 12 ten-base primers showed 63 loci. The percentage of polymorphic loci and the expected heterozygosity obtained by using allozyme analysis were lower than those obtained by RAPD. The index of similarity between these two populations were 0.955 and 0.932 as revealed by allozyme analysis and RAPD technology. There was gene flow between the above populations.

  19. Caffeine in tea Camellia sinensis--content, absorption, benefits and risks of consumption.

    PubMed

    Gramza-Michałowska, A

    2014-01-01

    Therapeutic properties of tea Camellia sinensis are of particular interest since it has been consumed for ages and was always regarded as safe beverage. Tea is most popular beverage in the world because of its attractive aroma, exceptional taste, health promoting and pharmaceutical potential. Current results showed that antioxidative, antibacterial and other health effects are attributed to its caffeine content and caffeine - polyphenols interactions. An overview is given on caffeine content in different tea leaves beverage. Special attention is drawn to caffeine physiological effect on human organism. Controversies concerning the possible caffeine influence on human physical and psychological health are briefly summarized and presented.

  20. Vanadium-Enriched Cordyceps sinensis, a Contemporary Treatment Approach to Both Diabetes and Depression in Rats

    PubMed Central

    Guo, Jianyou; Li, ChangYu; Wang, Jie; Liu, Yongmei; Zhang, Jiahui

    2011-01-01

    This article studies a contemporary treatment approach toward both diabetes and depression management by vanadium-enriched Cordyceps sinensis (VECS). Streptozotocin-induced hyperglycemic rats were used in the study. After the rats were administered with VECS, a significant reduction in blood glucose levels was seen (P < .05) and the levels of serum insulin increased significantly (P < .05). At the same time, the study revealed a significant decrease in immobility with a corresponding increase in the swimming and climbing behavior in hyperglycemic rats following VECS treatment. The results described herein demonstrate that VECS is a contemporary treatment approach that advocates an aggressive stance toward both diabetes and depression management. PMID:21799679

  1. Cordyceps sinensis Increases Hypoxia Tolerance by Inducing Heme Oxygenase-1 and Metallothionein via Nrf2 Activation in Human Lung Epithelial Cells

    PubMed Central

    Manickam, Manimaran; Misra, Kshipra

    2013-01-01

    Cordyceps sinensis, an edible mushroom growing in Himalayan regions, is widely recognized in traditional system of medicine. In the present study, we report the efficacy of Cordyceps sinensis in facilitating tolerance to hypoxia using A549 cell line as a model system. Treatment with aqueous extract of Cordyceps sinensis appreciably attenuated hypoxia induced ROS generation, oxidation of lipids and proteins and maintained antioxidant status similar to that of controls via induction of antioxidant gene HO1 (heme oxygenase-1), MT (metallothionein) and Nrf2 (nuclear factor erythroid-derived 2-like 2). In contrast, lower level of NFκB (nuclear factor kappaB) and tumor necrosis factor-α observed which might be due to higher levels of HO1, MT and transforming growth factor-β. Further, increase in HIF1 (hypoxia inducible factor-1) and its regulated genes; erythropoietin, vascular endothelial growth factor, and glucose transporter-1 was observed. Interestingly, Cordyceps sinensis treatment under normoxia did not regulate the expression HIF1, NFκB and their regulated genes evidencing that Cordyceps sinensis per se did not have an effect on these transcription factors. Overall, Cordyceps sinensis treatment inhibited hypoxia induced oxidative stress by maintaining higher cellular Nrf2, HIF1 and lowering NFκB levels. These findings provide a basis for possible use of Cordyceps sinensis in tolerating hypoxia. PMID:24063008

  2. Phylogenomic and MALDI-TOF MS analysis of Streptococcus sinensis HKU4T reveals a distinct phylogenetic clade in the genus Streptococcus.

    PubMed

    Teng, Jade L L; Huang, Yi; Tse, Herman; Chen, Jonathan H K; Tang, Ying; Lau, Susanna K P; Woo, Patrick C Y

    2014-10-20

    Streptococcus sinensis is a recently discovered human pathogen isolated from blood cultures of patients with infective endocarditis. Its phylogenetic position, as well as those of its closely related species, remains inconclusive when single genes were used for phylogenetic analysis. For example, S. sinensis branched out from members of the anginosus, mitis, and sanguinis groups in the 16S ribosomal RNA gene phylogenetic tree, but it was clustered with members of the anginosus and sanguinis groups when groEL gene sequences used for analysis. In this study, we sequenced the draft genome of S. sinensis and used a polyphasic approach, including concatenated genes, whole genomes, and matrix-assisted laser desorption ionization-time of flight mass spectrometry to analyze the phylogeny of S. sinensis. The size of the S. sinensis draft genome is 2.06 Mb, with GC content of 42.2%. Phylogenetic analysis using 50 concatenated genes or whole genomes revealed that S. sinensis formed a distinct cluster with Streptococcus oligofermentans and Streptococcus cristatus, and these three streptococci were clustered with the "sanguinis group." As for phylogenetic analysis using hierarchical cluster analysis of the mass spectra of streptococci, S. sinensis also formed a distinct cluster with S. oligofermentans and S. cristatus, but these three streptococci were clustered with the "mitis group." On the basis of the findings, we propose a novel group, named "sinensis group," to include S. sinensis, S. oligofermentans, and S. cristatus, in the Streptococcus genus. Our study also illustrates the power of phylogenomic analyses for resolving ambiguities in bacterial taxonomy.

  3. Evaluation of mercury and lead content in the liver of the cormorant (Phalacrocorax carbo sinensis) population of Kis-Balaton, Hungary.

    PubMed

    Lehel, József; Gál, János; Faragó, Sándor; Berta, Erzsébet; Andrásofszky, Emese; Fekete, Sándor György; Mándoki, Míra; Budai, Péter; Kormos, Eva; Marosán, Miklós

    2013-06-01

    Mercury and lead concentrations were measured in the livers of cormorant (Phalacrocorax carbo sinensis), an aquatic bird species living and nesting in the special, highly protected nature conservation area of Kis-Balaton, Hungary. The measurements of metal concentrations were performed by atomic absorption spectrometry using the cold vapour method for mercury and the electrothermal method for lead. Mercury concentrations in the livers were significantly higher in the adult population (4.479 ± 3.336 mg/kg dry matter, DM) than in the juvenile birds (2.682 ± 2.087 mg/kg DM), indicating an increase of bioaccumulation with age. A similar pattern was not observed for lead. There were no statistical differences between males and females either in mercury or in lead concentrations. The average levels of mercury (3.580 ± 2.906 mg/kg DM) and lead (0.746 ± 0.499 mg/kg DM) were statistically different in the liver. No correlation was found between the concentrations of the two heavy metals. Recently, the wild birds have been chronically exposed to subtoxic amounts of metals which have a tendency to accumulate especially in the soft tissues.

  4. Anti-Osteoporotic Effects of Angelica sinensis (Oliv.) Diels Extract on Ovariectomized Rats and Its Oral Toxicity in Rats

    PubMed Central

    Lim, Dong Wook; Kim, Yun Tai

    2014-01-01

    Angelica sinensis root is one of the herbs most commonly used in China; it is also often included in dietary supplements for menopause in Europe and North America. In the present study, we examined the anti-osteoporotic effects of A. sinensis extract in an ovariectomized (OVX) rat model of osteoporosis as well as toxicity of the extract after repeated oral administration. The OVX rats were treated with 17β-estradiol (10 μg/kg i.p. once daily) or A. sinensis extract (30, 100, and 300 mg/kg, p.o. once daily) for four weeks. The bone (femur) mineral density (BMD) of rats treated with the extract (300 mg/kg) was significantly higher than that of the OVX-control, reaching BMD of the estradiol group. Markers of bone turnover in osteoporosis, serum alkaline phosphatase, collagen type I C-telopeptide and osteocalcin, were significantly decreased in the extract group. The body and uterus weight and serum estradiol concentration were not affected, and no treatment-related toxicity was observed during extract administration in rats. The results obtained indicate that A. sinensis extract can prevent the OVX-induced bone loss in rats via estrogen-independent mechanism. PMID:25325255

  5. The first fossil skull of Alligator sinensis from the Pleistocene, Taiwan, with a paleogeographic implication of the species

    NASA Astrophysics Data System (ADS)

    Hsi-yin, Shan; Yen-nien, Cheng; Xiao-chun, Wu

    2013-06-01

    A nearly complete fossil skull of Alligatoridae from the Pleistocene, Penghu Channel, east of Taiwan, is reported. It can be referred to the most latest clade of Alligatorinae, which includes Alligator sinensis, Alligator mississippiensis and Alligator mefferdi, on the basis of the following features: the splenial is excluded from the mandibular symphysis; the anterior tip of the splenial passes dorsal to the Meckelian groove; and the mandible is gently curved between the fourth alveoli and the mid dentary. It differs from A. mississippiensis and A. mefferdi mainly in the following characters: the breadth between the supratemporal fenestrae is approximately equal to the interorbital width, the snout is about half the length of the skull; and the anterior part of the snout is subtriangular in dorsal view. These features suggest that the Penghu alligator is most probably referable to A. sinensis. This is the only fossil skull of A. sinensis known. The discovery of the skull in Penghu Channel not only provides the first solid fossil evidence to indicate that the geological distribution of A. sinensis extended farther southeast than the historical/archeological range of the species but also adds new information on the biodiversity of the Penghu fauna.

  6. Herbal Medicine Cordyceps sinensis Improves Health-Related Quality of Life in Moderate-to-Severe Asthma

    PubMed Central

    Wang, Ningqun; Li, Jie; Chen, Wenqiang; Chen, Yujing

    2016-01-01

    Moderate-to-severe asthma has a substantial impact on the health-related quality of life (HR-QOL) of the patients. Cordyceps sinensis is a traditional Chinese medicine that is evaluated clinically for the treatment of many diseases, such as chronic allograft nephropathy, diabetic kidney disease, and lung fibrosis. In order to investigate the effects of Cordyceps sinensis on patients with moderate-to-severe persistent asthma, 120 subjects were randomized to receive Corbin capsule containing Cordyceps sinensis for 3 months (treatment group, n = 60), whereas the control group (n = 60) did not receive treatment with Corbin capsule. Inhaled corticosteroid and as-needed β-agonists were used in the treatment of both groups. HR-QOL was measured with the Juniper's Asthma Quality of Life Questionnaire (AQLQ). The incidence of asthma exacerbation, pulmonary function testing, and serum measurements of inflammatory mediators were also evaluated. The results showed that the treatment group indicated a significant increase in AQLQ scores and lung function compared with the control group. The expression levels of the inflammation markers IgE, ICAM-1, IL-4, and MMP-9 in the serum were decreased and IgG increased in the treatment group compared with the control group. Therefore, the conclusion was reached that a formulation of Cordyceps sinensis improved the HR-QOL, asthma symptoms, lung function, and inflammatory profile of the patients with moderate-to-severe asthma. This trial is registered with ChiCTR-IPC-16008730. PMID:28050193

  7. Cordyceps sinensis protects against liver and heart injuries in a rat model of chronic kidney disease: a metabolomic analysis

    PubMed Central

    Liu, Xia; Zhong, Fang; Tang, Xu-long; Lian, Fu-lin; Zhou, Qiao; Guo, Shan-mai; Liu, Jia-fu; Sun, Peng; Hao, Xu; Lu, Ying; Wang, Wei-ming; Chen, Nan; Zhang, Nai-xia

    2014-01-01

    Aim: To test the hypothesis that the traditional Chinese medicine Cordyceps sinensis could improve the metabolic function of extrarenal organs to achieve its anti-chronic kidney disease (CKD) effects. Methods: Male SD rats were divided into CKD rats (with 5/6-nephrectomy), CKD rats treated with Cordyceps sinensis (4 mg•kg-1•d-1, po), and sham-operated rats. After an 8-week treatment, metabolites were extracted from the hearts and livers of the rats, and then subjected to 1H-NMR-based metabolomic analysis. Results: Oxidative stress, energy metabolism, amino acid and protein metabolism and choline metabolism were considered as links between CKD and extrarenal organ dysfunction. Within the experimental period of 8 weeks, the metabolic disorders in the liver were more pronounced than in the heart, suggesting that CKD-related extrarenal organ dysfunctions occurred sequentially rather than simultaneously. Oral administration of Cordyceps sinensis exerted statistically significant rescue effects on the liver and heart by reversely regulating levels of those metabolites that are typically perturbed in CKD. Conclusion: Oral administration of Cordyceps sinensis significantly attenuates the liver and heart injuries in CKD rats. The 1H NMR-based metabolomic approach has provided a systematic view for understanding of CKD and the drug treatment, which can also be used to elucidate the mechanisms of action of other traditional Chinese medicines. PMID:24632844

  8. Herbal Medicine Cordyceps sinensis Improves Health-Related Quality of Life in Moderate-to-Severe Asthma.

    PubMed

    Wang, Ningqun; Li, Jie; Huang, Xiaobo; Chen, Wenqiang; Chen, Yujing

    2016-01-01

    Moderate-to-severe asthma has a substantial impact on the health-related quality of life (HR-QOL) of the patients. Cordyceps sinensis is a traditional Chinese medicine that is evaluated clinically for the treatment of many diseases, such as chronic allograft nephropathy, diabetic kidney disease, and lung fibrosis. In order to investigate the effects of Cordyceps sinensis on patients with moderate-to-severe persistent asthma, 120 subjects were randomized to receive Corbin capsule containing Cordyceps sinensis for 3 months (treatment group, n = 60), whereas the control group (n = 60) did not receive treatment with Corbin capsule. Inhaled corticosteroid and as-needed β-agonists were used in the treatment of both groups. HR-QOL was measured with the Juniper's Asthma Quality of Life Questionnaire (AQLQ). The incidence of asthma exacerbation, pulmonary function testing, and serum measurements of inflammatory mediators were also evaluated. The results showed that the treatment group indicated a significant increase in AQLQ scores and lung function compared with the control group. The expression levels of the inflammation markers IgE, ICAM-1, IL-4, and MMP-9 in the serum were decreased and IgG increased in the treatment group compared with the control group. Therefore, the conclusion was reached that a formulation of Cordyceps sinensis improved the HR-QOL, asthma symptoms, lung function, and inflammatory profile of the patients with moderate-to-severe asthma. This trial is registered with ChiCTR-IPC-16008730.

  9. CHINESE MITTEN CRABS (ERIOCHEIR SINENSIS) IN THE ST. LAWRENCE RIVER (CANADA): NEW RECORDS AND RISK OF INVASION

    EPA Science Inventory

    The Chinese mitten crab, Eriocheir sinensis, is an internationally renowned aquatic invader. Native to China and North/South Korea, this catadromous crab has successfully invaded several rivers and estuaries in eleven countries in Western Europe as well as the San Francisco Bay ...

  10. Cloning, purification and characterization of a 90kDa heat shock protein from Citrus sinensis (sweet orange).

    PubMed

    Mendonça, Yuri A; Ramos, Carlos H I

    2012-01-01

    Protein misfolding is stimulated by stress, such as heat, and heat shock proteins (Hsps) are the first line of defense against these undesirable situations. Plants, which are naturally sessile, are perhaps more exposed to stress factors than some other organisms, and consequently, the role of Hsps is crucial to maintain homeostasis. Hsp90, because of its key role in infection and other stresses, is targeted in therapies that improve plant production by increasing resistance to both biotic and abiotic stress. In addition, Hsp90 is a primary factor in the maintenance of homeostasis in plants. Therefore, we cloned and purified Hsp90 from Citrus sinensis (sweet orange). Recombinant C. sinensis Hsp90 (rCsHsp90) was produced and measured by circular dichroism (CD), intrinsic fluorescence spectroscopy and dynamic light scattering. rCsHsp90 formed a dimer in solution with a Stokes radius of approximately 62Å. In addition, it was resistant to thermal unfolding, was able to protect citrate synthase from aggregation, and Western blot analysis demonstrated that CsHsp90 was constitutively expressed in C. sinensis cells. Our analysis indicated that CsHsp90 is conformationally similar to that of yeast Hsp90, for which structural information is available. Therefore, we showed that C. sinensis expresses an Hsp90 chaperone that has a conformation and function similar to other Hsp90s.

  11. Performance of 'Valencia' Orange (Citrus sinensis [L.] Osbeck) on 17 rootstocks in a trial severely affected by huanglongbing

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Valencia orange (Citrus sinensis L. Osbeck) was grown on 17 rootstocks through seven years of age and the first four harvest seasons in a central Florida field trial severely affected by huanglongbing (HLB) disease. All trees in the trial had huanglongbing symptoms and were shown by Polymerase chain...

  12. Fragile Sites of 'Valencia' Sweet Orange (Citrus sinensis) Chromosomes Are Related with Active 45s rDNA.

    PubMed

    Lan, Hong; Chen, Chun-Li; Miao, Yin; Yu, Chang-Xiu; Guo, Wen-Wu; Xu, Qiang; Deng, Xiu-Xin

    2016-01-01

    Citrus sinensis chromosomes present a morphological differentiation of bands after staining by the fluorochromes CMA and DAPI, but there is still little information on its chromosomal characteristics. In this study, the chromosomes in 'Valencia' C. sinensis were analyzed by fluorescence in situ hybridization (FISH) using telomere DNA and the 45S rDNA gene as probes combining CMA/DAPI staining, which showed that there were two fragile sites in sweet orange chromosomes co-localizing at distended 45S rDNA regions, one proximally locating on B-type chromosome and the other subterminally locating on D-type chromosome. While the chromosomal CMA banding and 45S rDNA FISH mapping in the doubled haploid line of 'Valencia' C. sinensis indicated six 45S rDNA regions, four were identified as fragile sites as doubled comparing its parental line, which confirmed the cytological heterozygosity and chromosomal heteromorphisms in sweet orange. Furthermore, Ag-NOR identified two distended 45S rDNA regions to be active nucleolar organizing regions (NORs) in diploid 'Valencia' C. sinensis. The occurrence of quadrivalent in meiosis of pollen mother cells (PMCs) in 'Valencia' sweet orange further confirmed it was a chromosomal reciprocal translocation line. We speculated this chromosome translocation was probably related to fragile sites. Our data provide insights into the chromosomal characteristics of the fragile sites in 'Valencia' sweet orange and are expected to facilitate the further investigation of the possible functions of fragile sites.

  13. Peculiar liquid-feeding and pathogen transmission behavior of Aedes togoi and comparison with Anopheles sinensis

    NASA Astrophysics Data System (ADS)

    Lee, Sang Joon; Kang, Dooho; Lee, Seung Chul; Ha, Young-Ran

    2016-02-01

    Female mosquitoes transmit various diseases as vectors during liquid-feeding. Identifying the determinants of vector efficiency is a major scientific challenge in establishing strategies against these diseases. Infection rate and transmission efficiency are interconnected with the mosquito-induced liquid-feeding flow as main indexes of vector efficiency. However, the relationship between liquid-feeding characteristics and pathogen remains poorly understood. The liquid-feeding behavior of Aedes togoi and Anopheles sinensis was comparatively investigated in conjunction with vector efficiency via micro-particle image velocimetry. The flow rates and ratio of the ejection volume of Aedes togoi were markedly higher than those of Anophels sinensis. These differences would influence pathogen re-ingestion. Wall shear stresses of these mosquito species were also clearly discriminatory affecting the infective rates of vector-borne diseases. The variations in volume of two pump chambers and diameter of proboscis of these mosquito species were compared to determine the differences in the liquid-feeding process. Liquid-feeding characteristics influence vector efficiency; hence, this study can elucidate the vector efficiency of mosquitoes and the vector-pathogen interactions and contribute to the development of strategies against vector-borne diseases.

  14. Comparing photosynthetic characteristics of Isoetes sinensis Palmer under submerged and terrestrial conditions

    PubMed Central

    Yang, Tao; Liu, Xing

    2015-01-01

    Crassulacean acid metabolism (CAM) is widespread in terrestrial and aquatic species, plastic in response to environmental changes. Isoetes L. is one of the earliest basal vascular plants and CAM is popular in this genus. Isoetes sinensis Palmer is an amphibious species, alternating frequently between terrestrial and aquatic environments. Given this, we investigated and compared photosynthetic characteristics over a diurnal cycle under submerged condition (SC) and terrestrial condition (TC). The results suggest that I. sinensis possesses a stronger CAM capacity under SC. Compared with under TC, titratable acidity levels and organic acid concentrations were more enriched under SC, whereas soluble sugar or starch and protein levels were lower under SC. Transcript analyses for nine photosynthetic genes revealed that CAM-associated genes possessed high transcripts under SC, but C3-related transcripts were highly expressed under TC. In addition, the enzyme activity measurements demonstrated that PEPC activity over a diurnal cycle was slightly higher under SC, whereas Rubisco activity during the daytime was greater under TC. This comprehensive study probably facilitates general understandings about the CAM photosynthetic characteristics of Isoetes in response to the environmental changes. PMID:26634994

  15. Molecular cloning, computational and expression analysis of anthocyanidin reductase in tea (Camellia sinensis).

    PubMed

    Thirugnanasambantham, Krishnaraj; Muralidaran, Senguttuvan; Mandal, Abul Kalam Azad

    2014-09-01

    Tea [Camellia sinensis (L.) O. Kuntze] is one of the most popular non-alcoholic beverages rich in phenolic compounds, which includes epigallocatechin gallate (EGCG), epigallocatechin (EGC), epicatechin gallate (ECG), epicatechin (EC) and catechin (C). Anthocyanidin reductase (ANR) is responsible for catechin biosynthesis in plants, and analysis of its protein sequences and structures will be valuable for further research in the field. We have screened our dormant bud-specific complementary DNA (cDNA) library and reported 1,322-bp cDNA encoding CsANR. Analysis of the sequence revealed the presence of 1,011-bp open reading frame with coding capacity for a polypeptide of 337 amino acids, flanked by 1,123- and 196-bp 5' and 3' untranslated regions, respectively. Theoretical molecular weight (MW) and isoelectric point (pI) of the deduced ANR protein were predicted (using ProtParam) to be 36.4 kDa and 6.54. For the first time, we have reported 3D model of ANR from C. sinensis. Quality of the predicted model was analysed with PROCHECK analysis. Molecular docking of modelled ANR revealed similar binding pockets for both substrates and products. Expression analyses of CsANR and accumulation pattern of catechins were observed to be varied with developmental age of tissue and seasonal condition. Variation in accumulation pattern of catechins and its fractions was found to be correlated with expression pattern of ANR.

  16. Effect of Cordyceps sinensis and taurine either alone or in combination on streptozotocin induced diabetes.

    PubMed

    El Zahraa Z El Ashry, Fatma; Mahmoud, Mona F; El Maraghy, Nabila N; Ahmed, Ahmed F

    2012-03-01

    The present study aimed to investigate the antidiabetic effects of Cordyceps sinensis, taurine and their combination in comparison with glibenclamide both in vivo and in vitro using streptozotocin rat model. The diabetic rats were orally given glibenclamide, C. sinensis, taurine or Cordyceps and taurine combination for 21 days. Their effects were studied both in vivo and in vitro. Oral administration of Cordyceps, taurine and their combination decreased serum glucose, fructosamine, total cholesterol, triglycerides levels, insulin resistance index and pancreatic malondialdehyde content. Cordyceps significantly increased serum insulin, HDL-cholesterol, total antioxidant capacity levels, β cell function percent, and pancreatic reduced glutathione (GSH) content. However, taurine was unable to elevate pancreatic GSH level to a significant level. These natural products and their combinations were more effective than glibenclamide in reducing insulin resistance index and they had stronger antioxidant properties. Cordyceps and taurine significantly enhanced glucose uptake by diaphragms of normal and diabetic rats in absence and presence of insulin. In conclusion, Cordyceps and taurine either alone or in combination have less potent hypoglycemic effects than glibenclamide; however, they have more ability to reduce insulin resistance and stronger antioxidant properties.

  17. Three-dimensional structures of the tracheal systems of Anopheles sinensis and Aedes togoi pupae

    PubMed Central

    Ha, Young-Ran; Yeom, Eunseop; Ryu, Jeongeun; Lee, Sang-Joon

    2017-01-01

    Mosquitoes act as a vector for the transmission of disease. The World Health Organization has recommended strict control of mosquito larvae because of their “few, fixed, and findable” features. The respiratory system of mosquito larvae and pupae in the water has a weak point. As aquatic organisms, mosquito larvae and pupae inhale atmosphere oxygen. However, the mosquito pupae have a non-feeding stage, unlike the larvae. Therefore, detailed study on the tracheal system of mosquito pupae is helpful for understanding their survival strategy. In this study, the three-dimensional (3D) structures of the tracheal systems of Anopheles sinensis and Aedes togoi pupae were comparatively investigated using synchrotron X-ray microscopic computed tomography. The respiratory frequencies of the dorsal trunks were also investigated. Interestingly, the pupae of the two mosquito species possess special tracheal systems of which the morphological and functional features are distinctively different. The respiratory frequency of Ae. togoi is higher than that of An. sinensis. These differences in the breathing phenomena and 3D structures of the respiratory systems of these two mosquito species provide an insight into the tracheal systems of mosquito pupae. PMID:28287649

  18. Comparative Transcriptome Analysis Reveals Sex-Biased Gene Expression in Juvenile Chinese Mitten Crab Eriocheir sinensis

    PubMed Central

    Liu, Yuan; Hui, Min; Cui, Zhaoxia; Luo, Danli; Song, Chengwen; Li, Yingdong; Liu, Lei

    2015-01-01

    Sex-biased genes are considered to account for most of phenotypic differences between males and females. In order to explore the sex-biased gene expression in crab, we performed the whole-body transcriptome analysis in male and female juveniles of the Chinese mitten crab Eriocheir sinensis using next-generation sequencing technology. Of the 23,349 annotated unigenes, 148 were identified as sex-related genes. A total of 29 candidate genes involved in primary sex determination pathways were detected, indicating the sex determination cascade of the mitten crab might be more complex than previously supposed. Differential expression analysis showed 448 differentially expressed genes (DEGs) between the two transcriptomes. Most of DEGs were involved in processes such as metabolism and immunity, and not associated with obvious sexual function. The pathway predominantly enriched for DEGs were related to lysosome, which might reflect the differences in metabolism between males and females. Of the immune DGEs, 18 up-regulated genes in females were humoral immune factors, and eight up-regulated genes in males were pattern recognition receptors, suggesting sex differences of immune defense might exist in the mitten crab. In addition, two reproduction-related genes, vitellogenin and insulin-like androgenic gland factor, were identified to express in both sexes but with significantly higher level in males. Our research provides the first whole-body RNA sequencing of sex-specific transcriptomes for juvenile E. sinensis and will facilitate further studies on molecular mechanisms of crab sexual dimorphism. PMID:26193085

  19. Hidden surface microstructures on Carboniferous insect Brodioptera sinensis (Megasecoptera) enlighten functional morphology and sensorial perception

    PubMed Central

    Prokop, Jakub; Pecharová, Martina; Ren, Dong

    2016-01-01

    Megasecoptera are insects with haustellate mouthparts and petiolate wings closely related to Palaeodictyoptera and one of the few insect groups that didn’t survive the Permian-Triassic mass extinction. Recent discovery of Brodioptera sinensis in early Pennsylvanian deposits at Xiaheyan in northern China has increased our knowledge of its external morphology using conventional optical stereomicroscopy. Environmental scanning electron microscopy (ESEM) of structures, such as antennae, mouthparts, wing surfaces, external copulatory organs and cerci have shed light on their micromorphology and supposed function. A comparative study has shown an unexpected dense pattern of setae on the wing membrane of B. sinensis. In addition, unlike the results obtained by stereomicroscopy it revealed that the male and female external genitalia clearly differ in their fine structure and setation. Therefore, the present study resulted in a closer examination of the microstructure and function of previously poorly studied parts of the body of Paleozoic insects and a comparison with homologous structures occurring in other Palaeodictyopteroida, Odonatoptera and Ephemerida. This indicates, that the role and presumptive function of these integumental protuberances is likely to have been a sensory one in the coordination of mouthparts and manipulation of stylets, escape from predators, enhancement of aerodynamic properties and copulatory behaviour. PMID:27321551

  20. Jasmonic acid is a crucial signal transducer in heat shock induced sesquiterpene formation in Aquilaria sinensis.

    PubMed

    Xu, Yan-Hong; Liao, Yong-Cui; Zhang, Zheng; Liu, Juan; Sun, Pei-Wen; Gao, Zhi-Hui; Sui, Chun; Wei, Jian-He

    2016-02-23

    Agarwood, a highly valuable resinous and fragrant heartwood of Aquilaria plants, is widely used in traditional medicines, incense and perfume. Only when Aquilaria trees are wounded by external stimuli do they form agarwood sesquiterpene defensive compounds. Therefore, understanding the signaling pathway of wound-induced agarwood formation is important. Jasmonic acid (JA) is a well-characterized molecule that mediates a plant's defense response and secondary metabolism. However, little is known about the function of endogenous JA in agarwood sesquiterpene biosynthesis. Here, we report that heat shock can up-regulate the expression of genes in JA signaling pathway, induce JA production and the accumulation of agarwood sesquiterpene in A. sinensis cell suspension cultures. A specific inhibitor of JA, nordihydroguaiaretic acid (NDGA), could block the JA signaling pathway and reduce the accumulation of sesquiterpene compounds. Additionally, compared to SA and H2O2, exogenously supplied methyl jasmonate has the strongest stimulation effect on the production of sesquiterpene compounds. These results clearly demonstrate the central induction role of JA in heat-shock-induced sesquiterpene production in A. sinensis.

  1. Jasmonic acid is a crucial signal transducer in heat shock induced sesquiterpene formation in Aquilaria sinensis

    PubMed Central

    Xu, Yan-Hong; Liao, Yong-Cui; Zhang, Zheng; Liu, Juan; Sun, Pei-Wen; Gao, Zhi-Hui; Sui, Chun; Wei, Jian-He

    2016-01-01

    Agarwood, a highly valuable resinous and fragrant heartwood of Aquilaria plants, is widely used in traditional medicines, incense and perfume. Only when Aquilaria trees are wounded by external stimuli do they form agarwood sesquiterpene defensive compounds. Therefore, understanding the signaling pathway of wound-induced agarwood formation is important. Jasmonic acid (JA) is a well-characterized molecule that mediates a plant’s defense response and secondary metabolism. However, little is known about the function of endogenous JA in agarwood sesquiterpene biosynthesis. Here, we report that heat shock can up-regulate the expression of genes in JA signaling pathway, induce JA production and the accumulation of agarwood sesquiterpene in A. sinensis cell suspension cultures. A specific inhibitor of JA, nordihydroguaiaretic acid (NDGA), could block the JA signaling pathway and reduce the accumulation of sesquiterpene compounds. Additionally, compared to SA and H2O2, exogenously supplied methyl jasmonate has the strongest stimulation effect on the production of sesquiterpene compounds. These results clearly demonstrate the central induction role of JA in heat-shock-induced sesquiterpene production in A. sinensis. PMID:26902148

  2. Contents and compositions of policosanols in green tea (Camellia sinensis) leaves.

    PubMed

    Choi, Sol Ji; Park, Su Yeon; Park, Ji Su; Park, Sang-Kyu; Jung, Mun Yhung

    2016-08-01

    Policosanol (PC) is a mixture of health promoting bioactive long-chain aliphatic alcohols. Here, we report that green tea (Camellia sinensis) leaves are the exceptionally rich plant-sources of PC. Young and tender leaves and old and turf leaves of C. sinensis were hand-picked in spring and autumn. The total contents of PC in the leaves were in the range of 726.2-1363.6mg/kg as determined by a GC-MS/MS. The compositions of PC in the leaves were different with harvest season and types. The total contents of PC in commercial green tea leaves were found to be in the range of 856.7-1435.1mg/kg. Interestingly, the infused green tea leaves contained the higher PC than the non-infused green tea product, reaching to 1629.4mg/kg. This represents the first report on the contents and compositions of PC in green tea leaves, showing unambiguous evidence of their potential as rich sources of PC.

  3. SSR-based genetic maps of Miscanthus sinensis and M. sacchariflorus, and their comparison to sorghum.

    PubMed

    Kim, Changsoo; Zhang, Dong; Auckland, Susan A; Rainville, Lisa K; Jakob, Katrin; Kronmiller, Brent; Sacks, Erik J; Deuter, Martin; Paterson, Andrew H

    2012-05-01

    We present SSR-based genetic maps from a cross between Miscanthus sacchariflorus Robustus and M. sinensis, the progenitors of the promising cellulosic biofuel feedstock Miscanthus × giganteus. cDNA-derived SSR markers were mapped by the two-way pseudo-testcross model due to the high heterozygosity of each parental species. A total of 261 loci were mapped in M. sacchariflorus, spanning 40 linkage groups and 1,998.8 cM, covering an estimated 72.7% of the genome. For M. sinensis, a total of 303 loci were mapped, forming 23 linkage groups and 2,238.3 cM, covering 84.9% of the genome. The use of cDNA-derived SSR loci permitted alignment of the Miscanthus linkage groups to the sorghum chromosomes, revealing a whole genome duplication affecting the Miscanthus lineage after the divergence of subtribes Sorghinae and Saccharinae, as well as traces of the pan-cereal whole genome duplication. While the present maps provide for many early research needs in this emerging crop, additional markers are also needed to improve map density and to further characterize the structural changes of the Miscanthus genome since its divergence from sorghum and Saccharum.

  4. Comparative Transcriptome Analysis Reveals Sex-Biased Gene Expression in Juvenile Chinese Mitten Crab Eriocheir sinensis.

    PubMed

    Liu, Yuan; Hui, Min; Cui, Zhaoxia; Luo, Danli; Song, Chengwen; Li, Yingdong; Liu, Lei

    2015-01-01

    Sex-biased genes are considered to account for most of phenotypic differences between males and females. In order to explore the sex-biased gene expression in crab, we performed the whole-body transcriptome analysis in male and female juveniles of the Chinese mitten crab Eriocheir sinensis using next-generation sequencing technology. Of the 23,349 annotated unigenes, 148 were identified as sex-related genes. A total of 29 candidate genes involved in primary sex determination pathways were detected, indicating the sex determination cascade of the mitten crab might be more complex than previously supposed. Differential expression analysis showed 448 differentially expressed genes (DEGs) between the two transcriptomes. Most of DEGs were involved in processes such as metabolism and immunity, and not associated with obvious sexual function. The pathway predominantly enriched for DEGs were related to lysosome, which might reflect the differences in metabolism between males and females. Of the immune DGEs, 18 up-regulated genes in females were humoral immune factors, and eight up-regulated genes in males were pattern recognition receptors, suggesting sex differences of immune defense might exist in the mitten crab. In addition, two reproduction-related genes, vitellogenin and insulin-like androgenic gland factor, were identified to express in both sexes but with significantly higher level in males. Our research provides the first whole-body RNA sequencing of sex-specific transcriptomes for juvenile E. sinensis and will facilitate further studies on molecular mechanisms of crab sexual dimorphism.

  5. Adaptive divergence with gene flow in incipient speciation of Miscanthus floridulus/sinensis complex (Poaceae).

    PubMed

    Huang, Chao-Li; Ho, Chuan-Wen; Chiang, Yu-Chung; Shigemoto, Yasumasa; Hsu, Tsai-Wen; Hwang, Chi-Chuan; Ge, Xue-Jun; Chen, Charles; Wu, Tai-Han; Chou, Chang-Hung; Huang, Hao-Jen; Gojobori, Takashi; Osada, Naoki; Chiang, Tzen-Yuh

    2014-12-01

    Young incipient species provide ideal materials for untangling the process of ecological speciation in the presence of gene flow. The Miscanthus floridulus/sinensis complex exhibits diverse phenotypic and ecological differences despite recent divergence (approximately 1.59 million years ago). To elucidate the process of genetic differentiation during early stages of ecological speciation, we analyzed genomic divergence in the Miscanthus complex using 72 randomly selected genes from a newly assembled transcriptome. In this study, rampant gene flow was detected between species, estimated as M = 3.36 × 10(-9) to 1.20 × 10(-6) , resulting in contradicting phylogenies across loci. Nevertheless, beast analyses revealed the species identity and the effects of extrinsic cohesive forces that counteracted the non-stop introgression. As expected, early in speciation with gene flow, only 3-13 loci were highly diverged; two to five outliers (approximately 2.78-6.94% of the genome) were characterized by strong linkage disequilibrium, and asymmetrically distributed among ecotypes, indicating footprints of diversifying selection. In conclusion, ecological speciation of incipient species of Miscanthus probably followed the parapatric model, whereas allopatric speciation cannot be completely ruled out, especially between the geographically isolated northern and southern M. sinensis, for which no significant gene flow across oceanic barriers was detected. Divergence between local ecotypes in early-stage speciation began at a few genomic regions under the influence of natural selection and divergence hitchhiking that overcame gene flow.

  6. Extraction and chemical characterization of Angelica sinensis polysaccharides and its antioxidant activity.

    PubMed

    Ai, Songtao; Fan, Xindong; Fan, Linfeng; Sun, Qi; Liu, Yu; Tao, Xiaofeng; Dai, Kerong

    2013-05-15

    In the present study, the Angelica sinensis polysaccharides (ASP) extraction procedure was optimized by an L9 (3(4)) orthogonal array experimental design (OAD) with four factors at three levels. Under the optimal extraction condition (extraction time 180 min, ratio of water to solid 6, extraction temperature 100°C, and extraction number 4), extraction yield of ASP was 5.6%. Rabbits were fed for 40 days with A. sinensis polysaccharides at a dose of 150 or 300 mg/kg body weight, respectively. At the end of 40 days, animals received cerebral ischemia reperfusion operation. CT perfusion imaging (CTP) analysis showed that rCBF and rCBV were significantly increased, whereas rMTT and rTTP were decreased in the ischemia cerebral tissue compared to CIR group rabbits. ASP significantly decreased oxidative damage, and increased antioxidant enzymes activities in brains of CIR animals. Moreover, ASP significantly enhanced the Ach, Na(+),K(+)-ATPase, Ca(2+),Mg2(+)-ATPase and glucose levels, decreased AChE activity in brain tissue of the experimental animals. These results suggest a potent role of ASP in protection of brain oxidative injury in CIR animals.

  7. Citrus sinensis Annotation Project (CAP): A Comprehensive Database for Sweet Orange Genome

    PubMed Central

    Chang, Ji-Wei; Hao, Bao-Hai; Xing, Feng; Li, Sen; Xu, Qiang; Deng, Xiu-Xin; Chen, Ling-Ling

    2014-01-01

    Citrus is one of the most important and widely grown fruit crop with global production ranking firstly among all the fruit crops in the world. Sweet orange accounts for more than half of the Citrus production both in fresh fruit and processed juice. We have sequenced the draft genome of a double-haploid sweet orange (C. sinensis cv. Valencia), and constructed the Citrus sinensis annotation project (CAP) to store and visualize the sequenced genomic and transcriptome data. CAP provides GBrowse-based organization of sweet orange genomic data, which integrates ab initio gene prediction, EST, RNA-seq and RNA-paired end tag (RNA-PET) evidence-based gene annotation. Furthermore, we provide a user-friendly web interface to show the predicted protein-protein interactions (PPIs) and metabolic pathways in sweet orange. CAP provides comprehensive information beneficial to the researchers of sweet orange and other woody plants, which is freely available at http://citrus.hzau.edu.cn/. PMID:24489955

  8. Peculiar liquid-feeding and pathogen transmission behavior of Aedes togoi and comparison with Anopheles sinensis

    PubMed Central

    Lee, Sang Joon; Kang, Dooho; Lee, Seung Chul; Ha, Young-Ran

    2016-01-01

    Female mosquitoes transmit various diseases as vectors during liquid-feeding. Identifying the determinants of vector efficiency is a major scientific challenge in establishing strategies against these diseases. Infection rate and transmission efficiency are interconnected with the mosquito-induced liquid-feeding flow as main indexes of vector efficiency. However, the relationship between liquid-feeding characteristics and pathogen remains poorly understood. The liquid-feeding behavior of Aedes togoi and Anopheles sinensis was comparatively investigated in conjunction with vector efficiency via micro-particle image velocimetry. The flow rates and ratio of the ejection volume of Aedes togoi were markedly higher than those of Anophels sinensis. These differences would influence pathogen re-ingestion. Wall shear stresses of these mosquito species were also clearly discriminatory affecting the infective rates of vector-borne diseases. The variations in volume of two pump chambers and diameter of proboscis of these mosquito species were compared to determine the differences in the liquid-feeding process. Liquid-feeding characteristics influence vector efficiency; hence, this study can elucidate the vector efficiency of mosquitoes and the vector-pathogen interactions and contribute to the development of strategies against vector-borne diseases. PMID:26839008

  9. Hirsutella sinensis mycelium attenuates bleomycin-induced pulmonary inflammation and fibrosis in vivo

    PubMed Central

    Huang, Tsung-Teng; Lai, Hsin-Chih; Ko, Yun-Fei; Ojcius, David M.; Lan, Ying-Wei; Martel, Jan; Young, John D.; Chong, Kowit-Yu

    2015-01-01

    Hirsutella sinensis mycelium (HSM), the anamorph of Cordyceps sinensis, is a traditional Chinese medicine that has been shown to possess various pharmacological properties. We previously reported that this fungus suppresses interleukin-1β and IL-18 secretion by inhibiting both canonical and non-canonical inflammasomes in human macrophages. However, whether HSM may be used to prevent lung fibrosis and the mechanism underlying this activity remain unclear. Our results show that pretreatment with HSM inhibits TGF-β1–induced expression of fibronectin and α-SMA in lung fibroblasts. HSM also restores superoxide dismutase expression in TGF-β1–treated lung fibroblasts and inhibits reactive oxygen species production in lung epithelial cells. Furthermore, HSM pretreatment markedly reduces bleomycin–induced lung injury and fibrosis in mice. Accordingly, HSM reduces inflammatory cell accumulation in bronchoalveolar lavage fluid and proinflammatory cytokines levels in lung tissues. The HSM extract also significantly reduces TGF-β1 in lung tissues, and this effect is accompanied by decreased collagen 3α1 and α-SMA levels. Moreover, HSM reduces expression of the NLRP3 inflammasome and P2X7R in lung tissues, whereas it enhances expression of superoxide dismutase. These findings suggest that HSM may be used for the treatment of pulmonary inflammation and fibrosis. PMID:26497260

  10. Hibiscus rosa- sinensis leaf extract as coagulant aid in leachate treatment

    NASA Astrophysics Data System (ADS)

    Awang, Nik Azimatolakma; Aziz, Hamidi Abdul

    2012-12-01

    Hibiscus rosa- sinensis is a biodegradable material that has remained untested for flocculating properties. The objective of this study is to examine the efficiency of coagulation-flocculation processes for the removal of color, iron (Fe3+), suspended solids, turbidity and ammonia nitrogen(NH3-N), from landfill leachate using 4,000 mg/L alum in conjunction with H. rosa- sinensis leaf extract (HBaqs). Hydroxyl (O-H) and (carboxyl) C=O functional groups along the HBaqs chain help to indulge flocculating efficiency of HBaqs via bridging. The experiments confirm the positive coagulation properties of HBaqs. The Fe3+ removal rate using 4,000 mg/L alum as sole coagulant was approximately 60 %, and increased to 100 % when 4,000 mg/L alum was mixed with 500 mg/L HBaqs. By mixing, 4,000 mg/L alum with 100-500 mg/L HBaqs, 72 % of SS was removed as compared with only 45 % reduction using 4,000 mg/L alum as sole coagulant.

  11. Anticancer and antimetastatic effects of cordycepin, an active component of Cordyceps sinensis.

    PubMed

    Nakamura, Kazuki; Shinozuka, Kazumasa; Yoshikawa, Noriko

    2015-01-01

    Cordyceps sinensis, a fungus that parasitizes on the larva of Lepidoptera, has been used as a valued traditional Chinese medicine. We investigated the effects of water extracts of Cordyceps sinensis (WECS), and particularly focused on its anticancer and antimetastatic actions. Based on in vitro studies, we report that WECS showed an anticancer action, and this action was antagonized by an adenosine A3 receptor antagonist. Moreover, this anticancer action of WECS was promoted by an adenosine deaminase inhibitor. These results suggest that one of the components of WECS with an anticancer action might be an adenosine or its derivatives. Therefore, we focused on cordycepin (3'-deoxyadenosine) as one of the active ingredients of WECS. According to our experiments, cordycepin showed an anticancer effect through the stimulation of adenosine A3 receptor, followed by glycogen synthase kinase (GSK)-3β activation and cyclin D1 suppression. Cordycepin also showed an antimetastatic action through inhibiting platelet aggregation induced by cancer cells and suppressing the invasiveness of cancer cells via inhibiting the activity of matrix metalloproteinase (MMP)-2 and MMP-9, and accelerating the secretion of tissue inhibitor of metalloproteinase (TIMP)-1 and TIMP-2 from cancer cells. In conclusion, cordycepin, an active component of WECS, might be a candidate anticancer and antimetastatic agent.

