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Sample records for clonorchis sinensis adult

  1. Cholecystolithiasis Is Associated with Clonorchis sinensis Infection

    PubMed Central

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

    2012-01-01

    Background The objective of this study was to analyze gallbladder stones for direct evidence of a relationship between Clonorchis sinensis infection and gallbladder stones formation. Methodology We investigated one hundred eighty-three gallbladder stones for the presence of Clonorchis sinensis eggs using microscopy, and analyzed their composition using Fourier transform infrared spectroscopy. We confirmed the presence of Clonorchis sinensis eggs in the gallbladder stones using real-time fluorescent PCR and scanning electron microscopy. Principal Findings Clonorchis sinensis eggs were detected in 122 of 183 gallbladder stones based on morphologic characteristics and results from real-time fluorescent PCR. The proportion of pigment stones, cholesterol stones and mixed gallstones in the egg-positive stones was 79.5% (97/122), 3.3% (4/122) and 17.2% (21/122), respectively, while 29.5% (18/61), 31.1% (19/61) and 39.3% (24/61) in the egg-negative stones. The proportion of pigment stone in the Clonorchis sinensis egg-positive stones was higher than in egg-negative stones (P<0.0001). In the 30 egg-positive stones examined by scanning electron microscopy, dozens or even hundreds of Clonorchis sinensis eggs were visible (×400) showing a distinct morphology. Many eggs were wrapped with surrounding particles, and in some, muskmelon wrinkles was seen on the surface of the eggs. Also visible were pieces of texture shed from some of the eggs. Some eggs were depressed or without operculum while most eggs were adhered to or wrapped with amorphous particles or mucoid matter (×3000). Conclusion Clonorchis sinensis eggs were detected in the gallbladder stones which suggests an association between Clonorchis sinensis infection and gallbladder stones formation, especially pigment stones. PMID:22905137

  2. 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.

  3. 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

  4. Chromosomes of the liver fluke, Clonorchis sinensis

    PubMed Central

    Park, Gab-Man; Im, Kyung-il; Huh, Sun

    2000-01-01

    A karyological study was carried out in order to compared the chromosome numbers, chromosome morphologies and karyotypes of the oriental liver fluke, Clonorchis sinensis (Trematoda: Opisthorchiidae), collected from Korea and China. Chromosome preparations were made by means of air-drying method. The chromosome number was 2n=56 in both Korean and Chinese flukes, and chromosomes were divided into two groups based on this size; consisting of 8 pairs of large and 20 pairs of small chromosomes. However, the karyotypes showed some differences between Korean and Chinese flukes. The karyotype of liver flukes from Korea consisted of three metacentric pairs, one meta-/submetacentric pair, 16 submetacentric pairs and eight subtelocentric pairs of chromosomes. On the other hand, liver flukes from China consisted of two metacentric pairs, two meta-/submetacentric pairs, 16 submetacentric pairs and eight subtelocentric pairs of chromosomes. PMID:11002660

  5. Molecular identification, immunolocalization, and characterization of Clonorchis sinensis triosephosphate isomerase.

    PubMed

    Zhou, Juanjuan; Liao, Hua; Li, Shan; Zhou, Chenhui; Huang, Yan; Li, Xuerong; Liang, Chi; Yu, Xinbing

    2015-08-01

    Clonorchis sinensis triosephosphate isomerase (CsTIM) is a key regulatory enzyme of glycolysis and gluconeogenesis, which catalyzes the interconversion of glyceraldehyde 3-phosphate to dihydroxyacetone phosphate. In this study, the biochemical characterizations of CsTIM have been examined. A full-length complementary DNA (cDNA; Cs105350) sequence encoding CsTIM was obtained from our C. sinensis cDNA library. The open reading frame of CsTIM contains 759 bp which encodes 252 amino acids. The amino acid sequence of CsTIM shares 60-65% identity with other species. Western blot analysis displayed that recombinant CsTIM (rCsTIM) can be probed by anti-rCsTIM rat serum and anti-C. sinensis excretory/secretory products (anti-CsESPs) rat serum. Quantitative reverse transcription (RT)-PCR and western blotting analysis revealed that CsTIM messenger RNA (mRNA) and protein were differentially expressed in development cycle stages of the parasite, including adult worm, metacercaria, excysted metacercaria, and egg. In addition, immunolocalization assay showed that CsTIM was located in the seminal vesicle, eggs, and testicle. Moreover, rCsTIM exhibited active enzyme activity in catalytic reactions. The Michaelis constant (K m) of rCsTIM was 0.33 mM, when using glyceraldehyde 3-phosphate as the substrate. The optimal temperature and pH of CsTIM were 37 °C and 7.5-9.5, respectively. Collectively, these results suggest that CsTIM is an important protein involved in glycometabolism, and CsTIM possibly take part in many biological functions in the growth and development of C. sinensis.

  6. [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

  7. 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.

  8. 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

  9. 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.

  10. 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.

  11. 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.

  12. 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

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

    PubMed

    Xu, Min-Jun; Liu, Quan; Nisbet, Alasdair J; Cai, Xian-Quan; Yan, Chao; Lin, Rui-Qing; Yuan, Zi-Guo; Song, Hui-Qun; He, Xian-Hui; Zhu, Xing-Quan

    2010-09-28

    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. 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. 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 Opisthorchis viverrini and

  14. 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.

  15. Comparison of two serpins of Clonorchis sinensis by bioinformatics, expression, and localization in metacercaria.

    PubMed

    Yang, Yabo; Hu, Dong; Wang, Lexun; Liang, Chi; Hu, Xuchu; Xu, Jin; Huang, Yan; Yu, Xinbing

    2014-06-01

    Clonorchiasis, which has been an important public health problem in China, is caused by ingestion of raw or undercooked fish contaminated by live metacercaria. Therefore, preventing fish from infecting is of great significance for controlling the disease. SERPINs (serine protease inhibitors) are well known as negative regulators of hemostasis, thrombolysis, and innate immune responses. In the present study, two full-length sequences encoding SERPIN were identified from metacercaria cDNA library of Clonorchis sinensis (C. sinensis) and were denominated as CsSERPIN and CsSERPIN3, respectively. Bioinformatics analysis showed that the two sequences shares 35.9% identity to each other. Both of the sequences have SERPIN domain and the greatest difference between the two domains is the reactive centre loop. Transmembrane region was found in CsSERPIN3 while not in CsSERPIN. The expression of the two CsSERPINs was significantly higher at the life stage of metacercaria than that of adult. The transcription levels of CsSERPIN and CsSERPIN3 at metacercaria stage were 3.249- and 11.314-fold of that at adult stage, respectively. Furthermore, the expression of CsSERPIN was 4.32-fold of that of CsSERPIN3 at metacercaria stage. Immunobiochemistry revealed that CsERPIN was dispersed at subtegument and oral sucker of metacercaria, while CsSERPIN3 localized intensely in the tegument of metacercaria of C. sinensis inside of the cyst wall. All these indicated that the CsSERPINs play important roles at metacercaria stage of the parasite. CsSERPIN may take part in regulation of endogenous serine proteinase and CsSERPIN3 may be involved in immune evasion and be a potential candidate for vaccine and drug target for clonorchiasis.

  16. [Cloning and prokaryotic expression of transcriptional co-activator gene of Clonorchis sinensis and functional analysis of the expressed protein].

    PubMed

    Zhang, Yong-li; Yu, Xin-bing; Wu, De; Wu, Zhong-dao; Bi, Hui-xiang

    2005-02-28

    To construct prokaryotic recombinant plasmids of transcriptional co-activator (TC) gene of Clonorchis sinensis, express and purify the recombinant protein and analyze its biological function. A pair of primers was designed according to the known sequence of TC gene. The TC gene fragment was amplified by PCR. After purification and digestion with BamH I and Sal I, the TC gene was connected to the prokaryotic expression vectors, pGEX-4T-1 and pET30a(+). By cloning target gene into these vectors, pGEX-4T-1 and pET30a(+), prokaryotic recombinant plasmids of TC gene were constructed and transferred into E. coli BL21. The positive expressed recombinants were detected by SDS-PAGE and Western blotting. Immobilized metal (Ni2+) chelation affinity chromatography was used to purify His-TC produced by the expression of the recombinant protein pET30a(+)-TC. The recombinant plasmids, pGEX-4T-1-TC and pET30a(+)-TC, were constructed successfully. SDS-PAGE testified that the molecular weight of the recombinant protein was correct. Western blot analysis of GST-TC recombinant protein testified that the recombinant protein could be recognized by immunized rabbit serum, which means the protein is GST-immune active and the clone can express recombinant Clonorchis sinensis antigen. After affinity chromatography of the pET-TC protein, there was only one protein band with expected size on the SDS-PAGE gel. The TC gene was screened from cDNA library of adult Clonorchis sinensis, cloned, expressed and purified. The purified protein of TC gene will be of importance for further research on the biological function of the gene.

  17. 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

  18. 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.

  19. Specific and common antigens of Clonorchis sinensis and Opisthorchis viverrini (Opisthorchidae, Trematoda)

    PubMed Central

    Choi, Min-Ho; Ryu, Jin-Sook; Lee, Mejeong; Li, Shunyu; Chung, Byung-Suk; Chai, Jong-Yil; Sithithaworn, Paiboon; Tesana, Smarn

    2003-01-01

    The antigenic characterizations and serological reactions of human liver flukes, Clonorchis sinensis and Opisthorchis viverrini, were analyzed by immunoblot. The antigenic profiles of the crude extract of Clonorchis contained major proteins of 8, 26-28, 34-37, 43, and 70 kDa, and those of Opisthorchis 34-37, 43, 70, and 100 kDa. Of these, the 8, 26-28 and 34-37 kDa bands of Clonorchis and the 100 kDa of Opisthorchis were major components of each excretory-secretory antigen. The 8 and 26-28 kDa bands were specific to Clonorchis but the 100 kDa of Opisthorchis cross-reacted with the sera of clonorchiasis, and the 34-37, 70 and 100 kDa bands cross-reacted with sera of other helminthiases. The frequency and intensity of the immunoblot reactions were positively correlated with the intensity of the liver fluke infection. PMID:12972729

  20. 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

  1. 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.

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

    PubMed

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

    2016-11-01

    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. [BMB Reports 2016; 49(11): 590-597].

  3. 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

  4. 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.

  5. 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.

  6. Production and Deformation of Clonorchis sinensis Eggs during In Vitro Maintenance

    PubMed Central

    Uddin, Md. Hafiz; Bae, Young Mee; Choi, Min-Ho; Hong, Sung-Tae

    2012-01-01

    Clonorchis sinensis is a carcinogenic human liver fluke. The present study monitored eggs produced by long-term maintained adult worms of C. sinensis to confirm their egg productivity in vitro. The worms from infected rabbits were incubated in vitro in 1× Locke’s solution and broth media (RPMI-1640, DMEM and IMDM). Numbers of expelled eggs were counted sequentially and their morphological changes were monitored by microscopy after 1, 30, 60, and 90 days of cultivation. On the 1–3 days of cultivation, the eggs counted maximum 4,756±202 eggs/worm/day in IMDM medium. The number of eggs gradually decreased less than 1,000 at 7–14 days and below 100 at 21days but continued to pass eggs after 56 days in all media. Length of the eggs were reduced about 1 µm at 30 days, and the length/width ratio was maintained around 1.8 at 30 days but decreased to 1.7 at 60 days and 1.5 at 90 days. Faust-Meleney index (FMI) decreased as the cultivation duration increased and lowest FMI (5662.9±974.7) observed in IMDM media at day 90 (P = 0.001). Microscopic findings of the eggs recognized the miracidium in most of eggs at 60 days but not in those at 90 days. Instead, the eggs contained dark granules or vacuoles in the deformed shell at 90 days. Scanning electron microscopy revealed partial loss of wrinkles on the deformed egg surface and prominent abopercular knob. Eggs viability decreased as the cultivation progressed and showed significant positive correlation with FMI and length/width ratio. In conclusion, the cultivated worms pass only the eggs which are preformed in their uterus before cultivation. One gravid C. sinensis contains about 37,000 eggs in its uterus and produces about 4,000 eggs every day. The deformed eggs with FMI less than 7,000 and length/width ratio lower than 1.7 are non-viable. PMID:23285144

  7. 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

  8. [Research on chemotherapy measures in different Clonorchis sinensis infectiosity endemic areas].

    PubMed

    Fang, Yue-yi; Ruan, Cai-wen; Gao, Xiang-xiong; Tan, Tan; Chen, Rao-rao; Hao, Yang

    2014-06-01

    To explore the suitable chemotherapy measures in different Clonorchis sinensis infectiosity endemic areas. The interventional groups and control groups were set up in the heavy, moderate and light C. sinensis infectiosity areas in Guangdong Province, respectively. In the intervention groups, the chemotherapy was administered among all the residents aged above 3 years in the heavy endemic areas, the chemotherapy was given among the focus populations in the moderate endemic areas, and the chemotherapy was administered among the residents infected with C. sinensis confirmed by stool examinations in the light endemic areas. No measures were carried out in the control groups. One year after the interventions, the C. sinensis infection rates of the interventional groups decreased by 47.90%, 86.52% and 100%, the abnormal rates of liver B ultrasonic examinations decreased by 26.50%, 31.00% and 100%, the ALT abnormal rates decreased by 48.70%, 62.30% and 100%, and the AST abnormal rates decreased by 55.50%, 39.90% and 100% in the heavy, moderate and light endemic areas, respectively. There was no decreasing trend of above-mentioned indexes in the control groups. In the different C. sinensis infectiosity endemic areas, the different chemotherapy measures should be carried out.