  12. Therapeutic efficacy of a polysaccharide isolated from Cordyceps sinensis on hypertensive rats.

    PubMed

    Xiang, Feixiang; Lin, Liming; Hu, Min; Qi, Xiangqian

    2016-01-01

    This study was aimed to investigate the antihypertensive effect of a polysaccharide fraction from Cordyceps sinensis on spontaneously hypertensive rats (SHR). The CSP1, one component of Cordyceps sinensis polysaccharides (CSP), was obtained after water extraction, deproteinization, de-colorization and purification with DEAE-cellulose 52. And a more homogeneous component CSP1-2 was obtained using Sepharose CL-6B chromatography. CSP1-2 mainly consisted of mannose, glucose and galactose in a molar ratio of about 2:2:1 and its average molecular weight was approximately 2.70×10(4)Da. Pharmacological tests showed that CSP1, in which the CSP1-2 was its main component, had antihypertensive effect by stimulating the secretion of vasodilator NO, decreasing the level of ET-1, epinephrine, noradrenaline and angiotensin II, inhibiting the increase of transforming growth factor β1 (TGF-β1) and lowering the level of inflammatory mediator of C-reactive protein (CRP). These results suggested that CSP1 may possess high potential in treating hypertension.

  13. Enhancement of cordyceps polysaccharide production via biosynthetic pathway analysis in Hirsutella sinensis.

    PubMed

    Lin, Shan; Liu, Zhi-Qiang; Baker, Peter James; Yi, Ming; Wu, Hui; Xu, Feng; Teng, Yi; Zheng, Yu-Guo

    2016-11-01

    The addition of various sulfates for enhanced cordyceps polysaccharide (CP) production in submerged cultivation of H. sinensis was investigated, and manganese sulfate was found the most effective. 2mM of manganese sulfate on 0day (d) was investigated as the optimal adding condition, and the CP production reached optimum with 5.33%, increasing by 93.3% compared with the control. Furthermore, the consumption of three main precursors of CP was studied over cultivation under two conditions. Intracellular mannose content decreased by 43.1% throughout 6days cultivation, which corresponded to CP accumulation rate sharply increased from 0 d to 6 d, and mannose was considered as the most preferred precursor for generating CP. Subsequently, mannose biosynthetic pathway was constructed and verified for the first time in H. sinensis, which constituted the important part of CP biosynthesis, and transcriptional levels of the biosynthetic genes were studied. Transcriptional level of gene cpsA was significantly up-regulated 5.35-fold and it was a key gene involved both in mannose and CP biosynthesis. This study demonstrated that manganese sulfate addition is an efficient and simple way to improve CP production. Transcriptional analysis based on biosynthetic pathway was helpful to find key genes and better understand CP biosynthesis.

  14. [Community structure and difference of endophtic bacteria in Aquilaria sinensis with and without agarwood].

    PubMed

    Huang, Ya-Li; Kuang, Zao-Yuan; Song, Meng-Wei; Zhang, Ren

    2015-01-01

    Aquilaria sinensis can generate agarwood, which is closely related with endophyte. Up to now, studies mainly focused on the effects of endophytic fungi on agarwood formation, but studies about endophytic bacteria are rarely reported. In our research, the T-RFs and Shannon index of endophytic bacteria in samples of agarwood increase. The number of distinctive T-RFs fragments of corresponding samples in the same group accounted for more than 60% the number of total T-RFs fragments. In samples of no-agarwood, the dominant bacterial population are Anoxybacillus, Clostridium, Candidatus endobugula, Lysinibacillus. In samples of agarwood, the dominant bacterial population are Clostridium, Lysinibacillus, Luteimonas, phytoplasma. Besides, there are. specific T-RFs fragment in samples of agarwood and no-agarwood respectively. When we perform cluster analysis, we found samples of agarwood highly gather together and samples of no-agarwood highly gather together. This means community of endophytic bacteria emerge significant and regular changes during agarwood formation, which may be result of agarwood production, or maybe it is important reason of agarwood production. In this paper, we obtain more comprehensive and accurate community of endophytic bacteria in Aquilaria sinensis and it's variation during agarwood formation using T-RFLP, which is first study of effects of endophytic bacteria on agarwood formation, and will help to exploit resource of endophytic bacteria more reasonably.

  15. Immunohistochemical localization of caffeine in young Camellia sinensis (L.) O. Kuntze (tea) leaves.

    PubMed

    van Breda, Shane V; van der Merwe, Chris F; Robbertse, Hannes; Apostolides, Zeno

    2013-03-01

    The anatomical localization of caffeine within young Camellia sinensis leaves was investigated using immunohistochemical methods and confocal scanning laser microscopy. Preliminary fixation experiments were conducted with young C. sinensis leaves to determine which fixation procedure retained caffeine the best as determined by high-performance liquid chromatography analysis. High pressure freezing, freeze substitution, and embedding in resin was deemed the best protocol as it retained most of the caffeine and allowed for the samples to be sectioned with ease. Immunohistochemical localization with primary anti-caffeine antibodies and conjugated secondary antibodies on leaf sections proved at the tissue level that caffeine was localized and accumulated within vascular bundles, mainly the precursor phloem. With the use of a pressure bomb, xylem sap was collected using a micro syringe. The xylem sap was analyzed by thin-layer chromatography and the presence of caffeine was determined. We hypothesize that caffeine is synthesized in the chloroplasts of photosynthetic cells and transported to vascular bundles where it acts as a chemical defense against various pathogens and predators. Complex formation of caffeine with chlorogenic acid is also discussed as this may also help explain caffeine's localization.

  16. Synergisms in Alpha-glucosidase Inhibition and Antioxidant Activity of Camellia sinensis L. Kuntze and Eugenia uniflora L. Ethanolic Extracts

    PubMed Central

    Vinholes, Juliana; Vizzotto, Márcia

    2017-01-01

    Background: Camellia sinensis, the most consumed and popular beverages worldwide, and Eugenia uniflora, a Brazilian native species, have been already confirmed to have beneficial effects in the treatment of diabetes mellitus. However, their potential acting together against an enzyme linked to this pathology has never been exploited. Objective: The aim of this study was to evaluate the inhibitory properties of individual and combined ethanolic extracts of the leaves of C. sinensis and E. uniflora over alpha-glucosidase, a key digestive enzyme used on the Type 2 diabetes mellitus (T2DM) control. In addition, their inhibitory activity against 2,2-diphenyl-1-picrylhydrazyl radical (DPPH•) and peroxyl radicals was also assayed. Materials and Methods: Enzyme inhibition and antioxidant potential were assessed based on in vitro assays. Total phenolic compounds, carotenoids, and chlorophylls A and B were achieved using spectrophotometric methods. Results: E. uniflora was almost 40 times more active on alpha-glucosidase than C. sinensis and combined extracts showed a significant synergistic effect with an obtained IC50 value almost 5 times lower than the theoretical value. C. sinensis extract was twice more active than E. uniflora concerning DPPH•, in contrast, E. uniflora was almost 10 times more effective than C. sinensis on inhibition of peroxyl radicals with a significant synergistic effect for combined extracts. The extracts activities may be related with their phytochemicals, mainly phenolic compounds, and chlorophylls. Conclusion: Combined C. sinensis and E. uniflora ethanolic extracts showed synergistic effect against alpha-glucosidase and lipid peroxidation. These herbal combinations can be used to control postprandial hyperglycemia and can also provide antioxidant defenses to patients with T2DM. SUMMARY Alfa-glucosidase and antioxidant Interaction between Camellia sinensis L. Kuntze and Eugenia uniflora L. ethanolic extracts was investigated.Extracts showed

  17. Comparative Transcriptome Analysis of the Accessory Sex Gland and Testis from the Chinese Mitten Crab (Eriocheir sinensis)

    PubMed Central

    He, Lin; Jiang, Hui; Cao, Dandan; Liu, Lihua; Hu, Songnian; Wang, Qun

    2013-01-01

    The accessory sex gland (ASG) is an important component of the male reproductive system, which functions to enhance the fertility of spermatozoa during male reproduction. Certain proteins secreted by the ASG are known to bind to the spermatozoa membrane and affect its function. The ASG gene expression profile in Chinese mitten crab (Eriocheir sinensis) has not been extensively studied, and limited genetic research has been conducted on this species. The advent of high-throughput sequencing technologies enables the generation of genomic resources within a short period of time and at minimal cost. In the present study, we performed de novo transcriptome sequencing to produce a comprehensive transcript dataset for the ASG of E. sinensis using Illumina sequencing technology. This analysis yielded a total of 33,221,284 sequencing reads, including 2.6 Gb of total nucleotides. Reads were assembled into 85,913 contigs (average 218 bp), or 58,567 scaffold sequences (average 292 bp), that identified 37,955 unigenes (average 385 bp). We assembled all unigenes and compared them with the published testis transcriptome from E. sinensis. In order to identify which genes may be involved in ASG function, as it pertains to modification of spermatozoa, we compared the ASG and testis transcriptome of E. sinensis. Our analysis identified specific genes with both higher and lower tissue expression levels in the two tissues, and the functions of these genes were analyzed to elucidate their potential roles during maturation of spermatozoa. Availability of detailed transcriptome data from ASG and testis in E. sinensis can assist our understanding of the molecular mechanisms involved with spermatozoa conservation, transport, maturation and capacitation and potentially acrosome activation. PMID:23342039

  18. The inhibitory mechanism of Cordyceps sinensis on cigarette smoke extract-induced senescence in human bronchial epithelial cells

    PubMed Central

    Liu, Ailing; Wu, Jinxiang; Li, Aijun; Bi, Wenxiang; Liu, Tian; Cao, Liuzhao; Liu, Yahui; Dong, Liang

    2016-01-01

    Objectives Cellular senescence is a state of irreversible growth arrest induced either by telomere shortening (replicative senescence) or stress. The bronchial epithelial cell is often injured by inhaled toxic substances, such as cigarette smoke. In the present study, we investigated whether exposure to cigarette smoke extract (CSE) induces senescence of bronchial epithelial cells; and Cordyceps sinensis mechanism of inhibition of CSE-induced cellular senescence. Methods Human bronchial epithelial cells (16HBE cells) cultured in vitro were treated with CSE and/or C. sinensis. p16, p21, and senescence-associated-galactosidase activity were used to detect cellular senescence with immunofluorescence, quantitative polymerase chain reaction, and Western blotting. Reactive oxygen species (ROS), PI3K/AKT/mTOR and their phosphorylated proteins were examined to testify the activation of signaling pathway by ROS fluorescent staining and Western blotting. Then, inhibitors of ROS and PI3K were used to further confirm the function of this pathway. Results Cellular senescence was upregulated by CSE treatment, and C. sinensis can decrease CSE-induced cellular senescence. Activation of ROS/PI3K/AKT/mTOR signaling pathway was enhanced by CSE treatment, and decreased when C. sinensis was added. Blocking ROS/PI3K/AKT/mTOR signaling pathway can attenuate CSE-induced cellular senescence. Conclusion CSE can induce cellular senescence in human bronchial epithelial cells, and ROS/PI3K/AKT/mTOR signaling pathway may play an important role in this process. C. sinensis can inhibit the CSE-induced senescence. PMID:27555762

  19. Evaluation of the wound-healing activity of Hibiscus rosa sinensis L (Malvaceae) in Wistar albino rats

    PubMed Central

    Bhaskar, Anusha; Nithya, V.

    2012-01-01

    Objective: To investigate the wound-healing potency of the ethanolic extract of the flowers of Hibiscus rosa sinensis. Materials and Methods: The wound-healing activity of H. rosa sinensis (5 and 10% w/w) on Wistar albino rats was studied using three different models viz., excision, incision and dead space wound. The parameters studied were breaking strength in incision model, granulation tissue dry weight, breaking strength and collagen content in dead space wound model, percentage of wound contraction and period of epithelization in excision wound model. The granulation tissue formed on days 4, 8, 12, and 16 (post-wound) was used to estimate total collagen, hexosamine, protein, DNA and uronic acid. Data were analyzed by Analysis of Variance (ANOVA) test. P<0.05 was considered statistically significant. Results: The extract increased cellular proliferation and collagen synthesis at the wound site, as evidenced by increase in DNA, total protein and total collagen content of granulation tissues. The extract-treated wounds were found to heal much faster as indicated by improved rates of epithelialization and wound contraction. The extract of H. rosa sinensis significantly (P<0.001) increased the wound-breaking strength in the incision wound model compared to controls. The extract-treated wounds were found to epithelialize faster, and the rate of wound contraction was significantly (P<0.001) increased as compared to control wounds. Wet and dry granulation tissue weights in a dead space wound model increased significantly (P<0.001). There was a significant increase in wound closure rate, tensile strength, dry granuloma weight, wet granuloma weight and decrease in epithelization period in H. rosa sinensis-treated group as compared to control and standard drug-treated groups. Conclusion: The ethanolic extract of H. rosa sinensis had greater wound-healing activity than the nitrofurazone ointment. PMID:23248396

  20. B-cell lymphoma-2 localization in the female reproductive tract of the Chinese soft-shelled turtle, Pelodiscus sinensis and its relationship with sperm storage.

    PubMed

    Le, Yuan; Chen, Shaofan; Hu, Lisi; Zhang, Linli; Ullah, Shakeeb; Liu, Tengfei; Yang, Ping; Liu, Yi; Chen, Qiusheng

    2015-12-01

    The aim of the present study was to investigate the expression and localization of B-cell lymphoma-2 (Bcl-2) in the oviduct of the Chinese soft-shelled turtle, Pelodiscus sinensis, during the reproductive cycle to analyze the relationship between Bcl-2 and sperm storage. Bcl-2 expression was confirmed in the P. sinensis oviduct by western blot analysis. Hematoxylin-eosin staining showed that female P. sinensis stored sperm from November to April of the following year. The oviduct showed positive immunostaining for Bcl-2 of epithelial ciliated cells, gland ducts, and gland cells. Bcl-2 expression in the oviduct was associated with sperm storage occurrence. This indicates that the survival factor Bcl-2 may play a role in P. sinensis sperm storage.

  1. Effects of Ilex latifolia and Camellia sinensis on cholesterol and circulating immune complexes in rats fed with a high-cholesterol diet.

    PubMed

    Luo, Xian-Yang; Li, Na-Na; Liang, Yue-Rong

    2013-01-01

    Hypercholesterolaemia is one of the risk factors for atherosclerosis and subsequent cardiovascular disease. Here, we investigated the effects of dietary supplementation with Ilex latifolia or green tea (Camellia sinensis) on the levels of plasma total cholesterol, high-density lipoprotein cholesterol and circulating immune complexes in Sprague Dawley rats fed with a high-cholesterol diet. We demonstrated that daily administration by gavage of I. latifolia or C. sinensis at doses of 1.0 or 2.0 g/kg body weight for 30 days resulted in a significant decrease in plasma total cholesterol levels and circulating immune complexes and an increase in high-density lipoprotein cholesterol in rats fed with a high-cholesterol diet compared with levels in the high-cholesterol diet control group. C. sinensis was more effective than I. latifolia. I. latifolia and C. sinensis could be used as food supplements to protect against the development of hypercholesterolaemia.

  2. Cordyceps sinensis biomass produced by submerged fermentation in high-fat diet feed rats normalizes the blood lipid and the low testosterone induced by diet

    PubMed Central

    Freire dos Santos, Leandro; Rubel, Rosália; Bonatto, Sandro José Ribeiro; Zanatta, Ana Lucia; Aikawa, Júlia; Yamaguchi, Adriana Aya; Torres, Maria Fernanda; Soccol, Vanete Thomaz; Habu, Sascha; Prado, Karin Braun; Soccol, Carlos Ricardo

    2012-01-01

    This study investigated the effect of Cordyceps sinensis biomass supplementation obtained from submerged fermentation on blood lipid and low testosterone induced by high-fat diet (HFD). The experiments were carried out using a long-term intake of HFD and HFD plus Simvastatin or C. sinensis (4 months). Our results show that plasma cholesterol, triglycerides and LDL were decreased by Cordyceps sinensis biomass supplementation (CSBS). A long-term intake of HFD caused a significant liver damage which has been reverted by CSBS. CSBS normalized decreasing testosterone levels observed in high-fat diet feed rats. All these findings lead us to suggest that C. sinensis was able to decrease blood lipid concentration, increase hepatoprotective activity and normalize testosterone levels. PMID:27847459

  3. Genetic structure of Miscanthus sinensis and Miscanthus sacchariflorus in Japan indicates a gradient of bidirectional but asymmetric introgression

    SciTech Connect

    Clark, Lindsay V.; Stewart, J. Ryan; Nishiwaki, Aya; Toma, Yo; Kjeldsen, Jens Bonderup; Jorgensen, Uffe; Zhao, Hua; Peng, Junhua; Yoo, Ji Hye; Heo, Kweon; Yu, Chang Yeon; Yamada, Toshihiko; Sacks, Erik J.

    2015-01-24

    Unilateral introgression from diploids to tetraploids has been hypothesized to be an important evolutionary mechanism in plants. However, few examples have been definitively identified, perhaps because data of sufficient depth and breadth were difficult to obtain before the advent of affordable high-density genotyping. Throughout Japan, tetraploid Miscanthus sacchariflorus and diploid Miscanthus sinensis are common, and occasionally hybridize. In this study, 667 M. sinensis and 78 M. sacchariflorus genotypes from Japan were characterized using 20 704 SNPs and ten plastid microsatellites. Similarity of SNP genotypes between diploid and tetraploid M. sacchariflorus indicated that the tetraploids originated through autopolyploidy. Structure analysis indicated a gradient of introgression from diploid M. sinensis into tetraploid M. sacchariflorus throughout Japan; most tetraploids had some M. sinensis DNA. Among phenotypically M. sacchariflorus tetraploids, M. sinensis ancestry averaged 7% and ranged from 1-39%, with introgression greatest in southern Japan. Unexpectedly, rare (~1%) diploid M. sinensis individuals from northern Japan were found with 6-27% M. sacchariflorus ancestry. Population structure of M. sinensis in Japan included three groups, and was driven primarily by distance, and secondarily by geographic barriers such as mountains and straits. Miscanthus speciation is a complex and dynamic process. In contrast to limited introgression between diploid M. sacchariflorus and M. sinensis in northern China, selection for adaptation to a moderate maritime climate probably favoured cross-ploidy introgressants in southern Japan. Ultimately, these results will help guide the selection of Miscanthus accessions for the breeding of biomass cultivars.

  4. Determination of nucleosides in Cordyceps sinensis and Ganoderma lucidum by high performance liquid chromatography method

    PubMed Central

    Khan, Masood Shah; Parveen, Rabea; Mishra, Kshipra; Tulsawani, Rajkumar; Ahmad, Sayeed

    2015-01-01

    Background: Nucleosides are supportive in the regulation and modulation of various physiological processes in body, they acts as precursors in nucleic acid synthesis, enhance immune response, help in absorption of iron and influence the metabolism of fatty acids. Cordyceps sinensis and Ganoderma lucidum are well-known for its use in traditional medicine of China, Nepal and India. They are rich in nucleosides such as adenine, adenosine, cordycepin, etc. Hence, a simple, economic and accurate high-performance liquid chromatography (HPLC) analytical method was proposed for determination of adenine and adenosine for the quality control of plants. Materials and Methods: Chromatographic experiments were conducted on YL9100 HPLC system (South Korea). Reversed-phase chromatography was performed on a C18 column with methanol and dihydrogen phosphate as the mobile phase in isocratic elution method at a flow rate of 1.0 mL/min. Detection was carried out at 254 nm, which gives a sharp peak of adenine and adenosine at a retention time of 6.53 ± 0.02 min and 12.41 ± 0.02, respectively. Results and Discussion: Linear regression analysis data for the calibration plot showed a good linear relationship between response and concentration in the range of 25–200 µg/mL for adenosine and 100–800 µg/mL for adenine with regression coefficient of 0.999 and 0.996, respectively. The adenine was found 0.16% and 0.71% w/w in G. lucidum and in C. sinensis, respectively, and adenosine was found to be 0.14% w/w in G. lucidum whereas absent in C. sinensis. Conclusion: The developed HPLC method for the quantification of adenosine and adenine can be used for the quality control and standardization of crude drug and for the different herbal formulations, in which adenine and adenosine are present as major constituents. The wide linearity range, sensitivity, accuracy, and simple mobile phase imply the method is suitable for routine quantification of adenosine and adenine with high precision and

  5. Altered Proteomic Polymorphisms in the Caterpillar Body and Stroma of Natural Cordyceps sinensis during Maturation

    PubMed Central

    Wu, Zi-Mei; Gao, Ling; Yao, Yi-Sang; Tan, Ning-Zhi; Wu, Jian-Yong; Ni, Luqun; Zhu, Jia-Shi

    2014-01-01

    Objective To examine the maturational changes in proteomic polymorphisms resulting from differential expression by multiple intrinsic fungi in the caterpillar body and stroma of natural Cordyceps sinensis (Cs), an integrated micro-ecosystem. Methods The surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS) biochip technique was used to profile the altered protein compositions in the caterpillar body and stroma of Cs during its maturation. The MS chromatograms were analyzed using density-weighted algorithms to examine the similarities and cluster relationships among the proteomic polymorphisms of the Cs compartments and the mycelial products Hirsutella sinensis (Hs) and Paecilomyces hepiali (Ph). Results: SELDI-TOF MS chromatograms displayed dynamic proteomic polymorphism alterations among samples from the different Cs compartments during maturation. More than 1,900 protein bands were analyzed using density-weighted ZUNIX similarity equations and clustering methods, revealing integral polymorphism similarities of 57.4% between the premature and mature stromata and 42.8% between the premature and mature caterpillar bodies. The across-compartment similarity was low, ranging from 10.0% to 18.4%. Consequently, each Cs compartment (i.e., the stroma and caterpillar body) formed a clustering clade, and the 2 clades formed a Cs cluster. The polymorphic similarities ranged from 0.51% to 1.04% between Hs and the Cs compartments and were 2.8- to 4.8-fold higher (1.92%–4.34%) between Ph and the Cs compartments. The Hs and Ph mycelial samples formed isolated clades outside of the Cs cluster. Conclusion Proteomic polymorphisms in the caterpillar body and stroma of Cs change dynamically during maturation. The proteomic polymorphisms in Hs and Ph differ from those in Cs, suggesting the presence of multiple Cs-associated fungi and multiple Ophiocordyceps sinensis genotypes with altered differential protein expression in the Cs compartments

  6. Seasonal trends, sampling plans and parasitoid complex of the Chinese wax scale, Ceroplastes sinensis Del Guercio (Hemiptera: Coccidae), in Mediterranean citrus groves.

    PubMed

    Martínez-Ferrer, M T; Campos-Rivela, J M; Verdú, M J

    2015-02-01

    Seasonal trends and the parasitoid complex of Chinese wax scale (Ceroplastes sinensis) was studied from July 2010 to February 2013. Six commercial citrus groves located in northeastern Spain were sampled fortnightly. Chinese wax scale completed a single annual generation. Egg oviposition started in May and continued until mid-July. Egg hatching began in mid-June, and in the first quarter of August, the maximum percentage of hatched eggs was reached. In the same groves, the parasitoid species of C. sinensis were determined together with their seasonal trends, relative abundance and occurrence on C. sinensis. Four hymenoptera were found parasitizing C. sinensis, mainly on third instars and females: Coccophagus ceroplastae (Aphelinidae), Metaphycus helvolus (Encyrtidae), Scutellista caerulea (Pteromalidae) and Aprostocetus ceroplastae (Eulophidae). The most abundant species was A. ceroplastae, corresponding to 54% of the parasitoids emerged. Coccophagus ceroplastae and M. helvolus represented 19%, whereas S. caerulea comprised 8% of the total. This study is the first published record of C. ceroplastae in Spain and the first record of M. helvolus on C. sinensis in Spain. Concerning the economical thresholds normally used, sampling plans developed for the management of C. sinensis in citrus groves should target population densities of around 12-20% of invaded twigs, equivalent to 0.2-0.5 females per twig. The sample size necessary to achieve the desired integrated pest management precision is 90-160 twigs per grove for the enumerative plan and about 160-245 twigs per grove for the binomial plan.

  7. Identification of conserved and novel microRNAs in Aquilaria sinensis based on small RNA sequencing and transcriptome sequence data.

    PubMed

    Gao, Zhi-Hui; Wei, Jian-He; Yang, Yun; Zhang, Zheng; Xiong, Huan-Ying; Zhao, Wen-Ting

    2012-08-15

    Agarwood is in great demand for its high value in medicine, incense, and perfume across Asia, Middle East, and Europe. As agarwood is formed only when the Aquilaria trees are wounded or infected by some microbes, overharvesting and habitat loss are threatening some populations of agarwood-producing species. Aquilaria sinensis is such a significant economic tree species. To promote the production efficiency and protect the resource of A. sinensis, it would be critical to reveal the regulation mechanisms of stress-induced agarwood formation. MicroRNAs (miRNAs), a key gene expression regulator involved in various plant stress response and metabolic processes, might function in agarwood formation, but no report concerning miRNAs in Aquilaria is available. In this study, the small RNA high-throughput sequencing and 454 transcriptome data were adopted to identify both conserved and novel miRNAs in A. sinensis. Deep sequencing showed that the small RNA (sRNA) population of A. sinensis was complex and the length of sRNAs varied. By in silico analysis of the small RNA deep sequencing data and transcriptome data, we discovered 27 novel miRNAs in A. sinensis. Based on the mature miRNA sequence conservation, we identified 74 putative conserved miRNAs from A. sinensis and 10 of them were confirmed with hairpin forming precursor. Interestingly, a novel miRNA sequence was determined to be the miRNA of asi-miR408, but with accumulation much higher than asi-miR408. The expression levels of ten stress-responsive miRNAs were examined during the time-course after wound treatment. Eight were shown to be wound-responsive. This not only shows the existence of miRNAs in this Asian economically significant tree species but also indicated its critical role in stress-induced agarwood formation. The highly accumulated miRNA of asi-miR408 implied miRNAs would be functional as well as miRNAs in plants.

  8. Genetic diversity of Ophiocordyceps sinensis, a medicinal fungus endemic to the Tibetan Plateau: Implications for its evolution and conservation

    PubMed Central

    2009-01-01

    Background Ophiocordyceps sinensis (syn. Cordyceps sinensis), endemic to alpine regions on the Tibetan plateau, is one of the most valuable medicinal fungi in the world. Huge commercial demand has led to excessive harvest and a dramatic decline in its numbers. The diversity of terrains and climates on the Tibetan Plateau and the broad insect host range (more than 50 species in the family Hepialidae) may have resulted in substantial intraspecific genetic diversity for this fungus. The objective of this study was to evaluate the population distribution of O. sinensis from geographically diverse regions of the Tibetan Plateau based on nrDNA ITS and MAT1-2-1 gene sequences. Understanding of the genetic diversity and genesis of O. sinensis will provide important information for the evolution and conservation of this fungus. Results Significant sequence variations in the ITS and MAT1-2-1 genes (27 and 23 informative sites, eight and seven haplotypes, respectively) were observed. Phylogenetic analysis based on ITS sequences, MAT1-2-1 sequences, or their combined data set, clustered isolates from northern regions in one clade (clade I), whereas isolates from southern regions were dispersed in all four clades (clade I-IV). Single-strand conformation polymorphism (SSCP) analyses of 2639 ITS clones from seven samples revealed 91 different SSCP patterns that were subsequently sequenced. ITS heterogeneity was found in XZ-LZ07-H1 (Nyingchi population), and 17 informative sites and five haplotypes were detected from 15 clones. The five haplotypes clustered into three clades (clade I, II, and IV). Conclusions Significant genetic divergence in O. sinensis was observed and the genetic diversification was greater among southern isolates than that among northern isolates. The polymorphism of nrDNA ITS sequences suggested that O. sinensis spread from a center of origin (the Nyingchi District) to southern regions and subsequently to northern areas. These results suggest that southern

  9. Quantitative Analysis of Major Phytochemicals in Orthodox tea (Camellia sinensis), Oxidized under Compressed Air Environment.

    PubMed

    Panda, Brajesh Kumar; Datta, Ashis Kumar

    2016-04-01

    This study describes major changes in phytochemical composition of orthodox tea (Camellia sinensis var. Assamica) oxidized under compressed air (CA). The experiments for oxidation were conducted under air pressure (101, 202, and 303 kPa) for 150 min. Relative change in the concentrations of caffeine, catechins, theaflavins (TF), and thearubigins (TR) were analyzed. Effect of CA pressure was found to be nonsignificant in regulating caffeine concentration during oxidation. But degradation in different catechins as well as formation of different TF was significantly affected by CA pressure. At high CA pressure, TF showed highest peak value. TR was found to have slower rate of formation during initial phase of oxidation than TF. Even though the rate of TR formation was significantly influenced by CA, a portion of catechins remained unoxidized at end of oxidation. Except caffeine, the percent change in rate of formation or degradation were more prominent at 202 kPa.

  10. Structural characterization, sulfation and antitumor activity of a polysaccharide fraction from Cyclina sinensis.

    PubMed

    Jiang, Changxing; Xiong, Qingping; Li, Songlin; Zhao, Xirong; Zeng, Xiaoxiong

    2015-01-22

    In the present study, we investigated the preliminary structure, sulfation and antitumor activity of a polysaccharide fraction from Cyclina sinensis (CSPS-1). Results of structural characterization showed that the backbone chain of CSPS-1 was composed of glucose linked by α-(1→4) glycosidic bond, and the branch chain was attached to backbone chain by (1→6) glycosidic bond. CSPS-1 was sulfated by chlorosulfonic acid-pyridine method under different modification conditions according to the orthogonal test L9(3(4)), affording nine sulfated polysaccharides (CSPS-1-S). The optimal sulfation conditions for CSPS-1 were reaction temperature of 65°C, reaction time of 2h and chlorosulfonic acid-pyridine ratio of 1:4. Structural analysis revealed that sulfation had occurred at position of C-6 in CSPS-1. In addition, CSPS-1-S exhibited significantly higher inhibitory activity in vitro against human gastric cancer BGC-823 cells.

  11. Hot-water extract from mycelia of Cordyceps sinensis as a substitute for antibiotic growth promoters.

    PubMed

    Koh, Jong Ho; Suh, Hyung Joo; Ahn, Tae Seok

    2003-04-01

    Broiler chicks were orally dosed with a hot-water extract of mycelia from Cordyceps sinensis (CS-HW) to assess possible substitution of Avilamycin as an antibiotic growth promoter (AGP). The growth performance (body weight gain and survivability) and the health index (the microflora in the small intestines and the antibody titer to Newcastle disease virus) of chicks were significantly improved in the CS-HW (600 mg/kg diet) and the Avilamycin (20 mg/kg diet) fed group in comparison with the control group (p < 0.05). The Avilamycin-fed group and the CS-HW-fed group had similar growth performances but the latter gave a better microbial flora in the small intestines. These results indicate that CS-HW enhances the physiological activity in chicks and can be used as a substitute for AGPs.

  12. 2-(2-phenylethyl)chromone derivatives in Chinese agarwood "Qi-Nan" from Aquilaria sinensis.

    PubMed

    Yang, De Lan; Mei, Wen Li; Zeng, Yan Bo; Guo, Zhi Kai; Zhao, You Xing; Wang, Hao; Zuo, Wen Jian; Dong, Wen Hua; Wang, Qing Huang; Dai, Hao Fu

    2013-09-01

    Five new 2-(2-phenylethyl)chromone derivatives, qinanones A-E (1-5), together with eight known 2-(2-phenylethyl)chromone derivatives (6-13), were isolated from the Et2O extract of high-quality Chinese agarwood "Qi-Nan" originating from Aquilaria sinensis. The structures of the new 2-(2-phenylethyl)chromones were elucidated by spectroscopic techniques (UV, IR, 1D and 2D NMR) and MS analyses. In the bioassay for acetylcholinesterase inhibitors, compounds 1-6, 10, and 12 exhibited weak inhibitory activities (inhibition percentage ranged from 10 % to 24 % at the concentration of 50 µg/mL). Compared with other agarwoods, "Qi-Nan" was different in containing 2-(2-phenylethyl)chromones with unsubstituted chromone rings.

  13. Effect of hydrogen peroxide and camellia sinensis extract on reduction of oxygen level in graphene oxide

    NASA Astrophysics Data System (ADS)

    Celina Selvakumari, J.; Dhanalakshmi, J.; Pathinettam Padiyan, D.

    2016-10-01

    The intention of this work is to reduce the oxygen level in graphene oxide. The reduction process was initiated while preparing graphene oxide using modified Hummer’s method. In this new method, increase in hydrogen peroxide concentration during the preparation process results in the oxygen content reduction. Adding green tea (camellia sinensis) extract with increased hydrogen peroxide results in further reduction of oxygen content and changed the graphene oxide to reduced graphene oxide. The structural and optical properties of the new found reduced graphene oxide was analysed using XRD, FTIR, TEM, Raman and UV-vis spectra. The overall observation reflects that the sp3 carbon network of graphene oxide changed into sp2 carbon lattice of graphene which is very handful in supercapacitor and biosensor fields.

  14. Anti-ulcer effect of the hot water extract of black tea (Camellia sinensis).

    PubMed

    Maity, S; Vedasiromoni, J R; Ganguly, D K

    1995-06-01

    The effect of the hot water extract of black tea (Camellia sinensis (L.) O. Kuntze, Theaceae) on ulceration induced by various ulcerogens and by cold restraint stress (CRS) was investigated in albino rats. While prior administration of tea extract for 7 days significantly reduced the incidence of ulcer, ulcer number and ulcer index produced by aspirin, indomethacin, ethanol, reserpine and CRS, it failed to inhibit the ulcers induced by serotonin and histamine. Tea extract also favourably altered the changes in acid and peptic activity of gastric secretion induced by aspirin, indomethacin, ethanol, reserpine and CRS. The observations suggest that the hot water extract of black tea possesses anti-ulcer activity, probably mediated through prostaglandins.

  15. Dyeing wool and cotton fibres with acidic extract of Hibiscus rosa sinensis flower.

    PubMed

    Hayat, Lamya; Jacob, Dangly Ann

    2016-05-02

    The focus of this work is to extract a natural dye for colouring camel wool as a substitute for synthetic dyes used in the Sadu House of Kuwait. Their target is to keep the tradition of tent and rug production natural in all its manifestations. Therefore, our task was to find an abundant source that provides a colour preferably red to purple. Hibiscus rosa sinensis (HRS) is an abundantly available plant in Kuwait that was explored for extraction of the red dye to colour camel wool permanently. The powdered petals of red flowers of HRS was extracted with 5% acetic acid which yielded a deep red colour that showed a great potential for woollen fibre dyeing. The use of mordants like alum and some metal salts manifested a wide range of fixed colours which intensified at 85 °C. The colours produced had excellent fastness and was accepted by the Sadu House.

  16. Hydroxylated polymethoxyflavones and methylated flavonoids in sweet orange (Citrus sinensis) peel.

    PubMed

    Li, Shiming; Lo, Chih-Yu; Ho, Chi-Tang

    2006-06-14

    Polymethoxyflavones (PMFs) from citrus genus have been of particular interest because of their broad spectrum of biological activities, including antiinflammatory, anticarcinogenic, and antiatherogenic properties. There have been increasing interests in the exploration of health beneficial properties of PMFs in citrus fruits. Therefore, the isolation and characterization of PMFs from sweet orange (Citrus sinensis) peel will lead to new applications of the byproducts from orange juice processes and other orange consumption in nutraceutical and pharmaceutical products. In our study, eight hydroxylated PMFs, six PMFs, one polymethoxyflavanone, one hydroxylated polymethoxyflavanone, and two hydroxylated polymethoxychalcones were isolated from sweet orange peel and their structures were elucidated by various MS, UV, and different NMR techniques. Some of the hydroxylated PMFs and chalcones are newly isolated from sweet orange peel.

  17. Biosynthesis, characterisation and antimicrobial activity of silver nanoparticles using Hibiscus rosa-sinensis petals extracts.

    PubMed

    Nayak, Debasis; Ashe, Sarbani; Rauta, Pradipta Ranjan; Nayak, Bismita

    2015-10-01

    Green synthesis of metallic nanoparticles has lured the world from the chemical and physical approaches owing to its rapid, non-hazardous and economic aspect of production mechanism. In this study, silver nanoparticles (AgNPs) were synthesised using petal extracts of Hibiscus rosa-sinensis. The AgNPs displayed characteristic surface plasmon resonance peak at around 421 nm having a mean particle size of 76.25±0.17 nm and carried a charge of -41±0.2 mV. The X-ray diffraction patterns displayed typical peaks of face centred cubic crystalline silver. The surface morphology was characterised by scanning electron microscopy and atomic force microscopy. Fourier transform infrared spectroscopy studies confirmed the surface modifications of the functional groups for the synthesis of AgNPs. Furthermore, the synthesised AgNPs displayed proficient antimicrobial activity against pathogenic strains of Vibrio cholerae, Escherichia coli, Klebsiella pneumoniae and Staphylococcus aureus.

  18. A novel green chemical route for synthesis of silver nanoparticles using camellia sinensis.

    PubMed

    Kamal, Sarika Srinivas Kalyan; Sahoo, Prasanta Kumar; Vimala, Johnson; Premkumar, Manda; Ram, Shanker; Durai, Loganathan

    2010-12-01

    The thrust to develop environmental friendly procedures for production of Nanoparticles arises from the very fact that current nanotechnology research uses a lot of chemicals, which are potential threat to both environment and public health. Tea (Camellia Sinensis) with its rich source of polyphenolic compounds has been exploited for the reduction and capping of silver nanoparticles (Ag-NPs), making it a complete green chemical route. The reduction of Ag+ to Ag0 was observed by the color change from pale yellow to dark yellow. The reaction was followed with the help of UV-Visible spectrometer. Crystal structure was obtained by carrying out X-ray diffraction studies and it showed face centered cubic (fcc) structure. The particle size and morphology were obtained from transmission electron microscopy (TEM) and small angle X-ray scattering (SAXS) studies. An average particle size of 25 nm silver particles could be obtained using this method and the TEM and SAXS data corroborate with each other.

  19. Physicochemical and sensory quality of yogurt incorporated with pectin from peel of Citrus sinensis.

    PubMed

    Arioui, Fatiha; Ait Saada, Djamel; Cheriguene, Abderrahim

    2017-03-01

    Industrial by-product like orange peel plays an important role in pectin manufacture. The objective of this article was to extract pectin from peel of Citrus sinensis and to study the effect of its incorporation on the quality of yogurt during the period of fermentation and postacidification. Physicochemical, organoleptic, and rheological properties of yogurt prepared with pectin were studied in order to determine the best preparation depending on the rate of pectin. The extraction pectin yield was estimated to more than 24%. The viscosity and acidity were increased with increasing of the pectin rate. The best viscosity value was obtained with 0.6% of pectin. Furthermore, the effect of the rate of pectin incorporation in the fermented milks was clearly observed on the number of Streptococcus thermophilus and Lactobacillus bulgaricus, the cohesiveness, the adhesiveness, the taste, and the whey exudation.

  20. Influence of Camellia sinensis extract on Zinc Oxide nanoparticle green synthesis

    NASA Astrophysics Data System (ADS)

    Nava, O. J.; Luque, P. A.; Gómez-Gutiérrez, C. M.; Vilchis-Nestor, A. R.; Castro-Beltrán, A.; Mota-González, M. L.; Olivas, A.

    2017-04-01

    This work addresses low cost, non-toxic green synthesis of Zinc Oxide nanoparticles prepared using different amounts of Camellia sinensis extract. The Synthesized material was studied and characterized through Fourier transform infrared spectroscopy (FTIR), x-ray diffraction (XRD), transmission electron microscopy (TEM). The Zinc Oxide nanoparticles presented the desired Znsbnd O bond at 618 cm-1, demonstrated growth in a purely hexagonal Wurtzite crystal structure, and, depending on the amount of extract used, they presented different size and shape homogeneity. The photocatalytic activity of the obtained Zinc Oxide nanoparticles was studied. The photocatalytic degradation studies were done at a 1:1 M ratio of methylene blue to Zinc Oxide nanoparticles under UV light. The obtained results presented a better degradation rate than commercially available Zinc Oxide nanoparticles.

  1. Effects of Fermented Mushroom of Cordyceps sinensis, Rich in Selenium, on Uterine Cervix Cancer

    PubMed Central

    Ji, Jing; Liu, Juan; Liu, Haijuan; Wang, Yueling

    2014-01-01

    The purpose of this study was to investigate the effect of fermented mushroom of Cordyceps sinensis (CS), rich in selenium (Se-CS), on uterine cervical cancer in mice. The methylcholanthrene- (MCA-) induced tumor model was used in this paper. After the mice were administered Se-CS, the animals showed 40% tumor incidence (P < 0.05). Se-CS also enhanced the immune functions. Se-CS treatment showed significant (P < 0.05–0.01) restoration in the level of glutathione content, lipid peroxidation, glutathione peroxidase activity, glutathione reductase activity, catalase activity, Na+/K+-ATPase activity, and glutathione S transferase activity. This finding suggested that the concomitant use of Se and CS could be a potential therapeutic approach to improve the efficacy of therapy for uterine cervical cancer. PMID:24971145

  2. Classical and molecular cytogenetics of Khawia sinensis (Cestoda: Caryophyllidea), invasive parasite of carp, Cyprinus carpio.

    PubMed

    Orosová, Martina; Oros, Mikuláš

    2012-05-01

    Chromosomes of the invasive tapeworm Khawia sinensis (Caryophyllidea), the specific parasite of common carp, were analyzed by means of conventional Giemsa staining and using fluorescent DAPI and YOYO-1 dyes, silver staining, and fluorescent in situ hybridization (FISH) with 18S rDNA probe. The karyotype is composed of eight pairs of metacentric and telocentric chromosomes (2n = 16, n = 3m + 5t, TCL = 42.54 μm). Constitutive heterochromatin was located at pericentromeric regions of all pairs, except for the largest metacentric pair (no. 1), which possessed no DAPI-positive band. FISH with rDNA probe revealed that both homologues of chromosome pair no. 6 carry a cluster of ribosomal arrays, which were located interstitially close to the centromere. Present results are compared with previous cytogenetic data on Khawia spp., and comments are made on the karyotypes with respect to their phylogenetic links.

  3. Spasmolytic and Antibacterial Activity of Two Citrus sinensis Osbeck Varieties Cultivated in Mexico.

    PubMed

    Sánchez-Recillas, Amanda; Arroyo-Herrera, Ana Ly; Araujo-León, Jesús Alfredo; Hernández Núñez, Emanuel; Ortiz Andrade, Rolffy

    2017-01-01

    Antibacterial activity on ATCC strains of Escherichia coli, Salmonella enterica, Salmonella enteritidis, and Salmonella choleraesuis and spasmolytic effect on contraction on rat ileum trips were determinate. Eight organic extracts (hexanic and methanolic) of albedo (mesocarp) and flavedo (pericarp) of two varieties (Valencian and National) of Citrus sinensis (L.) Osbeck of Yucatán, México, were studied. Additionally, chromatographic fingerprints were obtained and correlated with their pharmacological effects. MAN, MAV, and HFN extract caused inhibition against S. choleraesuis (MIC: 1000 µg/mL) and S. enteritidis (MIC: 1000 µg/mL). Regarding the spasmolytic effect, the Valencian extracts variety was more efficient on spontaneous contraction, HAV (Emax = 51.98 ± 1.98%), MAV (Emax = 35.98 ± 1.42%), HFV (Emax = 68.91 ± 4.14%), and MFV (Emax = 51.28 ± 2.59%), versus National variety, HAN (Emax = 43.80 ± 6.32%), MAN (Emax = 14.62 ± 1.69%), HFN (Emax = 64.87 ± 3.04%), and MFN (Emax = 31.01 ± 3.92%). Chromatographic fingerprints of HFV and HFN were found to have some similar signals that belong to monoterpenes, whereas for HAN and HAV similar signals were found belonging to fatty acids and triterpenoids. Methanolic extracts showed signals of (1) furfural, (2) furfural acetone (3) furfuraldehyde and (4) β-sitosterol compounds. Flavedo portion of C. sinensis possessed spasmolytic effect on rat ileum strips and antibacterial activity against Salmonella strains. This species is source for obtaining bioactive compounds with therapeutic potential in the treatment of infectious diarrhea.