  9. [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.

  10. [Genomics and transcriptomics of the Chinese liver fluke Clonorchis sinensis (Opisthorchiidae, Trematoda)].

    PubMed

    Chelomina, G N

    2017-01-01

    The review summarizes the results of first genomic and transcriptomic investigations of the liver fluke Clonorchis sinensis (Opisthorchiidae, Trematoda). The studies mark the dawn of the genomic era for opisthorchiids, which cause severe hepatobiliary diseases in humans and animals. Their results aided in understanding the molecular mechanisms of adaptation to parasitism, parasite survival in mammalian biliary tracts, and genome dynamics in the individual development and the development of parasite-host relationships. Special attention is paid to the achievements in studying the codon usage bias and the roles of mobile genetic elements (MGEs) and small interfering RNAs (siRNAs). Interspecific comparisons at the genomic and transcriptomic levels revealed molecular differences, which may contribute to understanding the specialized niches and physiological needs of the respective species. The studies in C. sinensis provide a basis for further basic and applied research in liver flukes and, in particular, the development of efficient means to prevent, diagnose, and treat clonorchiasis.

  11. Genome-wide characterization of microsatellites and marker development in the carcinogenic liver fluke Clonorchis sinensis.

    PubMed

    Nguyen, Thao T B; Arimatsu, Yuji; Hong, Sung-Jong; Brindley, Paul J; Blair, David; Laha, Thewarach; Sripa, Banchob

    2015-06-01

    Clonorchis sinensis is an important carcinogenic human liver fluke endemic in East and Southeast Asia. There are several conventional molecular markers that have been used for identification and genetic diversity; however, no information about microsatellites of this liver fluke is published so far. We here report microsatellite characterization and marker development for a genetic diversity study in C. sinensis, using a genome-wide bioinformatics approach. Based on our search criteria, a total of 256,990 microsatellites (≥12 base pairs) were identified from a genome database of C. sinensis, with hexanucleotide motif being the most abundant (51%) followed by pentanucleotide (18.3%) and trinucleotide (12.7%). The tetranucleotide, dinucleotide, and mononucleotide motifs accounted for 9.75, 7.63, and 0.14%, respectively. The total length of all microsatellites accounts for 0. 72% of 547 Mb of the whole genome size, and the frequency of microsatellites was found to be one microsatellite in every 2.13 kb of DNA. For the di-, tri-, and tetranucleotide, the repeat numbers redundant are six (28%), four (45%), and three (76%), respectively. The ATC repeat is the most abundant microsatellites followed by AT, AAT, and AC, respectively. Within 40 microsatellite loci developed, 24 microsatellite markers showed potential to differentiate between C. sinensis and Opisthorchis viverrini. Seven out of 24 loci showed to be heterozygous with observed heterozygosity that ranged from 0.467 to 1. Four primer sets could amplify both C. sinensis and O. viverrini DNA with different sizes. This study provides basic information of C. sinensis microsatellites, and the genome-wide markers developed may be a useful tool for the genetic study of C. sinensis.

  12. Prevalence of Clonorchis sinensis infection in freshwater fishes in northeastern China.

    PubMed

    Zhang, Y; Chang, Q C; Zhang, Y; Na, L; Wang, W T; Xu, W W; Gao, D Z; Liu, Z X; Wang, C R; Zhu, X Q

    2014-08-29

    The prevalence of Clonorchis sinensis infection in freshwater fishes was surveyed in Heilongjiang Province, northeastern China, between August 2011 and September 2013. Thirteen species of freshwater fish (n=3221) and one species of shrimp (n=93) were collected from Songhua river, Nenjiang river and other lakes or ponds in 37 sites of 15 representative cities in Heilongjiang Province. They were individually examined by digestion technique, and the C. sinensis metacercariae were identified morphologically followed by confirmation using sequences of the second internal transcribed spacer of ribosomal DNA. Ten of the 13 examined species of freshwater fishes were infected with C. sinensis metacercariae, while all shrimps were negative. The overall prevalence of C. sinensis infection in 3221 examined freshwater fishes was 19.96%, with 42.57% (272/639) in Pseudorasbora parva, 22.55% (83/368) in Hemicculter leuciclus, 20.44% (121/592) in Carassius auratus, 17.71% (68/384) in Saurogobio dabryi, 10.85% (23/212) in Rhodeus ocellatus, 10.54% (48/455) in Phoxinus lagowskii, 8.20% (21/256) in Perccottus glehnii, 6.25% (5/80) in Misgurnus anguillicaudatus, 4.55% (1/22) in Xenocypris davidi, and 1.49% (1/67) in Cyprinus carpio. The average infection intensity in P. parva was 103.3 encysted metacercariae per gram of fish meat in Zhaoyuan city. The average prevalence of C. sinensis infection in Songhua river, Nenjiang river and lakes or ponds were 31.96% (503/1574), 11.30% (102/903) and 7.93% (59/744), respectively. The prevalence of C. sinensis infection in Zhaoyuan city (43.68%) was the highest among all sampling locations. These results revealed a high-prevalence of C. sinensis infection in freshwater fishes in Heilongjiang Province, northeastern China, posing significant public health concern. Copyright © 2014 Elsevier B.V. All rights reserved.

  13. Prevalence of Clonorchis sinensis Infection among Residents along 5 Major Rivers in the Republic of Korea.

    PubMed

    Jeong, Young-Il; Shin, Hee-Eun; Lee, Sang-Eun; Cheun, Hyeng-Il; Ju, Jung-Won; Kim, Jung-Yeon; Park, Mi Yeoun; Cho, Shin-Hyeong

    2016-04-01

    Clonorchis sinensis is currently the most important parasite affecting public health problems in the Republic of Korea. We investigated the prevalence of C. sinensis infection among residents living along 5 major rivers in Korea. A total of 42,562 individual stool samples were collected from 37 localities and examined using the formalin-ether sedimentation technique. Helminth eggs were detected in 4,052 (9.5%) residents and 3,586 (8.4%) were infected with C. sinensis. The egg positive rate of C. sinensis in Nakdong, Seomjin, Geum, Yeongsan, and Han River was 11.7%, 9.9%, 6.5%, 3.1%, and 1.0%, respectively. The overall prevalence of clonorchiasis by sex was 11.2% in males and 6.2% in females. The age-prevalence was the highest in the 50-59 years band. It has been reconfirmed that the endemicity of clonorchiasis is higher in southern areas of Korea, especially along Nakdong and Seomjin Rivers. A combination of continuous control programs with health education initiatives is urgently required in these highly endemic areas of clonorchiasis in Korea.

  14. Unlocking the Transcriptomes of Two Carcinogenic Parasites, Clonorchis sinensis and Opisthorchis viverrini

    PubMed Central

    Young, Neil D.; Campbell, Bronwyn E.; Hall, Ross S.; Jex, Aaron R.; Cantacessi, Cinzia; Laha, Thewarach; Sohn, Woon-Mok; Sripa, Banchob; Loukas, Alex; Brindley, Paul J.; Gasser, Robin B.

    2010-01-01

    The two parasitic trematodes, Clonorchis sinensis and Opisthorchis viverrini, have a major impact on the health of tens of millions of humans throughout Asia. The greatest impact is through the malignant cancer ( = cholangiocarcinoma) that these parasites induce in chronically infected people. Therefore, both C. sinensis and O. viverrini have been classified by the World Health Organization (WHO) as Group 1 carcinogens. Despite their impact, little is known about these parasites and their interplay with the host at the molecular level. Recent advances in genomics and bioinformatics provide unique opportunities to gain improved insights into the biology of parasites as well as their relationships with their hosts at the molecular level. The present study elucidates the transcriptomes of C. sinensis and O. viverrini using a platform based on next-generation (high throughput) sequencing and advanced in silico analyses. From 500,000 sequences, >50,000 sequences were assembled for each species and categorized as biologically relevant based on homology searches, gene ontology and/or pathway mapping. The results of the present study could assist in defining molecules that are essential for the development, reproduction and survival of liver flukes and/or that are linked to the development of cholangiocarcinoma. This study also lays a foundation for future genomic and proteomic research of C. sinensis and O. viverrini and the cancers that they are known to induce, as well as novel intervention strategies. PMID:20582164

  15. 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.

  16. The draft genome of the carcinogenic human liver fluke Clonorchis sinensis.

    PubMed

    Wang, Xiaoyun; Chen, Wenjun; Huang, Yan; Sun, Jiufeng; Men, Jingtao; Liu, Hailiang; Luo, Fang; Guo, Lei; Lv, Xiaoli; Deng, Chuanhuan; Zhou, Chenhui; Fan, Yongxiu; Li, Xuerong; Huang, Lisi; Hu, Yue; Liang, Chi; Hu, Xuchu; Xu, Jin; Yu, Xinbing

    2011-10-24

    Clonorchis sinensis is a carcinogenic human liver fluke that is widespread in Asian countries. Increasing infection rates of this neglected tropical disease are leading to negative economic and public health consequences in affected regions. Experimental and epidemiological studies have shown a strong association between the incidence of cholangiocarcinoma and the infection rate of C. sinensis. To aid research into this organism, we have sequenced its genome. We combined de novo sequencing with computational techniques to provide new information about the biology of this liver fluke. The assembled genome has a total size of 516 Mb with a scaffold N50 length of 42 kb. Approximately 16,000 reliable protein-coding gene models were predicted. Genes for the complete pathways for glycolysis, the Krebs cycle and fatty acid metabolism were found, but key genes involved in fatty acid biosynthesis are missing from the genome, reflecting the parasitic lifestyle of a liver fluke that receives lipids from the bile of its host. We also identified pathogenic molecules that may contribute to liver fluke-induced hepatobiliary diseases. Large proteins such as multifunctional secreted proteases and tegumental proteins were identified as potential targets for the development of drugs and vaccines. This study provides valuable genomic information about the human liver fluke C. sinensis and adds to our knowledge on the biology of the parasite. The draft genome will serve as a platform to develop new strategies for parasite control.

  17. The draft genome of the carcinogenic human liver fluke Clonorchis sinensis

    PubMed Central

    2011-01-01

    Background Clonorchis sinensis is a carcinogenic human liver fluke that is widespread in Asian countries. Increasing infection rates of this neglected tropical disease are leading to negative economic and public health consequences in affected regions. Experimental and epidemiological studies have shown a strong association between the incidence of cholangiocarcinoma and the infection rate of C. sinensis. To aid research into this organism, we have sequenced its genome. Results We combined de novo sequencing with computational techniques to provide new information about the biology of this liver fluke. The assembled genome has a total size of 516 Mb with a scaffold N50 length of 42 kb. Approximately 16,000 reliable protein-coding gene models were predicted. Genes for the complete pathways for glycolysis, the Krebs cycle and fatty acid metabolism were found, but key genes involved in fatty acid biosynthesis are missing from the genome, reflecting the parasitic lifestyle of a liver fluke that receives lipids from the bile of its host. We also identified pathogenic molecules that may contribute to liver fluke-induced hepatobiliary diseases. Large proteins such as multifunctional secreted proteases and tegumental proteins were identified as potential targets for the development of drugs and vaccines. Conclusions This study provides valuable genomic information about the human liver fluke C. sinensis and adds to our knowledge on the biology of the parasite. The draft genome will serve as a platform to develop new strategies for parasite control. PMID:22023798

  18. 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.

  19. 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

  20. 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.

  1. 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.