  4. Characterization and expression pattern of p53 during spermatogenesis in the Chinese mitten crab Eriocheir sinensis.

    PubMed

    Hou, Cong-Cong; Yang, Wan-Xi

    2013-02-01

    p53, as a "Guardian of the Genome", plays an important role in cell cycle arrest, apoptosis, DNA repair and inhibition of angiogenesis in different tissues including testis. p53 gene and its protein perform many essential roles for mammalian spermatogenesis. To explore its functions during spermatogenesis in Eriocheir sinensis, we have cloned and sequenced the cDNA (1,218 bp) of p53 from the testis by degenerating primer PCR and rapid-amplification of cDNA ends. The protein alignment of p53 shows the conserved DNA binding domain, dimerization site and zinc binding site consisted of the predicted structures. Phylogenetic analysis revealed that p53 was more closer to Marsupenaeus japonicus and Tigriopus japonicus than other examined species. Tissue expression analysis of p53 mRNA showed p53 was distinctly expressed in accessory sexual gland, muscle, gill, heart, hepatopancreas and testis. In situ hybridization revealed that the p53 mRNA was weakly distributed around the nucleus, but stronger in the invaginated acrosomal tubule at the early stage. At the middle stage, p53 mRNA signal was increased than the early stage and the signal displayed dot-like pattern on the surface of cup-like nucleus. The signal on acrosomal cap is stronger than on the acrosomal tubule, despite acrosomal tubule signal was also distinct. At the late stage, the signal was still mainly located in acrosomal cap and acrosomal tubule. Sporadic signal were found surrounding the cup-like nucleus, but they were very weak. In the mature sperm, the signal was dramatically decreased. Even though the signal on cup-like nucleus and acrosomal tubule were distinct, they were weaker than those in middle stage. Based on these results, we concluded that p53 may play an important role in formation of acrosome biogenesis and nuclear shaping during spermiogenesis of E. sinensis.

  5. FOXL2 down-regulates vitellogenin expression at mature stage in Eriocheir sinensis

    PubMed Central

    Li, Qing; Xie, Jing; He, Lin; Wang, Yuanli; Yang, Hongdan; Duan, Zelin; Wang, Qun

    2015-01-01

    Ovarian development in crustaceans is characterized by rapid production of egg yolk protein in a process called vitellogenesis. In the present study, we investigated the involvement of a DEAD (Asp-Glu-Ala-Asp) box RNA helicase 20 (DDX20), forkhead transcription factor (FOXL)2 and fushi tarazu factor (FTZ-F)1 in the regulation of vitellogenesis. Based on ESTs from the testis and accessory gland of Eriocheir sinensis, we cloned the full-length cDNAs of foxl2 and fushitarazu factor 1 (ftz-f1), which include the conserved structural features of the forkhead family and nuclear receptor 5A (NR5A) family respectively. The expression of foxl2 mRNA surged at the mature stage of the ovary, when vtg mRNA swooped, suggesting that foxl2 negatively affects the vitellogenin (VTG) synthesis at this developmental stage. Etoposide (inducing germ cell apoptosis) treatment up-regulated FOXL2 and DDX20 at both the mRNA and the protein levels, primarily in the follicular cells as shown by immunofluorescence analysis. Furthermore, foxl2, ddx20 and ftz-f1 mRNA levels increased significantly with right-eyestalk ablation. Interactions between FOXL2 and DDX20 or FTZ-F1 were confirmed by co-immunoprecipitation and the forkhead domain of FOXL2 was identified as the specific structure interacting with FTZ-F1. In conclusion, FOXL2 down-regulates VTG expression by binding with DDX20 in regulation of follicular cell apoptosis and with FTZ-F1 to repress the synthesis of VTG at the mature stage. This report is the first to describe the molecular mechanism of VTG synthesis in E. sinensis and may shed new light on the regulation of cytochrome P450 enzyme by FOXL2 and FTZ-F1 in vitellogenesis. PMID:26430246

  6. Transcriptome-wide identification of Camellia sinensis WRKY transcription factors in response to temperature stress.

    PubMed

    Wu, Zhi-Jun; Li, Xing-Hui; Liu, Zhi-Wei; Li, Hui; Wang, Yong-Xin; Zhuang, Jing

    2016-02-01

    Tea plant [Camellia sinensis (L.) O. Kuntze] is a leaf-type healthy non-alcoholic beverage crop, which has been widely introduced worldwide. Tea is rich in various secondary metabolites, which are important for human health. However, varied climate and complex geography have posed challenges for tea plant survival. The WRKY gene family in plants is a large transcription factor family that is involved in biological processes related to stress defenses, development, and metabolite synthesis. Therefore, identification and analysis of WRKY family transcription factors in tea plant have a profound significance. In the present study, 50 putative C. sinensis WRKY proteins (CsWRKYs) with complete WRKY domain were identified and divided into three Groups (Group I-III) on the basis of phylogenetic analysis results. The distribution of WRKY family transcription factors among plantae, fungi, and protozoa showed that the number of WRKY genes increased in higher plant, whereas the number of these genes did not correspond to the evolutionary relationships of different species. Structural feature and annotation analysis results showed that CsWRKY proteins contained WRKYGQK/WRKYGKK domains and C2H2/C2HC-type zinc-finger structure: D-X18-R-X1-Y-X2-C-X4-7-C-X23-H motif; CsWRKY proteins may be associated with the biological processes of abiotic and biotic stresses, tissue development, and hormone and secondary metabolite biosynthesis. Temperature stresses suggested that the candidate CsWRKY genes were involved in responses to extreme temperatures. The current study established an extensive overview of the WRKY family transcription factors in tea plant. This study also provided a global survey of CsWRKY transcription factors and a foundation of future functional identification and molecular breeding.

  7. NMR Metabolic profiling of green tea (Camellia sinensis L.) leaves grown at Kemuning, Indonesia

    NASA Astrophysics Data System (ADS)

    Wahyuni, D. S. C.; Kristanti, M. W.; Putri, R. K.; Rinanto, Y.

    2017-01-01

    Green tea (Camellia sinensis L.) has been famous as a beverage and natural medicine. It contains a broad range of primary and secondary metabolites i.e. polyphenols. Nuclear Magnetic Resonance (NMR) has been widely used for metabolic profiling in medicinal plants. It provides a very fast and detailed analysis of the biomolecular composition of crude extracts. Moreover, an NMR spectrum is a physical characteristic of a compound and thus highly reproducible. Therefore, this study aims to profile metabolites of three different varieties of green tea C. Sinensis grown in Kemuning, Middle Java. Three varieties of green tea collected on Kemuning (TR1 2025, Gambung 4/5, and Chiaruan 143) were used in this study. 1H-NMR spectra were recorded at 230C on a 400 MHz Agilent WB (Widebore). The analysis was performed on dried green tea leaves and analyzed by 1H-NMR, 2D-J-resolved and 1H-1H correlated spectroscopy (COSY). MestRenova version 11.0.0 applied to identify metabolites in samples. A 1H-NMR spectrum of tea showed amino acids and organic acids signal at the area δ 0.8–4.0. These were theanine, alanine, threonine, succinic acid, aspartic acid, lactic acid. Anomeric protons of carbohydrate were shown by the region of β-glucose, α-glucose, fructose and sucrose. The phenolic region was depicted at area δ 5.5-8.5. Epigallocatechin derivates and caffeine were detected in the tea leaves. The detail compound identification was observed and discussed in the text.

  8. Antioxidant and antibacterial properties of green, black, and herbal teas of Camellia sinensis

    PubMed Central

    Chan, Eric W.C.; Soh, Eu Ying; Tie, Pei Pei; Law, Yon Peng

    2011-01-01

    Background: The role of non-polymeric phenolic (NP) and polymeric tannin (PT) constituents in the antioxidant and antibacterial properties of six brands of green, black, and herbal teas of Camellia sinensis were investigated. Materials and Methods: Total phenolic content (TPC) and ascorbic acid equivalent antioxidant capacity (AEAC) were assessed using the Folin-Ciocalteu and 2,2-diphenyl-1-picrylhydrazyl (DPPH) assays, respectively. Minimum inhibitory dose (MID) against Gram-positive Micrococcus luteus, Staphylococcus aureus, and Bacillus cereus, and Gram-negative. Escherichia coli, Salmonella typhi, and Pseudomonas aeruginosa was assessed using the disc-diffusion method. Teas were extracted with hot water successively three times for one hour each time. The extracts were fractionated using Sephadex LH-20 column chromatography to obtain the NP and PT constituents. Results: Extraction yields ranged from 12 to 23%. Yields of NP fractions (70–81%) were much higher than those of PT fractions (1–11%), suggesting that the former are the major tea components. Ranking of antioxidant properties of extracts was green tea>black tea>herbal tea. For all six teas, antioxidant properties of PT fractions were significantly higher than extracts and NP fractions. Extracts and fractions of all six teas showed no activity against the three Gram-negative bacteria. Green teas inhibited all three Gram-positive bacteria with S. aureus being the least susceptible. Black and herbal teas inhibited the growth of M. luteus and B. cereus, but not S. aureus. The most potent were the PT fractions of Boh Cameron Highlands and Ho Yan Hor with MID of 0.01 and 0.03 mg/disc against M. luteus. Conclusion: Results suggested that NP constituents are major contributors to the antioxidant and antibacterial properties of teas of C. sinensis. Although PT constituents have stronger antioxidant and antibacterial properties, they constitute only a minor component of the teas. PMID:22224051

  9. Does the morphology of the ear of the Chinese bamboo rat (Rhizomys sinensis) show "Subterranean" characteristics?

    PubMed

    Pleštilová, Lucie; Hrouzková, Ema; Burda, Hynek; Šumbera, Radim

    2016-05-01

    In spite of the growing interest in rodents with subterranean activity in general and the spalacids (Spalacidae) in particular, little is known about the biology of most members of this clade, such as the Chinese bamboo rat (Rhizomys sinensis). Here, we analyzed the ear morphology of R. sinensis with respect to hearing specialization for subterranean or aboveground modes of communication. It is well-known that ecology and style of life of a particular species can be reflected in morphology of its ear, its hearing and vocalization, so we expect that such information could provide us insight into its style of life and its sensory environment. The ratio between the eardrum and stapedial footplate areas, which influences the efficiency of middle ear sound transmission, suggests low hearing sensitivity, as is typical for subterranean species. The cochlea had 3.25 coils and resembled species with good low frequency hearing typical for subterranean mammals. The length of the basilar membrane was 18.9 ± 0.8 mm and its width slowly increased towards the cochlear apex from 60 to 85 μm. The mean density of outer hair cells was 344 ± 22 and of inner hair cells 114 ± 7.3 per 1 mm length of the organ of Corti, and increased apically. These values (except for relatively low hair cell density) usually characterize ears specialized for low frequency hearing. There was no evidence for an acoustic fovea. Apart of low hair cell density which is common in aboveground animals, this species has also relatively large auricles, suggesting the importance of sound localization during surface activity. The ear of the Chinese bamboo rat thus contains features typical for both aboveground and subterranean mammals and suggests that this spalacid has fossorial habits combined with regular aboveground activity.

  10. Preliminary Screening of Antibacterial Activity Using Crude Extracts of Hibiscus rosa sinensis

    PubMed Central

    Arullappan, Sangeetha; Zakaria, Zubaidah; Basri, Dayang Fredalina

    2009-01-01

    Hibiscus rosa sinensis, a member of the Malvaceae family, is widely cultivated in the tropics as an ornamental plant. It is often planted as a fence or hedge plant, and has several forms of flowers with varying colours. It is also used in traditional medicine to induce abortion, ease menstrual cramps, assist in childbirth and relieve headache, fever and inflammation. In this study, we evaluated the antibacterial activity of H. rosa sinesis extract using a disc diffusion method. Crude petroleum ether extract, ethyl acetate extract and methanol extract from the leaves, stems and flowers of the plant were prepared using a cold extraction technique. These extracts were tested at concentrations ranging from 4 mg/disc to 0.017 mg/disc against methicillin-resistant Staphylococcus aureus (MRSA), Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Klebsiella pneumonia. The petroleum ether extract from the leaves, stems and flowers and methanol extract from the leaves showed inhibition zones with diameters > 12 mm against MRSA. Overall, the petroleum ether extract from flowers at concentrations of 4 mg/disc and 2 mg/disc displayed the strongest inhibition zones of 18.6 ± 2.85 mm and 18.5 ± 0.29 mm, respectively, as compared to vancomycin (30 μg/ml), which did not differ significantly from the 18.0 ± 0.10 mm size of the vancomycin (30 μg/ml) inhibition zone (p < 0.05). In conclusion, H. rosa sinensis extract is a potential antibacterial agent for treating MRSA infection. PMID:24575183

  11. Molecular cloning, characterization and functional analysis of a heat shock protein 70 gene in Cyclina sinensis.

    PubMed

    Ren, Yipeng; Pan, Heting; Yang, Ying; Pan, Baoping; Bu, Wenjun

    2016-11-01

    Heat shock protein 70 (HSP70) is an important member of the heat shock protein superfamily and is involved in protecting organisms against various stressors. In the present study, we used RACE to clone a full-length Cyclina sinensis HSP70 cDNA termed CsHSP70. The full length of the CsHSP70 cDNA was 2308 bp, with a 5' untranslated region (UTR) of 42 bp, a 3' UTR of 268 bp, and an open reading frame (ORF) of 1998 bp encoding a polypeptide of 655 amino acids with an estimated molecular mass of 72.75 kDa and an estimated isoelectric point of 5.48. Quantitative real-time PCR was employed to analyze the tissue distribution and temporal expression of the CsHSP70 gene after bacterial challenge and cadmium (Cd) exposure. The CsHSP70 mRNA transcript was expressed ubiquitously in five examined tissues, with the highest expression in hemocytes (P < 0.05) and with the lowest expression in the hepatopancreas. Furthermore, the expression level of CsHSP70 in hemocytes at 3 h after Vibrio anguillarum challenge was extremely significantly up-regulated (P < 0.01). Moreover, the CsHSP70 transcript was up-regulated significantly following exposure to a safe Cd concentration (0.1 mg/L). Finally, after the CsHSP70 gene was silenced by RNA interference, the expression of the CsTLR13 and CsMyD88 genes were extremely significantly decreased (P < 0.01). The results indicated that CsHSP70 could play an important role in mediating the environmental stress and immune responses, and regulating TLR signaling pathway in C. sinensis.

  12. Spasmolytic and Antibacterial Activity of Two Citrus sinensis Osbeck Varieties Cultivated in Mexico

    PubMed Central

    Sánchez-Recillas, Amanda; Arroyo-Herrera, Ana Ly; Araujo-León, Jesús Alfredo; Hernández Núñez, Emanuel

    2017-01-01

    Antibacterial activity on ATCC strains of Escherichia coli, Salmonella enterica, Salmonella enteritidis, and Salmonella choleraesuis and spasmolytic effect on contraction on rat ileum trips were determinate. Eight organic extracts (hexanic and methanolic) of albedo (mesocarp) and flavedo (pericarp) of two varieties (Valencian and National) of Citrus sinensis (L.) Osbeck of Yucatán, México, were studied. Additionally, chromatographic fingerprints were obtained and correlated with their pharmacological effects. MAN, MAV, and HFN extract caused inhibition against S. choleraesuis (MIC: 1000 µg/mL) and S. enteritidis (MIC: 1000 µg/mL). Regarding the spasmolytic effect, the Valencian extracts variety was more efficient on spontaneous contraction, HAV (Emax = 51.98 ± 1.98%), MAV (Emax = 35.98 ± 1.42%), HFV (Emax = 68.91 ± 4.14%), and MFV (Emax = 51.28 ± 2.59%), versus National variety, HAN (Emax = 43.80 ± 6.32%), MAN (Emax = 14.62 ± 1.69%), HFN (Emax = 64.87 ± 3.04%), and MFN (Emax = 31.01 ± 3.92%). Chromatographic fingerprints of HFV and HFN were found to have some similar signals that belong to monoterpenes, whereas for HAN and HAV similar signals were found belonging to fatty acids and triterpenoids. Methanolic extracts showed signals of (1) furfural, (2) furfural acetone (3) furfuraldehyde and (4) β–sitosterol compounds. Flavedo portion of C. sinensis possessed spasmolytic effect on rat ileum strips and antibacterial activity against Salmonella strains. This species is source for obtaining bioactive compounds with therapeutic potential in the treatment of infectious diarrhea. PMID:28356916

  13. A novel integrin function in innate immunity from Chinese mitten crab (Eriocheir sinensis).

    PubMed

    Huang, Ying; Zhao, Ling-Ling; Feng, Jin-Ling; Zhu, Huan-Xi; Huang, Xin; Ren, Qian; Wang, Wen

    2015-10-01

    Integrins belong to a superfamily of conserved α β heterodimeric cell surface receptors that have critical function in cell migration, differentiation, and survival. In this study, an integrin called EsIntegrin was identified from Chinese mitten crab Eriocheir sinensis. EsIntegrin cDNA is 4415 bp long with a 2457 bp open reading frame that encodes an 818 amino acid protein. EsIntegrin contains a signal peptide, an integrin beta subunit (N-terminal portion of extracellular region) INB domain, an epidermal growth factor (hEGF) domain, an integrin B tail domain, a transmembrane region, and an integrin b cyt domain. EsIntegrin was mainly expressed in hemocytes and the heart, with a relatively lower expression level in gills, nerves, intestine, hepatopancreas, muscles, and eyestalk. When healthy crabs were challenged with LPS, PGN, Staphyloccocus aureus, or Vibrio parahaemolyticus, EsIntegrin expression level was upregulated significantly. Recombinant EsIntegrin has agglutination activity to Gram-positive (e.g., S. aureus and Bacillus subtilis) and Gram-negative bacteria (e.g., V. parahaemolyticus and Aeromonas hydrophila) in the presence of calcium. Furthermore, rEsIntegrin could not only bind to various bacteria such as S. aureus, Micrococcus luteus, B. subtilis, Bacillus megaterium, Bacillus thuringiensis, V. parahaemolyticus, Vibrio anguillarum, A. hydrophila, Vibrio natriegens, and Escherichia coli, but this compound also helped crabs in clearing virulent Gram-negative bacterium, V. parahaemolyticus, in vivo. These data suggested that EsIntegrin might function as cellular receptor that is involved in anti-bacterial immunity from E. sinensis.

  14. Genome wide characterization of short tandem repeat markers in sweet orange (Citrus sinensis).

    PubMed

    Biswas, Manosh Kumar; Xu, Qiang; Mayer, Christoph; Deng, Xiuxin

    2014-01-01

    Sweet orange (Citrus sinensis) is one of the major cultivated and most-consumed citrus species. With the goal of enhancing the genomic resources in citrus, we surveyed, developed and characterized microsatellite markers in the ≈347 Mb sequence assembly of the sweet orange genome. A total of 50,846 SSRs were identified with a frequency of 146.4 SSRs/Mbp. Dinucleotide repeats are the most frequent repeat class and the highest density of SSRs was found in chromosome 4. SSRs are non-randomly distributed in the genome and most of the SSRs (62.02%) are located in the intergenic regions. We found that AT-rich SSRs are more frequent than GC-rich SSRs. A total number of 21,248 SSR primers were successfully developed, which represents 89 SSR markers per Mb of the genome. A subset of 950 developed SSR primer pairs were synthesized and tested by wet lab experiments on a set of 16 citrus accessions. In total we identified 534 (56.21%) polymorphic SSR markers that will be useful in citrus improvement. The number of amplified alleles ranges from 2 to 12 with an average of 4 alleles per marker and an average PIC value of 0.75. The newly developed sweet orange primer sequences, their in silico PCR products, exact position in the genome assembly and putative function are made publicly available. We present the largest number of SSR markers ever developed for a citrus species. Almost two thirds of the markers are transferable to 16 citrus relatives and may be used for constructing a high density linkage map. In addition, they are valuable for marker-assisted selection studies, population structure analyses and comparative genomic studies of C. sinensis with other citrus related species. Altogether, these markers provide a significant contribution to the citrus research community.

  15. Molecular characterization of a cytosolic manganese superoxide dismutase from the Chinese mitten crab, Eriocheir sinensis.

    PubMed

    Zhao, D X; Chen, L Q; Qin, J G; Qin, C J; Zhang, H; Wu, P; Li, E C

    2014-11-11

    A cytosolic manganese superoxide dismutase gene (Es-cMnSOD) was cloned from the Chinese mitten crab Eriocheir sinensis, using reverse transcription-polymerase chain reaction and the rapid amplification of cDNA ends. The open reading frame of Es-cMnSOD is 867 bp in length and encodes a 288-amino acid protein without a signal peptide. The calculated molecular mass of the translated protein of Es-cMnSOD is 31.43 kDa, with an estimated isoelectric point of 6.30. The deduced amino acid sequence of Es-cMnSOD has similarities of 90, 89, 84, 87, and 81% to those of white shrimp Litopenaeus vannamei MnSOD, black tiger shrimp Penaeus monodon MnSOD, giant freshwater prawn Macrobrachium rosenbergii MnSOD, blue crab Callinectes sapidus MnSOD, and red swamp crayfish Procambarus clarkii MnSOD, respectively. Es-cMnSOD contains a manganese superoxide dismutase domain (DVWEHAYY) and 4 conserved amino acids responsible for binding manganese. Es-cMnSOD was expressed in the hemocytes, eyestalk, muscle, intestine, gill, and hepatopancreas. Es-cMnSOD transcripts in hemocytes of E. sinensis increased at 1.5 and 48 h after injection of Aeromonas hydrophila, indicating that the induction of the SOD system response occurred within a short period of time. This study suggests that MnSOD may play a critical role in crab immunity, allowing efficient activation of an early innate immune response in the crab.

  16. Improving Training Condition Assessment in Endurance Cyclists: Effects of Ganoderma lucidum and Ophiocordyceps sinensis Dietary Supplementation

    PubMed Central

    Rossi, Paola; Buonocore, Daniela; Altobelli, Elisa; Brandalise, Federico; Cesaroni, Valentina; Iozzi, Davide; Savino, Elena; Marzatico, Fulvio

    2014-01-01

    The main reasons for taking daily dietary supplements are to maintain good health, to improve homeostasis, and to create conditions for reducing the risk of disease. Due to growing market demand, the search for effective, nontoxic, natural compounds with antioxidant and ergogenic properties has increasingly become a matter of interest. This paper describes how a specific combination of fungal supplements can help improve the performance of endurance athletes. We report the effects of a brief 3-month trial of two fungal supplements, Ganoderma lucidum and Cordyceps sinensis (3 capsules of O. sinensis and 2 capsules of G. lucidum per day), in 7 healthy male volunteers, aged between 30 and 40 years, who are all amateur cyclists that participate in “Gran Fondo” cycling races. This trial investigated the effects of fungal supplements on the level of physical fitness of the athletes by monitoring and comparing the following biomarkers just before and after physical exertion: the testosterone/cortisol ratio in the saliva and oxidative stress (DPPH free radical scavenging activity). A decrease of more than 30% in the testosterone/cortisol ratio after race compared to before race was considered as a risk factor for nonfunctional overreaching (NFO) or the overtraining syndrome (OTS). The results show that, after 3 months of supplementation, the testosterone/cortisol ratio changed in a statistically significant manner, thereby protecting the athletes from NFO and OTS. Antioxidant activity was measured by quantifying the scavenging ability of the human serum on the synthetic free radical DPPH. After 3 months of fungal supplementation, the data demonstrate an increased scavenger capacity of free radicals in the athletes' serum after the race, thereby protecting the athletes from oxidative stress. PMID:24799948

  17. Final Report DE-SC0006634. Quantifying phenotypic and genetic diversity of Miscanthus sinensis as a resource for knowledge-based improvement of M. ×giganteus (M. sinensis × M. sacchariflorus)

    SciTech Connect

    Sacks, Erik

    2016-02-08

    Miscanthus is especially attractive as a bioenergy crop for temperate environments because it produces high yields, needs few inputs, and grows well during the cool weather of early spring and late fall when few warm-season grasses can. However, Miscanthus feedstock production for the emerging U.S. bioenergy industry and for existing demand in Europe is based on a single sterile, vegetatively propagated variety of M. ×giganteus. M. ×giganteus is an interspecific hybrid of the parental species M. sinensis and M. sacchariflorus. Prior to the current study, little information existed about the genetic diversity and breeding potential of either M. ×giganteus parental species. In the current project, we studied more than 600 accessions of M. sinensis from throughout its native range in China, Japan, and Korea, in addition to ornamental cultivars and U.S. naturalized populations. Using thousands of DNA markers, we identified seven geographically distinct genetic groups of M. sinensis. Notably, we found that the ornamental cultivars and U.S. naturalized populations were derived from only a subset of the Southern Japan group, indicating that our study greatly increased the genetic diversity available for breeding new biomass cultivars. Additionally, this new understanding of M. sinensis population structure could be used to predict which crosses may produce progeny with the greatest hybrid vigor. Replicated field trials were also established at multiple locations in North America and Asia. Data on traits of importance for biomass productivity, such as flowering time, yield and height, were taken. Analyses of the phenotypic data from the field trials along with the DNA markers allowed us to identify many marker-trait associations. These results will enable marker-assisted breeding, which will allow selection at the seedling stage rather than waiting two to three years to obtain phenotypic data. Thus, this study is expected to greatly increase the efficiency of breeding

  18. Polymyositis - adult

    MedlinePlus

    ... rash is a sign of a similar condition, dermatomyositis . Common symptoms include: Muscle weakness in the shoulders ... in the treatment of refractory adult and juvenile dermatomyositis and adult polymyositis: a randomized, placebo-phase trial. ...

  19. Characterisation of volatile and non-volatile metabolites in etiolated leaves of tea (Camellia sinensis) plants in the dark.

    PubMed

    Yang, Ziyin; Kobayashi, Eiji; Katsuno, Tsuyoshi; Asanuma, Toshimichi; Fujimori, Tamaki; Ishikawa, Takamasa; Tomomura, Miho; Mochizuki, Kazuo; Watase, Takaya; Nakamura, Yoriyuki; Watanabe, Naoharu

    2012-12-15

    Aroma is an essential factor affecting the quality of tea (Camellia sinensis) products. While changes of volatile compounds during tea manufacturing have been intensively studied, the effect of environmental factors on volatile contents of fresh tea leaves has received less attention. We found that C. sinensis var. Yabukita kept in darkness by shading treatment for 3 weeks developed etiolated leaves with significantly increased levels of volatiles, especially volatile phenylpropanoids/benzenoids (VPBs). Upstream metabolites of VPBs, in particular shikimic acid, prephenic acid, and phenylpyruvic acid, showed lower levels in dark treated than in control leaves, whereas the contents of most amino acids including l-phenylalanine, a key precursor of VPBs, were significantly enhanced. In addition, analysis by ultra performance liquid chromatography-time of flight mass spectrometry, capillary electrophoresis-time of flight mass spectrometry, high performance liquid chromatography, and gas chromatography-mass spectrometry indicated that volatile and non-volatile metabolite profiles differed significantly between dark treated and untreated leaves.

  20. Fine Structure of Bacteroids in Root Nodules of Vigna sinensis, Acacia longifolia, Viminaria juncea, and Lupinus angustifolius

    PubMed Central

    Dart, P. J.; Mercer, F. V.

    1966-01-01

    Dart, P. J. (University of Sydney, Sydney, Australia), and F. V. Mercer. Fine structure of bacteroids in root nodules of Vigna sinensis, Acacia longifolia, Viminaria juncea, and Lupinus angustifolius. J. Bacteriol. 91:1314–1319.—In nodules of Vigna sinensis, Acacia longifolia, and Viminaria juncea, membrane envelopes enclose groups of bacteroids. The bacteroids often contain inclusion granules and electron-dense bodies, expand little during development, and retain their rod form with a compact, central nucleoid area. The membrane envelope may persist around bacteroids after host cytoplasm breakdown. In nodules of Lupinus angustifolius, the membrane envelopes enclose only one or two bacteroids, which expand noticeably during development and change from their initial rod structure. Images PMID:5929757

  1. Hypothesis of potential active components in Angelica sinensis by using biomembrane extraction and high performance liquid chromatography.

    PubMed

    Dong, Z B; Li, S P; Hong, M; Zhu, Q

    2005-07-15

    The screening and analysis of bioactive components in traditional Chinese medicines (TCMs) is very important not only for the quality control of crude drugs but also for elucidating the therapeutic principle. In this study, a method for screening potential active components from TCMs was developed by using biomembrane extraction and high performance liquid chromatography. Based on the methodology, aqueous extract of Angelica sinensis (WEAS) was used, and four compounds were detected by HPLC in the desorption eluate of red cell membrane extraction for WEAS. The compounds were identified as ferulic acid, ligustilide, senkyunolide H and senkyunolide I based on their UV, MS and NMR spectra. Actually, ferulic acid and ligustilide are considered as major active components in Angelica sinensis. Therefore, this method may be applied to predict the potential bioactivities of multiple compounds in TCMs simultaneously.

  2. Fragile Sites of ‘Valencia’ Sweet Orange (Citrus sinensis) Chromosomes Are Related with Active 45s rDNA

    PubMed Central

    Lan, Hong; Chen, Chun-Li; Miao, Yin; Yu, Chang-Xiu; Guo, Wen-Wu; Xu, Qiang; Deng, Xiu-Xin

    2016-01-01

    Citrus sinensis chromosomes present a morphological differentiation of bands after staining by the fluorochromes CMA and DAPI, but there is still little information on its chromosomal characteristics. In this study, the chromosomes in ‘Valencia’ C. sinensis were analyzed by fluorescence in situ hybridization (FISH) using telomere DNA and the 45S rDNA gene as probes combining CMA/DAPI staining, which showed that there were two fragile sites in sweet orange chromosomes co-localizing at distended 45S rDNA regions, one proximally locating on B-type chromosome and the other subterminally locating on D-type chromosome. While the chromosomal CMA banding and 45S rDNA FISH mapping in the doubled haploid line of ‘Valencia’ C. sinensis indicated six 45S rDNA regions, four were identified as fragile sites as doubled comparing its parental line, which confirmed the cytological heterozygosity and chromosomal heteromorphisms in sweet orange. Furthermore, Ag-NOR identified two distended 45S rDNA regions to be active nucleolar organizing regions (NORs) in diploid ‘Valencia’ C. sinensis. The occurrence of quadrivalent in meiosis of pollen mother cells (PMCs) in ‘Valencia’ sweet orange further confirmed it was a chromosomal reciprocal translocation line. We speculated this chromosome translocation was probably related to fragile sites. Our data provide insights into the chromosomal characteristics of the fragile sites in ‘Valencia’ sweet orange and are expected to facilitate the further investigation of the possible functions of fragile sites. PMID:26977938

  3. Transcriptome Profiling Analysis on Whole Bodies of Microbial Challenged Eriocheir sinensis Larvae for Immune Gene Identification and SNP Development

    PubMed Central

    Cui, Zhaoxia; Li, Xihong; Liu, Yuan; Song, Chengwen; Hui, Min; Shi, Guohui; Luo, Danli; Li, Yingdong

    2013-01-01

    To study crab immunogenetics of individuals, newly hatched Eriocheir sinensis larvae were stimulated with a mixture of three pathogen strains (Gram-positive bacteria Micrococcus luteus, Gram-negative bacteria Vibrio alginolyticus and fungi Pichia pastoris; 108 cfu·mL-1). A total of 44,767,566 Illumina clean reads corresponding to 4.52 Gb nucleotides were generated and assembled into 100,252 unigenes (average length: 1,042 bp; range: 201-19,357 bp). 17,097 (26.09%) of 65,535 non-redundant unigenes were annotated in NCBI non-redundant protein (Nr) database. Moreover, 23,188 (35.38%) unigenes were assigned to three Gene Ontology (GO) categories, 15,071 (23.00%) to twenty-six Clusters of orthologous Groups (COG) and 8,574 (13.08%) to six Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, respectively. Numerous genes were further identified to be associated with multiple immune pathways, including Toll, immune deficiency (IMD), janus kinase (JAK)-signal transducers and activators of transcription (STAT) and mitogen-activated protein kinase (MAPK) pathways. Some of them, such as tumor necrosis factor receptor associated factor 6 (TRAF6), fibroblast growth factor (FGF), protein-tyrosine phosphatase (PTP), JNK-interacting protein 1 (JIP1), were first identified in E. sinensis. TRAF6 was even first discovered in crabs. Additionally, 49,555 single nucleotide polymorphisms (SNPs) were developed from over 13,309 unigenes. This is the first transcriptome report of whole bodies of E. sinensis larvae after immune challenge. Data generated here not only provide detail information to identify novel genes in genome reference-free E. sinensis, but also facilitate our understanding on host immunity and defense mechanism of the crab at whole transcriptome level. PMID:24324760

  4. Characterization of the glutathione S-transferase gene family through ESTs and expression analyses within common and pigmented cultivars of Citrus sinensis (L.) Osbeck

    PubMed Central

    2014-01-01

    Background Glutathione S-transferases (GSTs) represent a ubiquitous gene family encoding detoxification enzymes able to recognize reactive electrophilic xenobiotic molecules as well as compounds of endogenous origin. Anthocyanin pigments require GSTs for their transport into the vacuole since their cytoplasmic retention is toxic to the cell. Anthocyanin accumulation in Citrus sinensis (L.) Osbeck fruit flesh determines different phenotypes affecting the typical pigmentation of Sicilian blood oranges. In this paper we describe: i) the characterization of the GST gene family in C. sinensis through a systematic EST analysis; ii) the validation of the EST assembly by exploiting the genome sequences of C. sinensis and C. clementina and their genome annotations; iii) GST gene expression profiling in six tissues/organs and in two different sweet orange cultivars, Cadenera (common) and Moro (pigmented). Results We identified 61 GST transcripts, described the full- or partial-length nature of the sequences and assigned to each sequence the GST class membership exploiting a comparative approach and the classification scheme proposed for plant species. A total of 23 full-length sequences were defined. Fifty-four of the 61 transcripts were successfully aligned to the C. sinensis and C. clementina genomes. Tissue specific expression profiling demonstrated that the expression of some GST transcripts was 'tissue-affected' and cultivar specific. A comparative analysis of C. sinensis GSTs with those from other plant species was also considered. Data from the current analysis are accessible at http://biosrv.cab.unina.it/citrusGST/, with the aim to provide a reference resource for C. sinensis GSTs. Conclusions This study aimed at the characterization of the GST gene family in C. sinensis. Based on expression patterns from two different cultivars and on sequence-comparative analyses, we also highlighted that two sequences, a Phi class GST and a Mapeg class GST, could be involved in

  5. Genetic Divergence between Camellia sinensis and Its Wild Relatives Revealed via Genome-Wide SNPs from RAD Sequencing.

    PubMed

    Yang, Hua; Wei, Chao-Ling; Liu, Hong-Wei; Wu, Jun-Lan; Li, Zheng-Guo; Zhang, Liang; Jian, Jian-Bo; Li, Ye-Yun; Tai, Yu-Ling; Zhang, Jing; Zhang, Zheng-Zhu; Jiang, Chang-Jun; Xia, Tao; Wan, Xiao-Chun

    2016-01-01

    Tea is one of the most popular beverages across the world and is made exclusively from cultivars of Camellia sinensis. Many wild relatives of the genus Camellia that are closely related to C. sinensis are native to Southwest China. In this study, we first identified the distinct genetic divergence between C. sinensis and its wild relatives and provided a glimpse into the artificial selection of tea plants at a genome-wide level by analyzing 15,444 genomic SNPs that were identified from 18 cultivated and wild tea accessions using a high-throughput genome-wide restriction site-associated DNA sequencing (RAD-Seq) approach. Six distinct clusters were detected by phylogeny inferrence and principal component and genetic structural analyses, and these clusters corresponded to six Camellia species/varieties. Genetic divergence apparently indicated that C. taliensis var. bangwei is a semi-wild or transient landrace occupying a phylogenetic position between those wild and cultivated tea plants. Cultivated accessions exhibited greater heterozygosity than wild accessions, with the exception of C. taliensis var. bangwei. Thirteen genes with non-synonymous SNPs exhibited strong selective signals that were suggestive of putative artificial selective footprints for tea plants during domestication. The genome-wide SNPs provide a fundamental data resource for assessing genetic relationships, characterizing complex traits, comparing heterozygosity and analyzing putatitve artificial selection in tea plants.

  6. Metal transport protein 8 in Camellia sinensis confers superior manganese tolerance when expressed in yeast and Arabidopsis thaliana

    PubMed Central

    Li, Qinghui; Li, Yue; Wu, Xiayuan; Zhou, Lin; Zhu, Xujun; Fang, Wanping

    2017-01-01

    Manganese (Mn) is an important micronutrient element required for plant growth and development, playing catalytic roles in enzymes, membranes and DNA replication. The tea plant (Camellia sinensis) is able to accumulate high concentration of Mn without showing signs of toxicity, but the molecular mechanisms underlying this remain largely unknown. In this study, the C. sinensis cultivar ‘LJCY’ had higher Mn tolerance than cultivar ‘YS’, because chlorophyll content reduction was lower under the high Mn treatment. Proteomic analysis of the leaves revealed that C. sinensis Metal Tolerance Protein 8 (CsMTP8) accumulated in response to Mn toxicity in cultivar ‘LJCY’. The gene encoding CsMTP8, designated as CsMTP8 was also isolated, and its expression enhanced Mn tolerance in Saccharomyces cerevisiae. Similarly, the overexpression of CsMTP8 in Arabidopsis thaliana increased plant tolerance and reduced Mn accumulation in plant tissues under excess Mn conditions. Subcellular localization analysis of green florescence fused protein indicated that CsMTP8 was localized to the plasma membranes. Taken together, the results suggest that CsMTP8 is a Mn-specific transporter, which is localized in the plasma membrane, and transports excess Mn out of plant cells. The results also suggest that it is needed for Mn tolerance in shoots. PMID:28051151

  7. High-resolution imaging and in situ cutting of leaf epicuticular waxes of the biomass plant Miscanthus sinensis.

    PubMed

    Kim, Ki Woo

    2013-01-01

    The leaf surface features of the biomass plant Miscanthus sinensis were investigated by electron and ion beam microscopy. Fully developed leaves were collected from the plant, air-dried and examined by field emission scanning electron microscopy. Stomata and longitudinal stripes were present on both the adaxial and abaxial leaf surfaces. Longitudinally aggregated rodlets surrounded the stomata and formed hollow cylinders or chimney-like structures. With varying lengths up to ∼10 μm, the rodlets were curved (<1 μm in diameter) and almost occluded the stomata. As another type of epicuticular wax, platelets were observed in the vicinity of the stomata. The platelets were arranged into rather radially assembled clusters. As a novel approach for cross sectioning native epicuticular waxes, focused ion beam milling allowed precise in situ cutting of the stomata and epicuticular waxes. The longitudinally aggregated rodlets of the stomatal chimneys of M. sinensis were derived from the stomatal guard cells. These results suggest that the epicuticular waxes of M. sinensis can be categorized as stomatal chimneys (Strelitzia type) and rosettes (Fabales type). Combined electron and ion beam microscopy can help unravel the ultrastructure and vertical profile of epicuticular waxes in a range of plant taxa.

  8. The first detection of white spot syndrome virus in naturally infected cultured Chinese mitten crabs, Eriocheir sinensis in China.

    PubMed

    Ding, Zhengfeng; Yao, Yufeng; Zhang, Fengxiang; Wan, Jinjuan; Sun, Mengling; Liu, Hongyan; Zhou, Gang; Tang, Jianqing; Pan, Jianlin; Xue, Hui; Zhao, Ziming

    2015-08-01

    An epidemic with a high mortality rate (80-100%) recently occurred in the cultured Chinese mitten crab, Eriocheir sinensis, which is a very important economic crustacean species in China. Using negative stain, histopathology and nested PCR supplemented by sequencing we identified white spot syndrome virus (WSSV) in these crabs. Challenge experiments revealed that the disease was caused by WSSV and confirmed the crab's susceptibility to this virus, which was consistent with previous laboratory-based studies. A cumulative mortality of 100% was observed within 10 days post WSSV injection. This is the first report of WSSV-associated disease outbreaks in the Chinese mitten crab, which is normally reported as an important penaeid-shrimp viral pathogen. Furthermore, this is only the second report to describe a significant pathogen in pond-cultured E. sinensis. These results will enhance the early diagnosis of WSSV in the crab farms and help in monitoring efforts directed at determining the prevalence of the virus in E. sinensis.

  9. Agarwood Formation Induced by Fermentation Liquid of Lasiodiplodia theobromae, the Dominating Fungus in Wounded Wood of Aquilaria sinensis.

    PubMed

    Chen, Xuyu; Sui, Chun; Liu, Yangyang; Yang, Yun; Liu, Peiwei; Zhang, Zheng; Wei, Jianhe

    2017-04-01

    Agarwood is broadly used in incense and medicine. Traditionally, agarwood formation is induced by wounding the trunks and branches of some species of Aquilaria spp., including A. sinensis. As recently evidenced, some fungi or their fermentation liquid may have the potential of inducing agarwood formation. The present study aimed to analyze the fungi isolated from an agarwood-producing A. sinensis tree and subsequently identify the fungi capable of promoting agarwood formation. We identified a total of 110 fungi isolates based on their morphological characteristics and rDNA ITS sequences. These isolates came from four different layers (namely the decomposing layer, agarwood layer, transition layer, and normal layer) near the agarwood formation site of the trunk. According to the experimental results, most of them belonged to Dothideomycetes (81.82%), while the others to Sordariomycetes (13.64%) or Eurotiomycetes (4.55%). Of note, 88 isolates were shown belonging to the species of Lasiodiplodia theobromae that are most frequently isolated from different layers. In addition, when the fermentation liquid of two isolates of L. theobromae (AF4 and AF12) and one isolate of Fusarium solani (AF21) was inoculated into the A. sinensis wood using the Agar-Wit technique, promoted agarwood formation was observed; however, the effect of AF21 did not keep stable in the later test, while AF4 and AF12 still functioned 1 year later. This study may lay a foundation for exploring the underlying mechanism of agarwood formation as well as fungi application in agarwood production.

  10. [Cloning, prokaryotic expression, and functional identification of a sesquiterpene synthase gene (AsSS4) from Aquilaria sinensis].

    PubMed

    Liang, Liang; Guo, Qing-Mei; Zhang, Zheng; Xu, Yan-Hong; Han, Xiao-Min; Liu, Juan

    2014-12-01

    A sesquiterpene synthase (AsSS4) full-length open reading frame (ORF) cDNA was cloned from wounded stems of Aquilaria sinensis by RT-PCR method. The result showed that the ORF of AsSS4 was 1,698 bp encoding 565 amino acids. Prokaryotic expression vector pET28a-AsSS4 was constructed and transformed into E. coli BL21 (DE3) pLysS. Recombinant AsSS4 protein was obtained after induction by IPTG and SDS-PAGE analysis with a MW of 64 kD. Enzymatic reactions using farnesyl pyrophosphate showed that recombinant AsSS4 protein purified by Ni-agarose gel yielded five sesquiterpene compounds, cyclohexane, 1-ethenyl-1-methyl-2, 4-bis(1-methylethenyl)-, β-elemene, α-guaiene, α-caryophyllene and δ-guaiene. This paper reported the first cloning and functional characterization of AsSS4 gene from A. sinensis, which will establish a foundation for future studies on the molecular mechanisms of wound-induce agarwood formation in A. sinensis

  11. A Kazal-type serine proteinase inhibitor from Cyclina sinensis is involved in immune response and signal pathway initiation.

    PubMed

    Ren, Yipeng; Zhang, Hao; Pan, Baoping; Yan, Chuncai

    2015-11-01

    Serine protease inhibitors (SPIs) are an important group of protease inhibitors involved in a variety of biological processes. In the present study, a Kazal-type serine protease inhibitor homolog gene (designated as CsKPI) was identified from a Cyclina sinensis cDNA library. The open reading frame consists of 456 bp and encodes a protein of 151 amino acid residues with a theoretical molecular mass of 16.85 kDa and an isoelectric point of 5.74. Furthermore, using quantitative real-time PCR, we focused on the expression patterns of CsKPI found in tissues and on the stimulation of this gene's expression by bacteria. The results show that a higher-level mRNA expression of CsKPI was detected in hemocytes (P < 0.05) and was significantly upregulated at 3 h (P < 0.01) upon receiving bacterial challenges with Vibrio anguillarum. In addition, after the CsKPI gene was silenced by RNA interference, the expression of the CsTLR2 and CsMyD88 genes was extremely significantly decreased (P < 0.01) in C. sinensis. Finally, the recombinant CsKPI (rCsKPI) protein was purified and shown to exhibit less inhibitory activity than C-lyz against V. anguillarum in vitro. Hence, we propose that CsKPI plays an important role in the innate immunity and mediates TLR2 and MyD88-dependent pathway initiation in C. sinensis.

  12. Reduced testosterone production in TM3 Leydig cells treated with Aspalathus linearis (Rooibos) or Camellia sinensis (tea).

    PubMed

    Opuwari, C S; Monsees, T K

    2015-02-01

    Flavonoids are major compounds of Aspalathus linearis and Camellia sinensis. They are classified as endocrine disruptors and some have been shown to inhibit testosterone production. TM3 Leydig cell cultures were treated with 250-5000 μg mL(-1) A. linearis (unfermented or fermented rooibos) or Camellia sinensis (green or black tea) for 24 h in the absence or presence of 6 mIU/200 μl human chorionic gonadotropin (hCG). Under nonstimulated conditions, all teas tend to decrease testosterone production (3.9-31.8%). However, under hCG-stimulation, a significant reduction in testosterone production was observed at all concentrations by both rooibos and tea (16.3-37.9%). MTT assay and phase contrast microscopy, revealed that at 250-1000 μg ml(-1) , both plants maintained the viability, proliferation and morphology of the cells, while 5000 μg ml(-1) was cytotoxic to the cells (P < 0.05). In conclusion, the results here demonstrate the anti-androgenic property of A. linearis and C. sinensis.

  13. Toona Sinensis and Moschus Decoction Induced Cell Cycle Arrest in Human Cervical Carcinoma HeLa Cells

    PubMed Central

    Zhen, Hong; Zhang, Yifei; Fang, Zhijia; Huang, Zhiwei; Shi, Ping

    2014-01-01

    Toona sinensis and Moschus are two herb materials used in traditional Chinese medicine, most commonly for their various biological activities. In this study, we investigated the inhibitory effect of three decoctions from Toona sinensis, Moschus, and Toona sinensis and Moschus in combination on cell growth in several normal and cancer cell lines by cell viability assay. The results showed that the combined decoction exhibited the strongest anticancer effects, compared to two single decoctions. The observations indicated that the combined decoction did not induce cell apoptosis and autophagy in HeLa cells by fluorescence microscopy. Flow cytometry analysis revealed that the combined decoction arrested HeLa cell cycle progression in S-phase. After the decoction incubation, among 41 cell cycle related genes, eight were reduced, while five were increased in mRNA levels by real-time PCR assay. Western blotting showed that there were no apparent changes of protein levels of Cyclin E1, while P27 expression significantly declined and the levels of CDC7 and CDK7 obviously increased. The data suggest that the RB pathway is partially responsible for the decoction-induced S-phase cell cycle arrest in HeLa cells. Therefore, the combined decoction may have therapeutic potential as an anticancer formula for certain cancers. PMID:24511319

  14. Refugial isolation and range expansions drive the genetic structure of Oxyria sinensis (Polygonaceae) in the Himalaya-Hengduan Mountains.

    PubMed

    Meng, Lihua; Chen, Gang; Li, Zhonghu; Yang, Yongping; Wang, Zhengkun; Wang, Liuyang

    2015-05-27

    The formation of the Mekong-Salween Divide and climatic oscillations in Pleistocene were the main drivers for the contemporary diversity and genetic structure of plants in the Himalaya-Hengduan Mountains (HHM). To identify the relative roles of the two historical events in shaping population history of plants in HHM, we investigated the phylogeographic pattern of Oxyria sinensis, a perennial plant endemic to the HHM. Sixteen chloroplast haplotypes were identified and were clustered into three phylogenetic clades. The age of the major clades was estimated to be in the Pleistocene, falling into several Pleistocene glacial stages and postdating the formation of the Mekong-Salween Divide. Range expansions occurred at least twice in the early and middle Pleistocene, but the spatial genetic distribution rarely changed since the Last Glacial Maximum. Our results suggest that temporary mountain glaciers may act as barriers in promoting the lineage divergence in O. sinensis and that subsequential range expansions and secondary contacts might reshape the genetic distribution in geography and blur the boundary of population differentiation created in the earlier glacial stages. This study demonstrates that Pleistocene climatic change and mountain glaciers, rather than the Mekong-Salween Divide, play the primary role in shaping the spatial genetic structure of O. sinensis.

  15. Fast Simultaneous Determination of 13 Nucleosides and Nucleobases in Cordyceps sinensis by UHPLC-ESI-MS/MS.

    PubMed

    Zong, Shi-Yu; Han, Han; Wang, Bing; Li, Ning; Dong, Tina Ting-Xia; Zhang, Tong; Tsim, Karl W K

    2015-12-04

    A reliable ultra-high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UHPLC-ESI-MS/MS) method for the fast simultaneous determination of 13 nucleosides and nucleobases in Cordyceps sinensis (C. sinensis) with 2-chloroadenosine as internal standard was developed and validated. Samples were ultrasonically extracted in an ice bath thrice, and the optimum analyte separation was performed on an ACQUITY UPLC(TM) HSS C18 column (100 mm × 2.1 mm, 1.8 μm) with gradient elution. All targeted analytes were separated in 5.5 min. Furthermore, all calibration curves showed good linear regression (r > 0.9970) within the test ranges, and the limits of quantitation and detection of the 13 analytes were less than 150 and 75 ng/mL, respectively. The relative standard deviations (RSDs) of intra- and inter-day precisions were <6.23%. Recoveries of the quantified analytes ranged within 85.3%-117.3%, with RSD < 6.18%. The developed UHPLC-ESI-MS/MS method was successfully applied to determine nucleosides and nucleobases in 11 batches of C. sinensis samples from different regions in China. The range for the total content in the analyzed samples was 1329-2057 µg/g.

  16. Highly Selective Bioconversion of Ginsenoside Rb1 to Compound K by the Mycelium of Cordyceps sinensis under Optimized Conditions.

    PubMed

    Wang, Wei-Nan; Yan, Bing-Xiong; Xu, Wen-Di; Qiu, Ye; Guo, Yun-Long; Qiu, Zhi-Dong

    2015-10-23

    Compound K (CK), a highly active and bioavailable derivative obtained from protopanaxadiol ginsenosides, displays a wide variety of pharmacological properties, especially antitumor activity. However, the inadequacy of natural sources limits its application in the pharmaceutical industry. In this study, we firstly discovered that Cordyceps sinensis was a potent biocatalyst for the biotransformation of ginsenoside Rb1 into CK. After a series of investigations on the biotransformation parameters, an optimal composition of the biotransformation culture was found to be lactose, soybean powder and MgSO₄ without controlling the pH. Also, an optimum temperature of 30 °C for the biotransformation process was suggested in a range of 25 °C-50 °C. Then, a biotransformation pathway of Rb1→Rd→F2→CK was established using high performance liquid chromatography/quadrupole time-of-flight mass spectrometry (HPLC-Q-TOF-MS). Our results demonstrated that the molar bioconversion rate of Rb1 to CK was more than 82% and the purity of CK produced by C. sinensis under the optimized conditions was more than 91%. In conclusion, the combination of C. sinensis and the optimized conditions is applicable for the industrial preparation of CK for medicinal purposes.

  17. Refugial isolation and range expansions drive the genetic structure of Oxyria sinensis (Polygonaceae) in the Himalaya-Hengduan Mountains

    PubMed Central

    Meng, Lihua; Chen, Gang; Li, Zhonghu; Yang, Yongping; Wang, Zhengkun; Wang, Liuyang

    2015-01-01

    The formation of the Mekong-Salween Divide and climatic oscillations in Pleistocene were the main drivers for the contemporary diversity and genetic structure of plants in the Himalaya-Hengduan Mountains (HHM). To identify the relative roles of the two historical events in shaping population history of plants in HHM, we investigated the phylogeographic pattern of Oxyria sinensis, a perennial plant endemic to the HHM. Sixteen chloroplast haplotypes were identified and were clustered into three phylogenetic clades. The age of the major clades was estimated to be in the Pleistocene, falling into several Pleistocene glacial stages and postdating the formation of the Mekong-Salween Divide. Range expansions occurred at least twice in the early and middle Pleistocene, but the spatial genetic distribution rarely changed since the Last Glacial Maximum. Our results suggest that temporary mountain glaciers may act as barriers in promoting the lineage divergence in O. sinensis and that subsequential range expansions and secondary contacts might reshape the genetic distribution in geography and blur the boundary of population differentiation created in the earlier glacial stages. This study demonstrates that Pleistocene climatic change and mountain glaciers, rather than the Mekong-Salween Divide, play the primary role in shaping the spatial genetic structure of O. sinensis. PMID:26013161

  18. Genetic Divergence between Camellia sinensis and Its Wild Relatives Revealed via Genome-Wide SNPs from RAD Sequencing

    PubMed Central

    Liu, Hong-Wei; Wu, Jun-Lan; Li, Zheng-Guo; Zhang, Liang; Jian, Jian-Bo; Li, Ye-Yun; Tai, Yu-Ling; Zhang, Jing; Zhang, Zheng-Zhu; Jiang, Chang-Jun; Xia, Tao; Wan, Xiao-Chun

    2016-01-01

    Tea is one of the most popular beverages across the world and is made exclusively from cultivars of Camellia sinensis. Many wild relatives of the genus Camellia that are closely related to C. sinensis are native to Southwest China. In this study, we first identified the distinct genetic divergence between C. sinensis and its wild relatives and provided a glimpse into the artificial selection of tea plants at a genome-wide level by analyzing 15,444 genomic SNPs that were identified from 18 cultivated and wild tea accessions using a high-throughput genome-wide restriction site-associated DNA sequencing (RAD-Seq) approach. Six distinct clusters were detected by phylogeny inferrence and principal component and genetic structural analyses, and these clusters corresponded to six Camellia species/varieties. Genetic divergence apparently indicated that C. taliensis var. bangwei is a semi-wild or transient landrace occupying a phylogenetic position between those wild and cultivated tea plants. Cultivated accessions exhibited greater heterozygosity than wild accessions, with the exception of C. taliensis var. bangwei. Thirteen genes with non-synonymous SNPs exhibited strong selective signals that were suggestive of putative artificial selective footprints for tea plants during domestication. The genome-wide SNPs provide a fundamental data resource for assessing genetic relationships, characterizing complex traits, comparing heterozygosity and analyzing putatitve artificial selection in tea plants. PMID:26962860

  19. Mechanisms Involved in Toxicity of Liver Caused by Piroxicam in Mice and Protective Effects of Leaf Extract of Hibiscus rosa-sinensis L.

    PubMed Central

    Sahu, C. R.

    2016-01-01

    Piroxicam is one of the important therapeutic nonsteroidal anti-inflammatory class of drugs used mainly to suppress pain and inflammation in arthritis and other musculoskeletal disorders. Besides being anti-inflammatory, these drugs are analgesic and antipyretic often used for the relief of nonspecific fever condition. Recently, piroxicam has also gained attention as an effective therapy for tumors, colorectal, and invasive bladder cancers. The objective of the current study is to evaluate the protective effects of the alcoholic leaf extract of Hibiscus rosa-sinensis (AEH), Malvaceae, against piroxicam-induced toxicity in mice. Sixty adult Swiss albino mice (Mus musculus) were divided into four groups (n = 10), which included a control group, a group treated orally with AEH (30 mg kg−1 b.w.) for 15 days, a group treated orally with piroxicam (6.6 mg kg−1 b.w.) for 15 days, and another group treated orally with piroxicam and AEH for 15 days. The results indicated that treatment with piroxicam alone resulted in a significant increase in the activities of serum marker enzymes, namely, aspartate transaminase, alanine transaminase, and alkaline phosphatase with profound hepatic lipid peroxidation as evidenced by a marked increment in the level of thoibarbituric acid reactive substances along with a distinct diminution in reduced glutathoine content and various antioxidant enzymes such as superoxide dismutase, catalase, and glutathione peroxidase in the liver. However, treatment with AEH during piroxicam treatment retrieved or partially antagonized the effects induced by piroxicam toward the normal values of controls. Histopathological observations also corroborate with the above findings. It can be concluded that AEH exhibited a protective action against piroxicam toxicity and effective in combating oxidative stress-induced hepatic damage. PMID:26819562

  20. Temperature- and Relative Humidity-Dependent Life History Traits of Phenacoccus solenopsis (Hemiptera: Pseudococcidae) on Hibiscus rosa-sinensis (Malvales: Malvaceae).

    PubMed

    Chen, H S; Yang, L; Huang, L F; Wang, W L; Hu, Y; Jiang, J J; Zhou, Z S

    2015-08-01

    Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae), a worldwide distributive invasive pest, originated from the United States, and it was first reported in Guangdong province, China, in 2008. The effects of temperature and relative humidity (RH) on the life history traits of P. solenopsis on Hibiscus rosa-sinensis L. (Malvales: Malvaceae) were studied at seven constant temperatures (15, 20, 25, 27.5, 30, 32.5, and 35°C) and three RHs (45, 60, and 75%). The results showed that temperature, RH, and their interactions significantly influenced the life history traits of P. solenopsis. First instar was the most sensitive stage to extreme temperatures with very low survival rates at 15 and 35°C. At 25-32.5°C and the three RHs, the developmental periods of entire immature stage were shorter with values between 12.5-18.6 d. The minimum threshold temperature and the effective accumulative temperature for the pest to complete one generation were 13.2°C and 393.7 degree-days, respectively. The percentage and longevity of female adults significantly differed among different treatments. It failed to complete development at 15 or 35°C and the three RHs. Female fecundity reached the maximum value at 27.5°C and 45% RH. The intrinsic rate for increase (r), the net reproductive rate (R0), and the finite rate of increase (λ) reached the maximum values at 27.5°C and 45% RH (0.22 d(-1), 244.6 hatched eggs, and 1.25 d(-1), respectively). Therefore, we conclude that 27.5°C and 45% RH are the optimum conditions for the population development of the pest.

  1. Organochlorines and mercury in livers of great cormorants (Phalacrocorax carbo sinensis) wintering in northeastern Mediterranean wetlands in relation to area, bird age, and gender.

    PubMed

    Goutner, V; Becker, P H; Liordos, V

    2011-01-15

    Wild birds are exposed to pollutants in their habitats. Top consumers of aquatic environments such as the fish-eating great cormorant (Phalacrocorax carbo sinensis) are especially affected due to the bioaccumulation of toxic substances in their tissues. This study analysed the livers of 80 great cormorants from Greece to estimate the concentration of organochlorines and mercury and to examine their possible toxic effects and origin. The results showed that mercury (geometric mean 8089 ng g⁻¹ dw), p,p'-DDE (2628 ng g⁻¹ dw), ∑HCHs (47 ng g⁻¹ dw) and HCB (116 ng g⁻¹ dw) concentrations can be considered high compared with those found in great cormorant livers elsewhere except in highly polluted areas, whereas ∑PCBs occurred in relatively low concentrations (1091 ng g⁻¹ dw). β-HCH was the dominant HCH isomer. Pollutant levels were generally unrelated to area, age and gender. However, p,p'-DDE and p,p'-DDD showed intersite differences, whilst the proportion of PCBs with 8 chlorine atoms were significantly higher in adult than 1st year great cormorants. Pollution did not reflect local patterns but rather these along the Baltic and Black Seas, whilst differences in p,p'-DDE concentration and ∑DDTs/∑PCBs ratios between Evros, Axios or Amvrakikos, found on common migration route, suggested different bird origins. Most birds had toxic mercury concentrations; 83.7% above 4000 ng g⁻¹ dw and 16% above 17,000 ng g⁻¹ dw. Other pollutant levels were too low to have adverse effects.

  2. Mechanisms Involved in Toxicity of Liver Caused by Piroxicam in Mice and Protective Effects of Leaf Extract of Hibiscus rosa-sinensis L.

    PubMed

    Sahu, C R

    2016-01-01

    Piroxicam is one of the important therapeutic nonsteroidal anti-inflammatory class of drugs used mainly to suppress pain and inflammation in arthritis and other musculoskeletal disorders. Besides being anti-inflammatory, these drugs are analgesic and antipyretic often used for the relief of nonspecific fever condition. Recently, piroxicam has also gained attention as an effective therapy for tumors, colorectal, and invasive bladder cancers. The objective of the current study is to evaluate the protective effects of the alcoholic leaf extract of Hibiscus rosa-sinensis (AEH), Malvaceae, against piroxicam-induced toxicity in mice. Sixty adult Swiss albino mice (Mus musculus) were divided into four groups (n = 10), which included a control group, a group treated orally with AEH (30 mg kg(-1) b.w.) for 15 days, a group treated orally with piroxicam (6.6 mg kg(-1) b.w.) for 15 days, and another group treated orally with piroxicam and AEH for 15 days. The results indicated that treatment with piroxicam alone resulted in a significant increase in the activities of serum marker enzymes, namely, aspartate transaminase, alanine transaminase, and alkaline phosphatase with profound hepatic lipid peroxidation as evidenced by a marked increment in the level of thoibarbituric acid reactive substances along with a distinct diminution in reduced glutathoine content and various antioxidant enzymes such as superoxide dismutase, catalase, and glutathione peroxidase in the liver. However, treatment with AEH during piroxicam treatment retrieved or partially antagonized the effects induced by piroxicam toward the normal values of controls. Histopathological observations also corroborate with the above findings. It can be concluded that AEH exhibited a protective action against piroxicam toxicity and effective in combating oxidative stress-induced hepatic damage.

  3. Expression patterns of two heat-shock cognate 70 genes during immune responses and larval development of the Chinese mitten crab Eriocheir sinensis.

    PubMed

    Li, P; Jiang, X F; Guo, W B; Yan, J; Zhou, K Y

    2016-09-16

    Two heat-shock protein (HSP) 70 family transcripts, heat-shock protein 70 cognate 5 and heat-shock protein 70 cognate 3 (designated as EsHSC70-5 and EsHSC70-3, respectively), were isolated from the Chinese mitten crab Eriocheir sinensis and their expression profiles were evaluated for their responsiveness to larval development and immune challenge in adult crabs. The HSPs exhibited 45-89% identity with other heat-shock proteins, and they shared similar structural features. EsHSC70 mRNA expression was detected not only during infection but also during the developmental larval stages. The EsHSC70s were enriched, and their expression fluctuated during early development. EsHSC70 mRNA expression was significantly induced by Vibrio parahaemolyticus challenge in all of the tissues studied (P < 0.05). Expression of EsHSC70 mRNA in the hepatopancreas and at the early zoeal stages was particularly pronounced, and the two EsHSC70s exhibited differential expression patterns both chronologically and spatially. The EsHSC70-5 mRNA level was significantly downregulated in the intestine and gills compared to that in controls at nearly all time points, and was expressed at a lower level after the bacterial challenge, indicating that EsHSC70-5 and EsHSC70-3 respond to immune challenges. The stage-specific enrichment of EsHSC70 transcripts in crabs suggests that these stress proteins play an essential role during brachyurization events.

  4. Effect of green tea (camellia sinensis l.) leaf extract on reproductive system of adult male albino rats

    PubMed Central

    Das, Shyamal Kanti; Karmakar, Soumendra Nath

    2015-01-01

    Green tea leaf extract (GTLE), used in this experiment has shown great influence on male reproductive system functionally as well as morphologically. The extract was prepared according to the method of Wei. H. et al. The extract was given to two different experimental animal groups with two different doses during 26 consecutive days. After treatment it was found that, the weight of the testis was markedly reduced instead of normal weight gain of all the animals. The sperm count and motility were reduced for the treated groups as compared with control animal group. The enzymes like SGPT and SGOT were as usual and other blood parameters like glucose and protein were also as usual comparing with controlled group. Testosterone level was reduced in the treated groups. FSH and LH levels were also altered accordingly in treated groups. Histological examination showed inhibition of spermatogenesis as evidenced by disintegration of seminiferous tubules of testis. Result of this study showed that GTLE has potent castrative effect on male reproductive system in dose dependent manner. PMID:27073594

  5. Effect of green tea (camellia sinensis l.) leaf extract on reproductive system of adult male albino rats.

    PubMed

    Das, Shyamal Kanti; Karmakar, Soumendra Nath

    2015-01-01

    Green tea leaf extract (GTLE), used in this experiment has shown great influence on male reproductive system functionally as well as morphologically. The extract was prepared according to the method of Wei. H. et al. The extract was given to two different experimental animal groups with two different doses during 26 consecutive days. After treatment it was found that, the weight of the testis was markedly reduced instead of normal weight gain of all the animals. The sperm count and motility were reduced for the treated groups as compared with control animal group. The enzymes like SGPT and SGOT were as usual and other blood parameters like glucose and protein were also as usual comparing with controlled group. Testosterone level was reduced in the treated groups. FSH and LH levels were also altered accordingly in treated groups. Histological examination showed inhibition of spermatogenesis as evidenced by disintegration of seminiferous tubules of testis. Result of this study showed that GTLE has potent castrative effect on male reproductive system in dose dependent manner.

  6. Genome-wide comparative analysis reveals similar types of NBS genes in hybrid Citrus sinensis genome and original Citrus clementine genome and provides new insights into non-TIR NBS genes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In this study, we identified and compared nucleotide-binding site (NBS) domain-containing genes from three Citrus genomes (C. clementina, C. sinensis from USA and C. sinensis from China). Phylogenetic analysis of all Citrus NBS genes across these three genomes revealed that there are three approxima...

  7. Development of expressed sequence tag-based microsatellite markers for the critically endangered Isoëtes sinensis (Isoetaceae) based on transcriptome analysis.

    PubMed

    Gichira, A W; Long, Z C; Wang, Q F; Chen, J M; Liao, K

    2016-07-15

    Isoëtes sinensis is a critically endangered quillwort. To facilitate studies on the conservation genetics of this species, we developed expressed sequence tag-simple sequence repeat (EST-SSR) markers. A total of 50,063 unigenes were predicted by transcriptome sequencing, 5294 (10.6%) of which significantly matched 3011 Gene Ontology annotations and 2363 were assigned to Kyoto Encyclopedia of Genes and Genomes metabolic pathways. Most of these (2297) were involved in metabolism. A total of 1982 SSR motifs were identified, with trinucleotides being the dominant repeat motif, and 1438 (72.6%) SSR primers were designed. Eighteen randomly selected primer pairs were used to genotype 24 I. sinensis accessions, which confirmed the suitability of these novel markers for molecular studies of I. sinensis. The heterozygosity index value ranged between 0.0799 and 0.9106, while the Shannon-Wiener diversity index value ranged between 0.1732 and 2.5589. The EST-SSRs reported in this study are linked to genic sequences, and are therefore ideal for investigating the evolutionary history of I. sinensis. These markers, together with the large EST dataset generated in this study, will greatly facilitate conservation genetic studies of I. sinensis.

  8. De Novo Characterization of Flower Bud Transcriptomes and the Development of EST-SSR Markers for the Endangered Tree Tapiscia sinensis

    PubMed Central

    Zhou, Xiao-Jun; Wang, Yue-Yue; Xu, Ya-Nan; Yan, Rong-Shan; Zhao, Peng; Liu, Wen-Zhe

    2015-01-01

    Tapiscia sinensis Oliv (Tapisciaceae) is an endangered species native to China famous for its androdioecious breeding system. However, there is a lack of genomic and transcriptome data on this species. In this study, the Tapiscia sinensis transcriptomes from two types of sex flower buds were sequenced. A total of 97,431,176 clean reads were assembled into 52,169 unigenes with an average length of 1116 bp. Through similarity comparison with known protein databases, 36,662 unigenes (70.27%) were annotated. A total of 10,002 (19.17%) unigenes were assigned to 124 pathways using the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway database. Additionally, 10,371 simple sequence repeats (SSRs) were identified in 8608 unigenes, with 16,317 pairs of primers designed for applications. 150 pairs of primers were chosen for further validation, and the 68 pairs (45.5%) were able to produce clear polymorphic bands. Six polymorphic SSR markers were used to Bayesian clustering analysis of 51 T. sinensis individuals. This is the first report to provide transcriptome information and to develop large-scale SSR molecular markers for T. sinensis. This study provides a valuable resource for conservation genetics and functional genomics research on T. sinensis for future work. PMID:26057749

  9. Production of intracellular selenium-enriched polysaccharides from thin stillage by Cordyceps sinensis and its bioactivities

    PubMed Central

    Yang, Shengli; Zhang, Hui

    2016-01-01

    Background Thin stillage was used as the substrate to produce intracellular selenium-enriched polysaccharides (ISPS) from Cordyceps sinensis to increase the value of agricultural coproducts. Methods Fermentation parameters were optimized using response surface methodology (RSM) to improve the production of ISPS. Then, the effects of ISPS on the antioxidant activities in vitro, as well as the glycosylated serum protein concentration, malondialdehyde level, and total antioxidant capacity of streptozotocin-induced diabetic rats were studied. Results The optimized conditions were as follows: sodium selenite concentration, 33.78 µg/L; incubation time, 8.24 days; and incubation temperature, 26.69°C. A maximum yield of 197.35 mg/g ISPS was obtained from the validation experiments, which was quite close to the predicted maximum yield of 198.6839 mg/g. FT-IR spectra indicated that ISPS has been successfully selenylation modified with similar structure to polysaccharide of intracellular polysaccharides. The in vitro scavenging effects of 1.0 mg/mL ISPS on hydroxyl, superoxide, and 1,1-diphenyl-2-picrylhydrazyl radicals were 74.62±4.05, 71.45±3.63, and 79.48±4.75%, respectively. The reducing power of ISPS was 0.45±0.01 (absorbance at 700 nm). Fasting blood glucose and glycosylated serum protein of group C (rats with diabetes that received drinking water with ISPS) were significantly lower than those of group B (rats with diabetes) (P<0.01) after treatment was administered for 2 and 4 weeks. Serum malonaldehyde content of group C was significantly lower than that of group B at 4 weeks (P<0.01). At 4 weeks, malonaldehyde contents in heart, liver, and kidney tissues of group C were significantly lower than those of group B; however, malonaldehyde content in pancreas tissue of group C was not significantly different. Total antioxidant capacities in liver, pancreas and kidney tissues of group C were significantly higher than those of group B, but total antioxidant capacity

  10. Phylogeographic analysis reveals significant spatial genetic structure of Incarvillea sinensis as a product of mountain building

    PubMed Central

    2012-01-01

    Background Incarvillea sinensis is widely distributed from Southwest China to Northeast China and in the Russian Far East. The distribution of this species was thought to be influenced by the uplift of the Qinghai-Tibet Plateau and Quaternary glaciation. To reveal the imprints of geological events on the spatial genetic structure of Incarvillea sinensis, we examined two cpDNA segments ( trnH- psbA and trnS- trnfM) in 705 individuals from 47 localities. Results A total of 16 haplotypes was identified, and significant genetic differentiation was revealed (GST =0.843, NST = 0.975, P < 0.05). The survey detected two highly divergent cpDNA lineages connected by a deep gap with allopatric distributions: the southern lineage with higher genetic diversity and differentiation in the eastern Qinghai-Tibet Plateau, and the northern lineage in the region outside the Qinghai-Tibet Plateau. The divergence between these two lineages was estimated at 4.4 MYA. A correlation between the genetic and the geographic distances indicates that genetic drift was more influential than gene flow in the northern clade with lower diversity and divergence. However, a scenario of regional equilibrium between gene flow and drift was shown for the southern clade. The feature of spatial distribution of the genetic diversity of the southern lineage possibly indicated that allopatric fragmentation was dominant in the collections from the eastern Qinghai-Tibet Plateau. Conclusions The results revealed that the uplift of the Qinghai-Tibet Plateau likely resulted in the significant divergence between the lineage in the eastern Qinghai-Tibet Plateau and the other one outside this area. The diverse niches in the eastern Qinghai-Tibet Plateau created a wide spectrum of habitats to accumulate and accommodate new mutations. The features of genetic diversity of populations outside the eastern Qinghai-Tibet Plateau seemed to reveal the imprints of extinction during the Glacial and the interglacial and

  11. Successional development from stands of Miscanthus sinensis to stands of Pinus densiflora and elements of microclimates: The seed germination and seedling establishment conditions of P. densiflora

    NASA Astrophysics Data System (ADS)

    Kashiwagi, Y.

    1991-09-01

    The succession mechanism from stands of Miscanthus sinensis to stands of Pinus densiflora was studied in relation to elements of microclimates within those plant communities. Laboratory experiments indicated that the seed germination of P. densiflora occurred at cumulative soil temperature between 75 °C ṡ day and 200 °C ṡ day. It is assumed from field research that establishment of P. densiflora seedlings is impossible if the effective cumulative air temperature at a height of 6 cm is less than 2,000 °C day, and the cumulative solar radiation figure totals less than 633 MJ/m2. The seedlings thrived in a M. sinensis stand growing on gently convex topography, which fulfills the requisites mentioned above. The seedlings in a M. sinensis stand growing on gently concave topography and in a P. densiflora stand died completely by October.

  12. Biosynthesis of catechin components is differentially regulated in dark-treated tea (Camellia sinensis L.).

    PubMed

    Hong, Gaojie; Wang, Jie; Zhang, Yong; Hochstetter, Danielle; Zhang, Shuping; Pan, Yue; Shi, Yunlong; Xu, Ping; Wang, Yuefei

    2014-05-01

    Tea (Camellia sinensis L.) is a crop with both commercial and medicinal value with remarkably high polyphenol content in the form of catechins. To understand the molecular regulation of catechin biosynthesis in tea, we treated the tea plants with darkness. We used qRT-PCR to validate the expression of genes involved in catechin biosynthesis. It indicated that dark treatment displayed different effects on the genes participating in tea flavonoid (FL) pathway. The early genes of FL biosynthesis pathway, CHSI, F3H and DFR, remained at steady expression levels when treated by darkness. It is noteworthy that the expression level of LAR increased and the level of ANS decreased under dark conditions. The vanillin assay showed that the dark-treated plants contained lower levels of total catechins than those grown under normal conditions. The HPLC analysis further demonstrated the changes in biosynthesis of catechins under these conditions. In accordance with the gene expression pattern, the content of epicatechins (ECs) declined and that of catechins (Cs) was elevated in response to the darkness. Our study uncovered the molecular mechanisms and biochemical changes of shading in tea cultivation.

  13. The galloyl catechins contributing to main antioxidant capacity of tea made from Camellia sinensis in China.

    PubMed

    Zhao, Chunjian; Li, Chunying; Liu, Shuaihua; Yang, Lei

    2014-01-01

    Total polyphenol content, catechins content, and antioxidant capacities of green, dark, oolong, and black teas made from Camellia sinensis in China were evaluated. The total polyphenol content of 20 samples of tea was in the range of 7.82-32.36%. Total catechins content was in the range of 4.34-24.27%. The antioxidant capacity of tea extract was determined by the oxygen radical absorbance capacity (ORAC) test and the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging test. Total polyphenol content, catechins content, and antioxidant capacity decreased in the following order: green > oolong > black > dark tea. A positive correlation existed between the antioxidant capacity and total polyphenol content or catechins content (R (2) = 0.67-0.87). The antioxidant capacities of five major catechins (epigallocatechin gallate (EGCG), epicatechin gallate (ECG), epicatechin, epigallocatechin, and catechin) were determined by online HPLC DPPH radical-scavenging; the antioxidant activity of tea was mainly attributed to the esterified catechins (EGCG or ECG).

  14. Quantification of flavonol glycosides in Camellia sinensis by MRM mode of UPLC-QQQ-MS/MS.

    PubMed

    Wu, Yahui; Jiang, Xiaolan; Zhang, Shuxiang; Dai, Xinlong; Liu, Yajun; Tan, Huarong; Gao, Liping; Xia, Tao

    2016-04-01

    Phenolic compounds are major components of tea flavour, in which catechins and flavonol glycosides play important roles in the astringent taste of tea infusion. However, the flavonol glycosides are difficult to quantify because of the large variety, as well as the inefficient seperation on chromatography. In this paper, a total of 15 flavonol glycosides in the tea plant (Camellia sinensis) were identified by the high performance liquid chromatography (HPLC) coupled to a time-of-flight mass spectrometer (TOF-MS), and a quantitative method was established based on multiple reaction monitoring (MRM) mode of ultra-high performance liquid chromatography (UPLC) coupled to a triple quadrupole mass spectrometer (QQQ-MS/MS). It provided the limit of detection and quantification to the order of picogram, which was more sensitive than the HPLC detection of the order of nanogram. The relative standard deviations of the intra- and inter-day variations in retention time and signal intensity (peak area) of six analytes were less than 0.26% and 4%, respectively. The flavonol glycosides of four tea cultivars were relatively quantified using the signal intensity (peak area) of product ion, in which six flavonol glycosides were quantified by the authentic standards. The results showed that the flavonol mono-, di- and tri-glycoside mostly accumulated in young leaves of the four tea cultivars. Notably, the myricetin 3-O-galactoside was the major component among the six flavonol glycosides detected.

  15. Regulation of formation of volatile compounds of tea (Camellia sinensis) leaves by single light wavelength.

    PubMed

    Fu, Xiumin; Chen, Yiyong; Mei, Xin; Katsuno, Tsuyoshi; Kobayashi, Eiji; Dong, Fang; Watanabe, Naoharu; Yang, Ziyin

    2015-11-16

    Regulation of plant growth and development by light wavelength has been extensively studied. Less attention has been paid to effect of light wavelength on formation of plant metabolites. The objective of this study was to investigate whether formation of volatiles in preharvest and postharvest tea (Camellia sinensis) leaves can be regulated by light wavelength. In the present study, in contrast to the natural light or dark treatment, blue light (470 nm) and red light (660 nm) significantly increased most endogenous volatiles including volatile fatty acid derivatives (VFADs), volatile phenylpropanoids/benzenoids (VPBs), and volatile terpenes (VTs) in the preharvest tea leaves. Furthermore, blue and red lights significantly up-regulated the expression levels of 9/13-lipoxygenases involved in VFADs formation, phenylalanine ammonialyase involved in VPBs formation, and terpene synthases involved in VTs formation. Single light wavelength had less remarkable influences on formation of volatiles in the postharvest leaves compared with the preharvest leaves. These results suggest that blue and red lights can be promising technology for remodeling the aroma of preharvest tea leaves. Furthermore, our study provided evidence that light wavelength can activate the expression of key genes involved in formation of plant volatiles for the first time.

  16. Enhancement of Gleditsia sinensis gum rheological properties with pressure cell treatment in semi-solid state.

    PubMed

    Zhou, Zi-yuan; Zhang, Wei-an; Duan, Jiu-fang; Zhang, Wei-ming; Sun, Da-feng; Jiang, Jian-xin

    2016-03-01

    The apparent viscosity, molecular weight, and molecular weight distribution are important physical properties that determine the functional properties of galactomannan gum. Gleditsia sinensis gum (GSG) in semi-solid state was pressure cell treated over a range of temperature (30-110 °C) under nitrogen maintained at a pressure of 1.0-4.0 MPa. Physicochemical properties of GSG samples both before and after the pressure cell treatment were characterized. These include measurements of rheological properties by LVDV-III Ultra Rheometer, molecular weight and radius of gyration by light scattering, and changes in surface morphology by scanning electron microscopy. GSG had the highest apparent viscosity at a treatment temperature of 30 °C; further increase in temperature led to decrease in apparent viscosity. The apparent viscosity of GSG can be efficiently improved at room temperature and low pressure. The process of pressure cell treatment of GSG in semi-solid state could be industrialized for enhancement of rheological properties of galactomannan gum.

  17. Incubation temperature affects the immune function of hatchling soft-shelled turtles, Pelodiscus sinensis

    PubMed Central

    Dang, Wei; Zhang, Wen; Du, Wei-Guo

    2015-01-01

    Identifying how developmental temperature affects the immune system is critical for understanding how ectothermic animals defend against pathogens and their fitness in the changing world. However, reptiles have received little attention regarding this issue. We incubated eggs at three ecologically relevant temperatures to determine how incubation temperature affects the immune function of hatchling soft-shelled turtles, Pelodiscus sinensis. When exposed to bacterial infections, hatchlings from 24 °C had lower cumulative mortalities (55%, therefore, higher immunocompetence) than those from 28 °C (85%) or 32 °C (100%). Consistent with higher immunocompetence, hatchlings from low incubation temperature had higher IgM, IgD, and CD3γ expressions than their counterparts from the other two higher incubation temperatures. Conversely, the activity of immunity-related enzymes did not match the among-temperature difference in immune function. Specifically, enzyme activity was higher at intermediate temperatures (alkaline phosphatase) or was not affected by incubation temperature (acid phosphatase, lysozyme). Our study is the first to provide unequivocal evidence (at the molecular and organismal level) about the significant effect of incubation temperature on offspring immunity in reptiles. Our results also indicate that the reduced immunity induced by high developmental temperatures might increase the vulnerability of reptiles to the outbreak of diseases under global warming scenarios. PMID:26028216

  18. Effect of salt treatment on theanine biosynthesis in Camellia sinensis seedlings.

    PubMed

    Deng, Wei-Wei; Wang, Shuo; Chen, Qi; Zhang, Zheng-Zhu; Hu, Xiang-Yang

    2012-07-01

    Theanine synthetase (TS) is an enzyme involved in theanine biosynthesis in tea plants. Recent studies have revealed that theanine biosynthesis, derived from nitrogen metabolism in tea (Camellia sinensis L.) plants, could be influenced by salt treatment. We have characterized CsTS at the molecular and biochemical level. The expression pattern of CsTS protein was examined by western blot using a self-prepared polyclonal antibody with high specificity and sensitivity. The effect of salt treatment on the levels of theanine synthesis was investigated in this study. Levels of theanine and the total free amino acids were gradually increased in shoots, and reached the maximum on the 8th day after treatment (DAT). The immunoblotting analysis suggested the accumulation of CsTS protein had increased gently up to 8 DAT, and subsequently declined, both in roots and shoots, which is one of the main evidences that resulted in the variation of theanine concentration under salt treatment. Together, these data revealed that theanine synthesis takes place both in root and shoot and CsTS accumulation is positively affected by salt treatment.

  19. The in vivo antioxidant and antifibrotic properties of green tea (Camellia sinensis, Theaceae).

    PubMed

    Tsai, Chia-Fang; Hsu, Yu-Wen; Ting, Hung-Chih; Huang, Chun-Fa; Yen, Cheng-Chieh

    2013-02-15

    The in vivo antioxidant and antifibrotic properties of green tea (Camellia sinensis, Theaceae) were investigated with a study of carbon tetrachloride (CCl(4))-induced oxidative stress and hepatic fibrosis in male ICR mice. Oral administration of green tea extract at doses of 125, 625 and 1250 mg/kg for 8 weeks significantly reduced (p<0.05) the levels of thiobarbituric acid-reactive substances (TBARS) and protein carbonyls in the liver by at least 28% compared with that was induced by CCl(4) (1 mL/kg) in mice. Moreover, green tea extract administration significantly increased (p<0.05) the activities of catalase, glutathione peroxidase (GSH-Px) and glutathione reductase (GSH-Rd) in the liver. Our study found that oral administration of green tea extract prevented CCl(4)-induced hepatic fibrosis, as evidenced by a decreased hydroxyproline level in the liver and a reduced incidence of hepatic fibrosis by histological observations. These results indicate that green tea exhibits potent protective effects against CCl(4)-induced oxidative stress and hepatic fibrosis in mice by inhibiting oxidative damage and increasing antioxidant enzyme activities.

  20. Sulfation and enhanced antioxidant capacity of an exopolysaccharide produced by the medicinal fungus Cordyceps sinensis.

    PubMed

    Yan, Jing-Kun; Wang, Wen-Qiang; Ma, Hai-Le; Wu, Jian-Yong

    2012-12-24

    EPS-1 was an exopolysaccharide produced by the medicinal fungus Cordyceps sinensis (Cs-HK1). In the present study, EPS-1 was sulfated with chlorosulfonic acid (CSA)-pyridine (Pyr) at different volume ratios, yielding four sulfated derivatives, SEPS-1A, B, C and D, with different degrees of substitution (DS: 0.25-1.38) and molecular weights (17.1-4.1 kDa). The sulfation of EPS-1 occurred most frequently at the C-6 hydroxyl groups due to their higher reactivity. In aqueous solution, the native EPS-1 formed random coils or aggregated networks, but the sulfated derivatives formed single helices. The antioxidant activities of the sulfated EPS-1 derivatives for scavenging hydroxyl radicals (•OH) and 2,2-azinobis-3-ehtylbenzothiazolin-6-sulfonic acid radicals (ABTS•+) were significantly increased with increasing DS and decreasing molecular weight (MW). Sulfation has thus been shown to be an effective and favorable strategy for improving the physico-chemical properties and bioactivities of fungal polysaccharides.

  1. Effects of temperature acclimation on body mass and energy budget in the Chinese bulbul Pycnonotus sinensis

    PubMed Central

    WU, Yu-Nan; Lin, Lin; XIAO, Yu-Chao; Zhou, Li-Meng; WU, Meng-Si; Zhang, Hui-Ying; LIU, Jin-Song

    2014-01-01

    Chinese bulbuls (Pycnonotus sinensis) are small passerine birds that inhabit areas of central, southern and eastern China. Previous observations suggest that free-living individuals of this species may change their food intake in response to seasonal changes in ambient temperature. In the present study, we randomly assigned Chinese bulbuls to either a 30 ℃ or 10 ℃ group, and measured their body mass (BM), body temperature, gross energy intake (GEI), digestible energy intake (DEI), and the length and mass of their digestive tracts over 28 days of acclimation at these temperatures. As predicted, birds in the 30 ℃ group had lower body mass, GEI and DEI relative to those in the 10 ℃ group. The length and mass of the digestive tract was also lower in the 30 ℃ group and trends in these parameters were positively correlated with BM, GEI and DEI. These results suggest that Chinese bulbuls reduced their absolute energy demands at relatively high temperatures by decreasing their body mass, GEI and DEI, and digestive tract size. PMID:24470452

  2. Essential and toxic metals in tea (Camellia sinensis) imported and produced in Ethiopia.

    PubMed

    Ashenef, Ayenew

    2014-01-01

    Sixteen samples of packed tea leaves (Camellia sinensis) were purchased from supermarkets in Addis Ababa, Ethiopia for metal analysis. Elements were measured by FAAS and graphite furnace atomic absorption spectrometer (GFAAS) employing external calibration curves. The levels in mg/kg dried weight basis varied from Cu: 4.7-12.9; Cd: 0.02-2.83; Pb: <0.01-2.29; Zn: 8.6-198.3; Mn: 81.7-962.2; Al: 3376.4-10,369.3; K: 7667.7-10,775; Li: 0.2-0.62; Ba: 9.4-1407.1; Mg: 1145.6-1834.1; Fe: 286.4-880.9; Ca: 1414.2-2646.0; Na: 147.1-557.7. Levels of exposure to the investigated metals by drinking tea were checked with the recommended daily allowance (RDA) of the WHO/FAO. Considering the average daily consumption rate of tea alone, the possible daily intakes of Al, Ba and Mn surpass the amenability to the side effects associated with these elements like Alzheimer's disease, kidney damage and Parkinson's disease, respectively, for which drinking tea should cause awareness. The other investigated elements are in the acceptable range.

  3. Structure and mechanical properties of pincers of lobster (Procambarus clarkii) and crab (Eriocheir Sinensis).

    PubMed

    Zhou, Fei; Wu, Zhiwei; Wang, Meiling; Chen, Kangmin

    2010-08-01

    The structure and mechanical properties of the pincer exoskeletons (cuticles) of lobster (Procambarus clarkii) and crab (Eriocheir Sinensis) were investigated, respectively. The microstructures and inorganic materials of the pincer exoskeletons were observed and determined using a scanning electron microscope and X-ray diffraction. The mechanical properties of the pincer exoskeletons were evaluated by nano-indentation tests and tensile tests under different conditions. The results showed that the inorganic materials in the lobster claw exoskeleton exhibited an amorphous structure, while those in the crab claw exoskeleton were crystallized as calcium carbonate with a calcite crystal structure and were stable at the temperature below 250 degrees C. The surfaces of the pincers were biologically unsmooth. Many concave valleys with setae near the tip and many convex domes far off the tip were observed on the surface of the lobster pincer, while many micro-spines were seen on the surface of crab pincer. The microstructures of the pincer exoskeleton exhibited highly mineralized chitin-protein fibers arranged in a twisted plywood structure. The surface hardness and elastic modulus of the crab claws were 0.33 GPa and 8.18 GPa, respectively, higher than those of the lobster claw (0.27 and 5.44 GPa). The transition in the mechanical properties and structure between the exocuticle and the endocuticle was discontinuity. The tensile strength of the crab pincer was two times higher than that of the lobster pincer, and the dry specimen was fractured more easily than the fresh specimen.