  2. The involvement of the cysteine proteases of Clonorchis sinensis metacercariae in excystment.

    PubMed

    Li, Shunyu; Chung, Young-Bae; Chung, Byung-Suk; Choi, Min-Ho; Yu, Jae-Ran; Hong, Sung-Tae

    2004-05-01

    The effects of trypsin, bile, trypsin-bile, pepsin, dithiothreitol (DTT) and metacercarial excretory-secretory product (ESP) on the in vitro excystment of Clonorchis sinensis metacercariae were investigated. The majority of metacercariae excysted immediately in trypsin-bile in PBS solution, a process which was complete after 30 min of incubation. When incubated in metacercarial ESP in PBS, excystment was potentiated in the presence of 5 mM DTT, but was inhibited dose-dependently by a cysteine protease inhibitor, iodoacetic acid. Two active protease bands of 28 and 40 kDa were identified in the ESP of metacercariae by gelatin substrate SDS-PAGE. Scanning electron microscopy demonstrated that the larvae in solutions of DTT and ESP migrated through a small hole on the metacercarial wall, whereas larvae were liberated by entire wall disruption in trypsin solution. These results suggest that trypsin is a major extrinsic factor of the rapid excystment of C. sinensis metacercariae, and that endogenous cysteine proteases are also involved in metacercarial excystment. Copyright 2004 Springer-Verlag

  3. Cystatin Capture Enzyme-Linked Immunosorbent Assay for Serodiagnosis of Human Clonorchiasis and Profile of Captured Antigenic Protein of Clonorchis sinensis

    PubMed Central

    Kim, Tae Yun; Kang, Shin-Yong; Park, Sun Hyo; Sukontason, Kom; Sukontason, Kabkaew; Hong, Sung-Jong

    2001-01-01

    Enzyme-linked immunosorbent assay (ELISA) with crude extracts of adult Clonorchis sinensis has been reported to have a high degree of sensitivity with a moderate degree of specificity for the serodiagnosis of clonorchiasis. The cystatin capture ELISA was investigated for its usefulness for the serodiagnosis of human clonorchiasis. Cystatin bound specifically to cysteine proteinases in crude extracts of adult C. sinensis worms, and its binding capacity was not hindered competitively by the other proteinase inhibitors tested. The cystatin capture ELISA for clonorchiasis showed a higher degree of specificity than the conventional ELISA, which produced some cross-reactivities to sera from patients with cysticercosis, sparganosis, and opisthorchiasis. Immunoglobulin G antibodies to C. sinensis cysteine proteinases were produced in experimental rabbits at week 3, and their levels increased rapidly and remained at a plateau after 8 weeks of infection. Of the proteins from the C. sinensis crude extract captured with cystatin, seven proteins were reactive with the serum from patients with clonorchiasis. The cystatin capture ELISA is indicated to be a sensitive and highly specific immunodiagnostic assay for serodiagnosis of human clonorchiasis. PMID:11687443

  4. Clonorchis sinensis: life cycle, intermediate hosts, transmission to man and geographical distribution in Korea.

    PubMed

    Choi, D W

    1984-01-01

    The snail host of Clonorchis sinensis in Korea is referred to as Parafossarulus manchouricus, but it has been recorded also as Bulimus striatulus japonicus in literature published up to late 1950. The specific name of the snail was corrected such that Bulimus striatulus of China, Japan, and Korea should be incorporated in the species of P. manchouricus. In general, the snail exists in limited areas of the rivers Han, Kum, Mankyung, Naktong, and Yeongsan, and the infestation of the snail with the cercaria of C. sinensis is very low. In recent years, ten species of small fresh-water fish are known as second intermediate hosts of the fluke. Of these, the southern top-mouthed minnow, Pseudorasbora parva, was most heavily infected. The striped shiner, Pungtungia herzi, Korean shiner, Gnathopogon atromaculatus, gudgeon, Pseudogobio esocinus, oily shiner, Sarcocheilichthys sinensis, and Korean barbel, Hemibarbus labeo, were infected intermediately. Whereas, the flat bitterling, Paracheilognathus rhombea, oily bitterling, Acheilognathus limbata, and Korean rose bitterling, Acanthorhodeus taenianalis were infected with a few cysts. No infections were found in the pale chub, Zacco platypus, crussian carp, Carassius carassius, and carp, Cyprinus carpio. Most Koreans are usually infected by eating slices of raw fresh-water fish coated with hot bean paste. Raw fish is a common accompaniment in drinking the rice wine "Marcgulee" in rural communities and a health food for males. Numerous epidemiological studies revealed that the prevalence rate for the fluke among the residents in the vicinity of rivers is higher than those in the areas far from the rivers. The rate is higher among males than females and increases with age.

  5. 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.

  6. 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. Copyright © 2014 Australian Society for

  7. Prevalence of Clonorchis sinensis Metacercariae in Fish from Water Systems of Seomjin-gang (River).

    PubMed

    Sohn, Woon-Mok; Na, Byoung-Kuk; Cho, Shin-Hyeong; Park, Mi-Yeoun; Kim, Cheon-Hyeon; Hwang, Min-Ah; No, Kyeong-Woo; Yoon, Ki-Bok; Lim, Hyun-Cheol

    2017-06-01

    The prevalence of Clonorchis sinensis metacercariae (CsMc) was examined in freshwater fish from the water systems of Seomjin-gang (River), the Republic of Korea. Total 1,604 fish from 7 local sites of Seomjin-gang were examined by artificial digestion methods. The metacercariae of C. sinensis were detected in 102 (39.8%) out of 256 fish (14 species) from the upper reaches of Seomjin-gang, i.e., Osucheon (22.3% in 6 fish species) in Imsil-gun, and Seomjin-gang (63.9% in 9 fish species) in Sunchang-gun, Jeollabuk-do. Their average density was 9.0 per infected fish. They were also found in 132 (48.0%) out of 275 fish (12 spp.) from the middle reaches of Seomjin-gang, i.e., Songdaecheon (58.9% in 4 fish species) in Namwon-si, Jeollabuk-do, and Seomjin-gang (45.2% in 10 fish species) in Gokseong-gun, Jeollanam-do. Their average density was 21.0 per infected fish. CsMc were detected in 77 (56.6%) out of 136 fish (11 species) from the lower reaches of Seomjin-gang, i.e., Seomjin-gang (73.3% in 11 fish species) in Gurye-gun, Jeollanam-do, and Namsancheon (8.6% in 1 fish species) in Hadong-gun, Gyeongsangnam-do. Their average density was 64.9 per infected fish. The metacercariae of Metorchis orientalis were also detected in 6 fish species from 4 sites of Seomjin-gang. Conclusively, it has been confirmed that CsMc are more or less prevalent in fish from some water systems of Seomjin-gang in Korea.

  8. Prevalence of Clonorchis sinensis Metacercariae in Fish from Water Systems of Seomjin-gang (River)

    PubMed Central

    Sohn, Woon-Mok; Na, Byoung-Kuk; Cho, Shin-Hyeong; Park, Mi-Yeoun; Kim, Cheon-Hyeon; Hwang, Min-Ah; No, Kyeong-Woo; Yoon, Ki-Bok; Lim, Hyun-Cheol

    2017-01-01

    The prevalence of Clonorchis sinensis metacercariae (CsMc) was examined in freshwater fish from the water systems of Seomjin-gang (River), the Republic of Korea. Total 1,604 fish from 7 local sites of Seomjin-gang were examined by artificial digestion methods. The metacercariae of C. sinensis were detected in 102 (39.8%) out of 256 fish (14 species) from the upper reaches of Seomjin-gang, i.e., Osucheon (22.3% in 6 fish species) in Imsil-gun, and Seomjin-gang (63.9% in 9 fish species) in Sunchang-gun, Jeollabuk-do. Their average density was 9.0 per infected fish. They were also found in 132 (48.0%) out of 275 fish (12 spp.) from the middle reaches of Seomjin-gang, i.e., Songdaecheon (58.9% in 4 fish species) in Namwon-si, Jeollabuk-do, and Seomjin-gang (45.2% in 10 fish species) in Gokseong-gun, Jeollanam-do. Their average density was 21.0 per infected fish. CsMc were detected in 77 (56.6%) out of 136 fish (11 species) from the lower reaches of Seomjin-gang, i.e., Seomjin-gang (73.3% in 11 fish species) in Gurye-gun, Jeollanam-do, and Namsancheon (8.6% in 1 fish species) in Hadong-gun, Gyeongsangnam-do. Their average density was 64.9 per infected fish. The metacercariae of Metorchis orientalis were also detected in 6 fish species from 4 sites of Seomjin-gang. Conclusively, it has been confirmed that CsMc are more or less prevalent in fish from some water systems of Seomjin-gang in Korea. PMID:28719955

  9. Identification and molecular characterization of a novel signaling molecule 14-3-3 epsilon in Clonorchis sinensis excretory/secretory products.

    PubMed

    Wang, Xiaoyun; Chen, Wenjun; Li, Xuerong; Zhou, Chenhui; Deng, Chuanhuan; Lv, Xiaoli; Fan, Yongxiu; Men, Jingtao; Liang, Chi; Yu, Xinbing

    2012-04-01

    Increasing evidence shows that 14-3-3 proteins are involved in many biology events in addition to signal transduction. Extensive investigations on structural and biochemical features of these signaling molecules have implied their importance in the biological process. In the present study, we have identified and characterized the 14-3-3 epsilon (Cs14-3-3) in Clonorchis sinensis that causes human clonorchiasis. Recombinant protein was expressed in Escherichia coli (E. coli) and identified by MALDI-TOF/TOF. Immunoblot results revealed that Cs14-3-3 was a component of excretory/secretory products. Ligand blot assay indicated that 14-3-3 epsilon could bind C. sinensis MAPKAPK 2 in a nonphosphorylation-dependent manner. This protein could be detected at four stages of the life cycle by RT-PCR experiments and immunolocalization showed that Cs14-3-3 was extensively distributed in C. sinensis, especially at the outer surface and the sucker of adult worm and cyst wall of metacercaria. Taken together, 14-3-3 epsilon might play some roles in the development of the parasites. In addition, Cs14-3-3 epsilon should be addressed for the diagnostic value in C. sinensis infection in consideration of high sensitivity and specificity. As an immune stimulus, C. sinensis 14-3-3 epsilon was found to provoke a Th1/Th2 balanced immune response by inducing high levels of both IgG1 and IgG2a. Recombinant Cs14-3-3 conferred effective protection both in worm reduction rate and egg reduction rate, suggesting that the signaling molecule Cs14-3-3 was a promising vaccine candidate against C. sinensis infection.

  10. Development of a polymerase chain reaction applicable to rapid and sensitive detection of Clonorchis sinensis eggs in human stool samples

    PubMed Central

    Cho, Pyo Yun; Na, Byoung-Kuk; Mi Choi, Kyung; Kim, Jin Su; Cho, Shin-Hyeong; Lee, Won-Ja; Lim, Sung-Bin; Cha, Seok Ho; Park, Yun-Kyu; Pak, Jhang Ho; Lee, Hyeong-Woo; Hong, Sung-Jong; Kim, Tong-Soo

    2013-01-01

    Microscopic examination of eggs of parasitic helminths in stool samples has been the most widely used classical diagnostic method for infections, but tiny and low numbers of eggs in stool samples often hamper diagnosis of helminthic infections with classical microscopic examination. Moreover, it is also difficult to differentiate parasite eggs by the classical method, if they have similar morphological characteristics. In this study, we developed a rapid and sensitive polymerase chain reaction (PCR)-based molecular diagnostic method for detection of Clonorchis sinensis eggs in stool samples. Nine primers were designed based on the long-terminal repeat (LTR) of C. sinensis retrotransposon1 (CsRn1) gene, and seven PCR primer sets were paired. Polymerase chain reaction with each primer pair produced specific amplicons for C. sinensis, but not for other trematodes including Metagonimus yokogawai and Paragonimus westermani. Particularly, three primer sets were able to detect 10 C. sinensis eggs and were applicable to amplify specific amplicons from DNA samples purified from stool of C. sinensis-infected patients. This PCR method could be useful for diagnosis of C. sinensis infections in human stool samples with a high level of specificity and sensitivity. PMID:23916334

  11. 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.

  12. 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

  13. 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.

  14. 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.

  15. Prevalence of Clonorchis sinensis Metacercariae in Freshwater Fish from Three Latitudinal Regions of the Korean Peninsula

    PubMed Central

    Cho, Shin-Hyeong; Na, Byoung-Kuk; Kim, Tong-Soo; Kong, Yoon; Eom, Keeseon; Seok, Won-Seok; Lee, Taejoon

    2011-01-01

    A large-scale survey was conducted to investigate the infection status of fresh water fishes with Clonorchis sinensis metacercariae (CsMc) in 3 wide regions, which were tentatively divided by latitudinal levels of the Korean peninsula. A total of 4,071 freshwater fishes were collected from 3 regions, i.e., northern (Gangwon-do: 1,543 fish), middle (Chungcheongbuk-do and Gyeongsangbuk-do: 1,167 fish), and southern areas (Jeollanam-do, Ulsan-si, and Gyeongsangnam-do: 1,361 fish). Each fish was examined by the artificial digestion method from 2003 to 2010. In northern areas, only 11 (0.7%) fish of 2 species, Pungtungia herzi and Squalidus japonicus coreanus from Hantan-gang, Cheolwon-gun, Gangwon-do were infected with av. 2.6 CsMc. In middle areas, 149 (12.8%) fish were infected with av. 164 CsMc. In southern areas, 538 (39.5%) fish were infected with av. 159 CsMc. In the analysis of endemicity in 3 regions with an index fish, P. herzi, 9 (6.2%) of 146 P. herzi from northern areas were infected with av. 2.8 CsMc. In middle areas, 34 (31.8%) of 107 P. herzi were infected with av. 215 CsMc, and in southern areas, 158 (92.9%) of 170 P. herzi were infected with av. 409 CsMc. From these results, it has been confirmed that the infection status of fish with CsMc is obviously different among the 3 latitudinal regions of the Korean peninsula with higher prevalence and burden in southern regions. PMID:22355206

  16. Expression, immunolocalization, and serological reactivity of a novel sphingomyelin phosphodiesterase-like protein, an excretory/secretory antigen from Clonorchis sinensis.

    PubMed

    Huang, Yanwei; Zheng, Youwei; Li, Yuzhe; Yang, Mei; Li, Ting; Zeng, Suxiang; Yu, Xinbing; Huang, Huaiqiu; Hu, Xuchu

    2013-06-01

    Clonorchiasis, caused by Clonorchis sinensis infection, is a zoonotic parasitic disease of hepatobiliary system in which the proteins released by adult are major pathogenetic factors. In this study, we first characterized a putative sphingomyelin phosphodiesterase (CsSMPase) A-like secretory protein, which was highly expressed in the adult worm. The full-length gene was cloned. The putative protein is of relatively low homology comparing with SMPase from other species, and of rich T cell and B cell epitopes, suggesting that it is an antigen of strong antigenicity. The complete coding sequence of the gene was expressed in the Escherichia coli. The recombinant CsSMPase (rCsSMPase) can be recognized by C. sinensis-infected serum, and the protein immunoserum can recognize a specific band in excretory/secretory products (ESPs) of C. sinensis adult by western blotting. Immunolocalization revealed that CsSMPase was not only localized on tegument, ventral sucker of metacercaria, and the intestine of adult but also on the nearby epithelium of bile duct of the infected Sprague-Dawley rats, implying that CsSMPase was mainly secreted and excreted through adult intestine and directly interacted with bile duct epithelium. Although immunized rats evoked high level antibody response, the antigen level was low in clonorchiasis patients. And the sensitivity and specificity of rCsSMPase were 50.0 % (12/24) and 88.4 % (61/69), in sera IgG-ELISA, respectively. It is likely due to the fact that CsSMPase binding to the plasma membrane of biliary epithelium decreases the antigen immune stimulation.

  17. 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.

  18. 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.

  19. 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

  20. 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

  1. 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

  2. Clonorchis sinensis infection and co-infection with the hepatitis B virus are important factors associated with cholangiocarcinoma and hepatocellular carcinoma.

    PubMed

    Shi, Yunliang; Jiang, Zhihua; Yang, Yichao; Zheng, Peiqiu; Wei, Haiyan; Lin, Yuan; Lv, Guoli; Yang, Qingli

    2017-08-12

    To evaluate the contributions of Clonorchis sinensis and hepatitis B virus to the development of cholangiocarcinoma (ICC) and hepatocellular carcinoma (HCC), C. sinensis and hepatitis B virus infections in 20 clinical liver cancer cases from a C. sinensis- and hepatitis B virus-epidemic region were detected. Eight cases of ICC, 11 cases of HCC and one mixed ICC and HCC case were verified by CT, pathological section and (or) observations during surgery. The C. sinensis infection was detected by stool microscopy and ELISA, and the worms and eggs found during surgery and in pathological sections also allowed for diagnoses. Hepatitis B virus infections were detected by ELISA. In the 20 cases, 18 patients were diagnosed with C. sinensis infections. Eight of the 20 patients were infected with the hepatitis B virus, and seven were co-infected with C. sinensis. In the eight ICC patients, seven were diagnosed with C. sinensis infection, and two had mixed infections with the hepatitis B virus. In the 11 HCC patients, 10 were diagnosed with C. sinensis, four had mixed infections with the hepatitis B virus, and only one HCC patient presented a single infection by the hepatitis B virus. These clinical observations revealed that C. sinensis infection and C. sinensis co-infection with the hepatitis B virus are important factors in ICC and HCC.

  3. 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.

  4. 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

  5. Bile components and amino acids affect survival of the newly excysted juvenile Clonorchis sinensis in maintaining media.

    PubMed

    Li, Shunyu; Kim, Tae Im; Yoo, Won Gi; Cho, Pyo Yun; Kim, Tong-Soo; Hong, Sung-Jong

    2008-10-01

    Clonorchis sinensis thrives on bile juice. The effects of bile and bile acids on newly excysted juvenile C. sinensis (CsNEJ) were studied in terms of survival. Survival of CsNEJs maintained in 1x Locke's solution, Dulbecco's modified Eagle's medium, NCTC 109, Eagle's, RPMI 1640, and 0.1% glucose was high, but dropped rapidly in 2x Locke's, 0.85% NaCl, and phosphate-buffered saline. Most amino acids in the media favored CsNEJ survival; however, aspartic and glutamic acids and adenine reduced survival. Survival was also significantly lower in media containing more than 0.1% bile. CsNEJs preconditioned in low bile media survived longer in higher bile media. All bile acids and conjugated bile salts were found to favor CsNEJ survival, except for lithocholic acid (LCA) which was toxic. NCTC 109 medium was found to be optimal for the in vitro maintenance of CsNEJs and 1x Locke's solution to be suitable for analyzing the biological effects of bioactive compounds and molecules. Based on these results, we propose that bile acids enhance activity of CsNEJs, but LCA deteriorate CsNEJs.

  6. Phylogenetic characterization of Clonorchis sinensis proteins homologous to the sigma-class glutathione transferase and their differential expression profiles.

    PubMed

    Bae, Young-An; Kim, Jeong-Geun; Kong, Yoon

    2016-01-01

    Glutathione transferase (GST) is one of the major antioxidant proteins with diverse supplemental activities including peroxidase, isomerase, and thiol transferase. GSTs are classified into multiple classes on the basis of their primary structures and substrate/inhibitor specificity. However, the evolutionary routes and physiological environments specific to each of the closely related bioactive enzymes remain elusive. The sigma-like GSTs exhibit amino acid conservation patterns similar to the prostaglandin D synthases (PGDSs). In this study, we analyzed the phylogenetic position of the GSTs of the biocarcinogenic liver fluke, Clonorchis sinensis. We also observed induction profile of the GSTs in association with the parasite's maturation and in response to exogenous oxidative stresses, with special attention to sigma-class GSTs and PGDSs. The C. sinensis genome encoded 12 GST protein species, which were separately assigned to cytosolic (two omega-, one zeta-, two mu-, and five sigma-class), mitochondrial (one kappa-class), and microsomal (one membrane-associated proteins in eicosanoid and glutathione metabolism-like protein) GST families. Multiple sigma GST (or PGDS) orthologs were also detected in Opisthorchis viverrini. Other trematode species possessed only a single sigma-like GST gene. A phylogenetic analysis demonstrated that one of the sigma GST lineages duplicated in the common ancestor of trematodes were specifically expanded in the opisthorchiids, but deleted in other trematodes. The induction profiles of these sigma GST genes along with the development and aging of C. sinensis, and against various exogenous chemical stimuli strongly suggest that the paralogous sigma GST genes might be undergone specialized evolution to cope with the diverse hostile biochemical environments within the mammalian hepatobiliary ductal system. Copyright © 2016 Elsevier B.V. All rights reserved.

  7. 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. Copyright © 2016 Elsevier Ltd. All rights reserved.

  8. 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.

  9. 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.

  10. Clonorchis sinensis omega-class glutathione transferases play major roles in the protection of the reproductive system during maturation and the response to oxidative stress.

    PubMed

    Kim, Jeong-Geun; Ahn, Chun-Seob; Kim, Seon-Hee; Bae, Young-An; Kwon, Na-Young; Kang, Insug; Yang, Hyun-Jong; Sohn, Woon-Mok; Kong, Yoon

    2016-06-13

    Clonorchis sinensis causes a major food-borne helminthic infection. This species locates in mammalian hepatobiliary ducts, where oxidative stressors and hydrophobic substances are profuse. To adapt to the hostile micromilieu and to ensure its long-term survival, the parasite continuously produces a diverse repertoire of antioxidant enzymes including several species of glutathione transferases (GSTs). Helminth GSTs play pertinent roles during sequestration of harmful xenobiotics since most helminths lack the cytochrome P-450 detoxifying enzyme. We isolated and analyzed the biochemical properties of two omega-class GSTs of C. sinensis (CsGSTo1 and CsGSTo2). We observed spatiotemporal expression patterns in accordance with the maturation of the worm's reproductive system. Possible biological protective roles of CsGSTos in these organs under oxidative stress were investigated. The full-length cDNAs of CsGSTo1 and 2 constituted 965 bp and 1,061 bp with open reading frames of 737 bp (246 amino acids) and 669 bp (223 amino acids). They harbored characteristic N-terminal thioredoxin-like and C-terminal α-helical domains. A cysteine residue, which constituted omega-class specific active site, and the glutathione-binding amino acids, were recognized in appropriate positions. They shared 44 % sequence identity with each other and 14.8-44.8 % with orthologues/homologues from other organisms. Bacterially expressed recombinant proteins (rCsGSTo1 and 2) exhibited dehydroascorbate reductase (DHAR) and thioltransferase activities. DHAR activity was higher than thioltransferase activity. They showed weak canonical GST activity toward 1-chloro-2,4-dinitrobenzene. S-hexylglutathione potently and competitively inhibited the active-site at nanomolar concentrations (0.63 and 0.58 nM for rCsGSTo1 and 2). Interestingly, rCsGSTos exhibited high enzyme activity toward mu- and theta-class GST specific substrate, 4-nitrobenzyl chloride. Expression of CsGSTo transcripts and proteins

  11. 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.

  12. Infection rates of Enterobius vermicularis and Clonorchis sinensis of primary school children in Hamyang-gun, Gyeongsangnam-do (Province), Korea

    PubMed Central

    Kim, Bong Jin; Yeon, Je Wook

    2001-01-01

    The egg positive rate of Enterobius vermicularis and Clonorchis sinensis of school children in the rural area was studied in Hamyang-gun. Gyeongsangnam-do in Korea. Cellotape anal swab and formalin ether concentration methods were performed one time to 720 primary school children. The total egg positive rate of E. vermicularis was 12.6% in two schools (Baekjeon and Wiseong). In the Baekjeon and Wiseong primary school, the egg positive rate of E. vermicularis was 4.6% and 13.4%, respectively. Pinworm egg positive rate was 17.6% in the lower grades (1st. 2nd and 3rd), and 7.7% in higher grades (4th, 5th and 6th). The total egg positive rate of male and female was 12.6% and 12.7%, respectively. The egg positive rate of C. sinensis of Baekjeon and Wiseong primary school was 1.5% and 0.46%, respectively. The total egg positive rate of C. sinensis was 0.56%. This survey showed that continuous education and chemotherapy is necessary to treat and prevent reinfection of E. vermicularis. In the case of C. sinensis, health education for school children is recommended to prevent potential infection of adolescents. PMID:11775334

  13. 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

  14. [The Wormicidal Substances Of Fresh Water Fishes On Clonorchis Sinensis: II. Preliminary Research On The Wormicidal Substances From Mucous Substances Of Various Fresh Water Fishes

    PubMed

    Rhee, Jae Ku; Baek, Byeong Kirl; Ahn, Byung Zun; Park, Young Jun

    1980-06-01

    The present work which was investigated in July 1979, was to observe the wormicidal effects of the external mucous substances of 9 species of fresh water fishes (Cyprinus carpio, Parasilurus asotus, Anguilla japonica, Ophicephalus argus, Carassius carassius; golden crusian carp, Misgurnus anguillicaudatus, Zacco platypus, Pseudorasbora parva and Carassius carassius; crusian carp) on cercaria, liberated metacercaria and adult of Clonorchis sinensis. The mucous substances extracted by ether from the above 9 species of fishes were separated into many spots on the silica gel thin layer chromatography, and divided into many fractions in the silica gel column chromatography which used petroleum ether (30%) and chloroform (70%) as a solvent. 1. On the silica gel thin layer chromatography of ether extracts from 9 species of fresh water fishes, each of the Rf. values which had wormicidal effect on the cercaria of C. sinensis were different from others; Rf. value of C. carassius (crusian carp) was 0.937, 0.709 in O. argus, 0.612 in A. japonica, 0.576 in P. asotus, 0.451 in C. carpio, 0.701 in Z. platypus, 0.385 in C. carassius(golden crusian carp) and 0.15 in P. parva. Time for wormicide was different from each other, too. It took 14 min. in a case of C. carassius(crusian carp), 25 min. in Z. platypus, 26 min. in C. carassius(golden crusian carp), 28 min. in C. carpio, 30 min. in P. asotus, 35 min. in O. argus, 40 min. P. parva and 180 min. in A. japonica. But any of the spots of M. anguillicaudatus did not show wormicidal effect on the cercaria within 10 hours. 2. In the silica gel column chromatography of ether extracts from 9 species of fresh water fishes, the fractions which had wormicidal effect on the metacercaria of C. sienesis were different from each other; in a case of C. carassius(crusian carp), the first fraction only had wormicidal effect, the 2nd in P. asotus, the 3rd in O. argus, the 4th in C. carassius(golden crusian carp), the 2nd in C. carpio and the 4th in

  15. Clonorchis sinensis excretory/secretory products promote the secretion of TNF-alpha in the mouse intrahepatic biliary epithelial cells via Toll-like receptor 4.

    PubMed

    Yan, Chao; Wang, Yan-Hong; Yu, Qian; Cheng, Xiao-Dan; Zhang, Bei-Bei; Li, Bo; Zhang, Bo; Tang, Ren-Xian; Zheng, Kui-Yang

    2015-10-24

    Toll-like receptor 4 (TLR4), as one of the most important pathogen pattern recognitions (PPRs) plays a central role in elicitation of innate immunity and mediation of adaptive responses against foreign antigens. However, little is known of the roles of TLR4 in the immune responses of biliary epithelial cells (BECs) induced by Clonorchis sinensis, a parasite of significance in human health. In the present study, the primary mouse intrahepatic biliary epithelial cells (MIBECs) were pre-treated with TLR4 inhibitor peptide or control peptide and then stimulated by excretory/secretory products (ESP) of C. sinensis, respectively. The expressions of TLR4 and relative cytokines were determined using western blot and a bead-based analytic detection system, respectively. The results showed that ESP of C. sinensis significantly increased the expression of TLR4 which promoted the expression of MyD88 and NF-κB in BECs; the levels of TNF-α but not IL-6 from MIBECs stimulated by ESP alone were also considerably increased, compared with the group of the medium stimulated. However, the concentration of TNF-α was significantly decreased when MIBECs were pre-treated with TLR4 inhibitor. In addition, ESP could depress the level of IL-6 in MIBECs which was elevated by LPS. Our data for the first time demonstrate that ESP of C. sinensis can potently induce secretion of pro-inflammatory cytokines via TLR4 in MIBECs, which suggests that TLR4 plays an important role in host defenses against C. sinensis and the pathogenesis of clonorchiasis.

  16. 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

  17. 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.

  18. [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.

  19. Antibodies against Clonorchis sinensis LDH could cross-react with LDHB localizing on the plasma membrane of human hepatocarcinoma cell SMMC-7721 and induce apoptosis.