  4. Immunohistochemical analysis for G protein in the olfactory organs of soft-shelled turtle, Pelodiscus sinensis

    PubMed Central

    NAKAMUTA, Shoko; YOKOSUKA, Makoto; TANIGUCHI, Kazumi; YAMAMOTO, Yoshio; NAKAMUTA, Nobuaki

    2015-01-01

    In turtles, the epithelia lining the upper and lower chambers of the nasal cavity project axons to the ventral and dorsal parts of the olfactory bulbs, respectively. In a semi-aquatic soft-shelled turtle, Pelodiscus sinensis, more than 1,000 odorant receptor genes have been found, but it is not known where they are expressed. In this study, we aimed to clarify the distribution of cells expressing these genes in the olfactory organs of soft-shelled turtles. Immunoreactions for the Gαolf, the α subunit of G protein coupled to the odorant receptors, were detected on the surface of epithelia lining both the upper and lower chambers of the nasal cavity. The receptor cells in the epithelium of both chambers possessed cilia on the tip of their dendrites, whereas microvillous, non-ciliated, receptor cells were not found. These data suggest that the odorant receptor genes are expressed by the ciliated receptor cells in the upper and lower chamber epithelia. Precise location of the vomeronasal epithelium is not known at present. PMID:26440778

  5. Irradiance heterogeneity within crown affects photosynthetic capacity and nitrogen distribution of leaves in Cedrela sinensis.

    PubMed

    Yoshimura, Kenichi

    2010-05-01

    Because light conditions in the forest understory are highly heterogeneous, photosynthetic acclimation to spatially variable irradiance within a crown is important for crown-level carbon assimilation. The effect of variation in irradiance within the crown on leaf nitrogen content and photosynthetic rate was examined for pinnate compound leaves in saplings of Cedrela sinensis, a pioneer deciduous tree. Five shading treatments, in which 0, 25, 50, 75 and 100% of leaves were shaded, were established by artificial heavy shading using shade screen umbrellas with 25% transmittance. Although the nitrogen content of leaves was constant regardless of shading treatment, ribulose 1.5-bisphosphate carboxylase/oxygenase (Rubisco) content and light-saturated photosynthetic capacity were lower in shade leaves within partially shaded crowns than within fully shaded crowns. Shade leaves within partially shaded crowns contained higher amount of amino acids. Most shade leaves died in partially shaded crowns, whereas more than half of shade leaves survived in totally shaded crowns. Assumptions on photosynthetic acclimation to local light conditions cannot explain why shade leaves have different photosynthetic capacities and survival rates in between partially and totally shaded crowns. Irradiance heterogeneity within the crown causes a distinct variation in photosynthetic activity between sun and shaded leaves within the crown.

  6. [Cloning and expression analysis of HMG-CoA reductase from Aquilaria sinensis (Lour.) Gilg].

    PubMed

    Xu, Yan-Hong; Yang, Xin; Zhang, Zheng; Liang, Liang; Wei, Jian-He

    2013-06-01

    3-Hydroxy-3-methylglutaryl-coenzyme A reductase (HMGR) is the first rate-limiting enzyme for sesquiterpene synthesis in the mevalonate (MVA) pathway. The specific primers were designed according to the transcript sequence of AsHMGR2 from the Aquilaria sinensis (Lour.) Gilg transcriptome database. The full-length cDNA of AsHMGR2 was cloned by RT-PCR and rapid amplification of cDNA ends (RACE) technology, and was analyzed at bioinformatics levels; AsHMGR2 expression profiles in different tissues and in responds to different treatments were analyzed by real-time PCR. The length of AsHMGR2 Open Reading Frame (ORF) was 1 749 bp, encoding 582 amino acids. The GenBank accession number is KC140287. Tissue expression analysis indicated that AsHMGR2 was mainly expressed in root and shoot tips, followed by stem, and was lowest in leaves. Inducible-experiments showed that the genes were induced by mechanical wound as well as chemical liquid induction, and reached the highest expression level at 6 h and 8 h, separately. The full-length cDNA of AsHMGR2 and its expression patterns will provide a foundation for further research on its function in agarwood sesquiterpene biosynthesis.

  7. Antitumor and antimicrobial activities of endophytic fungi from medicinal parts of Aquilaria sinensis.

    PubMed

    Cui, Jin-long; Guo, Shun-xing; Xiao, Pei-gen

    2011-05-01

    The purpose of this study was to isolate and characterize endophytic fungi from the stem tissue which can produce fragrant ingredients in Aquilaria sinensis (also called agarwood) to determine their antitumor and antimicrobial activities. Twenty-eight fungal endophytes were isolated from agarwood by strict sterile sample preparation and were classified into 14 genera and 4 taxonomic classes (Sordariomycetes, Dothideomycetes, Saccharomycetes, and Zygomycetes) based on molecular identification. Of the 28 isolates, 13 (46.4%) showed antimicrobial activity against at least one of the test strains by the agar well diffusion method, and 23 isolates (82.1%) displayed antitumor activity against at least one of five cancer cell lines by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The diameters of inhibition zones of YNAS07, YNAS14, HNAS04, HNAS05, HNAS08, and HNAS11 were equal to or higher than 14.0 mm against Staphylococcus aureus, Escherichia coli, Bacillus subtilis, B. subtilis, Aspergillus fumigatus, and B. subtilis, respectively. The inhibition rates of YNAS06, YNAS08, and HNAS06 were not less than 60% to 293-T, 293-T, and SKVO3 cells, respectively. These results suggest that the endophytic fungi associated with agarwood will provide us with not only useful micro-ecological information, but also potential antimicrobial and antitumor agents.

  8. 2-(2-Phenylethyl)chromone derivatives in artificial agarwood from Aquilaria sinensis.

    PubMed

    Liao, Ge; Mei, Wen-Li; Dong, Wen-Hua; Li, Wei; Wang, Pei; Kong, Fan-Dong; Gai, Cui-Juan; Song, Xi-Qiang; Dai, Hao-Fu

    2016-04-01

    Seven new 2-(2-phenylethyl)chromone derivatives (1-7) including a chlorinated one (4), together with eight known ones (8-15), were isolated from the EtOAc extract of artificial agarwood originating from Aquilaria sinensis (Lour.) Gilg. All structures including the absolute configurations were unambiguously elucidated by spectroscopic (NMR, UV, IR, MS) methods, Mosher's method, and comparison with reported data in the literatures. Among those, compounds 8, 12, and 14 exhibited significant inhibition against α-glucosidase in vitro with IC50 values of 0.15, 0.05, and 0.09 mM, respectively (with acarbose as the positive control; IC50: 0.98 mM). In addition, compounds 3, 9, 11, and 14 showed weak inhibitory activity against AChE; and compounds 12 and 13 displayed weak cytotoxicity against human gastric cell line (SGC-7901) among three types of tested human cancer cell lines (BEL-7402, K562, and SGC-7901).

  9. [Establishment of a cell suspension culture system of endangered Aquilaria sinensis (Lour.) Gilg].

    PubMed

    Liu, Juan; Han, Xiao-Min; Liang, Liang; Liu, Qing-Chang; Xu, Yan-Hong; Yang, Cheng-Min; Zhang, Zheng; Sun, Jing; Wei, Jian-He

    2014-08-01

    Aquilaria sinensis callus induced by stem tips were used to establish the suspension cell system. The results showed that the most suitable medium for callus induction and subculture is MS + 2.0 mg x L(-1) NAA + 1.0 mg x L(-1) 6-BA. After 12 times of subculture, the energetic and loose callus, which were appropriate for cell suspension culture, were cultured and shook in liquid medium MS + 2.0 mg x L(-1) NAA + 1.0 mg x L(-1) 6-BA + 500.0 mg x L(-1) casein hydrolysate (CH) to establish the suspension cell system. The growth curve of suspension cells showed a "S" type. At the beginning of the culture, cell density increased slowly; during 4 to 6 days, suspension cells reached logarithmic growth period; during 7 to 12 days, suspension cells were in the platform period; but after 12 days, cell density and activity went down obviously. Agarwood sesquiterpenes were not detected in the suspension cells during the growth period, however, they could be detected in MeJA treated suspension cells. In this study, a stable and active growing suspension cell system was established, which was a proper system to study the mechanism of agarwood sesquiterpene formation, and additionally provided a potential way to generate agarwood sesquiterpenes through application of cell culture.

  10. Antitumor and antimicrobial activities of endophytic fungi from medicinal parts of Aquilaria sinensis *

    PubMed Central

    Cui, Jin-long; Guo, Shun-xing; Xiao, Pei-gen

    2011-01-01

    The purpose of this study was to isolate and characterize endophytic fungi from the stem tissue which can produce fragrant ingredients in Aquilaria sinensis (also called agarwood) to determine their antitumor and antimicrobial activities. Twenty-eight fungal endophytes were isolated from agarwood by strict sterile sample preparation and were classified into 14 genera and 4 taxonomic classes (Sordariomycetes, Dothideomycetes, Saccharomycetes, and Zygomycetes) based on molecular identification. Of the 28 isolates, 13 (46.4%) showed antimicrobial activity against at least one of the test strains by the agar well diffusion method, and 23 isolates (82.1%) displayed antitumor activity against at least one of five cancer cell lines by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The diameters of inhibition zones of YNAS07, YNAS14, HNAS04, HNAS05, HNAS08, and HNAS11 were equal to or higher than 14.0 mm against Staphylococcus aureus, Escherichia coli, Bacillus subtilis, B. subtilis, Aspergillus fumigatus, and B. subtilis, respectively. The inhibition rates of YNAS06, YNAS08, and HNAS06 were not less than 60% to 293-T, 293-T, and SKVO3 cells, respectively. These results suggest that the endophytic fungi associated with agarwood will provide us with not only useful micro-ecological information, but also potential antimicrobial and antitumor agents. PMID:21528493

  11. Camellia sinensis (L.) Kuntze Extract Ameliorates Chronic Ethanol-Induced Hepatotoxicity in Albino Rats.

    PubMed

    Lodhi, Poonam; Tandan, Neeraj; Singh, Neera; Kumar, Divyansh; Kumar, Monu

    2014-01-01

    The goal of this study was to investigate the hepatoprotective effects of aqueous extract of Camellia sinensis or green tea extract (AQGTE) in chronic ethanol-induced albino rats. All animals were divided into 4 groups in the study for a 5-week duration. 50% ethanol was given orally to the rats with two doses (5 mg/kg bw and 10 mg/kg bw) of AQGTE. Ethanol administration caused a significant increase in the levels of plasma and serum enzymatic markers, alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP), and nonenzymatic markers (cholesterol and triglycerides), lipid peroxidation contents, malondialdehyde (MDA), and glutathione-S-transferase (GST), and decreased the activities of total proteins, albumin, and cellular antioxidant defense enzymes such as superoxide dismutase (SOD). The elevation and reduction in these biochemical enzymes caused the damage in hepatocytes histologically due to the high production of ROS, which retards the antioxidant defense capacity of cell. AQGTE was capable of recovering the level of these markers and the damaged hepatocytes to their normal structures. These results support the suggestion that AQGTE was able to enhance hepatoprotective and antioxidant effects in vivo against ethanol-induced toxicity.

  12. Effect of green tea (Camellia sinensis) extract on healing process of surgical wounds in rat.

    PubMed

    Asadi, Sayyed Yazdan; Parsaei, Pouya; Karimi, Mehrdad; Ezzati, Sareh; Zamiri, Alaleh; Mohammadizadeh, Fereshteh; Rafieian-Kopaei, Mahmoud

    2013-01-01

    Green tea (Camellia sinensis) has anti-oxidant and anti-inflammatory properties and may enhance wound healing process. The present study, therefore, was aimed to examine the effect of green tea ethanolic extract on wound healing process. For this experimental study, 36 healthy male Wistar rats were randomly designated to three groups of A, B, and C which, respectively treated with, Vaseline + 0.6% green tea extract, Vaseline and normal saline for 21 days. Wounds' length and area were measured by caliper every other day and specimens were taken at 3rd, 12th, and 21st day for microscopical examinations. Data were analyzed by SPSS 16 using survival analysis (Breslow test), repeated measured ANOVA, one-way ANOVA and Mann-Whitney. P < 0.05 was considered as statistically significant. The mean healing duration of surgical wounds in groups A and B was 14.66 and 20.66 (P = 0.018), respectively. Decrease in healing duration in the group A was significantly higher within the first two weeks compared with control groups (P = 0.05). Microscopic examinations also indicated a significant difference in wound healing process between groups A and C throughout the whole study duration as well as groups A and B during the 3rd week of the study (P < 0.05). Green tea extract could help wound healing process, probably effective on surgical wounds healing.

  13. Molecular Cloning and Characterization of Hydroperoxide Lyase Gene in the Leaves of Tea Plant (Camellia sinensis).

    PubMed

    Deng, Wei-Wei; Wu, Yi-Lin; Li, Ye-Yun; Tan, Zhen; Wei, Chao-Ling

    2016-03-02

    Hydroperoxide lyase (HPL, E.C. 4.1.2.) is the major enzyme in the biosynthesis of natural volatile aldehydes and alcohols in plants, however, little was known about HPL in tea plants (Camellia sinensis). A unique cDNA fragment was isolated by suppressive subtractive hybridization (SSH) from a tea plant subjected to herbivory by tea geometrid Ectropis obliqua. This full length cDNA acquired by RACE was 1476 bp and encoded 491 amino acids. DNA and protein BLAST searches showed high homology to HPL sequences from other plants. The His-tag expression vector pET-32a(+)/CsHPL was constructed and transferred into Escherichia coli Rosetta (DE3). The expression product of recombinant CsHPL in E. coli was about 60 kDa. The enzyme activity of CsHPL was 0.20 μmol·min(-1)·mg(-1). Quantitative RT-PCR analysis indicated CsHPL was strongly up-regulated in tea plants after Ectropis obliqua attack, suggesting that it may be an important candidate for defense against insects in tea plants.

  14. Green synthesis of gold and silver nanoparticles using Hibiscus rosa sinensis

    NASA Astrophysics Data System (ADS)

    Philip, Daizy

    2010-03-01

    Biological synthesis of gold and silver nanoparticles of various shapes using the leaf extract of Hibiscus rosa sinensis is reported. This is a simple, cost-effective, stable for long time and reproducible aqueous room temperature synthesis method to obtain a self-assembly of Au and Ag nanoparticles. The size and shape of Au nanoparticles are modulated by varying the ratio of metal salt and extract in the reaction medium. Variation of pH of the reaction medium gives silver nanoparticles of different shapes. The nanoparticles obtained are characterized by UV-vis, transmission electron microscopy (TEM), X-ray diffraction (XRD) and FTIR spectroscopy. Crystalline nature of the nanoparticles in the fcc structure are confirmed by the peaks in the XRD pattern corresponding to (1 1 1), (2 0 0), (2 2 0) and (3 1 1) planes, bright circular spots in the selected area electron diffraction (SAED) and clear lattice fringes in the high-resolution TEM image. From FTIR spectra it is found that the Au nanoparticles are bound to amine groups and the Ag nanoparticles to carboxylate ion groups.

  15. Two antibacterial C-type lectins from crustacean, Eriocheir sinensis, stimulated cellular encapsulation in vitro.

    PubMed

    Jin, Xing-Kun; Li, Shuang; Guo, Xiao-Nv; Cheng, Lin; Wu, Min-Hao; Tan, Shang-Jian; Zhu, You-Ting; Yu, Ai-Qing; Li, Wei-Wei; Wang, Qun

    2013-12-01

    The first step of host fighting against pathogens is that pattern recognition receptors recognized pathogen-associated molecular patterns. However, the specificity of recognition within the innate immune molecular of invertebrates remains largely unknown. In the present study, we investigated how invertebrate pattern recognition receptor (PRR) C-type lectins might be involved in the antimicrobial response in crustacean. Based on our previously obtained completed coding regions of EsLecA and EsLecG in Eriocheir sinensis, the recombinant EsLectin proteins were produced via prokaryotic expression system and affinity chromatography. Subsequently, both rEsLecA and rEsLecG were discovered to have wide spectrum binding activities towards microorganisms, and their microbial-binding was calcium-independent. Moreover, the binding activities of both rEsLecA and rEsLecG induced the aggregation against microbial pathogens. Both microorganism growth inhibitory activities assays and antibacterial activities assays revealed their capabilities of suppressing microorganisms growth and directly killing microorganisms respectively. Furthermore, the encapsulation assays signified that both rEsLecA and rEsLecG could stimulate the cellular encapsulation in vitro. Collectively, data presented here demonstrated the successful expression and purification of two C-type lectins proteins in the Chinese mitten crab, and their critical role in the innate immune system of an invertebrate.

  16. Tea creaming in nonfermented teas from Camellia sinensis and Ilex vomitoria.

    PubMed

    Kim, Youngmok; Talcott, Stephen T

    2012-11-28

    Tea creaming is the development of a cloudy or hazy appearance in tea and ready-to-drink tea products on cooling and is highly undesirable in the tea beverage industry. Commonly associated with fermented black or oolong teas, the objective of this study was to investigate the physicochemical mechanism of the formation of tea cream in nonfermented green tea (Camellia sinensis) and a caffeine-containing botanical tea from yaupon holly (Ilex vomitoria) that is free of catechin-based polyphenolics. Four tea-creaming activators (phenolics, soluble protein, caffeine, and metal ions) were added to tea infusions as well as decaffeinated teas created by chloroform extraction. Tea-creaming activators increased the weight and turbidity of both teas with the exception of soluble protein addition (as bovine serum albumin) to green tea, whereas the greatest increase in turbidity occurred with the addition of metal ions in green tea. Tea creaming was equally developed at three incubation temperatures (4, 25, and 40 °C) in both teas, but tea-creaming compositions in each tea were different at the incubating temperatures. The antioxidant capacity of each tea was lowered after creaming due to the loss of antioxidants that participated in tea cream formation.

  17. Regulation of formation of volatile compounds of tea (Camellia sinensis) leaves by single light wavelength

    PubMed Central

    Fu, Xiumin; Chen, Yiyong; Mei, Xin; Katsuno, Tsuyoshi; Kobayashi, Eiji; Dong, Fang; Watanabe, Naoharu; Yang, Ziyin

    2015-01-01

    Regulation of plant growth and development by light wavelength has been extensively studied. Less attention has been paid to effect of light wavelength on formation of plant metabolites. The objective of this study was to investigate whether formation of volatiles in preharvest and postharvest tea (Camellia sinensis) leaves can be regulated by light wavelength. In the present study, in contrast to the natural light or dark treatment, blue light (470 nm) and red light (660 nm) significantly increased most endogenous volatiles including volatile fatty acid derivatives (VFADs), volatile phenylpropanoids/benzenoids (VPBs), and volatile terpenes (VTs) in the preharvest tea leaves. Furthermore, blue and red lights significantly up-regulated the expression levels of 9/13-lipoxygenases involved in VFADs formation, phenylalanine ammonialyase involved in VPBs formation, and terpene synthases involved in VTs formation. Single light wavelength had less remarkable influences on formation of volatiles in the postharvest leaves compared with the preharvest leaves. These results suggest that blue and red lights can be promising technology for remodeling the aroma of preharvest tea leaves. Furthermore, our study provided evidence that light wavelength can activate the expression of key genes involved in formation of plant volatiles for the first time. PMID:26567525

  18. Why do Chinese alligators (Alligator sinensis) form bellowing choruses: a playback approach.

    PubMed

    Wang, Xianyan; Wang, Ding; Zhang, Song; Wang, Chaolin; Wang, Renping; Wu, Xiaobing

    2009-10-01

    Crocodilians are quite vocal relative to other reptile groups, and the alligators are among the most vocal of the crocodilians. The Chinese alligator, Alligator sinensis, is usually solitary but engages in bellowing choruses in certain waters during the mating season. This paper reports the organization of Chinese alligator's bellowing choruses based upon field observations and playback experiments. Alligators of both genders engaged in the choruses, remaining immobile throughout and inclining toward bellowing synchronously (i.e., starting and finishing at about the same time). The choruses lasted about 10 min with abrupt onset and offset. Moreover, playback experiments revealed that both male and female alligators responded equally to bellowing stimuli from the same and opposite sexes and that none of the tested alligators approached the loudspeaker in spite of playback of male or female stimuli. These suggest that Chinese alligators may not bellow to compete for or attract mates during the choruses. Instead, when their ecological behaviors, namely, dispersed inhabitation, multi-copulation, restricted mating season, etc., are considered, we hypothesize that they may synchronize bellows to enhance group detectability for assembling individuals into certain waters for subsequent copulations.

  19. Molecular and functional characterization of BAFF from the Yangtze alligator (Alligator sinensis, Alligatoridae).

    PubMed

    Zhang, Jia-Xin; Song, Ren; Sang, Ming; Sun, Si-Qing; Ma, Lei; Zhang, Jie; Zhang, Shuang-Quan

    2015-10-01

    B-cell activating factor (BAFF) from the TNF family is critical for B-cell survival and maturation. In this study, we identified a Yangtze alligator (Alligator sinensis, Alligatoridae) BAFF cDNA, designated as asBAFF, using reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE). The open reading frame of this cDNA encodes a 287-amino acid protein containing a predicted transmembrane domain and a furin protease cleavage site, similar to mammalian and avian BAFF. The amino acid identity between biologically soluble asBAFF (assBAFF) and csBAFF, hsBAFF, and msBAFF is 94, 76, and 71%, respectively. Real-time quantitative PCR analysis showed that the asBAFF gene is strongly expressed in the spleen. Since BAFF is always expressed as inclusion bodies in bacteria, it is difficult to purify. To enhance the soluble expression of assBAFF in Escherichia coli, we fused the extracellular region of the asBAFF gene to a small ubiquitin-related modifier gene (SUMO). Purified assBAFF was able to promote the survival of splenic lymphocytes and co-stimulate the proliferation of mouse B cells with anti-mouse IgM. These findings suggest that asBAFF plays an important role in the survival and proliferation of Yangtze alligator B cells, and because it is evolutionarily highly conserved, functional cross-reactivity exists between mammalian and Yangtze alligator BAFF.

  20. Transcriptome analysis of a spontaneous mutant in sweet orange [Citrus sinensis (L.) Osbeck] during fruit development.

    PubMed

    Liu, Qing; Zhu, Andan; Chai, Lijun; Zhou, Wenjing; Yu, Keqin; Ding, Jian; Xu, Juan; Deng, Xiuxin

    2009-01-01

    Bud mutations often arise in citrus. The selection of mutants is one of the most important breeding channels in citrus. However, the molecular basis of bud mutation has rarely been studied. To identify differentially expressed genes in a spontaneous sweet orange [C. sinensis (L.) Osbeck] bud mutation which causes lycopene accumulation, low citric acid, and high sucrose in fruit, suppression subtractive hybridization and microarray analysis were performed to decipher this bud mutation during fruit development. After sequencing of the differentially expressed clones, a total of 267 non-redundant transcripts were obtained and 182 (68.2%) of them shared homology (E-value < or = 1x10(-10)) with known gene products. Few genes were constitutively up- or down-regulated (fold change > or = 2) in the bud mutation during fruit development. Self-organizing tree algorithm analysis results showed that 95.1% of the differentially expressed genes were extensively coordinated with the initiation of lycopene accumulation. Metabolic process, cellular process, establishment of localization, response to stimulus, and biological regulation-related transcripts were among the most regulated genes. These genes were involved in many biological processes such as organic acid metabolism, lipid metabolism, transport, and pyruvate metabolism, etc. Moreover, 13 genes which were differentially regulated at 170 d after flowering shared homology with previously described signal transduction or transcription factors. The information generated in this study provides new clues to aid in the understanding of bud mutation in citrus.

  1. Phylogeography of the inshore fish, Bostrychus sinensis, along the Pacific coastline of China.

    PubMed

    Qiu, Fan; Li, Hai; Lin, Hungdu; Ding, Shaoxiong; Miyamoto, Michael M

    2016-03-01

    This study assesses the phylogeography of the Chinese four-eyed sleeper (Bostrychus sinensis) with one mitochondrial and one nuclear genes and two morphological characters. Phylogenetic and population genetic analyses of the sequences reveals two phylogeographic lineages from the East and South China Seas, which are corroborated by the morphological data. The vicariance of the two lineages is attributed to the Pleistocene Ice Age exposure of the Taiwan Strait and consequent connection of Taiwan to the mainland, which thereby introduced an ecological barrier to gene flow between populations in the East and South China Seas. The distributions of the two lineages now overlap in the East China Sea and this secondary contact is attributed to biased northward migration along the two main currents of the Taiwan Strait following its interglacial re-flooding. In conclusion, this study reinforces the importance of "vicariance, then secondary contact" due to Late Pliocene and Pleistocene sea-level changes to the phylogeography of marine species. Specifically, it corroborates the importance of Pleistocene sea-level changes in the Taiwan Strait to the phylogeography of Chinese inshore species.

  2. Early exposure of rotating magnetic fields promotes central nervous regeneration in planarian Girardia sinensis.

    PubMed

    Chen, Qiang; Lin, Gui-miao; Wu, Nan; Tang, Sheng-wei; Zheng, Zhi-jia; Lin, Marie Chia-mi; Xu, Gai-xia; Liu, Hao; Deng, Yue-yue; Zhang, Xiao-yun; Chen, Si-ping; Wang, Xiao-mei; Niu, Han-ben

    2016-05-01

    Magnetic field exposure is an accepted safe and effective modality for nerve injury. However, it is clinically used only as a supplement or salvage therapy at the later stage of treatment. Here, we used a planarian Girardia sinensis decapitated model to investigate beneficial effects of early rotary non-uniform magnetic fields (RMFs) exposure on central nervous regeneration. Our results clearly indicated that magnetic stimulation induced from early RMFs exposure significantly promoted neural regeneration of planarians. This stimulating effect is frequency and intensity dependent. Optimum effects were obtained when decapitated planarians were cultured at 20 °C, starved for 3 days before head-cutting, and treated with 6 Hz 0.02 T RMFs. At early regeneration stage, RMFs exposure eliminated edema around the wound and facilitated subsequent formation of blastema. It also accelerated cell proliferation and recovery of neuron functionality. Early RMFs exposure up-regulated expression of neural regeneration related proteins, EGR4 and Netrin 2, and mature nerve cell marker proteins, NSE and NPY. These results suggest that RMFs therapy produced early and significant benefit in central nervous regeneration, and should be clinically used at the early stage of neural regeneration, with appropriate optimal frequency and intensity.

  3. Angelica sinensis improves exercise performance and protects against physical fatigue in trained mice.

    PubMed

    Yeh, Tzu-Shao; Huang, Chi-Chang; Chuang, Hsiao-Li; Hsu, Mei-Chich

    2014-03-31

    Angelica sinensis (AS) is a well-known medicinal herb and food material with antioxidative and multifunctional pharmacological activities. However, we lack evidence of the effect of AS on exercise performance and physical fatigue. We aimed to evaluate the potential beneficial effect of AS on ergogenic and anti-fatigue functions after physiological challenge. Male ICR strain mice were randomly assigned to four groups (n=10 per group) for treatment: (1) sedentary control and vehicle treatment (vehicle control); (2) exercise training with vehicle treatment (exercise control); (3) exercise training with AS treatment at 0.41 g/kg/day (Ex-AS1); and (4) 2.05 g/kg/day (Ex-AS5); both the vehicle and AS were orally administered for 6 weeks. Exercise performance and anti-fatigue function were evaluated by forelimb grip strength, exhaustive swimming time, and levels of serum lactate, ammonia, glucose, and creatine kinase (CK) after a 15-min swimming exercise. Trend analysis revealed that AS treatments significantly increased endurance swimming time and blood glucose level, and decreased serum lactate, ammonia and CK levels. Liver and muscle glycogen contents were higher for Ex-AS1 and Ex-AS5 groups than the exercise control. Therefore, AS supplementation improved exercise performance and had anti-fatigue properties in mice and may be an effective ergogenic aid in exercise training.

  4. Differential expression of fertility genes boule and dazl in Chinese sturgeon (Acipenser sinensis), a basal fish.

    PubMed

    Ye, Huan; Li, Chuang-Ju; Yue, Hua-Mei; Yang, Xiao-Ge; Wei, Qi-Wei

    2015-05-01

    The gene family DAZ (deleted in Azoospermia), including boule, dazl and DAZ, performs highly conserved functions in germ cell development and fertility across animal phyla. Differential expression patterns have been demonstrated for the family members in invertebrates and vertebrates including fish. Here, we report the identification of boule and dazl and their expression at both RNA and protein levels in developing and mature gonads of Chinese sturgeon (Acipenser sinensis). Firstly, the isolation of the boule and dazl genes in Chinese sturgeon and the observation of the two genes in coelacanth suggest that dazl originated after the divergence of bony fish from cartilaginous fish but before the emergence of the Actinistia. Quantitative real-time PCR and western blot analyses reveal that boule and dazl RNA and proteins are restricted to the testis and ovary. In situ hybridization and fluorescent immunohistochemistry show that the bisexual mitotic and meiotic germ cell expression of dazl RNA and protein is conserved in vertebrates, while Chinese sturgeon boule RNA and protein exhibit mitotic and meiotic expression in the testis, and also likely display mitotic and meiotic expression in female. Moreover, we directly demonstrate for the first time that sturgeon Balbiani body/mitochondrial cloud disperses in the cytoplasm of early developing oocytes and co-localizes with Dazl to some extent. Finally, urbilaterian boule may also have an ancestral function in oogenesis. Taken together, these results provide useful information on the evolution of DAZ family genes, expression patterns and functions in animal reproduction.

  5. Preparation and characterization of collagen from soft-shelled turtle (Pelodiscus sinensis) skin for biomaterial applications.

    PubMed

    Nagai, Nobuhiro; Kobayashi, Hatsumi; Katayama, Shizuka; Munekata, Masanobu

    2009-01-01

    Collagen was isolated from the skin of soft-shelled turtle (Pelodiscus sinensis) by acid solubilization with pepsin. The yield of soft-shelled turtle collagen (STC) was 12.1% on a dry weight basis. The electrophoresis assay showed that STC consisted of a alpha(1)alpha(2) heterodimer similar to porcine collagen (PC). Amino-acid composition analysis showed that the hydroxyproline content of STC was 7.8%, which was lower than that of PC (9.5%). The denaturation temperature of STC was 36 degrees C from optical rotation analysis. An accelerated fibrillogenesis of STC was observed in phosphate-buffered saline at 25 degrees C. The resulting STC fibrillar gel had microfibrillar network with fibril diameter of ca. 124 nm, as revealed by observation with scanning electron microscopy. The compressive moduli of the STC gel and the PC gel were 3.2 +/- 0.8 kPa and 3.6 +/- 0.3 kPa, respectively. The potential of the STC gel for biomaterial applications was investigated by in vitro cell culture. Human dermal fibroblasts were three-dimensionally cultured in the STC gel and their growth was evaluated by DNA content measurement. Steady growth was observed in the STC gel for a 6-day culture period, although the growth rate was slower than in the PC gel. In conclusion, STC could be used as a novel collagen source for biomaterial applications.

  6. Identification of native catechin fatty acid esters in green tea (Camellia sinensis).

    PubMed

    Myers, Richard A; Fuller, Ewa; Yang, Weichun

    2013-11-27

    Catechins are potent antioxidants and make up the primary class of polyphenols present in tea (Camellia sinensis). They are especially abundant in the less-fermented green teas that have been employed in various foods to enhance shelf life stability (Senanayake, N. J. Funct. Foods 2013, in press. Gramza, A.; Korczak, J. Trends Food Sci. 2005, 16, 351-358). The antioxidative activity of native (polar) catechins has proven to be useful in foods of relatively high polarity, while mixed results have been achieved in high-fat foods. However, the polarity of catechins can be attenuated by esterification with fatty acids, producing adducts that effectively partition into lipids and protect against rancidity even in high-fat foods (Cutler, S.; Fuller, E.; Rotberg, I.; Wray, C.; Troung, M.; Poss, M. International Patent WO 2013/036934 A1, March 14, 2013. Zhong, Y.; Shahidi, F. J. Agric. Food Chem. 2011, 59, 6526-6533). In this work, a search for the presence of naturally occurring lipid-conjugated catechins was undertaken in various green tea varieties. Rather than the traditional aqueous infusion, dried tea leaves were extracted with organic solvents followed by analysis for catechin adducts with both lower polarities and increased molecular weights as monitored by liquid chromatography and tandem mass spectrometry. Native catechin palmitates were identified and indirectly confirmed by synthesis and nuclear magnetic resonance as natural components of several Chinese green teas. Evidence of other fatty catechin esters was also observed.

  7. Behavior, metabolism and swimming physiology in juvenile Spinibarbus sinensis exposed to PFOS under different temperatures.

    PubMed

    Xia, Ji-Gang; Nie, Li-Juan; Mi, Xia-Mei; Wang, Wei-Zhen; Ma, Yi-Jie; Cao, Zhen-Dong; Fu, Shi-Jian

    2015-10-01

    The harmful effects of perfluorooctane sulfonate (PFOS) are of growing international concern. This paper aimed to gain an integrated understanding of fitness-related ecological end points, such as behavior, metabolism and swimming physiology, in juvenile Spinibarbus sinensis in response to PFOS toxicity at different temperatures. The fish were exposed to a range of PFOS concentrations (0, 0.32, 0.8, 2 and 5 mg/L) at different temperatures (18 and 28 °C) for 30 days. The effects on fish behavior, metabolic characteristics and aerobic swimming performance caused by PFOS at different temperatures were investigated. Our results showed that both PFOS and temperature had important influences on spontaneous swimming behavior, social interactions, routine metabolic rate (RMR), net energetic cost of transport (COTnet) and critical swimming speed (U crit) in fish. The lowest observed effect concentration for both U crit and RMR was 5 and 0.8 mg/L at 18 and 28 °C, respectively. We found that PFOS affected various behavioral and social end points and also appeared to affect metabolic rates and reduced U crit, likely as a result of increased COTnet, and that many of these effects also changed with respect to temperature. Our results further the understanding of the metabolic and behavioral toxicity of PFOS to aquatic organisms.

  8. Serotonergic Activity-Guided Phytochemical Investigation of the Roots of Angelica sinensis

    PubMed Central

    Deng, Shixin; Chen, Shao-Nong; Yao, Ping; Nikolic, Dejan; van Breemen, Richard B.; Bolton, Judy L.; Fong, Harry H.S.; Farnsworth, Norman R.; Pauli, Guido F.

    2006-01-01

    Serotonin receptor (5-HT7) binding assay-directed fractionation of a methanol extract of the dried roots of Angelica sinensis (Apiaceae) led to the isolation and identification of twenty-one compounds including a new phenolic ester, angeliferulate (1), and three new phthalides, 10-angeloylbutylphthalide (2), sinaspirolide (3) and ansaspirolide (4), along with seventeen known compounds, p-hydroxyphenethyl trans-ferulate (5), Z-ligustilide (6), Z-butylidenephthalide (7), senkyunolide I (8), Z-6-hydroxy-7-methoxy-dihydroligustilide (9), N-butylbenzenesulphonamide (10), 11(S),16(R)-dihydroxy-octadeca-9Z,17-dien-12,14-diyn-1-yl acetate (11), (3R,8S)-falcarindiol (12), heptadeca-1-ene-9,10-epoxy-4,6-diyne-3,8-diol (13), oplopandiol (14), 8-hydroxy-1-methoxy-, Z-9-heptadecene-4,6-diyn-3-one (15), imperatorin, ferulic acid, vanillin, stigmasterol, sucrose, and 1,3-di-linolenin. This is the first report of a sulfonamide (10) identified from a higher plant source, although its presence needs further investigation. Biosynthetic pathways for dimeric phthalides 3 and 4 are proposed. Compound 5, 7, 11, 12, 15, and imperatorin exhibited affinity toward 5-HT7 receptors in a competitive binding assay. PMID:16643021

  9. EST dataset of pituitary and identification of somatolactin and novel genes in Chinese sturgeon, Acipenser sinensis.

    PubMed

    Cao, Hong; Leng, Xiaoqian; Li, Chuangju; Wei, Qiwei; Gui, Jianfang; Cheng, Hanhua; Zhou, Rongjia

    2012-04-01

    Chinese sturgeon (Acipenser sinensis) is a rare and endangered species and also an important resource for the sturgeon aquaculture industry, however, a few genes have been identified in this species. We report here construction of a pituitary cDNA library from a 24 years old female Chinese sturgeon just after its spawning, and obtained 2,025 ESTs from the library. 885 unique sequences were identified, which were categorized into 12 functional groups. More than half of the unique sequences (57%) do not match with annotated sequences in the public databases. Three of these novel genes were further identified. Notably, a full-length of cDNA (1,143 bp) encoding somatolactin of 232 amino acids was identified. Phylogenetic analysis showed 97% amino acid identity with White sturgeon somatolactin. RT-PCR analysis indicated that the somatolactin mRNA was only detected in pituitary. Pituitary-specific expression of the somatolactin suggested that the protein may play important physiological functions in pituitary-endocrine system of the Chinese sturgeon.

  10. Detecting a novel Eriocheir sinensis reovirus by reverse transcription loop-mediated isothermal amplification assay.

    PubMed

    Ma, Y; Dai, T; Serwadda, A; Shen, H

    2016-11-01

    The novel Eriocheir sinensis reovirus (EsRV) is a pathogen that causes severe disease and high mortality rates in cultivated crabs. Here, we established a highly sensitive and specific rapid reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay that was cheaper and more suitable for field applications in crab aquaculture than those of traditional reverse transcription-polymerase chain reaction (RT-PCR) analysis. The amplification was completed within 45 min under isothermal conditions at 65°C. The RT-LAMP test for EsRV had a detection limit of 15 pg, and sensitivity was 100 times greater than that of conventional RT-PCR. The LAMP primers for EsRV were not amplified by other pathogen strains, indicating good specificity. In addition to detection by electrophoresis, RT-LAMP results were detectable by visual observations of reaction tube turbidity, and calcein was added to visually detect the amplification products. These results indicate that this highly convenient, rapid and sensitive RT-LAMP assay can be used to detect EsRV-infected aquatic organisms.

  11. Ammonia-oxidizing activity and microbial community structure in acid tea (Camellia sinensis) orchard soil

    NASA Astrophysics Data System (ADS)

    Okamura, K.; Takanashi, A.; Yamada, T.; Hiraishi, A.

    2012-03-01

    The purpose of this study was to determine the ammonia-oxidizing activity and the phylogentic composition of microorganisms involved in acid tea (Camellia sinensis) orchard soil. All soil samples were collected from three sites located in Tahara and Toyohashi, Aichi Prefecture, Japan. The potential nitrification rate (PNR) was measured by the chlorate inhibition method. The soil pH of tea orchards studied ranged from 2.78 to 4.84, differing significantly from sample to sample, whereas that of meadow and unplanted fields ranged from 5.78 to 6.35. The PNR ranged from 0.050 to 0.193 μg NO2--Ng-1 h-1 and were positively correlated with the soil pH (r2 = 0.382, p<0.001). Bulk DNA was extracted from a tea orchard soil (pH 4.8; PNR, 0.078 μg NO2--Ng-1 h-1) and subjected to PCR-aided clone library analyses targeting archaeal and bacterial amoA genes. The detected archaeal clones separated from the cluster of the 'Soil clones' and tightly clustered with the clones originating from other acidic soil environments including the Chinese tea orchard soil. These results suggest that the specific archaeal populations dominate as the ammonia oxidizers in acid tea-orchard soils and possibly other acid soils, independent of geographic locations, which results from the adaptation to specific ecological niches.

  12. Effects of Extract from Solid-State Fermented Cordyceps sinensis on Type 2 Diabetes Mellitus

    PubMed Central

    Kan, Wei-Chih; Wang, Hsien-Yi; Chien, Chih-Chiang; Li, Shun-Lai; Chen, Yu-Chun; Chang, Liang-Hao; Cheng, Chia-Hui; Tsai, Wan-Chen; Hwang, Jyh-Chang; Su, Shih-Bin; Huang, Li-Hsueh; Chuu, Jiunn-Jye

    2012-01-01

    Diabetes mellitus is the most common chronic disease in the world, and a wide range of drugs, including Chinese herbs, have been evaluated for the treatment of associated metabolic disorders. This study investigated the potential hypoglycemic and renoprotective effects of an extract from the solid-state fermented mycelium of Cordyceps sinensis (CS). We employed the KK/HIJ diabetic mouse model, in which the mice were provided with a high-fat diet for 8 weeks to induce hyperglycemia, followed by the administration of CS or rosiglitazone for 4 consecutive weeks. Several parameters were evaluated, including changes in body weight, plasma lipid profiles, oral glucose tolerance tests, insulin tolerance tests, and plasma insulin concentrations. Our results show that the CS extract significantly elevated HDL/LDL ratios at 4 weeks and decreased body weight gain at 8 weeks. Interestingly, CS treatment did not lead to obvious improvements in hyperglycemia or resistance to insulin, while in vitro MTT assays indicated that CS protects pancreatic beta cells against the toxic effects of STZ. CS also enhanced renal NKA activity and reduced the accumulation of mesangial matrix and collagen deposition. In conclusion, CS extract can potentially preserve β-cell function and offer renoprotection, which may afford a promising therapy for DM. PMID:22474523

  13. Effects of fermented Cordyceps sinensis on oxidative stress in doxorubicin treated rats

    PubMed Central

    Wu, Rong; Gao, Jian-Ping; Wang, Hui-Lin; Gao, Yan; Wu, Qian; Cui, Xiao-Hua

    2015-01-01

    Background: Cordyceps sinensis (CS) is one of the rare traditional Chinese herbs, only a very limited amount of natural CS is produced. Fermented CS, as a substitute for natural CS, is widely used in the field of supplementary medical treatment and health products. Its antagonistic effect on oxidative stress (OS) in vivo has not been investigated. Objective: Our aim was to investigate the antagonistic effect of fermented CS on OS in doxorubicin (DOX) treated rats and to compare the anti-OS effects in heart and liver tissues. Materials and Methods: OS rats were induced by tail-intravenous injection of DOX (total of 7.5 mg/kg), and then administered intragastrically with fermented CS (1.5 g/kg) for 4 weeks. At the end of the experiment, heart, liver and serum samples were taken for and biochemical analyses. Results: Fermented CS significantly increased the activities of glutathione peroxidase and catalase and the scavenging activity of O2− in serum, and the total superoxide dismutase activity in cardiac tissue; reduced the malondialdehyde content in liver and cardiac tissues. Conclusion: Fermented CS can inhibit DOX-induced OS reactions, and the anti-OS effects have high selectivity to heart and liver, especially to heart. Thus, fermented CS may be a candidate used for the prevention against various cardiac diseases induced by OS. PMID:26600716

  14. Construction of a Cordyceps sinensis exopolysaccharide-conjugated selenium nanoparticles and enhancement of their antioxidant activities.

    PubMed

    Xiao, Yidong; Huang, Qilin; Zheng, Zhaomin; Guan, Han; Liu, Shiyu

    2017-06-01

    A Cordyceps sinensis exopolysaccharide (EPS)-conjugated selenium nanoparticles (SeNPs) were successfully constructed through the reduction of SeO3(2-). The EPS-SeNPs were characterized in terms of formation, morphology, size, Se distribution and phase by UV-vis, FT-IR, transmission electron microscopy (TEM), dynamic light scattering (DLS), energy dispersive X-ray (EDX) and wide angle X-ray diffraction (WAXD) measurements. Results revealed that the SeNPs conjugated to EPS were amorphous and could be well dispersed at a size range of 80-125nm. The interactions between the OH groups of EPS and SeNPs substituted for intermolecular interaction in native EPS to form new CO⋯Se bonds, resulting in good dispersion of SeNPs in the EPS matrix. Besides, the EPS-SeNPs at different Se/P ratios exhibited significant scavenging ability on superoxide anion radical (O2(-)) and ABTS radical cation (ABTS(+)) when compared to pure EPS, indicating that the conjugated SeNPs reinforced antioxidant effect of EPS. This work not only provides a simple and efficient way to construct well-dispersed SeNPs in aqueous system, and demonstrates the vital role of the EPS as a biopolymer template for dispersion, stabilization and size control of SeNPs, but also finds the EPS-SeNPs can potentially serve as a good antioxidant towards O2(-) and ABTS(+).