    PubMed

    Song, Tianzhang; Gan, Wenjia; Chen, Jintao; Huang, Lilin; Yin, Hongling; He, Tailong; Huang, Huaiqiu; Hu, Xuchu

    2016-04-01

    Lactate dehydrogenase (LDH) is a terminal enzyme in anaerobic glycolytic pathway. It widely exists in various organisms and is in charge of converting the glycolysis product pyruvic acid to lactic acid. Most parasites, including Clonorchis sinensis, predominantly depend on glycolysis to provide energy. Bioinformatic analysis predicts that the LDHs from many species have more than one transmembrane region, suggesting that it may be a membrane protein. C. sinensis LDH (CsLDH) has been confirmed as a transmembrane protein mainly located in the tegument. The antibodies against CsLDH can inhibit the worm's energy metabolism, kill the worm, and may have the same effects on human cancer cells. In this study, we cloned and characterized human LDHA (HsLDHA), HsLDHB, and CsLDH. Semi-quantitative real-time RCP showed that HsLDHB only existed in hepatocarcinoma cell SMMC-7721. Confocal microscopy and Western blot experiments revealed that HsLDHB was localized in the plasma membrane of SMMC-7721 cells, and the antibodies against CsLDH could cross-react with it. This cross-reaction could inhibit the enzymatic activity of HsLDHB. The cancer cells co-cultured with anti-CsLDH sera showed a significant decrease in cell proliferation rate and increases in caspase 9 and reactive oxygen species (ROS) levels. Therefore, anti-CsLDH antibodies can induce the apoptosis of cancer cells SMMC-7721 and may serve as a new tool to inhibit tumor.

  20. 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.

  1. Lineage-specific expansion and loss of tyrosinase genes across platyhelminths and their induction profiles in the carcinogenic oriental liver fluke, Clonorchis sinensis.

    PubMed

    Kim, Seon-Hee; Bae, Young-An

    2017-09-01

    Tyrosinase provides an essential activity during egg production in diverse platyhelminths by mediating sclerotization of eggshells. In this study, we investigated the genomic and evolutionary features of tyrosinases in parasitic platyhelminths whose genomic information is available. A pair of paralogous tyrosinases was detected in most trematodes, whereas they were lost in cyclophyllidean cestodes. A pseudophyllidean cestode displaying egg biology similar to that of trematodes possessed an orthologous gene. Interestingly, one of the paralogous tyrosinases appeared to have been multiplied into three copies in Clonorchis sinensis and Opisthorchis viverrini. In addition, a fifth tyrosinase gene that was minimally transcribed through all developmental stages was further detected in these opisthorchiid genomes. Phylogenetic analyses demonstrated that the tyrosinase gene has undergone duplication at least three times in platyhelminths. The additional opisthorchiid gene arose from the first duplication. A paralogous copy generated from these gene duplications, except for the last one, seemed to be lost in the major neodermatans lineages. In C. sinensis, tyrosinase gene expressions were initiated following sexual maturation and the levels were significantly enhanced by the presence of O2 and bile. Taken together, our data suggest that tyrosinase has evolved lineage-specifically across platyhelminths related to its copy number and induction mechanism.

  2. 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.

  3. 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

  4. Involvement of PSMD10, CDK4, and Tumor Suppressors in Development of Intrahepatic Cholangiocarcinoma of Syrian Golden Hamsters Induced by Clonorchis sinensis and N-Nitrosodimethylamine.

    PubMed

    Uddin, Md Hafiz; Choi, Min-Ho; Kim, Woo Ho; Jang, Ja-June; Hong, Sung-Tae

    2015-01-01

    Clonorchis sinensis is a group-I bio-carcinogen for cholangiocarcinoma (CCA). Although the epidemiological evidence links clonorchiasis and CCA, the underlying molecular mechanism involved in this process is poorly understood. In the present study, we investigated expression of oncogenes and tumor suppressors, including PSMD10, CDK4, p53 and RB in C. sinensis induced hamster CCA model. Different histochemical/immunohistochemical techniques were performed to detect CCA in 4 groups of hamsters: uninfected control (Ctrl.), infected with C. sinensis (Cs), ingested N-nitrosodimethylamine (NDMA), and both Cs infected and NDMA introduced (Cs+NDMA). The liver tissues from all groups were analyzed for gene/protein expressions by quantitative PCR (qPCR) and western blotting. CCA was observed in all hamsters of Cs+NDMA group with well, moderate, and poorly differentiated types measured in 21.8% ± 1.5%, 13.3% ± 1.3%, and 10.8% ± 1.3% of total tissue section areas respectively. All CCA differentiations progressed in a time dependent manner, starting from the 8th week of infection. CCA stroma was characterized with increased collagen type I, mucin, and proliferative cell nuclear antigen (PCNA). The qPCR analysis showed PSMD10, CDK4 and p16INK4 were over-expressed, whereas p53 was under-expressed in the Cs+NDMA group. We observed no change in RB1 at mRNA level but found significant down-regulation of RB protein. The apoptosis related genes, BAX and caspase 9 were found downregulated in the CCA tissue. Gene/protein expressions were matched well with the pathological changes of different groups except the NDMA group. Though the hamsters in the NDMA group showed no marked pathological lesions, we observed over-expression of Akt/PKB and p53 genes proposing molecular interplay in this group which might be related to the CCA initiation in this animal model. The present findings suggest that oncogenes, PSMD10 and CDK4, and tumor suppressors, p53 and RB, are involved in the

  5. 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.

  6. [Effect of Excretory-secretory Products of Clonorchis sinensis on Nitric Oxide Production and NF-κB Activation in RAW264.7 Mouse Macrophages].

    PubMed

    Yang, Qing-li; Jiang, Zhi-hua; Shen, Ji-qing; Chen, Ying-dan; Zhou Xiao-nong

    2015-04-01

    To study the role of excretory-secretory products (ESPs) from Clonorchis sinensis in the production of nitric oxide (NO) and the activation of nuclear transcription factor kappa B (NF-κB) in the macrophages of RAW264.7 mouse. 20 µg/ml of C. sinensis EPSs, the organic solvent extracts of EPSs (ESP-ex), and 0.1 µg/ml of lipopolysaccharide from Salmonella minnesota (LPS-SM) were used as stimulators in co-culture with RAW264.7 mouse macrophages as experimental groups. The Hank's balanced salt solution (HBSS) served as control. At the same time the RAW264.7 macrophages were stimulated with EPSs, ESP-ex, and LPS-SM, and then added 0.3 mmol/L of SMT, a specific inhibitor of iNOS as the interference groups. After co-culture for 18 days, the concentrations of NO2- in the culture supernatants were detected with Griess regents, and the activation of NF-κB was determined by transfection with a NF-κB-inducible reporter plasmid, pNiFty2-SEAP. The activities of secreted embryonic alkaline phosphatase (SEAP) in culture supernatants were quantified by using HEK-Blue™ detection medium and expressed as the value of optical density at 620 nm (A620 value). The intercellular activities of SEAP were determined by microscopic observation. After stimulation with both ESPs-ex and LPS-SM, the concentrations of NO2- in culture supernatants were (14.30 ± 1.62) and (14.10 ± 2.17) µmol/L, respectively, which were significantly higher than that of the control [(7.70 ± 0.95) µmol/L] (P < 0.05), and significantly decreased to (8.97 ± 0.81) and (4.96 ± 1.36) µmol/L after adding SMT, respectively (P < 0.05). However, the concentration of NO2- in ESPs stimulation group [(4.06 ± 0.62) µmol/L] was lower than that of the control (P < 0.05), and almost unchanged [(3.99 ± 0.87) µmol/L] after adding SMT (P > 0.05). SEAP activity in ESP group (0.836 ± 0.005) was significantly higher than that of the control [(0.097 ± 0.009) µmol/L] (P < 0.05). A strong blue color reaction was observed

  7. 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.

  8. 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.

  9. 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

  10. 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

  11. 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. Copyright © 2014 Elsevier B.V. All rights reserved.

  12. 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.

  13. 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.

  14. Report: effects of Camellia sinensis L. (green tea) extract on the body and testicular weight changes in adult Wistar rate.

    PubMed

    Hijazi, Muhammad Mazhar; Khatoon, Nasira; Azmi, Muhammad Arshad; Rajput, Muhammad Tariq; Zaidi, Syed Ijaz Hussain; Azmi, Muhammad Ahmed; Perveen, Rehana; Naqvi, Syed Naimul Hassan; Rashid, Muhammad

    2015-01-01

    This research was aimed to study the effects of oral administration of Camellia sinensis L. on the testicular and body weights of adult Wistar rats for short and long time periods. The adult Wistar rats were divided into 3 groups (A, B and C). Every group had ten rats. Green tea extract 0.692% (w/v) was given to groups A and B on daily basis. The extracts were prepared fresh and given for a period of ten and thirty days, respectively, while distilled water was given to the group C rats only. The adult Wistar rats were sacrificed on eleventh and thirty-first day of experiment for the particular groups. The testes were dissected out cautiously, free from the supporter tissues and weighed to the adjacent 1 mg. There is no significant difference in the body weight in all 3 groups. Moreover, it was observed that Wistar rat's testicular weight was considerably increased in group B but no major changes were seen in group A. Our results indicated that green tea when given for short period of time may be effective to the testes but has no consequence on Wistar rat's body weight. However, it is indistinct if these alterations are reversible.

  15. 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.

  16. Carcinogenesis associated with parasites other than Schistosoma, Opisthorchis and Clonorchis: A systematic review.

    PubMed

    Machicado, Claudia; Marcos, Luis A

    2016-06-15

    Only three helminths (Schistosoma haematobium, Opisthorchis viverrini and Clonorchis sinensis) are directly associated with carcinogenesis in humans whereas the role of other parasites in cancer remains unclear. This study aimed to perform a systematic review to identify recent insights in the role of other parasite infections in carcinogenesis. We conducted systematic searches of MEDLINE and EMBASE on July 2015. Our primary outcome was the association between parasitic infections and carcinogenesis. Out of 1,266 studies, 19 were selected for detailed evaluation (eight for helminths and 11 for protozoa). The mechanisms of helminth-induced cancer included chronic inflammation, sustained proliferation, modulation of the host immune system, reprogramming of glucose metabolism and redox signaling, induction of genomic instability and destabilization of suppressor tumor proteins, stimulation of angiogenesis, resisting cell death, and activation of invasion and metastasis. In addition to the current knowledge, the following parasites were found in cancers or tumors: Echinococcus, Strongyloides, Fasciola, Heterakis, Platynosomum and Trichuris. Additional parasites were found in this systematic review that could potentially be associated with cancers or tumors but further evidence is needed to elaborate a cause-effect relationship. © 2016 UICC.

  17. [Epidemiological survey on clonorchiasis sinensis in an endemic area of South Hunan Province].

    PubMed

    Duan, Ji-Hui; Tang, Xiao-Yu; Wang, Qiao-Zhi; Tang, Yang; Zhang, Zong-Si; Li, Zheng-Xiang; Liu, Ai-Hua; Wu, Yan-Jun; Chen, Wen-Hua; Huang, Qi-Rong

    2009-12-01

    To make an epidemiological investigation on clonorchiasis sinensis and relevant factors in the south of Hunan Province. One village from each of Lengshuitan District and Qiyang County was selected for the survey from November to December in 2006. Stool samples from villagers were collected and examined by modified Kato-Katz method. Questionnairing was performed for relevant knowledge and attitude among residents. The infection rate in animal reservoirs and intermediate hosts were detected. A total of 586 cases with Clonorchis sinensis infection were found from 777 people with a prevalence of 75.4%. The average egg density was 451 eggs per gram (EPG) feces. Light, moderate and heavy infections occupied 85.5% (501/586), 14.0% (82/586), and 0.5% (3/586) respectively. Prevalence in males and females was 76.9% (316/411) and 73.8% (270/366) respectively with no significant difference (chi2 = 1.013, P > 0.05). Infections were found in all age groups, with the highest prevalence in the group of 70 to 79 years (85.7%, 30/35). By occupations, the prevalence was 82.5% (447/542) in peasants, 79.3% (42/53) in doctors, 73.7% (28/38) in teachers, and 73.5% (25/34) in local cadres. The infection rate was 17.4% (29/167) and 7.4% (2/27) in Parafossarulus seriatulus and Alocinma longicornis, and 69.2% (9/13) and 5.3% (1/19) in Carassius auratus and Cyprinus carpios respectively. Adult worms were found in all 3 dogs dissected. Over 80% inhabitants did not know that this disease can be acquired by eating raw fish. 95.6% (153/160) of the farmers and 56.7% (349/616) of the students had a history of eating raw fish. The water was contaminated with C. sinensis eggs by using untreated feces as fertilizer for farming and by scrubbing pail latrines in the ponds. The prevalence of clonorchiasis in human population is high in Lengshuitan District and Qiyang County of Hunan Province. Eating raw fish and using untreated feces as fertilizer are the most important epidemiological factors of the

  18. Fumigant toxicity of Citrus sinensis essential oil on Musca domestica L. adults in the absence and presence of a P450 inhibitor.