  15. Cordyceps sinensis polysaccharide inhibits PDGF-BB-induced inflammation and ROS production in human mesangial cells.

    PubMed

    Wang, Ying; Wang, Yan; Liu, Dan; Wang, Wang; Zhao, Huan; Wang, Min; Yin, Hongping

    2015-07-10

    CPS-F, a polysaccharide derived from Cordyceps sinensis, is a potential anti-inflammatory and anti-oxidative agent. We demonstrated that CPS-F not only inhibits platelet-derived growth factor BB (PDGF-BB)-induced intracellular reactive oxygen species (ROS) generation, and up-regulation of tumor necrosis factor-α (TNF-α), TNF-α receptor 1 (TNFR1), and monocyte chemotactic protein-1 (MCP-1), but also acts synergistically in combination with MAPK/ERK inhibitor U0126 and PI3K/Akt inhibitor LY294002. Additionally, up-regulation of pro-inflammatory factors was reversed by use of a combination of CPS-F and NADPH oxidase (NOX) inhibitor diphenyleneiodonium chloride (DPI) or silencing of NOX1. Furthermore, CPS-F prevents the PDGF receptor β (PDGFRβ) promoter activity induced by PDGF-BB in transfected cells and ameliorates increased levels of TNF-α, TNFR1, and MCP-1 when PDGFRβ is silenced, thereby suggesting that CPS-F possesses a bidirectional regulatory function. Our findings suggest CPS-F may exert its therapeutic effect for the treatment of glomerulonephritis related to human mesangial cells (HMCs) through the ERK1/2/Akt pathways.

  16. Hemidesmus indicus and Hibiscus rosa-sinensis Affect Ischemia Reperfusion Injury in Isolated Rat Hearts

    PubMed Central

    Khandelwal, Vinoth Kumar Megraj; Balaraman, R.; Pancza, Dezider; Ravingerová, Táňa

    2011-01-01

    Hemidesmus indicus (L.) R. Br. (HI) and Hibiscus rosa-sinensis L. (HRS) are widely used traditional medicine. We investigated cardioprotective effects of these plants applied for 15 min at concentrations of 90, 180, and 360 μg/mL in Langendorff-perfused rat hearts prior to 25-min global ischemia/120-min reperfusion (I/R). Functional recovery (left ventricular developed pressure—LVDP, and rate of development of pressure), reperfusion arrhythmias, and infarct size (TTC staining) served as the endpoints. A transient increase in LVDP (32%–75%) occurred at all concentrations of HI, while coronary flow (CF) was significantly increased after HI 180 and 360. Only a moderate increase in LVDP (21% and 55%) and a tendency to increase CF was observed at HRS 180 and 360. HI and HRS at 180 and 360 significantly improved postischemic recovery of LVDP. Both the drugs dose-dependently reduced the numbers of ectopic beats and duration of ventricular tachycardia. The size of infarction was significantly decreased by HI 360, while HRS significantly reduced the infarct size at all concentrations in a dose-dependent manner. Thus, it can be concluded that HI might cause vasodilation, positive inotropic effect, and cardioprotection, while HRS might cause these effects at higher concentrations. However, further study is needed to elucidate the exact mechanism of their actions. PMID:20953394

  17. Effect of hydroalcoholic extract of Hibiscus rosa sinensis Linn. leaves in experimental colitis in rats

    PubMed Central

    Kandhare, Amit D; Raygude, Kiran S; Ghosh, Pinaki; Ghule, Arvindkumar E; Gosavi, Tejas P; Badole, Sachin L; Bodhankar, Subhash L

    2012-01-01

    Objective To elucidate the ameliorative effect of hydroalcoholic extract of leaves of Hibiscus rosa sinensis (HRS) in acetic acid induced experimental colitis in male wistar rats. Methods The animals were administered with 2 mL acetic acid (4%) via intra rectal. The animals were divided into various treatment groups (n=6). Prednisolone was used as standard drug and HRS was administered at a dose of 50, 100 and 200 mg/kg p.o. The control group of animals received 1 mL of vehicle (distilled water). Ulcer area, ulcer index, spleen weight, colon weight to length ratio, macroscopic score, haematological parameters, colonic superoxide dismutase (SOD), glutathione (GSH), myeloperoxidase (MPO), malondialdehyde (MDA), tumor necrosis factor-α (TNF-α), nitric oxide (NO) and histological changes were recorded after the treatment regimen of 11 days. Results Intrarectal instillation of acetic acid caused enhanced ulcer area, ulcer index, spleen weight, colon weight to length ratio, colonic MPO, MDA, NO and TNF-α It caused significant decreased level of SOD and GSH. Pretreatment with HRS for 7 days exhibited significant effect in lowering of oxidative stress, colonic NO, TNF-α and elevation of SOD and GSH at a dose of 100 and 200 mg/kg in acetic acid induced colitis. Conclusions The present investigation demonstrates HRS is of potent therapeutic value in the amelioration of experimental colitis in laboratory animals by inhibiting the proinflammatory mediator like NO and TNF-α. PMID:23569927

  18. Incubation temperature affects the immune function of hatchling soft-shelled turtles, Pelodiscus sinensis.

    PubMed

    Dang, Wei; Zhang, Wen; Du, Wei-Guo

    2015-06-01

    Identifying how developmental temperature affects the immune system is critical for understanding how ectothermic animals defend against pathogens and their fitness in the changing world. However, reptiles have received little attention regarding this issue. We incubated eggs at three ecologically relevant temperatures to determine how incubation temperature affects the immune function of hatchling soft-shelled turtles, Pelodiscus sinensis. When exposed to bacterial infections, hatchlings from 24 °C had lower cumulative mortalities (55%, therefore, higher immunocompetence) than those from 28 °C (85%) or 32 °C (100%). Consistent with higher immunocompetence, hatchlings from low incubation temperature had higher IgM, IgD, and CD3γ expressions than their counterparts from the other two higher incubation temperatures. Conversely, the activity of immunity-related enzymes did not match the among-temperature difference in immune function. Specifically, enzyme activity was higher at intermediate temperatures (alkaline phosphatase) or was not affected by incubation temperature (acid phosphatase, lysozyme). Our study is the first to provide unequivocal evidence (at the molecular and organismal level) about the significant effect of incubation temperature on offspring immunity in reptiles. Our results also indicate that the reduced immunity induced by high developmental temperatures might increase the vulnerability of reptiles to the outbreak of diseases under global warming scenarios.

  19. Cloning, characterization and localization of CHS gene from blood orange, Citrus sinensis (L.) Osbeck cv. Ruby.

    PubMed

    Lu, Xu; Zhou, Wei; Gao, Feng

    2009-09-01

    Chalcone synthase (CHS) is involved in the biosynthesis of anthocyanin. In this study, a full-length DNA of CHS gene (named as CsCHS-bo) was cloned from the blood orange, Citrus sinensis (L.) Osbeck cv. Ruby. The gene was 1,512 bp in size containing an open reading frame (1,176 bp) encoding 391 amino acids. Comparative and bioinformatic analyses revealed that the deduced protein of CsCHS-bo was highly homologous to CHS from other plant species. The protein of CsCHS-bo had four CHS-specific conserved motifs and a CHS-family signature sequence GFGPG. Phylogenetic analysis indicated that the protein of CsCHS-bo was in a subgroup with CHS of Ruta Palmatum. The CsCHS-bo was localized to the chromosomes 2p, 4p and 6p by an improved fluorescence in situ hybridization technique, indicating that at least three copies of CsCHS-bo were present in the genome.

  20. Proteomic analysis of somatic embryogenesis in Valencia sweet orange (Citrus sinensis Osbeck).

    PubMed

    Pan, Zhiyong; Guan, Rui; Zhu, Shiping; Deng, Xiuxin

    2009-02-01

    Two dimensional gel electrophoresis combined with matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) was employed to study the somatic embryogenesis (SE) in Valencia sweet orange (Citrus sinensis Osbeck). Twenty-four differentially expressed proteins were identified at five time points of citrus SE (0, 1, 2, 3, 4 weeks after embryo initiation) covering globular, heart/torpedo and cotyledon-shaped embryo stages. The general expression patterns for these proteins were consistent with those appeared at 4 weeks of citrus SE. The most striking feature of our study was that five proteins were predicted to be involved in glutathione (GSH) metabolism and anti-oxidative stress, and they exhibited different expression patterns during SE. Based on that oxidative stress has been validated to enhance SE, the preferential representation for anti-oxidative proteins suggests that they could have a developmental role in citrus SE. Some proteins involved in cell division, photosynthesis and detoxification were also identified, and their possible roles in citrus SE were discussed.

  1. Clinical evaluation of Moro (Citrus sinensis (L.) Osbeck) orange juice supplementation for the weight management.

    PubMed

    Cardile, Venera; Graziano, Adriana Carol Eleonora; Venditti, Alessandro

    2015-01-01

    In the last years, several studies have recently evaluated the beneficial effects of red orange juice (Citrus sinensis (L.) Osbeck) and its active components in weight management and obesity. Moro orange is a cultivar of red orange, particularly rich in active compounds such as anthocyanins, hydroxycinnamic acids, flavone glycosides and ascorbic acid, which displays anti-obesity effects in in vitro and in vivo studies. In this clinical study, the effect of a Moro juice extract (Morosil(®), 400 mg/die) supplementation was evaluated in overweight healthy human volunteers for 12 weeks. Results showed that Moro juice extract intake was able to induce a significant reduction in body mass index (BMI) after 4 weeks of treatment (p < 0.05). Moreover, in subjects treated with Moro extract, body weight, BMI, waist and hip circumference were significantly different from the placebo group (p < 0.05). In conclusion, it could be suggested that the active compounds contained in Moro juice have a synergistic effect on fat accumulation in humans and Moro juice extract can be used in weight management and in the prevention of human obesity.

  2. Variety and Harvesting Season Effects on Antioxidant Activity and Vitamins Content of Citrus sinensis Macfad.

    PubMed

    Cardeñosa, Vanessa; Barreira, João C M; Barros, Lillian; Arenas-Arenas, Francisco J; Moreno-Rojas, José M; Ferreira, Isabel C F R

    2015-05-07

    Five sweet orange (Citrus sinensis Osbeck) varieties cultivated in Huelva (Spain) and picked at two seasons during two consecutive years, were characterized for their antioxidant activity (free radicals scavenging activity, reducing power and lipid peroxidation inhibition) and vitamin content (vitamin E and vitamin C). The effects induced by sweet orange variety and stage of maturity were comprehensively compared by applying 2-way ANOVA and linear discriminant analysis. The results indicated higher differences in antioxidant activity and vitamin contents in response to the effect of the harvesting season, when compared to the effect of sweet orange variety. Nevertheless, the results observed in 2012 showed less marked differences among the assayed sweet orange varieties. Either way, it might be concluded that oranges sampled in January show the highest antioxidant activity and vitamin contents. Furthermore, concerning the properties evaluated in this work, all sweet orange varieties represent good alternatives, except for Rhode Summer, which would not be the preferable choice as a target to enhance sweet orange overall characteristics.

  3. Protective effects of sweet orange (Citrus sinensis) peel and their bioactive compounds on oxidative stress.

    PubMed

    Chen, Zong-Tsi; Chu, Heuy-Ling; Chyau, Charng-Cherng; Chu, Chin-Chen; Duh, Pin-Der

    2012-12-15

    Protective effects of sweet orange (Citrus sinensis) peel and their bioactive compounds on oxidative stress were investigated. According to HPLC-DAD and HPLC-MS/MS analysis, hesperidin (HD), hesperetin (HT), nobiletin (NT), and tangeretin (TT) were present in water extracts of sweet orange peel (WESP). The cytotoxic effect in 0.2mM t-BHP-induced HepG2 cells was inhibited by WESP and their bioactive compounds. The protective effect of WESP and their bioactive compounds in 0.2mM t-BHP-induced HepG2 cells may be associated with positive regulation of GSH levels and antioxidant enzymes, decrease in ROS formation and TBARS generation, increase in the mitochondria membrane potential and Bcl-2/Bax ratio, as well as decrease in caspase-3 activation. Overall, WESP displayed a significant cytoprotective effect against oxidative stress, which may be most likely because of the phenolics-related bioactive compounds in WESP, leading to maintenance of the normal redox status of cells.

  4. Selection of reference genes for expression analyses of red-fleshed sweet orange (Citrus sinensis).

    PubMed

    Pinheiro, T T; Nishimura, D S; De Nadai, F B; Figueira, A; Latado, R R

    2015-12-28

    Red-fleshed oranges (Citrus sinensis) contain high levels of carotenoids and lycopene. The growing consumer demand for products with health benefits has increased interest in these types of Citrus cultivars as a potential source of nutraceuticals. However, little is known about the physiology of these cultivars under Brazilian conditions. Transcriptome and gene expression analyses are important tools in the breeding and management of red-fleshed sweet orange cultivars. Reverse transcription quantitative polymerase chain reaction is a method of quantifying gene expression, but various standardizations are required to obtain precise, accurate, and specific results. Among the standardizations required, the choice of suitable stable reference genes is fundamental. The objective of this study was to evaluate the stability of 11 candidate genes using various tissue and organ samples from healthy plants or leaves from citrus greening disease (Huanglongbing)-symptomatic plants of a Brazilian red-fleshed cultivar ('Sanguínea de Mombuca'), in order to select the most suitable reference gene for investigating gene expression under these conditions. geNorm and NormFinder identified genes that encoded translation initiation factor 3, ribosomal protein L35, and translation initiation factor 5A as the most stable genes under the biological conditions tested, and genes coding actin (ACT) and the subunit of the PSI reaction center subunit III were the least stable. Phosphatase, malate dehydrogenase, and ACT were the most stable genes in the leaf samples of infected plants.

  5. Characterization of three terpenoid glycosyltransferase genes in 'Valencia' sweet orange (Citrus sinensis L. Osbeck).

    PubMed

    Fan, Jing; Chen, Chunxian; Yu, Qibin; Li, Zheng-Guo; Gmitter, Frederick G

    2010-10-01

    Three putative terpenoid UDP-glycosyltransferase (UGT) genes, designated CsUGT1, CsUGT2, and CsUGT3, were isolated and characterized in 'Valencia' sweet orange (Citrus sinensis L. Osbeck). CsUGT1 consisted of 1493 nucleotides with an open reading frame encoding 492 amino acids, CsUGT2 consisted of 1727 nucleotides encoding 504 amino acids, and CsUGT3 consisted of 1705 nucleotides encoding 468 amino acids. CsUGT3 had a 145 bp intron at 730-874, whereas CsUGT1 and CsUGT2 had none. The three deduced glycosyltransferase proteins had a highly conserved plant secondary product glycosyltransferase motif in the C terminus. Phylogenetic analysis showed that CsUGT1 and CsUGT3 were classified into group L of glycosyltransferase family 1, and CsUGT2 was classified into group D. Through Southern blotting analysis, CsUGT1 was found to have two copies in the sweet orange genome, whereas CsUGT2 and CsUGT3 had at least seven and nine copies, respectively. CsUGT1, CsUGT2, and CsUGT3 were constitutively expressed in leaf, flower, and fruit tissues. The results facilitate further investigation of the function of terpenoid glycosyltransferases in citrus and the biosynthesis of terpenoid glycosides in vitro.

  6. Brassinolide alleviates salt stress and increases antioxidant activity of cowpea plants (Vigna sinensis).

    PubMed

    El-Mashad, Ali Abdel Aziz; Mohamed, Heba Ibrahim

    2012-07-01

    Soil salinity is one of the most severe factors limiting growth and physiological response in Vigna sinensis plants. Plant salt stress tolerance requires the activation of complex metabolic activities including antioxidative pathways, especially reactive oxygen species and scavenging systems within the cells which can contribute to continued growth under water stress. The present investigation was carried out to study the role of brassinolide in enhancing tolerance of cowpea plants to salt stress (NaCl). Treatment with 0.05 ppm brassinolide as foliar spray mitigated salt stress by inducing enzyme activities responsible for antioxidation, e.g., superoxide dismutase, peroxidase, polyphenol oxidase, and detoxification as well as by elevating contents of ascorbic acid, tocopherol, and glutathione. On the other hand, total soluble proteins decreased with increasing NaCl concentrations in comparison with control plants. However, lipid peroxidation increased with increasing concentrations of NaCl. In addition to, the high concentrations of NaCl (100 and 150 mM) decreased total phenol of cowpea plants as being compared with control plants. SDS-PAGE of protein revealed that NaCl treatments alone or in combination with 0.05 ppm brassinolide were associated with the disappearance of some bands or appearance of unique ones in cowpea plants. Electrophoretic studies of α-esterase, β-esterase, polyphenol oxidase, peroxidase, acid phosphatase, and superoxide dismutase isoenzymes showed wide variations in their intensities and densities among all treatments.

  7. Metabolomic unveiling of a diverse range of green tea (Camellia sinensis) metabolites dependent on geography.

    PubMed

    Lee, Jang-Eun; Lee, Bum-Jin; Chung, Jin-Oh; Kim, Hak-Nam; Kim, Eun-Hee; Jung, Sungheuk; Lee, Hyosang; Lee, Sang-Jun; Hong, Young-Shick

    2015-05-01

    Numerous factors such as geographical origin, cultivar, climate, cultural practices, and manufacturing processes influence the chemical compositions of tea, in the same way as growing conditions and grape variety affect wine quality. However, the relationships between these factors and tea chemical compositions are not well understood. In this study, a new approach for non-targeted or global analysis, i.e., metabolomics, which is highly reproducible and statistically effective in analysing a diverse range of compounds, was used to better understand the metabolome of Camellia sinensis and determine the influence of environmental factors, including geography, climate, and cultural practices, on tea-making. We found a strong correlation between environmental factors and the metabolome of green, white, and oolong teas from China, Japan, and South Korea. In particular, multivariate statistical analysis revealed strong inter-country and inter-city relationships in the levels of theanine and catechin derivatives found in green and white teas. This information might be useful for assessing tea quality or producing distinct tea products across different locations, and highlights simultaneous identification of diverse tea metabolites through an NMR-based metabolomics approach.

  8. Quick and selective extraction of Z-ligustilide from Angelica sinensis using magnetic multiwalled carbon nanotubes.

    PubMed

    Zeng, Qiong; Jia, Yan-Wei; Xu, Pei-Li; Xiao, Meng-Wei; Liu, Yi-Ming; Peng, Shu-Lin; Liao, Xun

    2015-12-01

    A facile and highly efficient magnetic solid-phase extraction method has been developed for Z-ligustilide, the major therapeutic agent in Angelica sinensis. The solid-phase adsorbent material used was prepared by conjugating carbon nanotubes with magnetic Fe3 O4 nanoparticles via a hydrothermal reaction. The magnetic material showed a high affinity toward Z-ligustilide due to the π-π stacking interaction between the carbon nanotubes and Z-ligustilide, allowing a quick and selective exaction of Z-ligustilide from complex sample matrices. Factors influencing the magnetic solid-phase extraction such as the amount of the added adsorbent, adsorption and desorption time, and desorption solvent, were investigated. Due to its high extraction efficiency, this method was proved highly useful for sample cleanup/enrichment in quantitative high-performance liquid chromatography analysis. The proposed method had a linear calibration curve (R(2) = 0.9983) over the concentration between 4 ng/mL and 200 μg/mL Z-ligustilide. The accuracy of the method was determined by the recovery, which was from 92.07 to 104.02%, with the relative standard deviations >4.51%.

  9. Estimating Biochemical Parameters of Tea (camellia Sinensis (L.)) Using Hyperspectral Techniques

    NASA Astrophysics Data System (ADS)

    Bian, M.; Skidmore, A. K.; Schlerf, M.; Liu, Y.; Wang, T.

    2012-07-01

    Tea (Camellia Sinensis (L.)) is an important economic crop and the market price of tea depends largely on its quality. This research aims to explore the potential of hyperspectral remote sensing on predicting the concentration of biochemical components, namely total tea polyphenols, as indicators of tea quality at canopy scale. Experiments were carried out for tea plants growing in the field and greenhouse. Partial least squares regression (PLSR), which has proven to be the one of the most successful empirical approach, was performed to establish the relationship between reflectance and biochemical concentration across six tea varieties in the field. Moreover, a novel integrated approach involving successive projections algorithms as band selection method and neural networks was developed and applied to detect the concentration of total tea polyphenols for one tea variety, in order to explore and model complex nonlinearity relationships between independent (wavebands) and dependent (biochemicals) variables. The good prediction accuracies (r2 > 0.8 and relative RMSEP < 10 %) achieved for tea plants using both linear (partial lease squares regress) and nonlinear (artificial neural networks) modelling approaches in this study demonstrates the feasibility of using airborne and spaceborne sensors to cover wide areas of tea plantation for in situ monitoring of tea quality cheaply and rapidly.

  10. Transcriptome analysis of a spontaneous mutant in sweet orange [Citrus sinensis (L.) Osbeck] during fruit development

    PubMed Central

    Liu, Qing; Zhu, Andan; Chai, Lijun; Zhou, Wenjing; Yu, Keqin; Ding, Jian; Xu, Juan; Deng, Xiuxin

    2009-01-01

    Bud mutations often arise in citrus. The selection of mutants is one of the most important breeding channels in citrus. However, the molecular basis of bud mutation has rarely been studied. To identify differentially expressed genes in a spontaneous sweet orange [C. sinensis (L.) Osbeck] bud mutation which causes lycopene accumulation, low citric acid, and high sucrose in fruit, suppression subtractive hybridization and microarray analysis were performed to decipher this bud mutation during fruit development. After sequencing of the differentially expressed clones, a total of 267 non-redundant transcripts were obtained and 182 (68.2%) of them shared homology (E-value ≤1×10−10) with known gene products. Few genes were constitutively up- or down-regulated (fold change ≥2) in the bud mutation during fruit development. Self-organizing tree algorithm analysis results showed that 95.1% of the differentially expressed genes were extensively coordinated with the initiation of lycopene accumulation. Metabolic process, cellular process, establishment of localization, response to stimulus, and biological regulation-related transcripts were among the most regulated genes. These genes were involved in many biological processes such as organic acid metabolism, lipid metabolism, transport, and pyruvate metabolism, etc. Moreover, 13 genes which were differentially regulated at 170 d after flowering shared homology with previously described signal transduction or transcription factors. The information generated in this study provides new clues to aid in the understanding of bud mutation in citrus. PMID:19218315

  11. Cognitive enhancing and antioxidant activity of ethyl acetate soluble fraction of the methanol extract of Hibiscus rosa sinensis in scopolamine-induced amnesia

    PubMed Central

    Nade, Vandana S.; Kanhere, Sampat V.; Kawale, Laxman A.; Yadav, Adhikrao V.

    2011-01-01

    Objective: The objective of the present study was to evaluate the cognitive enhancing and antioxidant activity of Hibiscus rosa sinensis. Materials and Methods: The learning and memory was impaired by administration of scopolamine (1 mg/kg, i.p.) in mice which is associated with altered brain oxidative status. The object recognition test (ORT) and passive avoidance test (PAT) were used to assess cognitive enhancing activity. Animals were treated with an ethyl acetate soluble fraction of the methanol extract of H. sinensis (25, 50 and 100 mg/kg, p.o). Results: The ethyl acetate soluble fraction of the methanol extract of H. sinensis (EASF) attenuated amnesia induced by scopolamine and aging. The discrimination index (DI) was significantly decreased in the aged and scopolamine group in ORT. Pretreatment with EASF significantly increased the DI. In PAT, scopolamine-treated mice exhibited significantly shorter step-down latencies (SDL). EASF treatment showed a significant increase in SDL in young, aged as well as in scopolamine-treated animals. The biochemical analysis of brain revealed that scopolamine treatment increased lipid peroxidation and decreased levels of superoxide dismutase (SOD) and glutathione reductase (GSH). Administration of extract significantly reduced LPO and reversed the decrease in brain SOD and GSH levels. The administration of H. sinensis improved memory in amnesic mice and prevented the oxidative stress associated with scopolamine. The mechanism of such protection of H. sinensis may be due to augmentation of cellular antioxidants. Conclusion: The results of the present study suggested that H. sinensis had a protective role against age and scopolamine-induced amnesia, indicating its utility in management of cognitive disorders. PMID:21572646

  12. Cordyceps sinensis: Genotoxic Potential in Human Peripheral Blood Cells and Antigenotoxic Properties Against Hydrogen Peroxide by Comet Assay.

    PubMed

    Vasiljevic, Jovana D; Zivkovic, Lada P; Cabarkapa, Andrea M; Bajic, Vladan P; Djelic, Ninoslav J; Spremo-Potparevic, Biljana M

    2016-06-01

    Context • Cordyceps sinensis (C sinensis) is a well-known, traditional, Chinese medicinal mushroom, valued for its beneficial properties for human health. C sinensis has been reported to have immunomodulatory, anticancer, antiaging, antioxidant and anti-inflammatory activity. Despite potential medicinal benefits, no previously published reports are available about the genotoxicity or antigenotoxicity of C sinensis, as detected by comet assay. Objective • The objective of the study was to evaluate both the genotoxic and antigenotoxic potential of an extract of C sinensis (CS extract) in human peripheral blood cells. Design • The research team designed a pilot study. Setting •The study was conducted at the Center for Biological Research, University of Belgrade, in Belgrade, Serbia. Participants • Participants were 6 healthy individuals (2 males and 4 females), between the ages of 20 and 45 y, recruited on a voluntary basis, who provided heparinized, peripheral blood samples. Intervention • Four concentrations of the CS extract-125 μg/mL, 250 μg/mL, 500 μg/mL, and 1000 μg/mL-were used in the treatment of tested blood cells from the blood samples. Three independent procedures were performed: (1) a genotoxicity assessment, (2) an antigenotoxicity assessment for pretreatment of human cells with the CS extract prior to their exposure to hydrogen peroxide (H2O2) (ie, an evaluation of the benefits of the CS extract as a preventive agent); and (3) posttreatment of human cells with the CS extract after their exposure to H2O2 (ie, an evaluation of the benefits of the CS extract as an interventional agent). Outcome Measures • Cells were graded by eye inspection into 5 classes, depending on the extent of DNA damage, representing: (1) class A-undamaged cells with no tail (<5% damaged DNA); (2) class B-low-level damage (5%-20%); (3) class C-medium-level damage (20%-40%); (4) class D-high-level damage (40%-95%), and (5) class E-total destruction (>95%).Results

  13. Angelica sinensis polysaccharides promotes apoptosis in human breast cancer cells via CREB-regulated caspase-3 activation.

    PubMed

    Zhou, Wei-Jie; Wang, Sheng; Hu, Zhuang; Zhou, Zhen-Yu; Song, Cai-Juan

    2015-11-20

    Angelica sinensis polysaccharide (ASP) is purified from the fresh roots of Angelica sinensis (AS). This traditional Chinese medicine has been used for thousands of years for treating gynecological diseases and used in functional foods for the prevention and treatment of various diseases, such as inflammation and cancer. The antitumor activity of ASP is related to its biological activities, because it suppresses a variety of pro-proliferative or anti-apoptotic factors that are dramatically expressed in cancer cells of given types. In this study, we show that angelica sinensis polysaccharide induced apoptosis in breast cancer cells of T47D over-expressing the Cyclic AMP response element binding protein (CREB), inducing apoptosis-related signaling pathway activity. The result also found that ASP caused cell death was linked to caspase activity, accompanied by the loss of mitochondrial membrane potential, cytochrome c release, and Bax translocation from the cytosol to the mitochondria. We found that ASP significantly affected the poly-ADP-ribose polymerase (PARP), Bcl-2 Associated X Protein (Bax), Bcl-2, Bcl-xL and apoptotic protease activating facter-1 (Apaf1) protein expression in a dose- and time-dependent manner. DAPI staining and Flow cytometry were used to analyze apoptosis. The nude mice xenograft model was used to evaluate the antitumor effect of ASP in vivo. ASP has profound antitumor effect on T47D cells, probably by inducing apoptosis through CREB signaling pathway. Thus, these results suggest that ASP would be a promising therapeutic agent for breast cancer.

  14. Isolation and dynamic expression of four genes involving in shikimic acid pathway in Camellia sinensis 'Baicha 1' during periodic albinism.

    PubMed

    Zhu, Xu-Jun; Zhao, Zhen; Xin, Hua-Hong; Wang, Ming-Le; Wang, Wei-Dong; Chen, Xuan; Li, Xing-Hui

    2016-10-01

    Flavonoids are the main flavor components and functional ingredients in tea, and the shikimic acid pathway is considered as one of the most important pathways in flavonoid biosynthesis, but little was known about the function of regulatory genes in the metabolism phenolic compounds in tea plant (Camellia sinensis), especially related genes in shikimic acid pathway. The dynamic changes of catechin (predominant flavonoid) contents were analyzed in this study, and four genes (CsPPT, CsDAHPS, CsSDH and CsCS) involving in shikimic acid pathway in C. sinensis albino cultivar 'Baicha 1' were cloned and characterized. The full-length cDNA sequences of these genes were obtained using reverse transcription-PCR and rapid amplification of cDNA ends. At the albinistic stage, the amounts of all catechins decreased to the lowest levels, when epigallocatechin gallate was the highest, whereas gallocatechin-3-O-gallate the lowest. Gene expression patterns analyzed by qRT-PCR showed that CsPPT and CsDAHPS were highly expressed in flowers and buds, while CsSDH and CsCS showed high expression levels in buds and leaves. It was also found that the transcript abundance of shikimic acid biosynthetic genes followed a tightly regulated biphasic pattern, and was affected by albinism. The transcript levels of CsPPT and CsDAHPS were decreased at albinistic stage followed elevated expression, whereas CsSDH and CsCS were increased only at re-greening stage. Taken together, these findings suggested that these four genes in C. sinensis may play different roles in shikimic acid biosynthesis and these genes may have divergent functions.

  15. Knockdown resistance in Anopheles vagus, An. sinensis, An. paraliae and An. peditaeniatus populations of the Mekong region

    PubMed Central

    2010-01-01

    Background In the Mekong region (Vietnam, Cambodia and Laos), a large investigation was conducted to assess the susceptibility of Anopheles species against DDT and pyrethroids. In this study, the resistance status of the potential malaria vectors An. vagus, An. sinensis, An. paraliae and An. peditaeniatus was assessed. Methods Bioassays were performed on field collected unfed female mosquitoes using the standard WHO susceptibility tests. In addition, the DIIS6 region of the para-type sodium channel gene was amplified and sequenced and four allele-specific PCR assays were developed to assess the kdr frequencies. Results In Southern Vietnam all species were DDT and pyrethroid resistant, which might suggest the presence of a kdr resistance mechanism. Sequence-analysis of the DIIS6 region of the para-type sodium channel gene revealed the presence of a L1014S kdr mutation in An. vagus, An. sinensis and An. paraliae. In An. peditaeniatus, a low frequency L1014S kdr mutation was found in combination with a high frequency L1014F kdr mutation. For pyrethroids and DDT, no genotypic differentiation was found between survivors and non-survivors for any of these species. In the two widespread species, An. vagus and An. sinensis, kdr was found only in southern Vietnam and in Cambodia near the Vietnamese border. Conclusions Different levels of resistance were measured in Laos, Cambodia and Vietnam. The kdr mutation in different Anopheles species seems to occur in the same geographical area. These species breed in open agricultural lands where malaria endemicity is low or absent and vector control programs less intensive. It is therefore likely that the selection pressure occurred on the larval stages by insecticides used for agricultural purposes. PMID:20646327

  16. Microwave-Assisted Extraction, Chemical Structures, and Chain Conformation of Polysaccharides from a Novel Cordyceps Sinensis Fungus UM01.

    PubMed

    Cheong, Kit-Leong; Wang, Lan-Ying; Wu, Ding-Tao; Hu, De-Jun; Zhao, Jing; Li, Shao-Ping

    2016-09-01

    Cordyceps sinensis is a well-known tonic food with broad medicinal properties. The aim of the present study was to investigate the optimization of microwave-assisted extraction (MAE) and characterize chemical structures and chain conformation of polysaccharides from a novel C. sinensis fungus UM01. Ion-exchange and gel filtration chromatography were used to purify the polysaccharides. The chemical structure of purified polysaccharide was determined through gas chromatography-mass spectrometry. Moreover, high performance size exclusion chromatography combined with refractive index detector and multiangle laser light scattering were conducted to analyze the molecular weight (Mw ) and chain conformation of purified polysaccharide. Based on the orthogonal design L9 , optimal MAE conditions could be obtained through 1300 W of microwave power, with a 5-min irradiation time at a solid to water ratio of 1:60, generating the highest extraction yield of 6.20%. Subsequently, the polysaccharide UM01-S1 was purified. The UM01-S1 is a glucan-type polysaccharide with a (1→4)-β-d-glucosyl backbone and branching points located at O-3 of Glcp with a terminal-d-Glcp. The Mw , radius of gyration (Rg ) and hydrodynamic radius (Rh ) of UM01-S1 were determined as 5.442 × 10(6)  Da, 21.8 and 20.2 nm, respectively. Using the polymer solution theory, the exponent (ν) value of the power law function was calculated as 0.38, and the shape factor (ρ = Rg /Rh ) was 1.079, indicating that UM01-S1 has a sphere-like conformation with a branched structure in an aqueous solution. These results provide fundamental information for the future application of polysaccharides from cultured C. sinensis in health and functional food area.

  17. Identification of UDP-glycosyltransferases involved in the biosynthesis of astringent taste compounds in tea (Camellia sinensis)

    PubMed Central

    Cui, Lilan; Yao, Shengbo; Dai, Xinlong; Yin, Qinggang; Liu, Yajun; Jiang, Xiaolan; Wu, Yahui; Qian, Yumei; Pang, Yongzhen; Gao, Liping; Xia, Tao

    2016-01-01

    Galloylated catechins and flavonol 3-O-glycosides are characteristic astringent taste compounds in tea (Camellia sinensis). The mechanism involved in the formation of these metabolites remains unknown in tea plants. In this paper, 178 UGT genes (CsUGTs) were identified in C. sinensis based on an analysis of tea transcriptome data. Phylogenetic analysis revealed that 132 of these genes were clustered into 15 previously established phylogenetic groups (A to M, O and P) and a newly identified group R. Three of the 11 recombinant UGT proteins tested were found to be involved in the in vitro biosynthesis of β-glucogallin and glycosylated flavonols. CsUGT84A22 exhibited catalytic activity toward phenolic acids, in particular gallic acid, to produce β-glucogallin, which is the immediate precursor of galloylated catechin biosynthesis in tea plants. CsUGT78A14 and CsUGT78A15 were found to be responsible for the biosynthesis of flavonol 3-O-glucosides and flavonol 3-O-galactosides, respectively. Site-directed mutagenesis of the Q373H substitution for CsUGT78A14 indicated that the Q (Gln) residue played a catalytically crucial role for flavonoid 3-O-glucosyltransferase activity. The expression profiles of the CsUGT84A22, CsUGT78A14, and CsUGT78A15 genes were correlated with the accumulation patterns of β-glucogallin and the glycosylated flavonols which indicated that these three CsUGT genes were involved in the biosynthesis of astringent compounds in C. sinensis. PMID:26941235

  18. Green synthesis of gold nanoparticles using Citrus fruits (Citrus limon, Citrus reticulata and Citrus sinensis) aqueous extract and its characterization.

    PubMed

    Sujitha, Mohanan V; Kannan, Soundarapandian

    2013-02-01

    This study reports the biological synthesis of gold nanoparticles by the reduction of HAuCl(4) by using citrus fruits (Citrus limon, Citrus reticulata and Citrus sinensis) juice extract as the reducing and stabilizing agent. A various shape and size of gold nanoparticles were formed when the ratio of the reactants were altered with respect to 1.0mM chloroauric acid solution. The gold nanoparticles obtained were characterized by UV-visible spectra, transmission electron microscopy (TEM) and X-ray diffraction (XRD). TEM studies showed the particles to be of various shapes and sizes and particle size ranges from 15 to 80 nm. Selected-area electron diffraction (SAED) pattern confirmed fcc phase and crystallinity of the particles. The X-ray diffraction analysis revealed the distinctive facets (111, 200, 220 and 222 planes) of gold nanoparticles. Dynamic light scattering (DLS) studies revealed that the average size for colloid gp(3) of C. limon, C. reticulata and C. sinensis are 32.2 nm, 43.4 nm and 56.7 nm respectively. The DLS graph showed that the particles size was larger and more polydispersed compared to the one observed by TEM due to the fact that the measured size also includes the bio-organic compounds enveloping the core of the Au NPs. Zeta potential value for gold nanoparticles obtained from colloid gp(3) of C. limon, C. reticulata and C. sinensis are -45.9, -37.9 and -31.4 respectively indicating the stability of the synthesized nanoparticles. Herein we propose a novel, previously unexploited method for the biological syntheses of polymorphic gold nanoparticles with potent biological applications.

  19. Antidepressant-like activity of anthocyanidins from Hibiscus rosa-sinensis flowers in tail suspension test and forced swim test

    PubMed Central

    Shewale, Pallavi B.; Patil, Rupali A.; Hiray, Yogesh A.

    2012-01-01

    Aim: Flowers of Hibiscus rosa-sinensis Linn (Malvaceae) popularly known as “China-rose flowers” contain flavonoids. Flavonoids have been found to have antidepressant activity. The aim of the present study is to evaluate the antidepressant activity of flavonoids in H. rosa-sinensis flowers with possible involvement of monoamines. Materials and Methods: Anti-depressant activity of methanol extract containing anthocyanins (MHR) (30 and 100 mg/kg) and anthocyanidins (AHR) (30 and 100 mg/ kg) of H. rosa-sinensis flowers were evaluated in mice using behavioral tests such as tail suspension test (TST) and forced swim test (FST). The mechanism of action involved in antidepressant activity was investigated by observing the effect of extract after pre-treatment with low dose haloperidol, prazosin and para-chlorophenylalanine (p-CPA). Results: Present study exhibited significant decrease in immobility time in TST and FST, similar to that of imipramine (10 mg/kg, i.p.) which served as a positive control. The extract significantly attenuated the duration of immobility induced by Haloperidol (50 μg/ kg, i.p., a classical D2-like dopamine receptor antagonist), Prazosin (62.5 μg/kg, i.p., an α1-adrenoceptor antagonist) and p-chlorophenylalanine (100 mg/kg, i.p., × 3 days; an inhibitor of serotonin synthesis) in both TST and FST. Conclusion: It can be concluded that MHR and AHR possess potential antidepressant activity (through dopaminergic, noradrenergic and serotonergic mechanisms) and has therapeutic potential in the treatment of CNS disorders and provides evidence at least at preclinical levels. PMID:23087504

  20. Green synthesis of gold nanoparticles using Citrus fruits (Citrus limon, Citrus reticulata and Citrus sinensis) aqueous extract and its characterization

    NASA Astrophysics Data System (ADS)

    Sujitha, Mohanan V.; Kannan, Soundarapandian

    2013-02-01

    This study reports the biological synthesis of gold nanoparticles by the reduction of HAuCl4 by using citrus fruits (Citrus limon, Citrus reticulata and Citrus sinensis) juice extract as the reducing and stabilizing agent. A various shape and size of gold nanoparticles were formed when the ratio of the reactants were altered with respect to 1.0 mM chloroauric acid solution. The gold nanoparticles obtained were characterized by UV-visible spectra, transmission electron microscopy (TEM) and X-ray diffraction (XRD). TEM studies showed the particles to be of various shapes and sizes and particle size ranges from 15 to 80 nm. Selected-area electron diffraction (SAED) pattern confirmed fcc phase and crystallinity of the particles. The X-ray diffraction analysis revealed the distinctive facets (1 1 1, 2 0 0, 2 2 0 and 2 2 2 planes) of gold nanoparticles. Dynamic light scattering (DLS) studies revealed that the average size for colloid gp3 of C. limon, C. reticulata and C. sinensis are 32.2 nm, 43.4 nm and 56.7 nm respectively. The DLS graph showed that the particles size was larger and more polydispersed compared to the one observed by TEM due to the fact that the measured size also includes the bio-organic compounds enveloping the core of the Au NPs. Zeta potential value for gold nanoparticles obtained from colloid gp3 of C. limon, C. reticulata and C. sinensis are -45.9, -37.9 and -31.4 respectively indicating the stability of the synthesized nanoparticles. Herein we propose a novel, previously unexploited method for the biological syntheses of polymorphic gold nanoparticles with potent biological applications.

  1. Multi-class mycotoxins analysis in Angelica sinensis by ultra fast liquid chromatography coupled with tandem mass spectrometry.

    PubMed

    Liu, Qiutao; Kong, Weijun; Guo, Weiying; Yang, Meihua

    2015-04-15

    An ultra fast liquid chromatography coupled with tandem mass spectrometry (UFLC-MS/MS) method was developed and validated for simultaneous analysis of multi-class mycotoxins including aflatoxins (AFB1, AFB2, AFG1 and AFG2), ochratoxin A (OTA), fumonisins (FB1 and FB2) and zearalanone (ZEN) in 20 batches of Angelica sinensis samples collected from different markets and stores in China. The eight mycotoxins were extracted and cleaned up by using QuEChERS-based procedure, and then were quantified under the multiple reaction monitoring (MRM) together with positive and negative ionization modes. Focusing on the optimization of extraction and clean-up conditions, as well as UFLC separation and MS/MS parameters of targeted analytes, the developed method expressed good linearity for the eight mycotoxins within their respective linear ranges with correlation coefficients all higher than 0.9974. The limits of detection (LODs) and quantification (LOQs) ranged from 0.005 to 0.125 μg/kg and from 0.0625 to 0.25 μg/kg, respectively. Recoveries for spiked A. sinensis sample at three different levels were all above 78.9% with relative standard deviations (RSDs) below 6.36% for all analytes. Analysis of real samples demonstrated that two visibly moldy A. sinensis samples were detected with AFB1 of 2.07 and 2.92 μg/kg, and AFG1 of 2.84 and 1.53 μg/kg. The proposed quantitative method with significant advantages including simple pretreatment, rapid determination and high sensitivity would be the preferred candidate for the determination and quantification of multi-class mycotoxin contaminants in complex matrixes, which well fulfilled the maximum residue limits (MRLs) from various countries.

  2. Transcriptome Analysis and Discovery of Genes Involved in Immune Pathways from Hepatopancreas of Microbial Challenged Mitten Crab Eriocheir sinensis

    PubMed Central

    Li, Xihong; Cui, Zhaoxia; Liu, Yuan; Song, Chengwen; Shi, Guohui

    2013-01-01

    Background The Chinese mitten crab Eriocheir sinensis is an important economic crustacean and has been seriously attacked by various diseases, which requires more and more information for immune relevant genes on genome background. Recently, high-throughput RNA sequencing (RNA-seq) technology provides a powerful and efficient method for transcript analysis and immune gene discovery. Methods/Principal Findings A cDNA library from hepatopancreas of E. sinensis challenged by a mixture of three pathogen strains (Gram-positive bacteria Micrococcus luteus, Gram-negative bacteria Vibrio alginolyticus and fungi Pichia pastoris; 108 cfu·mL−1) was constructed and randomly sequenced using Illumina technique. Totally 39.76 million clean reads were assembled to 70,300 unigenes. After ruling out short-length and low-quality sequences, 52,074 non-redundant unigenes were compared to public databases for homology searching and 17,617 of them showed high similarity to sequences in NCBI non-redundant protein (Nr) database. For function classification and pathway assignment, 18,734 (36.00%) unigenes were categorized to three Gene Ontology (GO) categories, 12,243 (23.51%) were classified to 25 Clusters of Orthologous Groups (COG), and 8,983 (17.25%) were assigned to six Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. Potentially, 24, 14, 47 and 132 unigenes were characterized to be involved in Toll, IMD, JAK-STAT and MAPK pathways, respectively. Conclusions/Significance This is the first systematical transcriptome analysis of components relating to innate immune pathways in E. sinensis. Functional genes and putative pathways identified here will contribute to better understand immune system and prevent various diseases in crab. PMID:23874555

  3. A new 2-(2-phenylethyl)chromone derivative in Chinese agarwood 'Qi-Nan' from Aquilaria sinensis.

    PubMed

    Yang, De-Lan; Wang, Hao; Guo, Zhi-Kai; Dong, Wen-Hua; Mei, Wen-Li; Dai, Hao-Fu

    2014-01-01

    Phytochemical analysis of the high quality Chinese agarwood 'Qi-Nan' originating from Aquilaria sinensis (Lour.) Glig led to the isolation of a new 2-(2-phenylethyl)chromone derivative, qinanones G (1), and four known 2-(2-phenylethyl)chromones (2-5). Their structures were elucidated by spectroscopic techniques (UV, IR, 1D and 2D NMR) and MS analyses. The NMR data of chromones 1-3 were first reported, and chromones 2 and 3 showed weak inhibitory activity against acetylcholinesterase.