    PubMed

    Rossi, Yanina Estefanía; Palacios, Sara María

    2013-07-01

    Essential oils (EOs) are potential tools for controlling Musca domestica L. In a fumigant assay, M. domestica adults treated with Citrus sinensis EO (LC50=3.9mg/dm(3)), with (4R)(+)-limonene (95.1%) being its main component, died within 15min or less. The terpenes absorbed by the flies and their metabolites, analyzed using SPME fiber, were (4R)(+)-limonene (LC50=6.2mg/dm(3)), α-pinene (LC50=11.5mg/dm(3)), β-pinene (LC50=6.4mg/dm(3)), and two new components, carveol (LC50=1122mg/dm(3)) and carvone (LC50=19mg/dm(3)), in a proportion of 50, 6.2, 12.5, 6.3 and 25%, respectively. Carveol and carvone were formed by oxidation of (4R)(+)-limonene mediated by cytochrome P450, as was suggested by a fumigation assay on flies previously treated with piperonyl butoxide, a P450 inhibitor. In this experiment, an increase in the toxicity of the EO and (4R)(+)-limonene was observed, as well as a lower production of carveol and carvone. Copyright © 2013 Elsevier B.V. All rights reserved.

  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. Molecular Changes in Opisthorchis viverrini (Southeast Asian Liver Fluke) during the Transition from the Juvenile to the Adult Stage

    PubMed Central

    Sripa, Jittiyawadee; Hall, Ross S.; Scheerlinck, Jean-Pierre; Laha, Thewarach; Sripa, Banchob; Gasser, Robin B.

    2012-01-01

    Background The Southeast Asian liver fluke (Opisthorchis viverrini) chronically infects and affects tens of millions of people in regions of Asia, leading to chronic illness and, importantly, inducing malignant cancer ( = cholangiocarcinoma). In spite of this, little is known, at the molecular level, about the parasite itself, its interplay with its hosts or the mechanisms of disease and/or carcinogenesis. Methodology/Principal Findings Here, we generated extensive RNA-Seq data (Illumina) representing adult and juvenile stages of O. viverrini, and combined these sequences with previously published transcriptomic data (454 technology) for this species, yielding a combined assembly of significantly increased quality and allowing quantitative assessment of transcription in the juvenile and adult stage. Conclusions This enhanced assembly reveals that, despite the substantial biological similarities between the human liver flukes, O. viverinni and Clonorchis sinensis, there are previously unrecognized differences in major aspects of their molecular biology. Most notable are differences among the C13 and cathepsin L-like cysteine peptidases, which play key roles in tissue migration, immune evasion and feeding, and, thus, represent potential drug and/or vaccine targets. Furthermore, these data indicate that major lineages of cysteine peptidases of socioeconomically important trematodes have evolved through a process of gene loss rather than independent radiation, contrasting previous proposals. PMID:23209858

  1. 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

  2. [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

  3. 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

  4. Meroterpenoid enantiomers from Ganoderma sinensis.

    PubMed

    Cao, Wen-Wen; Luo, Qi; Cheng, Yong-Xian; Wang, Shu-Mei

    2016-04-01

    Zizhines A-F (1-6), six pairs of new meroterpenoid enantiomers and a known meroterpenoid (7) were isolated from the fruiting bodies of Ganoderma sinensis. The structures and absolute configurations of the new substances were assigned by spectroscopic and computational methods. All the compounds apart from 7 were evaluated for their inhibition on extracellular matrix component (fibronectin) generation by using TGF-β1-induced rat kidney tubular epithelial cells. Although none of them was found to be active in these cells, the present findings add new facets for the chemistry of Ganoderma. Copyright © 2016. Published by Elsevier B.V.

  5. 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. Copyright AULA MEDICA EDICIONES 2014. Published by AULA MEDICA. All rights reserved.

  6. [Indiscriminate use of Latin name for natural Cordyceps sinensis insect-fungi complex and multiple Ophiocordyceps sinensis fungi].

    PubMed

    Yao, Yi-Sang; Zhu, Jia-Shi

    2016-04-01

    Natural Cordyceps sinensis(Dongchongxiacao) is an insect-fungi complex containing multiple Ophiocordyceps sinensis(≡Cordyceps sinensis) fungi and dead body of larva of the family of Hepialidae. But natural C. sinensis and O. sinensis fungi use the same Latin name, resulting in uncertainty of the specific meaning, even disturbing the formulation and implementation of governmental policies and regulations, and influencing consumer psychology onthe market. This paper reviews the history and current status of the indiscriminate use of the Latin name O. sinensis for both the natural insect-fungi complex C. sinensis and O. sinensis fungi and lists the rename suggetions. Some scholars suggested using the term O. sinensis for the fungi and renaming the natural C. sinensis "Chinese cordyceps". Others suggested renaming the natural C. sinensis "Ophiocordyceps & Hepialidae". Both suggestions have not reached general consensus due to various academic concerns. This paper also reviews the exacerbation of the academic uncertainties when forcing implementing the 2011 Amsterdam Declaration "One Fungus=One Name" under the academic debate. Joint efforts of mycological, zoological and botany-TCM taxonomists and properly initiating the dispute systems offered by International Mycology Association may solve the debate on the indiscriminate use of the Latin name O.sinensis for the natural insect-fungi complex,the teleomorph and anamorph(s) of O. sinensis fungi. Copyright© by the Chinese Pharmaceutical Association.

  7. Insecticide resistance of Anopheles sinensis and An. vagus in Hainan Island, a malaria-endemic area of China.

    PubMed

    Qin, Qian; Li, Yiji; Zhong, Daibin; Zhou, Ning; Chang, Xuelian; Li, Chunyuan; Cui, Liwang; Yan, Guiyun; Chen, Xiao-Guang

    2014-03-03

    Malaria is one of the most important public health problems in Southeast Asia, including Hainan Island, China. Vector control is the main malaria control measure, and insecticide resistance is a major concern for the effectiveness of chemical insecticide control programs. The objective of this study is to determine the resistance status of the main malaria vector species to pyrethroids and other insecticides recommended by the World Health Organization (WHO) for indoor residual sprays. The larvae and pupae of Anopheles mosquitoes were sampled from multiple sites in Hainan Island, and five sites yielded sufficient mosquitoes for insecticide susceptibility bioassays. Bioassays of female adult mosquitoes three days after emergence were conducted in the two most abundant species, Anopheles sinensis and An. vagus, using three insecticides (0.05% deltamethrin, 4% DDT, and 5% malathion) and following the WHO standard tube assay procedure. P450 monooxygenase, glutathione S-transferase and carboxylesterase activities were measured. Mutations at the knockdown resistance (kdr) gene and the ace-1 gene were detected by DNA sequencing and PCR-RFLP analysis, respectively. An. sinensis and An. vagus were the predominant Anopheles mosquito species. An. sinensis was found to be resistant to DDT and deltamethrin. An. vagus was susceptible to deltamethrin but resistant to DDT and malathion. Low kdr mutation (L1014F) frequency (<10%) was detected in An. sinensis, but no kdr mutation was detected in An. vagus populations. Modest to high (45%-75%) ace-1 mutation frequency was found in An. sinensis populations, but no ace-1 mutation was detected in An. vagus populations. Significantly higher P450 monooxygenase and carboxylesterase activities were detected in deltamethrin-resistant An. sinensis, and significantly higher P450 monooxygenase, glutathione S-transferase and carboxylesterase activities were found in malathion-resistant An. vagus mosquitoes. Multiple insecticide resistance was

  8. 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

  9. 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.

  10. 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.

  11. Beneficial effects of Hibiscus rosa-sinensis L. flower aqueous extract in pregnant rats with diabetes.

    PubMed

    Afiune, Luana Alves Freitas; Leal-Silva, Thaís; Sinzato, Yuri Karen; Moraes-Souza, Rafaianne Queiroz; Soares, Thaigra Sousa; Campos, Kleber Eduardo; Fujiwara, Ricardo Toshio; Herrera, Emilio; Damasceno, Débora Cristina; Volpato, Gustavo Tadeu

    2017-01-01

    The Hibiscus rosa-sinensis flower is widely used in Brazilian traditional medicine for the treatment of diabetes and has shown antifertility activity in female Wistar rats. However, there is no scientific confirmation of its effect on diabetes and pregnancy. The aim of this study was evaluate the effect of aqueous extract of H. rosa-sinensis flowers on maternal-fetal outcome in pregnant rats with diabetes. Diabetes was induced by streptozotocin (STZ, 40 mg/kg) in virgin, adult, female Wistar rats. After diabetes induction, the rats were mated. The pregnant rats were distributed into four groups (n minimum = 11 animals/group): non-diabetic, non-diabetic treated, diabetic, and diabetic treated. Oral aqueous extract of Hibiscus rosa-sinensis was administered to rats in the treatment groups during pregnancy. At term pregnancy, maternal reproductive outcomes, fetal parameters, and biochemical parameters were analyzed. The non-diabetic treated group showed decreased high density lipoprotein cholesterol, increased atherogenic index (AI) and coronary artery risk index (CRI), and increased preimplantation loss rate compared to the non-diabetic group. Although treatment with H. rosa-sinensis led to no toxicity, it showed deleterious effects on cardiac and reproductive functions. However, the diabetic treated group showed increased maternal and fetal weights, reduced AI and CRI, and reduced preimplantation loss rate compared to the untreated diabetic group. Our results demonstrate beneficial effects of this flower only in pregnant rats with diabetes and their offspring. Although these findings cannot be extrapolated to human clinical use, they show that the indiscriminate intake of H. rosa-sinensis may be harmful to healthy individuals and its use should be completely avoided in pregnancy.

  12. [AFLP analysis on genetic diversity of Angelica sinensis].

    PubMed

    Zhang, Hong-Yi; Liao, Wen-Bo

    2014-04-01

    To explore the genetic diversity and relationship of different germplasm of Angelica sinensis. Amplified fragment length polymorphism (AFLP) markers were developed to analyze genetic polymorphism in 12 populations 117 samples of Angelica sinensis. The amplified fragments were used for cluster analysis among the different germplasm of Angelica sinensis and to construct the genetic phylogenetic tree with NTSYSpc 2. 11 software. Eight primer combinations selected from 64 primer combinations were used for amplification and a total of 815 fragments were obtained. Analysis identified 812 polymorphic fragments, accounting for 99.63% of the total detected fragments. Different phenotypes and germplasm of Angelica sinensis could be divided by genetic phylogenetic tree analysis. AFLP molecular markers can indicate the significant polymorphism and genetic diversity among germplasm resources of Angelica sinensis. The cultivated purple-stemmed and green-stemmed phenotypes of Angelica sinensis may have different genotypes. The results can provide theoretical evidence for reasonable utilization and breeding new cultivar in molecular level.

  13. Potential anthelmintics: polyphenols from the tea plant Camellia sinensis L. are lethally toxic to Caenorhabditis elegans.

    PubMed

    Mukai, Daisuke; Matsuda, Noriko; Yoshioka, Yu; Sato, Masashi; Yamasaki, Toru

    2008-04-01

    A novel gallate of tannin, (-)-epigallocatechin-(2 beta-->O-->7',4 beta-->8')-epicatechin-3'-O-gallate (8), together with (-)-epicatechin-3-O-gallate (4), (-)-epigallocatechin (5), (-)-epigallocatechin-3-O-gallate (6), and (+)-gallocatechin-(4 alpha-->8')-epigallocatechin (7), were isolated from the tea plant Camellia sinensis (L.) O. Kuntze var. sinensis (cv., Yabukita). The structure of 8, including stereochemistry, was elucidated by spectroscopic methods and hydrolysis. The compounds, along with commercially available pyrogallol (1), (+)-catechin (2), and (-)-epicatechin (3), were examined for toxicity towards egg-bearing adults of Caenorhabditis elegans. The anthelmintic mebendazole (9) was used as a positive control. Neither 2 nor 3 were toxic but the other compounds were toxic in the descending order 8, 7 approximately 6, 9, 4, 5, 1. The LC(50) (96 h) values of 8 and 9 were evaluated as 49 and 334 micromol L(-1), respectively. These data show that many green tea polyphenols may be potential anthelmintics.

  14. 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

  15. A new sesquilignan glucoside from Uraria sinensis.

    PubMed

    Yang, Yingda; Hu, Zhengxi; Luo, Zengwei; Xue, Yongbo; Yao, Guangmin; Wang, Yanyan; Zhang, Yonghui

    2014-01-17

    A new sesquilignan glucoside, urariasinoside A (1), together with eight known compounds, including two lignans, a sesquilignan, a dilignan, and four flavonoid derivatives were isolated from the aerial parts of Uraria sinensis. Their structures were determined on the basis of extensive spectroscopic analyses and comparison with literature data. Compound 1 was evaluated for in vitro cytotoxicity activity against HL-60, SMMC-7721, A549, MCF-7, SW480, and BEAS-2B cell lines.

  16. 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

  17. 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

  18. 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

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

    PubMed

    Mallick, Neelam; Khan, Rafeeq Alam

    2016-01-01

    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. 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. 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. 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.

  20. Morphology of immature stages of blow fly, Lucilia sinensis Aubertin (Diptera: Calliphoridae), a potential species of forensic importance.