  4. A new 2-(2-phenylethyl)chromone glycoside in Chinese agarwood "Qi-Nan" from Aquilaria sinensis.

    PubMed

    Shao, Hang; Mei, Wen-Li; Kong, Fan-Dong; Dong, Wen-Hua; Li, Wei; Zhu, Guo-Peng; Dai, Hao-Fu

    2017-01-01

    A new 2-(2-phenylethyl)chromone glycoside, 2-[2-(4-glucosyloxy-3-methoxyphenyl)ethyl]chromone (1), was isolated from the high-quality Chinese agarwood "Qi-Nan" originating from Aquilaria sinensis (Lour.) Glig. The structure including the absolute configuration of the sugar moiety was elucidated by spectroscopic techniques (UV, IR, 1D and 2D NMR), MS analysis, PMP-labeling HPLC analysis methods, as well as comparison with literature data. To the best of our knowledge, it is the first time that chromone glycoside was discovered in agarwood, or even in the whole Aquilaria plants.

  5. Extraction process optimization of polyphenols from Indian Citrus sinensis – as novel antiglycative agents in the management of diabetes mellitus

    PubMed Central

    2014-01-01

    Background Diabetes mellitus is a chronic metabolic disorder characterized by increased blood glucose level. It has become an epidemic disease in the 21st century where, India leads the world with largest number of diabetic subjects. Non-enzymatic glycosylation (glycation) is severe form of diabetes, occurs between reducing sugar and proteins which results in the formation of advanced glycation end products (AGEs) that leads to the other complicated secondary disorders. In this context, Mangifera indica (Mango), Syzygium cumini (Jambul), Vitis vinifera (Grapes), Citrus sinensis (Orange), Artocarpus heterophyllus (Jackfruit), Manilkara zapota (Sapodilla) seeds were evaluated for their antiglyation activity. Attempts were made to isolate the polyphenols in the seeds that have recorded the maximum activity. Methods Different extraction methods (shake flask, centrifugation and pressurized hot water) using various extractants (organic solvents, hot water and pressurized hot water) were adopted to investigate the in vitro antiglycation activity. Central composite (CCD) design based Response Surface Methodology (RSM) was espoused to optimize the extraction process of polyphenols from the fruit seeds that have recorded poor antiglycation activity. The PTLC analysis was performed to isolate the polyphenols (Flavonoids and phenolic acids) and LC-PDA-MS analysis was done for structure prediction. Results Pressurized hot water extraction of Artocarpus heterophyllus (87.52%) and Citrus sinensis seeds (74.79%) was found to possess high and low antiglycation activity, respectively. The RSM mediated optimization process adopted for the Citrus sinensis seeds have revealed that 1:15 solvent ratio (hexane to heptane), 6 minutes and 1:20 solid to liquid ratio as the optimal conditions for the extraction of polyphenols with a maximum antiglycation activity (89.79%). The LC-PDA-MS analysis of preparative thin layer chromatography (PTLC) eluates of Artocarpus heterophyllus seed has

  6. Deep sequencing of the Camellia sinensis transcriptome revealed candidate genes for major metabolic pathways of tea-specific compounds

    SciTech Connect

    Shi, CY; Yang, H; Wei, CL; Yu, O; Zhang, ZZ; Sun, J; Wan, XC

    2011-01-01

    Tea is one of the most popular non-alcoholic beverages worldwide. However, the tea plant, Camellia sinensis, is difficult to culture in vitro, to transform, and has a large genome, rendering little genomic information available. Recent advances in large-scale RNA sequencing (RNA-seq) provide a fast, cost-effective, and reliable approach to generate large expression datasets for functional genomic analysis, which is especially suitable for non-model species with un-sequenced genomes. Using high-throughput Illumina RNA-seq, the transcriptome from poly (A){sup +} RNA of C. sinensis was analyzed at an unprecedented depth (2.59 gigabase pairs). Approximate 34.5 million reads were obtained, trimmed, and assembled into 127,094 unigenes, with an average length of 355 bp and an N50 of 506 bp, which consisted of 788 contig clusters and 126,306 singletons. This number of unigenes was 10-fold higher than existing C. sinensis sequences deposited in GenBank (as of August 2010). Sequence similarity analyses against six public databases (Uniprot, NR and COGs at NCBI, Pfam, InterPro and KEGG) found 55,088 unigenes that could be annotated with gene descriptions, conserved protein domains, or gene ontology terms. Some of the unigenes were assigned to putative metabolic pathways. Targeted searches using these annotations identified the majority of genes associated with several primary metabolic pathways and natural product pathways that are important to tea quality, such as flavonoid, theanine and caffeine biosynthesis pathways. Novel candidate genes of these secondary pathways were discovered. Comparisons with four previously prepared cDNA libraries revealed that this transcriptome dataset has both a high degree of consistency with previous EST data and an approximate 20 times increase in coverage. Thirteen unigenes related to theanine and flavonoid synthesis were validated. Their expression patterns in different organs of the tea plant were analyzed by RT-PCR and quantitative real

  7. Improvement of innate immune responses and defense activity in mitten crab (Eriocheir sinensis) by oral administration of beta-glucan.

    PubMed

    Zhou, Jin; Wang, Yong-Hong; Chu, Ju; Luo, Ling-Zhi

    2008-10-01

    The beta-glucan (BG), extracted from Saccharomyces cerevisiae cell wall, was orally administrated to the mitten crab (Eriocheir sinensis) at 0, 1, 5 and 10 g BG/kg diet for 28 days, followed by a challenge with Vibrio mimicus by intramuscular injection. Growth, phenoloxidase, superoxide dismutase, acid phosphatase and alkaline phosphatase activity were monitored after 14 and 28 days. The results showed an immunomodulatory effect and protection against V. mimicus by dietary supplementation of BG. The recommended concentration is 5 g BG/kg diet.

  8. CPR: Adult

    MedlinePlus

    Refresher Center Home FIRST AID, CPR and AED LIFEGUARDING Refresher Putting It All Together: CPR—Adult (2:03) Refresher videos only utilize this player QUICK LINKS Home RedCross.org Purchase Course ...

  9. A Systematic Review of the Mysterious Caterpillar Fungus Ophiocordyceps sinensis in Dong-ChongXiaCao (冬蟲夏草 Dōng Chóng Xià Cǎo) and Related Bioactive Ingredients

    PubMed Central

    Lo, Hui-Chen; Hsieh, Chienyan; Lin, Fang-Yi; Hsu, Tai-Hao

    2013-01-01

    The caterpillar fungus Ophiocordyceps sinensis (syn.† Cordyceps sinensis), which was originally used in traditional Tibetan and Chinese medicine, is called either “yartsa gunbu” or “DongChongXiaCao (冬蟲夏草 Dōng Chóng Xià Cǎo)” (“winter worm-summer grass”), respectively. The extremely high price of DongChongXiaCao, approximately USD $20,000 to 40,000 per kg, has led to it being regarded as “soft gold” in China. The multi-fungi hypothesis has been proposed for DongChongXiaCao; however, Hirsutella sinensis is the anamorph of O. sinensis. In Chinese, the meaning of “DongChongXiaCao” is different for O. sinensis, Cordyceps spp.,‡ and Cordyceps spƒ. Over 30 bioactivities, such as immunomodulatory, antitumor, anti-inflammatory, and antioxidant activities, have been reported for wild DongChongXiaCao and for the mycelia and culture supernatants of O. sinensis. These bioactivities derive from over 20 bioactive ingredients, mainly extracellular polysaccharides, intracellular polysaccharides, cordycepin, adenosine, mannitol, and sterols. Other bioactive components have been found as well, including two peptides (cordymin and myriocin), melanin, lovastatin, γ-aminobutyric acid, and cordysinins. Recently, the bioactivities of O. sinensis were described, and they include antiarteriosclerosis, antidepression, and antiosteoporosis activities, photoprotection, prevention and treatment of bowel injury, promotion of endurance capacity, and learning-memory improvement. H. sinensis has the ability to accelerate leukocyte recovery, stimulate lymphocyte proliferation, antidiabetes, and improve kidney injury. Starting January 1st, 2013, regulation will dictate that one fungus can only have one name, which will end the system of using separate names for anamorphs. The anamorph name “H. sinensis” has changed by the International Code of Nomenclature for algae, fungi, and plants to O. sinensis. PMID:24716152

  10. Identification, quantification and antioxidant activity of acylated flavonol glycosides from sea buckthorn (Hippophae rhamnoides ssp. sinensis).

    PubMed

    Chen, Chu; Xu, Xue-Min; Chen, Yang; Yu, Meng-Yao; Wen, Fei-Yan; Zhang, Hao

    2013-12-01

    A novel acylated flavonol glycoside: isorhamnetin (3-O-[(6-O-E-sinapoyl)-β-D-glucopyranosyl-(1→2)]-β-D-glucopyranosyl-7-O-α-L-rhamnopyranoside) (1), together with two known acylated flavonol glycosides: quercetin (3-O-[(6-O-E-sinapoyl)-β-D-glucopyranosyl-(1→2)]-β-D-glucopyranosyl-7-O-α-L-rhamnopyranoside) (2) and kaempferol (3-O-[(6-O-E-sinapoyl)-β-D-glucopyranosyl-(1→2)]-β-D-glucopyranosyl-7-O-α-L-rhamnopyranoside) (3) were isolated from the n-butanol fraction of sea buckthorn (Hippophae rhamnoides ssp. sinensis) berries for the first time by chromatographic methods, and their structures were elucidated using UV, MS, (1)H and (13)C NMR, and 2D NMR. Compounds 1-3 showed good scavenging activities, with respective IC50 values of 8.91, 4.26 and 30.90 μM toward the 2,2'-diphenyl-1-picrylhydrazyl (DPPH) radical; respective Trolox equivalent antioxidant capacities of 2.89, 4.04 and 2.44 μM μM(-1) toward 2,2'-azino-bis-3-ethyl-benzothiazoline-6-sulphonate (ABTS) radical. The quantitative analysis of the isolated acylated flavonol glycosides was performed by HPLC-DAD method. The contents of compounds 1-3 were in the range of 12.2-31.4, 4.0-25.3, 7.5-59.7 mg/100 g dried berries and 9.1-34.5, 75.1-182.1, 29.2-113.4 mg/100 g dried leaves, respectively.

  11. The molecular characterization of a catalase from Chinese mitten crab Eriocheir sinensis.

    PubMed

    Wang, M; Wang, L; Zhou, Z; Gao, Y; Wang, L; Shi, X; Gai, Y; Mu, C; Song, L

    2013-06-01

    Catalase (CAT) is an antioxidant enzyme and plays a significant role in the protection against oxidative stress by reducing hydrogen peroxide. The CAT cDNA of Eriocheir sinensis (EsCAT) was cloned via RACE technique. The complete sequence of EsCAT cDNA consisted of a 5' untranslated regions (UTR) of 224 bp, a 3' UTR of 1287 bp with a poly (A) tail and an open reading frame (ORF) of 1542 bp, which encoded a polypeptide of 513 amino acid residues with a calculated molecular mass of approximately 58.86 kDa and a theoretical isoelectric point of 6.880. The deduced amino acid sequence of EsCAT contained a highly conserved proximal active-site signature motif ((60)FDRERIPERVVHAKGAL(76)) and a proximal heme-ligand signature motif ((350)RLFSYNDTH(358)) and exhibited high similarity with other reported CATs. In the phylogenetic tree, EsCAT was clustered with the CATs from Scylla serrata and Portunus trituberculatus. The EsCAT transcripts were constitutively expressed in haepatopancreas, haemocytes, gill, gonad, muscle and heart, with highest expression level in haepatopancreas. The relative expression level of EsCAT mRNA in haemocytes was continuously up-regulated and reached the peak level at 48 h post-Vibrio anguillarum challenge. The purified recombinant EsCAT protein displayed antioxidant activity against hydrogen peroxide with high thermal stability and broad spectrum of pH values. All these results demonstrated that EsCAT was an efficient antioxidant enzyme and potentially involved in the regulation of redox and innate immune response of crabs.

  12. Malformations of the endangered Chinese sturgeon, Acipenser sinensis, and its causal agent

    PubMed Central

    Hu, Jianying; Zhang, Zhaobin; Wei, Qiwei; Zhen, Huajun; Zhao, Yanbin; Peng, Hui; Wan, Yi; Giesy, John P.; Li, Luoxin; Zhang, Bo

    2009-01-01

    The anadromous Chinese sturgeon (Acipenser sinensis) is endangered and listed among the first class of protected animals in China. The possible causes for the decline of this species are the effects of synthetic chemicals, and loss of critical habitat. Chinese sturgeon in the Yangtze River have accumulated triphenyltin (TPT) to 31–128 ng/g wet weigh (ww) in liver, which is greater than the concentrations of tributyltin (<1.0 ng/g ww). Maternal transfer of TPT has resulted in concentrations of 25.5 ± 13.0 ng/g ww in eggs of wild Chinese sturgeon, which poses a significant risk to the larvae naturally fertilized or hatched in the Yangtze River. The incidence of deformities in fry was 7.5%, with 1.2% of individuals exhibiting ocular abnormal development, and 6.3% exhibited skeletal/morphological deformations. The incidences of both ocular and skeletal/morphological deformations were directly proportional to the TPT concentration in the eggs of both the Chinese sturgeon and the Siberian sturgeon (Acipenser baerii) in controlled laboratory studies. The rates of deformities in the controlled studies were consistent with the rates caused at the similar concentrations in eggs collected from the field. Thus, TPT is the causal agent to induce the malformation of larvae of Chinese sturgeon. The incidence of deformed larvae of Chinese sturgeon is an indicator of overall population-level effects of TPT on Chinese sturgeon, because TPT at environmentally relevant concentrations can result in significantly decrease both quality and quantity of eggs and spawning frequency of fish. PMID:19470453

  13. A footprint of past climate change on the diversity and population structure of Miscanthus sinensis

    PubMed Central

    Clark, Lindsay V.; Brummer, Joe E.; Głowacka, Katarzyna; Hall, Megan C.; Heo, Kweon; Peng, Junhua; Yamada, Toshihiko; Yoo, Ji Hye; Yu, Chang Yeon; Zhao, Hua; Long, Stephen P.; Sacks, Erik J.

    2014-01-01

    Background and Aims Miscanthus is a perennial C4 grass that is a leading potential feedstock crop for the emerging bioenergy industry in North America, Europe and China. However, only a single, sterile genotype of M. × giganteus (M×g), a nothospecies derived from diploid M. sinensis (Msi) and tetraploid M. sacchariflorus (Msa), is currently available to farmers for biomass production. To facilitate breeding of Miscanthus, this study characterized genetic diversity and population structure of Msi in its native range of East Asia. Methods A total of 767 accessions were studied, including 617 Msi from most of its native range in China, Japan and South Korea, and 77 ornamental cultivars and 43 naturalized individuals from the USA. Accessions were evaluated with 21 207 restriction site-associated DNA sequencing single nucleotide polymorphism (SNP) markers, 424 GoldenGate SNPs and ten plastid microsatellite markers. Key Results Six genetic clusters of Msi from geographically distinct regions in Asia were identified. Genetic data indicated that (1) south-eastern China was the origin of Msi populations found in temperate eastern Asia, which is consistent with this area probably having been a refugium during the last glacial maximum (LGM); (2) Msi migrated directly from south-eastern China to Japan before migrating to the same latitudes in China and Korea, which is consistent with the known sequence of warming post-LGM; (3) ornamental Msi cultivars were derived from the southern Japan population, and US naturalized populations were derived from a sub-set of the ornamental cultivars; and (4) many ornamental cultivars previously described as Msi have hybrid ancestry from Msa and Msi, whereas US naturalized populations of Msi do not. Conclusions Population structure of Msi was driven by patterns of warming since the LGM, and secondarily by geographical barriers. This study will facilitate germplasm conservation, association analyses and identification of potential heterotic

  14. Biochemical properties of alpha-amylase from peel of Citrus sinensis cv. Abosora.

    PubMed

    Mohamed, Saleh Ahmed; Drees, Ehab A; El-Badry, Mohamed O; Fahmy, Afaf S

    2010-04-01

    alpha-Amylase activity was screened in the peel, as waste fruit, of 13 species and cultivars of Egyptian citrus. The species Citrus sinensis cv. Abosora had the highest activity. alpha-Amylase AI from Abosora peel was purified to homogeneity using anion and cation-exchange, and gel filtration chromatographies. Molecular weight of alpha-amylase AI was found to be 42 kDa. The hydrolysis properties of alpha-amylase AI toward different substrates indicated that corn starch is the best substrate. The alpha-amylase had the highest activity toward glycogen compared with amylopectin and dextrin. Potato starch had low affinity toward alpha-amylase AI but it did not hydrolyze beta-cyclodextrin and dextran. Apparent Km for alpha-amylase AI was 5 mg (0.5%) starch/ml. alpha-Amylase AI showed optimum activity at pH 5.6 and 40 degrees C. The enzyme was thermally stable up to 40 degrees C and inactivated at 70 degrees C. The effect of mono and divalent metal ions were tested for the alpha-amylase AI. Ba2+ was found to have activating effect, where as Li+ had negligible effect on activity. The other metals caused inhibition effect. Activity of the alpha-amylase AI was increased one and half in the presence of 4 mM Ca2+ and was found to be partially inactivated at 10 mM Ca2+. The reduction of starch viscosity indicated that the enzyme is endoamylase. The results suggested that, in addition to citrus peel is a rich source of pectins and flavanoids, alpha-amylase AI from orange peel could be involved in the development and ripening of citrus fruit and may be used for juice processing.

  15. Plant regeneration from protoplasts of immature Vigna sinensis cotyledons via somatic embryogenesis.

    PubMed

    Li, X B; Xu, Z H; Wei, Z M

    1995-12-01

    Protoplasts were isolated from immature cotyledons of Vigna sinensis and cultured in a modified MS Liquid medium containing 0. 2 mg/l 2, 4-dichlorophenoxyacetic acid (2, 4-D), 1 mg/l naphthaleneacetic acid (NAA) and 0. 5 mg/l 6-benzylaminopurine (BAP) in the dark at a density of 1 × 10(5)/ml. The protoplasts began to divide in 3-5 days. Sustained cell division resulted in formation of cell clusters and small calli, with the cell division frequency and plating efficiency of cell colonies reaching 27. 7% and 1. 7% respectively. When calli of 2 mm in size were transferred onto MSB medium (MS salts and B5 vitamins) containing 500mg/l NaCl, 500 mg/ 1 casein hydrolysate (CH), 2 mg/l 2,4-D and 0. 5 mg/l BAP for further growth, approximately 5% of the calli developed embryogenically. The embryogenic calli were selected and subcultured on the same composition of MSB medium and were able to maintain somatic embryogenesis capacity in subculture for a long time. When the calli were moved to MSB medium with 0. 1 mg/l indole-3-acetic acid (IAA), 0. 5mg/l kinetin(KT), 3-5% mannitol and 2% sucrose in the light, many somatic embryos formed from the calli. Only part of the embryoids developed further to the cotyledonary stage, and the others died at the globular, heart-shaped or torpedo stages. Finally, some cotyledonary embryoids germinated and developed into plants or shoots. The shoots were readily rooted on 1/2 strength MS medium with 0. 1-0.3 mg/l indole-3-butyric acid (IBA). The plants grew well in soil and were fertile.

  16. Gene Expression Profiling of Dendritic Cells in Different Physiological Stages under Cordyceps sinensis Treatment

    PubMed Central

    Li, Chia-Yang; Chiang, Chi-Shiun; Cheng, Wei-Chung; Wang, Shu-Chi; Cheng, Hung-Tsu; Chen, Chaang-Ray; Shu, Wun-Yi; Tsai, Min-Lung; Hseu, Ruey-Shyang; Chang, Cheng-Wei; Huang, Chao-Ying; Fang, Shih-Hua; Hsu, Ian C.

    2012-01-01

    Cordyceps sinensis (CS) has been commonly used as herbal medicine and a health supplement in China for over two thousand years. Although previous studies have demonstrated that CS has benefits in immunoregulation and anti-inflammation, the precise mechanism by which CS affects immunomodulation is still unclear. In this study, we exploited duplicate sets of loop-design microarray experiments to examine two different batches of CS and analyze the effects of CS on dendritic cells (DCs), in different physiology stages: naïve stage and inflammatory stage. Immature DCs were treated with CS, lipopolysaccharide (LPS), or LPS plus CS (LPS/CS) for two days, and the gene expression profiles were examined using cDNA microarrays. The results of two loop-design microarray experiments showed good intersection rates. The expression level of common genes found in both loop-design microarray experiments was consistent, and the correlation coefficients (Rs), were higher than 0.96. Through intersection analysis of microarray results, we identified 295 intersecting significantly differentially expressed (SDE) genes of the three different treatments (CS, LPS, and LPS/CS), which participated mainly in the adjustment of immune response and the regulation of cell proliferation and death. Genes regulated uniquely by CS treatment were significantly involved in the regulation of focal adhesion pathway, ECM-receptor interaction pathway, and hematopoietic cell lineage pathway. Unique LPS regulated genes were significantly involved in the regulation of Toll-like receptor signaling pathway, systemic lupus erythematosus pathway, and complement and coagulation cascades pathway. Unique LPS/CS regulated genes were significantly involved in the regulation of oxidative phosphorylation pathway. These results could provide useful information in further study of the pharmacological mechanisms of CS. This study also demonstrates that with a rigorous experimental design, the biological effects of a complex

  17. Effect of Cordyceps sinensis and Tripterygium wilfordii polyglycosidium on podocytes in rats with diabetic nephropathy

    PubMed Central

    HAO, LI; PAN, MENG-SHU; ZHENG, YUN; WANG, RUI-FENG

    2014-01-01

    The aim of the present study was to investigate the effects of Cordyceps sinensis (CS) and Tripterygium wilfordii polyglycosidium (TWP) on podocytes in rats with diabetic nephropathy (DN). DN rat models were established and divided randomly into normal control (group A), DN (group B), CS (group C), TWP (group D) and CS and TWP groups (group E). After 12 weeks, levels of 24-h urinary protein, blood urea nitrogen (BUN), serum creatinine (SCR), white blood cells, blood glucose (GLU), aspartate aminotransferase, alanine aminotransferase and kidney weight (KW)/body weight (BW) were determined. Renal pathological changes were evaluated using hematoxylin and eosin staining, whereas the structural changes in the podocytes were observed under a transmission electron microscope. The expression levels of nephrin and podocin were evaluated using immunofluorescence staining. Compared with group A, the SCR and BUN levels in group B were higher (P<0.05) and the GLU, KW/BW and the 24-h urine protein were markedly higher (P<0.01). Moreover, incidences of glomerular disorders, chronic tubulointerstitial damage and glomerular podocyte lesions in groups B, C, D and E were observed, compared with group A. The high cortical expression of nephrin and podocin protein decreased. Compared with group B, the KW/BW and 24-h urinary protein level in groups C, D and E were lower (P<0.01). The glomeruli, tubules and podocytes exhibited pathomorphological improvements and the nephrin and podocin protein expression levels were higher in the nephridial tissue. A decrease in KW/BW and the 24-h urinary protein level, as well as improvements in glomerular disorder, chronic tubulointerstitial damage and glomerular podocyte lesions, were observed in groups C, D and E. Therefore, the results demonstrated that CS and TWP exhibited a protective effect on the podocytes of rats with DN. Moreover, CS combined with TWP increased this protective effect. PMID:24926327

  18. Ergosterol Is the Active Compound of Cultured Mycelium Cordyceps sinensis on Antiliver Fibrosis

    PubMed Central

    Peng, Yuan; Tao, Yanyan; Wang, Qinglan; Shen, Li; Yang, Tao; Liu, Zulong; Liu, Chenghai

    2014-01-01

    Cultured mycelium Cordyceps sinensis (CMCS) is a Chinese herbal medicine, which is widely used for a variety of diseases including liver injury in clinic. The current study aims to investigate the protective effects of CMCS against liver fibrosis and to exploit its active antifibrotic substances in vivo and in vitro. For evaluating the antifibrotic effect of CMCS and ergosterol, male C57BL/6 mice were injected intraperitoneally with carbon tetrachloride (CCl4) and treated with CMCS (120 mg/kg/d) or ergosterol (50 mg/kg/d). Four weeks later, serum liver function, hepatic hydroxyproline (Hyp) content, liver inflammation, collagen deposition, and expression of alpha smooth muscle actin (α-SMA) in liver tissue were evaluated. Besides, toxicological effects of active compounds of CMCS on hepatocytes and hepatic stellate cells (HSCs) were detected and expressions of permeability of the lysosomal membrane, EdU, F-actin, and α-SMA of activated HSCs were analyzed to screen the antifibrotic substance in CMCS in vitro. Our results showed that CMCS could significantly alleviate levels of serum liver functions, attenuate hepatic inflammation, decrease collagen deposition, and relieve levels of α-SMA in liver, respectively. Ergosterol, the active compound in CMCS that was detected by HPLC, played a dose-dependent inhibition role on activated HSCs via upregulating expressions of permeability of the lysosomal membrane and downregulating levels of EdU, F-actin, and α-SMA on activated HSCs in vitro. Moreover, ergosterol revealed the antifibrotic effect alike in vivo. In conclusion, CMCS is effective in alleviating liver fibrosis induced by CCl4 and ergosterol might be the efficacious antifibrotic substance in CMCS in vivo and in vitro. PMID:25386220

  19. Evaluation of Safety and Antioxidant Activity of Yellow Tea (Camellia sinensis) Extract for Application in Food.

    PubMed

    Kujawska, Małgorzata; Ewertowska, Małgorzata; Ignatowicz, Ewa; Adamska, Teresa; Szaefer, Hanna; Gramza-Michałowska, Anna; Korczak, Józef; Jodynis-Liebert, Jadwiga

    2016-03-01

    The article presents an evaluation of the safety of yellow tea (Camellia sinensis) extract consumption and its antioxidant activity in an animal model. Wistar rats were exposed through diet to 2, 6, and 10 g yellow tea extract/kg feed for 90 days. No signs of toxicity and no differences in mean body weight gain in the treated and control rats were recorded throughout the experiment. No statistically significant differences in hematology findings and clinical chemistry parameters were observed between controls and treated groups. Microscopic examination of tissue sections revealed no pathology attributable to yellow tea extract intake. Lipid peroxidation level in the liver was slightly increased in medium-dose males and high-dose females and decreased in two female groups receiving 2 and 6 g/kg of the extract tested. Content of carbonyl groups in protein, as well as the basal level of DNA damage, was not changed. In a majority of rats, the activity of antioxidant enzymes was increased except superoxide dismutase in high-dose groups, glutathione peroxidase in high-dose females, glutathione reductase in low- and mid-dose groups, and glutathione S-transferase in mid-dose females and high-dose males. It could be concluded that rats tolerated well dietary treatment with yellow tea extract up to 0.8 g/kg b.w./day for 90 days. Results showed that yellow tea extract at the doses tested did not demonstrate adverse effects and improved the antioxidant status in the liver of rats.

  20. Exogenous abscisic acid significantly affects proteome in tea plant (Camellia sinensis) exposed to drought stress.

    PubMed

    Zhou, Lin; Xu, Hui; Mischke, Sue; Meinhardt, Lyndel W; Zhang, Dapeng; Zhu, Xujun; Li, Xinghui; Fang, Wanping

    2014-01-01

    Tea [Camellia sinensis (L.) O. Kuntze] is an important economic crop, and drought is the most important abiotic stress affecting yield and quality. Abscisic acid (ABA) is an important phytohormone responsible for activating drought resistance. Increased understanding of ABA effects on tea plant under drought stress is essential to develop drought-tolerant tea genotypes, along with crop management practices that can mitigate drought stress. The objective of the present investigation is evaluation of effects of exogenous ABA on the leaf proteome in tea plant exposed to drought stress. Leaf protein patterns of tea plants under simulated drought stress [(polyethylene glycol (PEG)-treated] and exogenous ABA treatment were analyzed in a time-course experiment using two-dimensional electrophoresis (2-DE), followed by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS). Among the 72 protein spots identified by MALDI-TOF MS, 16 proteins were downregulated and two were upregulated by exogenous ABA. The upregulated proteins have roles in glycolysis and photosystem II stabilization. Twenty-one protein spots were responsive to drought stress and most participate in carbohydrate and nitrogen metabolism, control of reactive oxygen species (ROS), defense, signaling or nucleic acid metabolism. The combined treatments of exogenous ABA and drought showed upregulation of 10 protein spots at 12 h and upregulation of 11 proteins at 72 h after initiation of drought stress. The results support the importance of the role that ABA plays in the tea plant during drought stress, by improving protein transport, carbon metabolism and expression of resistance proteins.

  1. Total polyphenol content and antioxidant capacity of commercially available tea (Camellia sinensis) in Argentina.

    PubMed

    Anesini, Claudia; Ferraro, Graciela E; Filip, Rosana

    2008-10-08

    Tea, Camellia sinensis (L.) O. Kuntze (Theaceae) is cultivated in Argentina in the northeastern region (provinces of Misiones and Corrientes), between 26 degrees and 28 degrees south latitude, the southernmost area of the world where tea is cultivated. The objective of this work was to determine the total polyphenol content and the in vitro antioxidant capacity of green and black tea cultivated and industrialized in Argentina. Twelve samples of eight brands were analyzed. The total polyphenol content was determined according to the International Organization for Standardization method (ISO) 14502-1 for the determination of substances characteristic of green and black tea. The antioxidant capacity was determined by the ferric thiocyanate method (FTC) and the 1,1-diphenyl-2-picrylhydrazyl (DPPH) free-radical scavenging assay. Green tea showed a higher polyphenol content than black tea. The total polyphenol concentration in green tea was found to vary from 21.02 +/- 1.54 to 14.32 +/- 0.45% of gallic acid equivalents (GAE), whereas in black tea, the polyphenol content ranged from 17.62 +/- 0.42 to 8.42 +/- 0.55% of GAE (P < 0.05). A similar profile was observed for the antioxidant capacity determined by both methods. The antioxidant activities were well correlated with the total polyphenol content (r (2) = 0.9935 for the ferric thiocyanate method and r (2) = 0.9141 for the 1,1-diphenyl-2-picrylhydrazyl free-radical scavenging assay). This is the first systematic screening for the quantification of polyphenols and antioxidant activity in tea commercialized in Argentine markets. The results obtained herein allow one to conclude that Argentine tea is of very good quality when compared to teas from other sources.

  2. Assessment of genetic diversity of Tunisian orange, Citrus sinensis (L.) Osbeck using microsatellite (SSR) markers.

    PubMed

    Mahjbi, A; Oueslati, A; Baraket, G; Salhi-Hannachi, A; Zehdi Azouzi, S

    2016-05-20

    Citrus are one of the most cultivated crops in the world. Economically, they are very important fruit trees in Tunisia. Little is known about the genetic diversity of the Tunisian Citrus germplasm. Exploring this diversity is a prerequisite for the identification and characterization of the local germplasm to circumvent and controlling genetic erosion caused by biotic and abiotic stress to aid its conservation and use. In the present study, we explored the genetic diversity of 20 Tunisian orange cultivars [Citrus sinensis (L.) Osbeck] and established their relationships by using seven simple sequence repeat (SSR) loci. In total, 37 alleles and 44 genotypes were scored. The sizes of alleles ranged from 90 to 280 bp. The number of alleles per locus was from 4 to 7, with an average of 5.28. Polymorphic information content value changed from 0.599 to 0.769 with an average of 0.675. Analysis of the genotypes revealed a heterozygote deficiency across all the genotypes. The observed heterozygosity varied from 0 to 1 (average of 0.671). Cluster analysis showed that three groups could be distinguished and the polymorphism occurred independently of the geographical origin of the studied orange cultivars. The detected SSR genotypes allowed the establishment of an identification key with a discriminating power of 100%. Multivariate analysis and the neighbor-joining phylogenetic tree indicated a narrow genetic base for the orange cultivars. The usefulness of SSR markers for orange fingerprinting and evaluation of the genetic diversity in the Tunisian germplasm are discussed in this paper.

  3. Involvement of polyamine biosynthesis in somatic embryogenesis of Valencia sweet orange (Citrus sinensis) induced by glycerol.

    PubMed

    Wu, Xiao-Ba; Wang, Jing; Liu, Ji-Hong; Deng, Xiu-Xin

    2009-01-01

    Culture of Citrus sinensis embryogenic callus on the embryo-inducing medium (EIM) containing glycerol gave rise to a large number of embryos, whereas very few embryos were observed on the callus growth medium (CGM). In the current paper, attempts were made to investigate whether polyamine biosynthesis was involved in glycerol-mediated somatic embryogenesis. Quantification of free polyamines by high-performance liquid chromatography showed that the cultures on EIM had less putrescine than those on CGM. However, increase in spermidine and spermine was detected in cultures on EIM during the first 20d of culture, coincident with abundant somatic embryogenesis. The globular embryos contained more polyamines than embryos at other stages. Semi-quantitative reverse transcriptase-polymerase chain reaction assay showed that expression levels of all of the five key genes involved in polyamine biosynthesis, with the exception of S-adenosylmethionine decarboxylase, were induced in cultures on EIM, and that their transcriptional levels were increased with maturation of the embryos. Addition of alpha-difluoromethylornithine, a polyamine biosynthesis inhibitor, to EIM resulted in remarkable inhibition of somatic embryogenesis, concurrent with notable reduction of endogenous putrescine and spermidine, particularly at higher concentrations. Exogenous application of 1mM putrescine to EIM together with 5mM alpha-difluoromethylornithine led to dramatic enhancement of endogenous polyamines, which successfully restored somatic embryogenesis. All of these, collectively, demonstrated that free polyamines, at least spermidine and spermine herein, were involved in glycerol-mediated promotion of somatic embryogenesis, which will open a new avenue for establishing a sophisticated system for somatic embryogenesis based on the modulation of endogenous polyamines.

  4. Prey detection by great cormorant (Phalacrocorax carbo sinensis) in clear and in turbid water.

    PubMed

    Strod, Tamir; Izhaki, Ido; Arad, Zeev; Katzir, Gadi

    2008-03-01

    The scattering and absorption of light by water molecules and by suspended and dissolved matter (turbidity) degrade image transmission and, thus, underwater perception. We tested the effects on visual detection of prey size and distance (affecting apparent prey size) and of low-level water turbidity in hand-reared great cormorants (Phalacrocorax carbo sinensis) diving for natural prey (fish) in a forced-choice situation. The cormorants' detection of underwater prey relied on vision. The minimal tested subtending visual angle of the prey at detection ranged between approximately 34.2' (prey size constant; distance varied) and 9.5' (distance constant; prey size varied). For all tested distances (0.8-3.1 m) the mean detection success was significantly higher than the chance level. The probability of a correct choice declined significantly with increased distance, with Detection success=-0.034D+1.021 (where D is distance, r(2)=0.5, N=70, P<0.001). The combined effect of turbidity and distance on the probability of detection success was significant, with both variables having a negative effect: Detection success=-0.286D-0.224Tu+1.691 (where Tu is turbidity, r(2)=0.68, N=144, P<0.001). At prey detection threshold, the relationship between distance and turbidity was: D=3.79e(-4.55Tu). It is concluded that (i) the subtending angle of natural prey at detection was lower than that of resolution of square-wave, high-contrast grating and (ii) turbidity, at levels significantly lower than commonly used in behavioural experiments, had a pronounced effect on visually mediated behaviour patterns.

  5. Effect of food restriction on the energy metabolism of the Chinese bulbul (Pycnonotus sinensis)

    PubMed Central

    LIANG, Qing-Jian; ZHAO, Lei; WANG, Jia-Qi; CHEN, Qian; ZHENG, Wei-Hong; LIU, Jin-Song

    2015-01-01

    Food resources play an important role in the regulation of animals’ physiology and behavior. We investigated the effect of short-term food restriction on metabolic thermogenesis of Chinese bulbuls (Pycnonotus sinensis) by measuring changes in body mass, body fat, basic metabolic rate (BMR), and organ mass of wild-caught Chinese bulbuls from Wenzhou, China. Short-term food restriction induced a significant decrease in body mass and body fat but body mass returned to normal levels soon after food was no longer restricted. Food restriction caused a significant reduction in BMR after 7 days (P<0.05), which returned to normal levels after food restriction ceased. Log total BMR was positively correlated with log body mass (r2=0.126, P<0.05). The dry masses of livers and the digestive tract were higher in birds that had been subject to temporary food restriction than in control birds and those subject to continual food restriction (P<0.001 and P<0.05, respectively). There was also significant differences in the dry mass of the lungs (P<0.05), heart (P<0.01), and spleen (P<0.05) in birds subject to short-term food restriction compared to control birds and those subject to continual food restriction. BMR was positively correlated with body and organ (heart, kidney and stomach) mass. These results suggest that the Chinese bulbul adjusts to restricted food availability by utilizing its energy reserves, lowering its BMR and changing the weight of various internal organs so as to balance total energy requirements. These may all be survival strategies that allow birds to cope with unpredictable variation in food abundance. PMID:25855226

  6. Camellia sinensis Ameliorates the Efficacy of Last Line Antibiotics Against Carbapenem Resistant Escherichia coli.

    PubMed

    Thakur, Pallavi; Chawla, Raman; Chakotiya, Ankita Singh; Tanwar, Ankit; Goel, Rajeev; Narula, Alka; Arora, Rajesh; Sharma, Rakesh Kumar

    2016-02-01

    Aquo-ethanolic extract of Camellia sinensis (PTRC-31911-A), standardized using Fourier transform infrared analysis, was found to have seven common functional groups in comparison with pre-identified marker compound 'quercetin'. Phyto-chemical quantitation analysis revealed the presence of 10.65 µg/mg of flavonoids. The bioactivity fingerprint profile of PTRC-31911-A includes IC50 (Hydroxyl radical site specific scavenging)  = 11.36 ± 0.5 µg/mL, IC80 (Hydroxyl radical non-site specific scavenging)  = 26.44 ± 0.5 µg/mL and IC50 (Superoxide ion scavenging)  = 10.141 ± 0.5 µg/mL. The drug combination analysis of PTRC-31911-A with five third-line antibiotics was carried out against carbapenem-resistant Escherichia coli. The analysis of combination of PTRC-31911-A (6.25-1000 µg/mL) and antibiotics (6.25-1000 µg/mL) revealed synergistic behaviour (fractional inhibitory concentration indices < 1) with tigecycline, ertapenem, meropenem, colistin and augmentin. The lead combination of PTRC-31911-A + ertapenem or meropenem showed maximum augmentative potential at 50 and 100 µg/mL, respectively, with nearly five-fold decrease in minimum inhibitory concentrations as compared with respective antibiotics alone. The synergistic effects implied that the antibacterial combinations of PTRC-31911-A and ertapenem, meropenem, colistin, tigecycline or augmentin would be more effective than a single monotherapy with either of the antibacterial agent.

  7. [Effects of agarwood formation induced by Fusarium sp. A2 on distribution and community composition of endophytic fungi in leaves of Aquilaria sinensis].

    PubMed

    Gao, Xiao-Xia; Zhou, Wei-Ping; Wang, Lei; Zhang, Wei-Min; Yan, Han-Jing

    2014-01-01

    The aim of the paper is to observe the distribution of the endophytic fungi in leaves of Aquilaria sinensis by using permanent paraffin-cut section, optical microscope photography and histochemistry. Total DNA was extracted with modified CTAB method and rDNA ITS regions of plant and endophytic fungi were amplified with eukaryotic universal primers. The rDNA ITS amplicon was characterized by RFLP analysis, sequencing of rDNA ITS library and phylogenetic analyses using PAUP by maximum parsimony. Fusarium sp. A2 was used to induce the formation of resinous in A. sinensis trees. As a result, endophytic fungi mainly distributed in spongy and phloem in leaf. Endophytic fungi distributed in the phloem in agarwood-producing tree and had a relatively high abundance. Phoma sp. and Collectrotrichum sp. were the absolute advantage species in the leaf tissues of non-resinous and agarwood-producing tree, respectively. Collectrotrichum sp. was the only fungal species detected both in the two types of A. sinensis with different levels of abundance. The culture-independent molecular method can be used to identify fungal species directly and rapidly from the plant tissues. Endophytic fungal communities in non-resinous and agarwood-producing A. sinensis leaf tissues were quite different.

  8. Secondary metabolite profiles of leaves of healthy and huanglongbing-infected orange (Citrus sinensis L.) seedlings measured by HPLC-fluorescence detection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Leaves of greenhouse-grown ‘Hamlin’ and ‘Valencia’ orange (Citrus sinensis L.) seedlings were analyzed by high performance liquid chromatography in a study of the progression of changes in secondary metabolite profiles resulting from infection by Candidatus Liberibacter asiaticus (CLas), the Huanglo...

  9. Medicinal flowers. XXII. Structures of chakasaponins V and VI, chakanoside I, and chakaflavonoside A from flower buds of Chinese tea plant (Camellia sinensis).

    PubMed

    Yoshikawa, Masayuki; Sugimoto, Sachiko; Nakamura, Seikou; Matsuda, Hisashi

    2008-09-01

    Two acylated oleanane-type triterpene oligoglycosides, chakasaponins V and VI, an aromatic glycoside, chakanoside I, and an acylated flavonol oligoglycoside, chakaflavonoside A, were isolated from the flower buds of Chinese tea plant [Camellia sinensis (L.) O. KUNTZE]. The chemical structures of those new glycosides were elucidated on the basis of chemical and physicochemical evidence.

  10. Laser therapy in a soft-shelled turtle (Pelodiscus sinensis) for the treatment of skin and shell ulceration. A case report.

    PubMed

    Kraut, S; Fischer, D; Heuser, W; Lierz, M

    2013-01-01

    Skin and shell diseases in aquatic turtles are often associated with several underlying causes. The presented case report describes aetiology including differential diagnoses, diagnostic procedures and therapy of a soft-shelled turtle (Pelodiscus sinensis) suffering from a septicaemic ulcerative dermatitis. Central aspect hereby is the positive curing effect of laser therapy on skin and shell lesions.

  11. Draft Genome Sequence of Curtobacterium sp. Strain ER1/6, an Endophytic Strain Isolated from Citrus sinensis with Potential To Be Used as a Biocontrol Agent.

    PubMed

    Garrido, Leandro Maza; Alves, João Marcelo Pereira; Oliveira, Liliane Santana; Gruber, Arthur; Padilla, Gabriel; Araújo, Welington Luiz

    2016-11-17

    Herein, we report a draft genome sequence of the endophytic Curtobacterium sp. strain ER1/6, isolated from a surface-sterilized Citrus sinensis branch, and it presented the capability to control phytopathogens. Functional annotation of the ~3.4-Mb genome revealed 3,100 protein-coding genes, with many products related to known ecological and biotechnological aspects of this bacterium.