    PubMed

    Sanit, Sangob; Sukontason, Kom; Kurahashi, Hiromu; Tomberlin, Jeffery K; Wannasan, Anchalee; Kraisittipanit, Rungroj; Sukontason, Kabkaew L

    2017-09-20

    Lucilia sinensis Aubertin (Diptera: Calliphoridae) is a blow fly species of potential forensic importance since adults are attracted to, and colonize, decomposing vertebrate remains. Blow fly larvae associated with human corpses can be useful evidence in forensic investigations; however, their use is dependent in most cases on proper species identification and availability of developmental data. For identification, morphological information on each life stage is traditionally used. We used scanning electron microscopy (SEM) to examine the ultrastructure of eggs, all instars, and puparia, of L. sinensis. The important characteristics used to differentiate L. sinensis from other species are provided. Distinctive features of the eggs are the slight widening median area extending almost the entire length. The last abdominal segment of the first instar bears elongated outer ventral tubercles along the rim of the last abdominal segment. These tubercles, as well as the well developed median and outer dorsal tubercles, are more prominent in the second and third instars. The surface integument of the tubercles is equipped with circular rows of microtrichia. Pairs of inner dorsal tubercle are absent. Each anterior spiracle is comprised of 9-12 papillae arrange in a single row in the second and third instars. As for the third instar, the dorsal spines between the first and second thoracic segments are delicate, narrow, small, and close together (as row or set). The peristigmatic tufts adjacent to the posterior spiracle of the third instar are moderately branches of short, fine hairs, but minute in puparia. In conclusion, the prominent outer ventral tubercle in all instars and puparia is a new diagnostic feature of L. sinensis and helpful in differentiating it from other Lucilia species that are forensically important. The description of immature L. sinensis in this study will be useful for forensic entomologists in countries where this species exists. Copyright © 2017

  1. [Molecular phylogenetic analysis of Paecilomyces hepiali and Cordyceps sinensis].

    PubMed

    Yang, Jin-Ling; Xiao, Wei; He, Hui-Xia; Zhu, Hui-Xin; Wang, Shu-Fang; Cheng, Ke-Di; Zhu, Ping

    2008-04-01

    Phylogenetic relationship between Paecilomyces hepiali and Cordyceps sinensis was studied by analyzing the sequence of rDNA-ITS. The samples of C. sinensis were collected from Hualong County in Qinghai Province and Kangding County in Sichuan Province in May and June, respectively. The rDNA-ITS fragments were obtained by PCR amplification with the template genomic DNA of the fresh stroma or caterpillar body of the collected samples and the cultured mycelium of P. hepiali, with the universal fungal primers ITS1/ITS4. The amplified fragments were cloned into pMD18-T Vector and sequenced. Phylogenetic analysis was performed with these sequences and those from GenBank. The result showed that all of the 46 clones randomly chosen from the amplification of C. sinensis shared identical or almost identical rDNA-ITS regions and had over 99% identity with some rDNA-ITS sequences of Hirsutella sinensis and C. sinensis registered in GenBank, but all of them had only about 72% identity with that of P. hepiali. Two pairs of specific primers were designed based on the rDNA-ITS sequence of P. hepiali, then PCR and Nest-PCR were performed with the template genomic DNA of the stroma or caterpillar body of C. sinensis samples mentioned above. The apparent bands amplified by Nest-PCR were obtained from all of the samples, and the sequences showed 100% identity with the rDNA-ITS sequence of P. hepiali. In addition, another pair of specific primers were designed based on the rDNA-ITS sequence registered in GenBank as the marker of C. sinensis (accession no. AB067740) but the latter only shared 87.3% identity with that of H. sinensis (accession no. AJ309353). This pair of primers was used to amplify the C. sinensis samples by PCR, and the amplified sequence showed 100% identity with that of AB067740. The result indicated that H. sinensis is the main body of C. sinensis, while some other endoparasitic fungi such as P. hepiali commonly exist in the natural C. sinensis.

  2. Structural analysis and immunostimulatory activity of glycopeptides from Paecilomyces sinensis.

    PubMed

    Zhu, Zhen-Yuan; Meng, Meng; Sun, Huiqing; Li, Yang; Yu, Na; Zhang, Yong-Min

    2016-03-01

    The parasitic fungus, Paecilomyces sinensis, is used to produce Cordyceps materials as a succedaneum of natural Cordyceps sinensis (C. sinensis) in China. In this work, a glycopeptide (CPS-II) was isolated and purified from Paecilomyces sinensis. The result of HPLC indicated that CPS-II was a glycopeptide. The estimated average molecular weight of CPS-II was 2 × 10(6) Da. FTIR, methylation, periodate oxidation, Smith degradation, (1)H NMR, (13)C NMR and CD were used for its structural analysis. The glycopeptide CPS-II was mainly composed of (1 → 3), (1 → 4) connected glucose and galactose as the backbone, there are (1 → 2,3,6) connected glucose, (1 → 3,6) connected mannose, and (1 → 6) connected galactose. Cell proliferation assay and morphological observations indicated that in a certain range of concentrations and time, CPS-II can significantly improve the proliferation activity of RAW264.7 cells.

  3. 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.

  4. 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

  5. [Studies on the chemical constituents of Angelica sinensis].

    PubMed

    Huang, Wei-hui; Song, Chun-qing

    2003-09-01

    To investigate the chemical components of Agelica sinensis. Chromatographic technologies were used for separation and purification, while spectral analysis was measured for structure elucidation. Five compounds were isolated and their structures were identified as Homosenkyunolide H (1), Homosenkyunolide I(2), Neoligustilide (3), 6-methoxycoumarin (4), Hypoxanthine-9-beta-D-ribofuranoside (5). Compounds 1 and 2 are new compounds. Compounds 3, 4 and 5 were isolated from Agelica sinensis for the first time.

  6. 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.

  7. 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

  8. [NIR Fingerprints of Different Medicinal Parts of Angelicae Sinensis Radix].

    PubMed

    Zhang, Ya-ya; Gu, Zhi-rong; Ding, Jun-xia; Wang, Yao-peng; Sun, Yu-jing; Wang, Ya-li

    2015-07-01

    To investigate the spectrum characteristics of near-intrared dittuse retlectance spectroscopy (NIR) fingerprint of different medicinal parts of Angelicae Sinensis Radix. 96 batches of samples were collected from 14 counties of Gansu Province and Yunnan Province. The NIR fingerprints were collected by integrated sphere. Similarity analysis and partial least square discriminant analysis(PLS-DA) were used to analyze the fingerprint. The average spectrum of NIR fingerprint of different medicinal parts of Angelicae Sinensis Radix showed some differences; the absorbance in characteristic absorption was in a decreasing order of body > tail > head > whole. Most NIR fingerprint similarities of different medicinal parts of Angelicae Sinensis Radix exceeded 0. 95. The established model of PLS-DA could be used to accurately classify the medicinal parts of Angelicae Sinensis Radix. The differences of NIR fingerprints of different medicinal parts of Angelicae Sinensis Radix were mainly existing in the wave number ranges of 8,443 - 8,284 cm -1, 7,003 - 6,896 cm-1, 6,102 - 5,864 cm-1, 4,847 - 4,674 cm-1, and 4,386 - 4,208 cm-1. The different medicinal parts of Angelicae Sinensis Radix have some differences in chemical components.

  9. 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.

  10. Development time and body size in Eupolyphaga sinensis along a latitudinal gradient from China.

    PubMed

    Hu, Yuwei; Zhu, Fen; Wang, Xiaoping; Lei, Chaoliang

    2011-02-01

    The responses of organisms to temperature variations may be via short term responses of the phenotype (phenotypic plasticity), or they could involve long-term evolutionary change and adaptation (via selection) to the genotype. These could involve changes to the mean size of the animal or to the thermal reaction norm. We examined the effects of various temperatures (of 22, 25, 28, and 31 °C) on development time, adult body size and preadult survivorship in three populations of the cockroach, Eupolyphaga sinensis (Walker), collected at different latitudes. We found substantial temperature-induced plasticity in development time, body size, and preadult survivorship, indicating that developmental temperatures have strong impacts on growth and life history traits of E. sinensis. Genetic differences for development time, body size, and preadult survivorship were detected among populations, and the three traits exhibited highly significant variations in the responses of different populations to various temperature conditions, indicating genetic differences among populations in terms of thermal reaction norms. We also found that two populations seem to support the beneficial acclimation hypothesis whereas the third mid-latitude population does not. The results are likely because of differences in season length and voltinism, indicating that not only temperature regime but also its interactions with generation time (and development time), voltinism, and season length are likely to have considerable effects on insect development time and body size. Overall, changes in development time, body size, and preadult survivorship in E. sinensis can all be regarded as adaptations to changing thermal regimes. © 2011 Entomological Society of America

  11. Haematopoietic effects of Angelica sinensis root cap polysaccharides against lisinopril-induced anaemia in albino rats.

    PubMed

    Younas, Fryal; Aslam, Bilal; Muhammad, Faqir; Mohsin, Mashkoor; Raza, Ahmad; Faisal, Muhammad Naeem; Hassan, Shamshud-Ul-; Majeed, Wafa

    2017-12-01

    Angelica sinensis L. (Umbelliferae) has medicinal properties. The present study evaluates the haematopoietic effects of A. sinensis polysaccharides (ASP) against lisinopril-induced anaemia. Thirty healthy adult male albino rats were randomly divided into five groups (n = 6). Group I was control group. Group II was treated with angiotensin-converting enzyme inhibitor (ACEI, 20 mg/kg/day) to induce anaemia. In group III, erythropoietin (EPO, 100 IU/kg/each) was administered in combination with ACEI. Group IV was treated with ASP (1 g/kg/day), extracted from A. sinensis root caps. In Group V, ASP (1 g/kg/day) was treated with ACEI. After 28 days, blood and tissue samples were collected for haematological and histopathological analysis, respectively. The results showed that ACEI significantly reduced the haemoglobin (Hb, 10.0 g/dL), packed cell volume (PCV, 39.5%), red blood cells (RBCs, 6.2 million/mm(3)), mean corpuscular volume (MCV, 53.5 fL) and mean corpuscular haemoglobin (MCH, 16.2 pg/cell) values. In the group treated with ASP, the Hb (13.7 g/dL) and RBCs (7.8 million/mm(3)) increased significantly (p < 0.05). The combination of ASP and ACEI led to the significant (p < 0.05) reduction in Hb (10.7 g/dL), PCV (33.3%), RBCs (6.0 million/mm(3)), MCV (54.42 fL) and MCH (16.44 pg/cell) values. While histopathological examination of the liver and kidney cells showed a mild degree of toxicity in the ASP-treated group. ASP has a potentiating effect on haematological parameters when given alone. However, when administered simultaneously with lisinopril, it showed an unfavourable effect with more complicated anaemia so it should not be used with ACEIs.

  12. 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

  13. Ultrastructure of the nephron in the soft-shelled turtle, Pelodiscus sinensis (Reptilia, Chelonia, Trionychidae).

    PubMed

    Xu, Chun-Sheng; Yang, Ping; Bao, Hui-Jun; Bian, Xun-Guang; Chen, Qiu-Sheng

    2013-01-01

    The structure of the nephron in adult soft-shelled turtles (Pelodiscus sinensis) was studied by light microscopy, transmission and scanning electron microscopy. The kidney contained 5-6 renal lobes. Nephrons of P. sinensis are composed of a renal corpuscle (RC) and of a renal tubule that appears divided morphologically into five distinct segments: neck segment (NS) (This segment is only present in approximately 10% of the nephrons), proximal tubule (PT), intermediate segment (IS), distal tubule (DT), and collecting duct (CD). The RCs and most of the convoluted DTs lie in the central zone, while the PTs and the CDs lie in the peripheral zone of the renal lobe. The renal corpuscle is relatively large with especial processes in podocytes and a thick basement membrane. The podocyte processes covering a large capillary area can be observed by TEM, and the major podocyte processes formed a very specific pattern in SEM. The podocyte processes expand to form a flattened network over the whole capillary loops surface, and only may observe little filtration slits in glomerular area. The neck segment when presentis short and has a relatively narrow lumen, consisting of cuboidal or squamous cells. There is a well-developed endocytic-lysosomal apparatus in the apical cytoplasm of the PT. The proximal tubule and intermediate segment cells show some differences between male and female. It showed that proximal tubule cells of male soft-shelt turtle contain lateral intercellular spaces, into which extensions of the cell membrane protrude, and the basal cell membrane forms a conspicuous labyrinth. Whereas, the basal and lateral cell membranes of the female are smooth, and no later-basal intercellular spaces. The differences between male and female in the middle segment cells is similar to proximal tubule cells. Not previously reported in vertebrate kidneys. The IS is the narrowest nephron segment, formed by multiciliated as well as nonciliated cells. In DT cells, basolateral

  14. 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

  15. Cloning and expression of prostaglandin E2 receptor subtype 1 (ep 1 ) in Bostrichthys sinensis.

    PubMed

    Lai, Xiao Jian; Hong, Wan Shu; Liu, Fang; Zhang, Yu Ting; Chen, Shi Xi

    2014-08-01

    Our previous studies suggested that prostaglandin E2 (PGE2) is a putative sex pheromone in Chinese black sleeper Bostrichthys sinensis, a fish species that inhabits intertidal zones and mates and spawns inside a muddy burrow. We found immunoreactivities of PGE2 receptor subtypes (Ep1-3) expressed in the olfactory sac, but only Ep1 presented higher density of immunoreactivity in mature fish than that in immature fish in both sexes. To gain a better understanding of the underlying molecular mechanism for the detection of PGE2 in the olfactory system, we cloned an ep 1 cDNA from the adult olfactory sac. The open-reading frame of the ep 1 consisted of 1,134-bp nucleotides that encoded a 378-amino acid-long protein with a seven-transmembrane domain, typical for the G protein-coupled receptors superfamily. Expression of ep 1 mRNA was observed in all tissues examined, with higher levels obtained in the olfactory sacs and testes. The expression of ep 1 mRNA in the olfactory sacs and gonads was significantly higher in both sexes of mature fish than in those of immature ones. Taken together, our results suggested that Ep1, which is highly expressed in the olfactory sacs and gonads of mature fish, is important for the control of reproduction and may be involved in PGE2-initiated spawning behavior in B. sinensis.