  12. Systems Pharmacology-based strategy to screen new adjuvant for hepatitis B vaccine from Traditional Chinese Medicine Ophiocordyceps sinensis

    PubMed Central

    Wang, Jingbo; Liu, Rui; Liu, Baoxiu; Yang, Yan; Xie, Jun; Zhu, Naishuo

    2017-01-01

    Adjuvants are common component for many vaccines but there are still few licensed for human use due to low efficiency or side effects. The present work adopted Systems Pharmacology analysis as a new strategy to screen adjuvants from traditional Chinese medicine. Ophiocordyceps sinensis has been used for many years in China and other Asian countries with many biological properties, but the pharmacological mechanism has not been fully elucidated. First in this study, 190 putative targets for 17 active compounds in Ophiocordyceps sinensis were retrieved and a systems pharmacology-based approach was applied to provide new insights into the pharmacological actions of the drug. Pathway enrichment analysis found that the targets participated in several immunological processes. Based on this, we selected cordycepin as a target compound to serve as an adjuvant of the hepatitis B vaccine because the existing vaccine often fails to induce an effective immune response in many subjects. Animal and cellular experiments finally validated that the new vaccine simultaneously improves the humoral and cellular immunity of BALB/c mice without side effects. All this results demonstrate that cordycepin could work as adjuvant to hepatitis b vaccine and systems-pharmacology analysis could be used as a new method to select adjuvants. PMID:28317886

  13. Identification and Functional Analysis of Interleukin-1β in the Chinese Soft-Shelled Turtle Pelodiscus sinensis

    PubMed Central

    Liang, Quan; Li, Weifen; Guo, Ningning; Tong, Chao; Zhou, Yingshan; Fang, Weihuan; Li, Xiaoliang

    2016-01-01

    Chinese soft-shelled turtle (Pelodiscus sinensis) is commercially cultured in East and Southeast Asia for its nutritional and medicinal values. In this study, we identified interleukin-1β (IL-1β) from Chinese soft-shelled turtle. The full-length cDNA of Pelodiscus sinensis IL-1β (tIL-1β) consists of 1529 base pairs with an 831-base-pair open reading frame, encoding 277 amino acids. The guanine-cytosine (GC) content in the coding sequence and 3’ untranslated region of tIL-1β is considerably higher than that of other vertebrates. Its mRNA expression level increased significantly during Aeromonas hydrophila infection. The tIL-1β lacks the typical IL-1β-converting enzyme (ICE) cut site found in mammalian IL-1β, but still could be cleaved by turtle caspase-1. By mutating the potential cleavage sites, we identified aspartic acid (Asp/D) 130 as the ICE cut site in tIL-1β. The peptide truncated at D130 was expressed using the baculovirus expression system; its bioactivity is confirmed by the ability to induce cyclooxygenase-2 (COX-2) and tIL-1β itself in peripheral blood monocytes. In conclusion, we characterized IL-1β from Chinese soft-shelled turtle and identified its D130 as a non-typical ICE cut size. PMID:27153094

  14. Ultrastructure of the Rust Fungus Puccinia miscanthi in the Teliospore Stage Interacting with the Biofuel Plant Miscanthus sinensis

    PubMed Central

    Kim, Ki Woo

    2015-01-01

    Interaction of the the rust fungus Puccinia miscanthi with the biofuel plant Miscanthus sinensis during the teliospore phase was investigated by light and electron microscopy. P. miscanthi telia were oval-shaped and present on both the adaxial and abaxial leaf surfaces. Teliospores were brown, one-septate (two-celled), and had pedicels attached to one end. Transmission electron microscopy revealed numerous electron-translucent lipid globules in the cytoplasm of teliospores. Extensive cell wall dissolution around hyphae was not observed in the host tissues beneath the telia. Hyphae were found between mesophyll cells in the leaf tissues as well as in host cells. Intracellular hyphae, possibly haustoria, possessed electron-dense fungal cell walls encased by an electron-transparent fibrillar extrahaustorial sheath that had an electron-dense extrahaustorial membrane. The infected host cells appeared to maintain their membrane-bound structures such as nuclei and chloroplasts. These results suggest that the rust fungus maintains its biotrophic phase with most mesophyll cells of M. sinensis. Such a nutritional mode would permit the rust fungus to obtain food reserves for transient growth in the course of host alteration. PMID:26361478

  15. Two endoplasmic reticulum proteins (calnexin and calreticulin) are involved in innate immunity in Chinese mitten crab (Eriocheir sinensis)

    PubMed Central

    Huang, Ying; Hui, Kaimin; Jin, Min; Yin, Shaowu; Wang, Wen; Ren, Qian

    2016-01-01

    Calnexin (Cnx) and calreticulin (Crt), which are important chaperones in the endoplasmic reticulum (ER), participate in the folding and quality control of client proteins. Cnx and Crt identified from Chinese mitten crab (Eriocheir sinensis) are designated as EsCnx and EsCrt, respectively. EsCnx and EsCrt are expressed in the hemocyte, hepatopancrea, gill, and intestine at the mRNA and protein level. Immunofluorescence analysis indicated that EsCnx and EsCRT are located in the ER. Moreover, the mRNA and protein expression levels of EsCnx and EsCrt were altered by challenge with lipopolysaccharides (LPS), peptidoglycans (PGN), Staphyloccocus aureus, and Vibrio parahaemolyticus. Recombinant EsCnx and EsCrt (rEsCnx and rEsCrt, respectively) proteins can bind to various Gram-positive and Gram-negative bacteria, as well as to different polysaccharides (LPS and PGN). rEsCnx and rEsCrt assisted in the clearance of V. parahaemolyticus in vivo, and the clearance efficiency was impaired after silencing of EsCnx and EsCrt. Our results suggest that the two ER proteins are involved in anti-bacterial immunity in E. sinensis. PMID:27279413

  16. Two Kazal-type protease inhibitors from Macrobrachium nipponense and Eriocheir sinensis: comparative analysis of structure and activities.

    PubMed

    Qian, Ye-Qing; Li, Ye; Yang, Fan; Yu, Yan-Qin; Yang, Jin-Shu; Yang, Wei-Jun

    2012-03-01

    Kazal-type inhibitors (KPIs) play important roles in many biological and physiological processes, such as blood clotting, the immune response and reproduction. In the present study, two male reproductive tract KPIs, termed Man-KPI and Ers-KPI, were identified in Macrobrachium nipponense and Eriocheir sinensis, respectively. The inhibitory activities of recombinant Man-KPI and Ers-KPI against chymotrypsin, elastase, trypsin and thrombin were determined. The results showed that both of them strongly inhibit chymotrypsin and elastase. Kinetic studies were performed to elucidate their inhibition mechanism. Furthermore, individual domains were also expressed to learn further which domain contributes to the inhibitory activities of intact KPIs. Only Man-KPI_domain3 is active in the inhibition of chymotrypsin and elastase. Meanwhile, Ers-KPI_domain2 and 3 are responsible for inhibition of chymotrypsin, and Ers-KPI_domains2, 3 and 4 are responsible for the inhibition of elastase. Meanwhile, the inhibitory activities of these two KPIs toward Macrobrachium rosenbergii, M. nipponense and E. sinensis sperm were compared with that of the Kazal-type peptidase inhibitor (MRPINK) characterized from the M. rosenbergii reproductive tract in a previous study. The results demonstrated that KPIs can completely inhibit the gelatinolytic activities of sperm proteases from their own species, while different levels of cross-inhibition were observed between KPI and proteases from different species. These results may provide new perspective to further clarify the mechanism of KPI-proteases interaction in the male reproductive system.

  17. Al-Fe interactions and growth enhancement in Melastoma malabathricum and Miscanthus sinensis dominating acid sulphate soils.

    PubMed

    Watanabe, Toshihiro; Jansen, Steven; Osaki, Mitsuru

    2006-12-01

    Plants growing in acid sulphate soils are subject to high levels of Al availability, which may have effects on the growth and distribution of these species. Although Fe availability is also high in acid sulphate soils, little is known about the effect of Fe on the growth of native plants in these soils. Two species dominating this soil type in Asia, viz. Melastoma malabathricum and Miscanthus sinensis were grown hydroponically in a nutrient solution with different concentrations of Al and Fe. Melastoma malabathricum is found to be sensitive to Fe (40 and 100 microm). Application of 500 microm Al, however, completely ameliorates Fe toxicity and is associated with a decrease of Fe concentration in shoots and roots. The primary reason for the Al-induced growth enhancement of M. malabathricum is considered to be the Al-induced reduction of toxic Fe accumulation in roots and shoots. Therefore, Al is nearly essential for M. malabathricum when growing in acid sulphate soils. In contrast, application of both Fe and Al does not reduce the growth of M. sinensis, and Al application does not result in lower shoot concentrations of Fe, suggesting that this grass species has developed different mechanisms for adaptation to acid sulphate soils.

  18. A second generation SNP and SSR integrated linkage map and QTL mapping for the Chinese mitten crab Eriocheir sinensis

    PubMed Central

    Qiu, Gao-Feng; Xiong, Liang-Wei; Han, Zhi-Ke; Liu, Zhi-Qiang; Feng, Jian-Bin; Wu, Xu-Gan; Yan, Yin-Long; Shen, Hong; Huang, Long; Chen, Li

    2017-01-01

    The Chinese mitten crab Eriocheir sinensis is the most economically important cultivated crab species in China, and its genome has a high number of chromosomes (2n = 146). To obtain sufficient markers for construction of a dense genetic map for this species, we employed the recently developed specific-locus amplified fragment sequencing (SLAF-seq) method for large-scale SNPs screening and genotyping in a F1 full-sib family of 149 individuals. SLAF-seq generated 127,677 polymorphic SNP markers, of which 20,803 valid markers were assigned into five segregation types and were used together with previous SSR markers for linkage map construction. The final integrated genetic map included 17,680 SNP and 629 SSR markers on the 73 linkage groups (LG), and spanned 14,894.9 cM with an average marker interval of 0.81 cM. QTL mapping localized three significant growth-related QTL to a 1.2 cM region in LG53 as well as 146 sex-linked markers in LG48. Genome-wide QTL-association analysis further identified four growth-related QTL genes named LNX2, PAK2, FMRFamide and octopamine receptors. These genes are involved in a variety of different signaling pathways including cell proliferation and growth. The map and SNP markers described here will be a valuable resource for the E. sinensis genome project and selective breeding programs. PMID:28045132

  19. CYP74B24 is the 13-hydroperoxide lyase involved in biosynthesis of green leaf volatiles in tea (Camellia sinensis).

    PubMed

    Ono, Eiichiro; Handa, Taiki; Koeduka, Takao; Toyonaga, Hiromi; Tawfik, Moataz M; Shiraishi, Akira; Murata, Jun; Matsui, Kenji

    2016-01-01

    Green leaf volatiles (GLVs) are C6-aliphatic aldehydes/alcohols/acetates, and biosynthesized from the central precursor fatty acid 13-hydroperoxides by 13-hydroperoxide lyases (HPLs) in various plant species. While GLVs have been implicated as defense compounds in plants, GLVs give characteristic grassy note to a bouquet of aroma in green tea, which is manufactured from young leaves of Camellia sinensis. Here we identify three HPL-related genes from C. sinensis via RNA-Sequencing (RNA-Seq) in silico, and functionally characterized a candidate gene, CYP74B24, as a gene encoding tea HPL. Recombinant CYP74B24 protein heterologously expressed in Escherichia coli specifically produced (Z)-3-hexenal from 13-HPOT with the optimal pH 6.0 in vitro. CYP74B24 gene was expressed throughout the aerial organs in a rather constitutive manner and further induced by mechanical wounding. Constitutive expression of CYP74B24 gene in intact tea leaves might account for low but substantial and constitutive formation of a subset of GLVs, some of which are stored as glycosides. Our results not only provide novel insights into the biological roles that GLVs play in tea plants, but also serve as basis for the improvement of aroma quality in tea manufacturing processes.

  20. Five 2-(2-Phenylethyl)chromones from Sodium Chloride-Elicited Aquilaria sinensis Cell Suspension Cultures.

    PubMed

    Zhang, Zhongxiu; Wang, Xiaohui; Yang, Wanqing; Wang, Juan; Su, Cong; Liu, Xiao; Li, Jun; Zhao, Yunfang; Shi, Shepo; Tu, Pengfei

    2016-04-27

    Five 2-(2-phenylethyl)chromones including a new one, (5S,6R,7S,8R)-5,8-dichloro-6,7-dihydroxy-2-phenylethyl-5,6,7,8-tetrahydro-4H-chromen-4-one (1), and four known ones (2-5), were isolated from 150 mM NaCl-elicited Aquilaria sinensis cell suspension cultures. In addition, three feruloyl amides (6-8), six nucleosides (9-14), (+)-syringaresinol (15), indole-3-carboxaldehyde (16), and two glycosides (17-18) were also obtained. The structures were unambiguously identified by analysis of their UV, IR, NMR, and HRESIMS data. The absolute configuration of the new 2-(2-phenylethyl)chromone (1) was established by a dimolybdenum tetraacetate-induced circular dichroism experiment. Compared to un-elicited cell lines, the appearance of 2-(2-phenylethyl)chromones in NaCl-treated cells occurred on the 3rd and 5th days of their treatment. 2-(2-Phenylethyl)chromones, feruloyl amides, nucleosides, and lignins have been reported to be closely related to plant defense; therefore, the identification of these compounds from NaCl-elicited A. sinensis cell suspension cultures would be useful for further exploring the mechanism of agarwood formation.

  1. Morphological and ultrastructural study of the efferent ductules in the Chinese soft-shelled turtle Pelodiscus sinensis.

    PubMed

    Waqas, Muhammad Yasir; Liu, Tengfei; Yang, Ping; Ahmed, Nisar; Zhang, Qian; Hu, Lisi; Hong, Chen; Chen, Qiusheng

    2016-02-01

    Comparative study of the turtle excurrent duct system increases our understanding the evolution of sperm motility and fertility maintenance in higher vertebrates. Therefore, in this study we observed the histology and ultrastructure organization of efferent ductules in the Pelodiscus sinensis using light and transmission electron microscopy. The efferent ductules are extra- testicular and 22-28 in number originate from rete testis. The epithelium is entirely composed of two types of cells, the predominant non-ciliated and ciliated cells. The ciliated cells have long cilia that protrude into the lumen to form a meshwork. These cells associated with clusters of mitochondria in the supranuclear cytoplasm and possess coated vesicles, vacuole, intracellular spaces, and junction complexes. Ciliated cells in the proximal portion of the ductules contain an endocytic apparatus with coated pits and tubules in the apical cytoplasm. Interdigitations and lipid droplets are predominantly present around the nuclei of these cells. The non-ciliated cells have clusters of mitochondria present in both the supranuclear and perinuclear cytoplasm whereas, the nuclei of these cells are lightly stained. Moreover, the contour of the epithelium towards lumen is irregular as it has a deep indentation. The apical cytoplasm goes deep into the lumen to form cytoplasmic processes. This is the first study to describe the detailed features of efferent ductules in Pelodiscus sinensis with, special focus on the morphology of ciliated cells, as these cells are involved in the mixing of luminal fluid and transport of spermatozoa towards the distal region.

  2. Aluminium dynamics from soil to tea plant (Camellia sinensis L.): is it enhanced by municipal solid waste compost application?

    PubMed

    Karak, Tanmoy; Sonar, Indira; Paul, Ranjit K; Frankowski, Marcin; Boruah, Romesh K; Dutta, Amrit K; Das, Dilip K

    2015-01-01

    Application of municipal solid waste compost (MSWC) in tea (Camellia sinensis L.) cultivation can increase the fertility status of soils and thus enhance the plant growth. The present study attempts at application of MSWC in tea (TV1 and TV23 clones) cultivation to assess the effect of different doses of MSWC on growth and translocation potential of Al on this plant as well as fate of Al in soil, through the calculation of a risk assessment code (RAC). The sequential extraction of Al in MSWC amended soils showed that the fractionation of Al in soil changed after compost application, with an overall increase of the fractions associated to with Fe-Mn oxides, organic and of the residual fraction. The accumulation of Al in different parts ofC. sinensisL., grown on MSWC amended soil effected an overall increased growth of the plant with increasing doses of MSWC. According to RAC, Al falls in medium to high risk, though no adverse effect on plant health was observed. Tea plants were found to adapt well to MSWC amended soils. However, long term field trials are necessary to completely assess the risk of Al accumulation in soils upon MSWC application. Hierarchical cluster analysis was applied aiming to check for the presence of homogenous groups among different treatments. It was found that in both TV1 and TV23, treatments formed two different groups.

  3. Citrus Functional Genomics and Molecular Modeling in Relation to Citrus sinensis (Sweet Orange) Infection with Xylella fastidiosa (Citrus Variegated Chlorosis).

    PubMed

    Dwivedi, Upendra N; Tiwari, Sameeksha; Prasanna, Pragya; Awasthi, Manika; Singh, Swati; Pandey, Veda P

    2016-08-01

    Citrus are among the economically most important fruit tree crops in the world. Citrus variegated chlorosis (CVC), caused by Xylella fastidiosa infection, is a serious disease limiting citrus production at a global scale. With availability of citrus genomic resources, it is now possible to compare citrus expressed sequence tag (EST) data sets and identify single-nucleotide polymorphisms (SNPs) within and among different citrus cultivars that can be exploited for citrus resistance to infections, citrus breeding, among others. We report here, for the first time, SNPs in the EST data sets of X. fastidiosa-infected Citrus sinensis (sweet orange) and their functional annotation that revealed the involvement of eight C. sinensis candidate genes in CVC pathogenesis. Among these genes were xyloglucan endotransglycosylase, myo-inositol-1-phosphate synthase, and peroxidase were found to be involved in plant cell wall metabolism. These have been further investigated by molecular modeling for their role in CVC infection and defense. Molecular docking analyses of the wild and the mutant (SNP containing) types of the selected three enzymes with their respective substrates revealed a significant decrease in the binding affinity of substrates for the mutant enzymes, thus suggesting a decrease in the catalytic efficiency of these enzymes during infection, thereby facilitating a favorable condition for infection by the pathogen. These findings offer novel agrigenomics insights in developing future molecular targets and strategies for citrus fruit cultivation in ways that are resistant to X. fastidiosa infection, and by extension, with greater harvesting efficiency and economic value.

  4. Fermentation behavior of osmophilic yeast Candida tropicalis isolated from the nectar of Hibiscus rosa sinensis flowers for xylitol production.

    PubMed

    Misra, Swati; Raghuwanshi, Shailendra; Gupta, Pritesh; Dutt, Kakoli; Saxena, R K

    2012-02-01

    Eighteen yeast species belonging to seven genera were isolated from ten samples of nectar from Hibiscus rosa sinensis and investigated for xylitol production using D-xylose as sole carbon source. Amongst these isolates, no. 10 was selected as the best xylitol producer and identified as Candida tropicalis on the basis of morphological, biochemical and 26S rDNA sequencing. C. tropicalis produced 12.11 gl(-1) of xylitol in presence of 50 gl(-1) of xylose in 72 h at pH 5, 30°C and 200 rpm. The strain of C. tropicalis obtained through xylose enrichment technique has resulted in a yield of 0.5 gg(-1) with a xylitol volumetric productivity of 1.07 gl(-1)h(-1) in the presence of 300 gl(-1) of xylose through batch fermentation. This organism has been reported for the first time from Hibiscus rosa sinensis flowers. Realizing, the importance of this high valued compound, as a sugar substitute, xylose enrichment technique was developed in order to utilize even higher concentrations of xylose as substrate for maximum xylitol production.

  5. Transcriptome-Based Analysis of Dof Family Transcription Factors and Their Responses to Abiotic Stress in Tea Plant (Camellia sinensis).

    PubMed

    Li, Hui; Huang, Wei; Liu, Zhi-Wei; Wang, Yong-Xin; Zhuang, Jing

    2016-01-01

    Tea plant (Camellia sinensis (L.) O. Kuntze) is affected by abiotic stress during its growth and development. DNA-binding with one finger (Dof) transcription factors (TFs) play important roles in abiotic stress tolerance of plants. In this study, a total of 29 putative Dof TFs were identified based on transcriptome of tea plant, and the conserved domains and common motifs of these CsDof TFs were predicted and analyzed. The 29 CsDof proteins were divided into 7 groups (A, B1, B2, C1, C2.1, C2.2, and D2), and the interaction networks of Dof proteins in C. sinensis were established according to the data in Arabidopsis. Gene expression was analyzed in "Yingshuang" and "Huangjinya" under four experimental stresses by qRT-PCR. CsDof genes were expressed differentially and related to different abiotic stress conditions. In total, our results might suggest that there is a potential relationship between CsDof factors and tea plant stress resistance.

  6. P38 participates in spermatogenesis and acrosome reaction prior to fertilization in Chinese mitten crab Eriocheir sinensis.

    PubMed

    Zhu, Ming; Sun, Wen-Juan; Wang, Yuan-Li; Li, Qing; Yang, Hong-Dan; Duan, Ze-Lin; He, Lin; Wang, Qun

    2015-04-01

    P38 mitogen-activated protein kinases (MAPKs) comprise a family of serine/threonine protein kinases that play important roles in cellular responses to inflammatory cytokines and environmental stresses. These kinases are involved in controlling cell division, differentiation and death in mammalian testes and therefore are critical to spermatogenesis. To explore their functions in male reproduction of Chinese mitten crabs, Eriocheir sinensis p38 (Es-p38) protein expression was determined in different tissues including testes at different developmental stages by Western blot. Es-p38 was expressed in various tissues, with higher levels in the heart, stomach, gills and testes. Total Es-p38 protein levels increased gradually during spermatogenesis, but phosphorylated Es-p38 was much higher in the spermatid (August-October) than the spermatocyte (July-August) and sperm (October-January) stages. Trypan blue staining and hematoxylin/eosin staining were both used to detect sperm motility and changes in sperm morphology during the acrosome reaction (AR) induced by pre-incubation with A23187 in vitro, activated Es-p38 proteins detected by fluorescent microscopy were translocated gradually to nuclear and apical cap regions, accumulating at the anterior of the acrosomal tubule. The results suggest the involvement of p38 MAPK in spermatogenesis and the AR in E. sinensis.

  7. Genetic connectivity of the broadcast spawning reef coral Platygyra sinensis on impacted reefs, and the description of new microsatellite markers

    NASA Astrophysics Data System (ADS)

    Tay, Y. C.; Noreen, A. M. E.; Suharsono; Chou, L. M.; Todd, P. A.

    2015-03-01

    As tropical coral reef habitats continue to be lost or degraded, understanding the genetic diversity and connectivity among populations is essential for making informed management decisions. This is particularly important in rapidly developing, land-scarce nations (such as Singapore) that require targeted conservation efforts. Sixty percentage of Singapore's coral cover has been lost over the past five decades, and with further coastal reclamation underway, it is imperative to understand the effects of development on coral connectivity. In this study, we used seven microsatellite markers, of which six are newly described here, to investigate the genetic diversity and connectivity of the massive hard coral Platygyra sinensis at nine sites in Singapore and three in the nearby Indonesian island of Bintan. Our results show that P. sinensis currently retains large effective population sizes, high genetic diversity, as well as high connectivity among sites within each locality, which suggest that these populations have good potential for continued survival provided that there are no island-wide disturbances. However, the Singapore Strait appears to be a mild barrier to gene flow, which may lead to an increased reliance on self-seeding at either location. We suggest some directions for their management based on these potential population boundaries, which can help pave the path for marine conservation planning in Singapore.

  8. Transcriptome-Based Analysis of Dof Family Transcription Factors and Their Responses to Abiotic Stress in Tea Plant (Camellia sinensis)

    PubMed Central

    Li, Hui; Huang, Wei; Liu, Zhi-Wei; Wang, Yong-Xin

    2016-01-01

    Tea plant (Camellia sinensis (L.) O. Kuntze) is affected by abiotic stress during its growth and development. DNA-binding with one finger (Dof) transcription factors (TFs) play important roles in abiotic stress tolerance of plants. In this study, a total of 29 putative Dof TFs were identified based on transcriptome of tea plant, and the conserved domains and common motifs of these CsDof TFs were predicted and analyzed. The 29 CsDof proteins were divided into 7 groups (A, B1, B2, C1, C2.1, C2.2, and D2), and the interaction networks of Dof proteins in C. sinensis were established according to the data in Arabidopsis. Gene expression was analyzed in “Yingshuang” and “Huangjinya” under four experimental stresses by qRT-PCR. CsDof genes were expressed differentially and related to different abiotic stress conditions. In total, our results might suggest that there is a potential relationship between CsDof factors and tea plant stress resistance. PMID:27872842

  9. Hibiscus rosa sinensis Linn. Petals Modulates Glycogen Metabolism and Glucose Homeostasis Signalling Pathway in Streptozotocin-Induced Experimental Diabetes.

    PubMed

    Pillai, Sneha S; Mini, S

    2016-03-01

    The prevalence of diabetes mellitus is becoming more and more serious and reaches epidemic proportions worldwide. Scientific research is constantly looking for new agents that could be used as dietary functional ingredients in the fight against diabetes. The objective of the present study was to evaluate the effect of ethyl acetate fraction of Hibiscus rosa sinensis Linn. petals on experimental diabetes at a dose of 25 mg/kg body weight and it was compared with standard anti-diabetic drug metformin. The elevated levels of serum glucose (398.56 ± 35.78) and glycated haemoglobin (12.89 ± 1.89) in diabetic rats were significantly decreased (156.89 ± 14.45 and 6.12 ± 0.49, respectively) by Hibiscus rosa sinensis petals (EHRS) administration. Hepatotoxicity marker enzyme levels in serum were normalized. The fraction supplementation restored the glycogen content by regulating the activities of glycogen metabolizing enzymes. It significantly modulated the expressions of marker genes involved in glucose homeostasis signalling pathway. Histopathological analysis of liver and pancreas supported our findings. The overall effect was comparable with metformin. Hence, our study reveals the role of hibiscus petals for alleviation of diabetes complications, thus it can be propagated as a nutraceutical agent.

  10. Spatial and temporal transcriptome changes occurring during flower opening and senescence of the ephemeral hibiscus flower, Hibiscus rosa-sinensis.

    PubMed

    Trivellini, Alice; Cocetta, Giacomo; Hunter, Donald A; Vernieri, Paolo; Ferrante, Antonio

    2016-10-01

    Flowers are complex systems whose vegetative and sexual structures initiate and die in a synchronous manner. The rapidity of this process varies widely in flowers, with some lasting for months while others such as Hibiscus rosa-sinensis survive for only a day. The genetic regulation underlying these differences is unclear. To identify key genes and pathways that coordinate floral organ senescence of ephemeral flowers, we identified transcripts in H. rosa-sinensis floral organs by 454 sequencing. During development, 2053 transcripts increased and 2135 decreased significantly in abundance. The senescence of the flower was associated with increased abundance of many hydrolytic genes, including aspartic and cysteine proteases, vacuolar processing enzymes, and nucleases. Pathway analysis suggested that transcripts altering significantly in abundance were enriched in functions related to cell wall-, aquaporin-, light/circadian clock-, autophagy-, and calcium-related genes. Finding enrichment in light/circadian clock-related genes fits well with the observation that hibiscus floral development is highly synchronized with light and the hypothesis that ageing/senescence of the flower is orchestrated by a molecular clock. Further study of these genes will provide novel insight into how the molecular clock is able to regulate the timing of programmed cell death in tissues.

  11. Spatial and temporal transcriptome changes occurring during flower opening and senescence of the ephemeral hibiscus flower, Hibiscus rosa-sinensis

    PubMed Central

    Trivellini, Alice; Cocetta, Giacomo; Hunter, Donald A.; Vernieri, Paolo; Ferrante, Antonio

    2016-01-01

    Flowers are complex systems whose vegetative and sexual structures initiate and die in a synchronous manner. The rapidity of this process varies widely in flowers, with some lasting for months while others such as Hibiscus rosa-sinensis survive for only a day. The genetic regulation underlying these differences is unclear. To identify key genes and pathways that coordinate floral organ senescence of ephemeral flowers, we identified transcripts in H. rosa-sinensis floral organs by 454 sequencing. During development, 2053 transcripts increased and 2135 decreased significantly in abundance. The senescence of the flower was associated with increased abundance of many hydrolytic genes, including aspartic and cysteine proteases, vacuolar processing enzymes, and nucleases. Pathway analysis suggested that transcripts altering significantly in abundance were enriched in functions related to cell wall-, aquaporin-, light/circadian clock-, autophagy-, and calcium-related genes. Finding enrichment in light/circadian clock-related genes fits well with the observation that hibiscus floral development is highly synchronized with light and the hypothesis that ageing/senescence of the flower is orchestrated by a molecular clock. Further study of these genes will provide novel insight into how the molecular clock is able to regulate the timing of programmed cell death in tissues. PMID:27591432

  12. An LDLa domain-containing C-type lectin is involved in the innate immunity of Eriocheir sinensis.

    PubMed

    Huang, Ying; An, Liang; Hui, Kai-Min; Ren, Qian; Wang, Wen

    2014-02-01

    C-type lectins (CTLs) have crucial functions in recognizing and eliminating pathogens in innate immunity. This study identified a novel low-density lipoprotein receptor class A (LDLa) domain-containing CTL, designated as EsCTLDcp, from the Chinese mitten crab Eriocheir sinensis. The EsCTLDcp cDNA is 1258 bp long, with a 975 bp open reading frame that encodes a 324-amino acid protein. EsCTLDcp contains a signal peptide, an LDLa, and a single C-type lectin-like domain. EsCTLDcp was only expressed in the hepatopancreas of normal crabs, and its expression was regulated following crab challenge with pathogen-associated molecular patterns and with bacteria. The recombinant EsCTLDcp agglutinates Gram-positive (Staphylococcus aureus) and Gram-negative bacteria (Vibrio parahaemolyticus and Aeromonas hydrophila) in the presence of calcium. rEsCTLDcp also binds to various bacteria including S. aureus, Bacillus thuringiensis, Bacillus subtilis, Escherichia coli, Vibrio natriegens, V. parahaemolyticus, and A. hydrophila. The rEsCTLDcp protein helped the crabs clear the virulent Gram-negative bacterium V. parahaemolyticus in vivo, as well as interacted with VP24, an envelope protein of white spot syndrome virus (WSSV). These data suggest that EsCTLDcp functions as a pattern-recognition receptor involved in the innate immunity of E. sinensis.

  13. [Identification of different Citrus sinensis (L.) Osbeck trees varieties using Fourier transform infrared spectroscopy and hierarchical cluster analysis].

    PubMed

    Yi, Shi-Lai; Deng, Lie; He, Shao-Lan; Shi, You-Ming; Zheng, Yong-Qiang; Lu, Qiang; Xie, Rang-Jin; Wei, Xian-Guoi; Li, Song-Wei; Jian, Shui-Xian

    2012-11-01

    Researched on diversity of the spring leaf samples of seven different Citrus sinensis (L.) Osbeck varieties by Fourier transform infrared (FTIR) spectroscopy technology, the results showed that the Fourier transform infrared spectra of seven varieties leaves was composited by the absorption band of cellulose and polysaccharide mainly, the wave number of characteristics absorption peaks were similar at their FTIR spectra. However, there were some differences in shape of peaks and relatively absorption intensity. The conspicuous difference was presented at the region between 1 500 and 700 cm(-1) by second derivative spectra. Through the hierarchical cluster analysis (HCA) of second derivative spectra between 1 500 and 700 cm(-1), the results showed that the clustering of the different varieties of Citrus sinensis (L.) Osbeck varieties was classification according to genetic relationship. The results showed that FTIR spectroscopy combined with hierarchical cluster analysis could be used to identify and classify of citrus varieties rapidly, it was an extension method to study on early leaves of varieties orange seedlings.

  14. Fertile organs and in situ spores of a new dipteridaceous fern Hausmannia sinensis from the Jurassic of northern China

    PubMed Central

    Wang, Yongdong; Zhang, Hong

    2010-01-01

    As a representative fossil member of the dipteridaceous fern, genus Hausmannia was reported worldwide from the Mesozoic strata; however, little is known about the fertile structures, including sporangia and in situ spores, of this genus. In this study, a new species Hausmannia sinensis was identified from the Middle Jurassic of Nei Mongol (Inner Mongolia), northern China. The specimens are compressions and are well preserved with details of sporangia and in situ spores. The leaf laminae are broadly fan-shaped, with an almost entire margin. Primary and lateral veins dichotomously branch to form square or polygonal meshes. Each ultimate mesh bears one to two circular sori of 0.4 mm in diameter. Sori are exindusiate; each sorus contains three to six round to ovoid sporangia. The annulus is developed and oblique, with stomial region present in proximal position. Spores are trilete, circular to oval in shape. Both proximal and distal surfaces are covered with baculate to subverrucate sculptures. Spores range from 20 to 30 μm in diameter (average 28 μm), and are comparable to the dispersed genera Baculatisporites Thomas and Pflug and Apiculatisporis Potonié and Kremp. Hausmannia sinensis represents the first compression species of genus Hausmannia form Eurasia, which shows the combination of well-preserved sori, sporangia, annuli and in situ spore characters, and is therefore helpful for further understanding the diversity and evolution of the Dipteridaceae fern lineage through time. PMID:19364738

  15. The Japanese ladybirds, Coccinula crotchi and Coccinula sinensis, are infected with very closely related strains of male-killing Flavobacterium.

    PubMed

    Elnagdy, Sherif; Messing, Susan; Majerus, Michael E N

    2014-12-01

    Male-killing is 1 of 4 known strategies that inherited parasitic endosymbionts have evolved to manipulate their host's reproduction. In early male-killing, infected male offspring are killed early in embryogenesis. Within the Insecta, male-killing bacteria have been found in a wide range of hosts. The Coccinellidae families of beetles, better known as ladybirds, are particularly prone to male-killer invasion. In samples of the coccinellid, Coccinula crotchi, from Japan, a new male-killing bacterium was revealed by phenotypic assay. Molecular genetic analysis revealed the identity to be a tetracycline-sensitive Flavobacterium that causes female-biased offspring sex ratio. Furthermore, that Flavobacterium strain was found to be closely related to the Flavobacterium causing male-killing in the congeneric Japanese coccinellid, Coccinula sinensis, which was collected from the same region. However, we found slightly different Flavobacterium strains infecting C. sinensis from regions with different environmental conditions. This may be an indication of horizontal transmission of male-killing Flavobacterium between these 2 ladybird spices. Finally, environmental conditions may affect the spread of male-killing bacteria among their hosts.

  16. Systems Pharmacology-based strategy to screen new adjuvant for hepatitis B vaccine from Traditional Chinese Medicine Ophiocordyceps sinensis.

    PubMed

    Wang, Jingbo; Liu, Rui; Liu, Baoxiu; Yang, Yan; Xie, Jun; Zhu, Naishuo

    2017-03-20

    Adjuvants are common component for many vaccines but there are still few licensed for human use due to low efficiency or side effects. The present work adopted Systems Pharmacology analysis as a new strategy to screen adjuvants from traditional Chinese medicine. Ophiocordyceps sinensis has been used for many years in China and other Asian countries with many biological properties, but the pharmacological mechanism has not been fully elucidated. First in this study, 190 putative targets for 17 active compounds in Ophiocordyceps sinensis were retrieved and a systems pharmacology-based approach was applied to provide new insights into the pharmacological actions of the drug. Pathway enrichment analysis found that the targets participated in several immunological processes. Based on this, we selected cordycepin as a target compound to serve as an adjuvant of the hepatitis B vaccine because the existing vaccine often fails to induce an effective immune response in many subjects. Animal and cellular experiments finally validated that the new vaccine simultaneously improves the humoral and cellular immunity of BALB/c mice without side effects. All this results demonstrate that cordycepin could work as adjuvant to hepatitis b vaccine and systems-pharmacology analysis could be used as a new method to select adjuvants.

  17. A Comprehensive Analysis of the Phylogeny, Genomic Organization and Expression of Immunoglobulin Light Chain Genes in Alligator sinensis, an Endangered Reptile Species.

    PubMed

    Wang, Xifeng; Cheng, Gang; Lu, Yan; Zhang, Chenglin; Wu, Xiaobing; Han, Haitang; Zhao, Yaofeng; Ren, Liming

    2016-01-01

    Crocodilians are evolutionarily distinct reptiles that are distantly related to lizards and are thought to be the closest relatives of birds. Compared with birds and mammals, few studies have investigated the Ig light chain of crocodilians. Here, employing an Alligator sinensis genomic bacterial artificial chromosome (BAC) library and available genome data, we characterized the genomic organization of the Alligator sinensis IgL gene loci. The Alligator sinensis has two IgL isotypes, λ and κ, the same as Anolis carolinensis. The Igλ locus contains 6 Cλ genes, each preceded by a Jλ gene, and 86 potentially functional Vλ genes upstream of (Jλ-Cλ)n. The Igκ locus contains a single Cκ gene, 6 Jκs and 62 functional Vκs. All VL genes are classified into a total of 31 families: 19 Vλ families and 12 Vκ families. Based on an analysis of the chromosomal location of the light chain genes among mammals, birds, lizards and frogs, the data further confirm that there are two IgL isotypes in the Alligator sinensis: Igλ and Igκ. By analyzing the cloned Igλ/κ cDNA, we identified a biased usage pattern of V families in the expressed Vλ and Vκ. An analysis of the junctions of the recombined VJ revealed the presence of N and P nucleotides in both expressed λ and κ sequences. Phylogenetic analysis of the V genes revealed V families shared by mammals, birds, reptiles and Xenopus, suggesting that these conserved V families are orthologous and have been retained during the evolution of IgL. Our data suggest that the Alligator sinensis IgL gene repertoire is highly diverse and complex and provide insight into immunoglobulin gene evolution in vertebrates.

  18. A Comprehensive Analysis of the Phylogeny, Genomic Organization and Expression of Immunoglobulin Light Chain Genes in Alligator sinensis, an Endangered Reptile Species

    PubMed Central

    Lu, Yan; Zhang, Chenglin; Wu, Xiaobing; Han, Haitang; Zhao, Yaofeng; Ren, Liming

    2016-01-01

    Crocodilians are evolutionarily distinct reptiles that are distantly related to lizards and are thought to be the closest relatives of birds. Compared with birds and mammals, few studies have investigated the Ig light chain of crocodilians. Here, employing an Alligator sinensis genomic bacterial artificial chromosome (BAC) library and available genome data, we characterized the genomic organization of the Alligator sinensis IgL gene loci. The Alligator sinensis has two IgL isotypes, λ and κ, the same as Anolis carolinensis. The Igλ locus contains 6 Cλ genes, each preceded by a Jλ gene, and 86 potentially functional Vλ genes upstream of (Jλ-Cλ)n. The Igκ locus contains a single Cκ gene, 6 Jκs and 62 functional Vκs. All VL genes are classified into a total of 31 families: 19 Vλ families and 12 Vκ families. Based on an analysis of the chromosomal location of the light chain genes among mammals, birds, lizards and frogs, the data further confirm that there are two IgL isotypes in the Alligator sinensis: Igλ and Igκ. By analyzing the cloned Igλ/κ cDNA, we identified a biased usage pattern of V families in the expressed Vλ and Vκ. An analysis of the junctions of the recombined VJ revealed the presence of N and P nucleotides in both expressed λ and κ sequences. Phylogenetic analysis of the V genes revealed V families shared by mammals, birds, reptiles and Xenopus, suggesting that these conserved V families are orthologous and have been retained during the evolution of IgL. Our data suggest that the Alligator sinensis IgL gene repertoire is highly diverse and complex and provide insight into immunoglobulin gene evolution in vertebrates. PMID:26901135

  19. Cloning and Functional Analysis of Histones H3 and H4 in Nuclear Shaping during Spermatogenesis of the Chinese Mitten Crab, Eriocheir sinensis.

    PubMed

    Wu, Jiang-Li; Kang, Xian-Jiang; Guo, Ming-Shen; Mu, Shu-Mei; Zhang, Zhao-Hui

    2015-01-01

    During spermatogenesis in most animals, the basic proteins associated with DNA are continuously changing and somatic-typed histones are partly replaced by sperm-specific histones, which are then successively replaced by transition proteins and protamines. With the replacement of sperm nuclear basic proteins, nuclei progressively undergo chromatin condensation. The Chinese Mitten Crab (Eriocheir sinensis) is also known as the hairy crab or river crab (phylum Arthropoda, subphylum Crustacea, order Decapoda, and family Grapsidae). The spermatozoa of this species are aflagellate, and each has a spherical acrosome surrounded by a cup-shaped nucleus, peculiar to brachyurans. An interesting characteristic of the E. sinensis sperm nucleus is its lack of electron-dense chromatin. However, its formation is not clear. In this study, sequences encoding histones H3 and H4 were cloned by polymerase chain reaction amplification. Western blotting indicated that H3 and H4 existed in the sperm nuclei. Immunofluorescence and ultrastructural immunocytochemistry demonstrated that histones H3 and H4 were both present in the nuclei of spermatogonia, spermatocytes, spermatids and mature spermatozoa. The nuclear labeling density of histone H4 decreased in sperm nuclei, while histone H3 labeling was not changed significantly. Quantitative real-time PCR showed that the mRNA expression levels of histones H3 and H4 were higher at mitotic and meiotic stages than in later spermiogenesis. Our study demonstrates that the mature sperm nuclei of E. sinensis contain histones H3 and H4. This is the first report that the mature sperm nucleus of E. sinensis contains histones H3 and H4. This finding extends the study of sperm histones of E. sinensis and provides some basic data for exploring how decapod crustaceans form uncondensed sperm chromatin.

  20. Adult Psychology.

    ERIC Educational Resources Information Center

    Bischof, Ledford J.

    This volume comprehensively reviews the research on the psychology of the middle aged (ages 40-65). Topics include the concept of maturity and maturation models, the measurement and influences of adult self image; marriage and sexual patterns; intergenerational relationships between and children; vocations and avocations (work, retirement, play,…

  1. The effect of temperature on repeat swimming performance in juvenile qingbo (Spinibarbus sinensis).

    PubMed

    Pang, Xu; Yuan, Xing-Zhong; Cao, Zhen-Dong; Zhang, Yao-Guang; Fu, Shi-Jian

    2015-02-01

    To investigate the effect of temperature on the repeat constant acceleration swimming performance and on the metabolic recovery capacity in juvenile qingbo (Spinibarbus sinensis), their constant acceleration test speed (U(CAT)) and excess post-exercise oxygen consumption (EPOC) recovery process were measured twice with 1-h intervals at different acclimation temperatures (10, 15, 20, 25 and 30 °C). Temperature significantly affected U(CAT), the pre-exercise metabolic rate (MO(2)), metabolic peak values (MO(2peak)), the metabolic scope (MS, MO(2peak)--pre-exercise MO(2)) and the magnitude of the EPOC (P < 0.05). These parameters significantly increased as the temperature increased from 15 to 25 °C and significantly decreased (U(CAT) and EPOC magnitude) or did not change (MO(2peak) and MS) when the temperature increased from 25 to 30 °C in the first test (P < 0.05). The relationships between temperature (T) and these parameters (U(CAT), MO(2peak), MS and EPOC magnitude) in the first test were as follows: U(CAT) = 62.14/{1 + [(T - 25.1)/21.1](2)} (r = 0.847, P < 0.001, n = 40); MO(2peak) = 1,052.11/{1 + [(T - 29.2)/18.9](2)} (r = 0.901, P < 0.001, n = 39); MS = 753.74/{1 + [(T - 27.1)/18.6](2)} (r = 0.768, P < 0.001, n = 39); and EPOC = 195.42/{1 + [(T - 25.6)/8.7](2)} (r = 0.752, P < 0.001, n = 39). The optimal temperatures for U(CAT), MO(2peak), MS and EPOC magnitude in juvenile qingbo were 25.1, 29.2, 27.1 and 28.6 °C, respectively. Repeat exercise had different effect on U(CAT) and EPOC magnitude at different temperature (interaction effect, P < 0.05). There was no difference in U(CAT) and in EPOC magnitude between the first and second tests at low temperatures (10-20 °C). However, both U(CAT) and EPOC magnitude decreased significantly during the second test compared with the first test at high temperatures (25 and 30 °C) (P < 0.05). The present study showed that the recovery of the constant acceleration swimming performance was poorer at higher temperatures

  2. Cloning and Characterization of a Flavonoid 3′-Hydroxylase Gene from Tea Plant (Camellia sinensis)

    PubMed Central

    Zhou, Tian-Shan; Zhou, Rui; Yu, You-Ben; Xiao, Yao; Li, Dong-Hua; Xiao, Bin; Yu, Oliver; Yang, Ya-Jun

    2016-01-01

    Tea leaves contain abundant flavan-3-ols, which include dihydroxylated and trihydroxylated catechins. Flavonoid 3′-hydroxylase (F3′H: EC 1.14.13.21) is one of the enzymes in the establishment of the hydroxylation pattern. A gene encoding F3′H, designated as CsF3′H, was isolated from Camellia sinensis with a homology-based cloning technique and deposited in the GenBank (GenBank ID: KT180309). Bioinformatic analysis revealed that CsF3′H was highly homologous with the characterized F3′Hs from other plant species. Four conserved cytochrome P450-featured motifs and three F3′H-specific conserved motifs were discovered in the protein sequence of CsF3′H. Enzymatic analysis of the heterologously expressed CsF3′H in yeast demonstrated that tea F3′H catalyzed the 3′-hydroxylation of naringenin, dihydrokaempferol and kaempferol. Apparent Km values for these substrates were 17.08, 143.64 and 68.06 μM, and their apparent Vmax values were 0.98, 0.19 and 0.44 pM·min−1, respectively. Transcription level of CsF3′H in the new shoots, during tea seed germination was measured, along with that of other key genes for flavonoid biosynthesis using real-time PCR technique. The changes in 3′,4′-flavan-3-ols, 3′,4′,5′-flavan-3-ols and flavan-3-ols, were consistent with the expression level of CsF3′H and other related genes in the leaves. In the study of nitrogen supply for the tea plant growth, our results showed the expression level of CsF3′H and all other tested genes increased in response to nitrogen depletion after 12 days of treatment, in agreement with a corresponding increase in 3′,4′-catechins, 3′,4′,5′-catechins and flavan 3-ols content in the leaves. All these results suggest the import