  16. 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.

  17. 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.

  18. 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.

  19. 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

  20. 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

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

    PubMed

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

    2012-02-01

    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). 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. 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. 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.

  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. First Report on Co-Occurrence Knockdown Resistance Mutations and Susceptibility to Beta-Cypermethrin in Anopheles sinensis from Jiangsu Province, China

    PubMed Central

    Tan, Wei-Long; Wang, Zhong-Ming; Li, Chun-Xiao; Chu, Hong-Liang; Xu, Yan; Dong, Yan-De; Wang, Zhong-can; Chen, Dong-Ya; Liu, Hui; Liu, Da-Peng; Liu, Nannan; Sun, Jun; Zhao, Tongyan

    2012-01-01

    The increasing prevalence of insecticide resistance in Anopheles sinensis, a major vector of malaria in Jiangsu province in eastern China, threatens to compromise the successful use of insecticides in malaria control strategies. It is therefore vital to understand the insecticide resistance status of An. sinensis in the region. This study examined the nucleotide diversity of the para-sodium channel and knockdown resistance (kdr) in five field populations of adult An. sinensis mosquitoes collected in Jiangsu province, identifying the L1014F and L1014C substitutions for the first time. Competitive polymerase chain reaction (PCR) amplification of specific allele (cPASA) and polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) for resistance diagnosis were developed and validated. Comparing the results with direct sequencing revealed that the PCR-RFLP method was more sensitive and specific whereas the cPASA method was more convenient and suitable. The significant positive correlation between kdr allele frequency and bioassay-based resistance phenotype demonstrates that the frequency of L1014F and L1014C substitutions in the kdr gene provides a useful molecular marker for monitoring beta-cypermethrin resistance in natural populations of An. sinensis. Our results point to the L1014F substitution as the key mutation associated with beta-cypermethrin resistance. The high resistance and mutation frequency detected in the five populations also suggest cross-resistance with other pyrethroids may occur in An. sinensis, highlighting the need for further surveys to map insecticide resistance in China and the adoption of a rational management of insecticide application for resistance management and mosquito vector control. PMID:22272229

  4. 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.

  5. [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.

  6. Three phenylethanoid glucosides of unusual structure from Chirita sinensis (Gesneriaceae).

    PubMed

    Damtoft, S; Jensen, S R

    1994-09-01

    From Chirita sinensis (Gesneriaceae) was isolated six phenylethanoid glucosides, namely the previously known positional isomers plantainoside A, calceolarioside A and calceolarioside B together with three new compounds named chiritoside A, B and C. It was proved by analysis of the NMR data that the chiritosides were the 4"-O-beta-glucopyranosyl-derivatives of the first three compounds.

  7. Alernaria undulata, a new species from Citrus sinensis

    USDA-ARS?s Scientific Manuscript database

    A new species belonging to the Alternaria infectoria species-group, A undulata, is described from imported fruit of Citrus sinensis (navel Orange). A. undulata was associated with visible lesions that appeared to originate from the peduncular scars of several fruits purchased from a retail display ...

  8. Bioavailable Citrus sinensis Extract: Polyphenolic Composition and Biological Activity.

    PubMed

    Pepe, Giacomo; Pagano, Francesco; Adesso, Simona; Sommella, Eduardo; Ostacolo, Carmine; Manfra, Michele; Chieppa, Marcello; Sala, Marina; Russo, Mariateresa; Marzocco, Stefania; Campiglia, Pietro

    2017-04-15

    Citrus plants contain large amounts of flavonoids with beneficial effects on human health. In the present study, the antioxidant and anti-inflammatory potential of bioavailable polyphenols from Citrus sinensis was evaluated in vitro and ex vivo, using the murine macrophages cell line J774A.1 and primary peritoneal macrophages. Following simulated gastro-intestinal digestion, the in vitro bioavailability of Citrus sinensis polyphenolic extract was assessed using the human cell line Caco-2 grown as monolayers on a transwell membrane. Data demonstrated a relative permeation of its compounds (8.3%). Thus, the antioxidant and anti-inflammatory effect of polyphenolic Citrus sinensis fraction (Cs) was compared to the bioavailable one (CsB). Results revealed that Citrus extract were able to reduce macrophages pro-inflammatory mediators, including nitric oxide, iNOS, COX-2 and different cytokines. Moreover, the effect of Citrus sinensis polyphenols was associated with antioxidant effects, such as a reduction of reactive oxygen species (ROS) and heme-oxygenase-1 (HO-1) increased expression. Our results provide evidence that the bioavailable polyphenolic constituents of the Citrussinensis extract accumulate prevalently at intestinal level and could reach systemic circulation exerting their effect. The bioavailable fraction showed a higher anti-inflammatory and antioxidant potential compared to the initial extract, thus highlighting its potential nutraceutical value.

  9. 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.

  10. 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

  11. 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

  12. 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

  13. 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.

  14. 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.

  15. Regulatory mechanism of Toona sinensis on mouse leydig cell steroidogenesis.

    PubMed

    Poon, Song Ling; Leu, Sew-Fen; Hsu, Hseng-Kuang; Liu, Ming-Yie; Huang, Bu-Miin

    2005-02-11

    Toona sinensis (TS), a kind of arbor, widely distributes nowadays in Asia. The leaves of TS have been used as an effective nutritious food in Chinese society for a long time. It was reported that Toona sinensis can induce apoptosis of cancer cells, reduce plasma glucose in diabetic rats, and improve lipolysis of differentiated 3T3-L1 adipocyte and its uptake of glucose. It has also been shown that TS may increase dynamic activity of human sperm. Thus, we are interested to investigate whether Toona sinensis has any effect on mouse Leydig cell testosterone production, which correlates to sperm activity. Primary mouse Leydig cells were purified to conduct the in vitro experiments. Different concentrations of crude Toona sinensis were added to primary mouse Leydig cells and the testosterone production was determined. The results showed that crude TS significantly inhibited both basal and human chorionic gonadotropin (hCG)-stimulated testosterone productions in dose dependent manner, respectively (P<0.05). Crude TS also reduced the forskolin- and dibutyryl-cAMP (dbcAMP)-stimulated testosterone production (P<0.05), which indicated that crude TS might affect protein kinase A (PKA) signal transduction pathway at the site after the formation of cyclic AMP. Moreover, TS inhibited Leydig cell steroidogenesis by suppressing the activity of steroidogenic enzymes including P450 side chain cleavage enzyme, 3 beta-hydroxysteroid dehydrogenase, 17 alpha-hydroxylase, 20 alpha-hydroxylase and 17 beta-hydroxysteroid dehydrogenase (P<0.05). In summary, these results suggested that TS inhibited steroidogenesis by suppressing the cAMP-PKA signaling pathway and the activities of steroidogenic enzymes in normal mouse Leydig cells.

  16. 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.

  17. 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

  18. 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

  19. 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

  20. 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

  1. 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.

  2. Characterization of the complete chloroplast genome of the endangered species Carya sinensis (Juglandaceae)

    Treesearch

    Yiheng Hu; Xi Chen; Xiaojia Feng; Keith E. Woeste; Peng Zhao

    2016-01-01

    Carya sinensis (Chinese Hickory, beaked walnut, or beaked hickory) is an endangered species that needs urgent conservation action. Here, we reported the complete chloroplast (cp) genome sequence and the genomic features of the C. sinensis cp, which is the first complete cp genome of any member of Carya. The...

  3. The complete mitochondrial genome of Sesarmops sinensis reveals gene rearrangements and phylogenetic relationships in Brachyura

    PubMed Central

    Liu, Yu; Zhang, Dai-Zhen; Wang, Zheng-Fei; Zhang, Hua-Bin; Chai, Xin-Yue; Zhou, Chun-Lin

    2017-01-01

    Mitochondrial genome (mitogenome) is very important to understand molecular evolution and phylogenetics. Herein, in this study, the complete mitogenome of Sesarmops sinensis was reported. The mitogenome was 15,905 bp in size, and contained 13 protein-coding genes (PCGs), two ribosomal RNA (rRNA) genes, 22 transfer RNA (tRNA) genes, and a control region (CR). The AT skew and the GC skew are both negative in the mitogenomes of S. sinensis. The nucleotide composition of the S. sinensis mitogenome was also biased toward A + T nucleotides (75.7%). All tRNA genes displayed a typical mitochondrial tRNA cloverleaf structure, except for the trnS1 gene, which lacked a dihydroxyuridine arm. S. sinensis exhibits a novel rearrangement compared with the Pancrustacean ground pattern and other Brachyura species. Based on the 13 PCGs, the phylogenetic analysis showed that S. sinensis and Sesarma neglectum were clustered on one branch with high nodal support values, indicating that S. sinensis and S. neglectum have a sister group relationship. The group (S. sinensis + S. neglectum) was sister to (Parasesarmops tripectinis + Metopaulias depressus), suggesting that S. sinensis belongs to Grapsoidea, Sesarmidae. Phylogenetic trees based on amino acid sequences and nucleotide sequences of mitochondrial 13 PCGs using BI and ML respectively indicate that section Eubrachyura consists of four groups clearly. The resulting phylogeny supports the establishment of a separate subsection Potamoida. These four groups correspond to four subsections of Raninoida, Heterotremata, Potamoida, and Thoracotremata. PMID:28622362

  4. Metabolic characterization of natural and cultured Ophicordyceps sinensis from different origins by 1H NMR spectroscopy.

    PubMed

    Zhang, Jianshuang; Zhong, Xin; Li, Shaosong; Zhang, Guren; Liu, Xin

    2015-11-10

    Ophicordyceps sinensis is a well-known traditional Chinese medicine and cultured mycelium is a substitute for wild O. sinensis. Metabolic profiles of wild O. sinensis from three geographical locations and cultivated mycelia derived from three origins were investigated using (1)H nuclear magnetic resonance (NMR) analysis combined with multivariate statistical analysis. A total of 56 primary metabolites were identified and quantified from O. sinensis samples. The principle component analysis (PCA) showed significant differences between natural O. sinensis and fermentation mycelia. Seven metabolites responsible for differentiation were screened out by orthogonal partial least squares discriminant analysis (OPLS-DA). The concentrations of mannitol, trehalose, arginine, trans-4-hydroxyproline, alanine and glucitol were significantly different between wild and cultured groups. The variation in metabolic profiling among artificial mycelia was greater than that among wild O. sinensis. Furthermore, wild samples from different origins were clearly distinguished by the levels of mannitol, trehalose and some amino acids. This study indicates that (1)H NMR-based metabolomics is useful for fingerprinting and discriminating O. sinensis of different geographical regions and cultivated mycelia of different strains. The present study provided an efficient approach for investigating chemical compositions and evaluating the quality of medicine and health food derived from O. sinensis. Copyright © 2015 Elsevier B.V. All rights reserved.

  5. Characterization of the complete mitochondrial genome of Khawia sinensis belongs among platyhelminths, cestodes.

    PubMed

    Feng, Yan; Feng, Han-Li; Fang, Yi-Hui; Su, Ying-Bing

    2017-06-01

    Khawia sinensis is an important species in freshwater fish causing considerable economic losses to the breeding industry. This is the first mt genome of a caryophyllidean cestode characterised. The entire mt genome of K. sinensis is 13,759 bp in length. This mt genome contains 12 protein-coding genes, 22 transfer RNA genes, two ribosomal RNA genes and two non-coding regions. The arrangement of the K. sinensis mt genome is the same as other tapeworms, however, the incomplete stop codon (A) is more frequent that other species. Phylogenetic analyses based on concatenated amino-acid sequences of the 12 protein-coding genes of 17 tapeworms including K. sinensis were conducted to assess the relationship of K. sinensis with other species, the result indicated K. sinensis was closely related with cestode species. This complete mt genome of K. sinensis will enrich the mitochondrial genome databases of tapeworms and provide important molecular markers for ecology, diagnostics, population variation and evolution of K. sinensis and other species. Copyright © 2017 Elsevier Inc. All rights reserved.

  6. [Effects of Different Altitudes and Sowing Dates on Direct Sowing Angelica sinensis Yield and Quality].

    PubMed

    Ji, Ying; Lin, Hai-ming; Deng, Ji-cheng; Qi, Ju-tao; Ma, Zhan-chuan; Jin, Ling; Wang, Wan-sheng

    2015-09-01

    To study the effects of different altitudes and sowing dates on direct sowing Angelica sinensis biomass, yield and quality, and to provide a theoretical basis for Angelica sinensis direct sowing cultivation techniques. Two factors trials were used to research the influence of altitude and sowing dates on yield and quality of direct sowing Angelica sinensis. The altitudes were located at 2500, 2000 and 1500 m, and the sowing dates were set up at autumn August 29, and Spring April 3 and April 24. The experiments were designed with split plot. Under the same altitude, roots and aboveground biomass of direct sowing Angelica sinensis were higher when sowing earlier. In the same sowing date, the root and aboveground biomass was the maximum at 2 000 m altitude, followed by elevation of 1 500 m. At 2 500 m altitude, Angelica sinensis root and aboveground biomass was the minimum. Sowing at 2 000 m altitude at August 29 direct sowing Angelica sinensis showed the highest biomass and yield, reaching 13 840.95 kg/hm2, significantly higher than the other treatments. Compared with transplanting Angelica sinensis in this region, the production of direct sowing Angelica sinensis was also 15. 3% higher. Angelica sinensis medicinal grade was significantly higher than the rest of the process. Angelica sinensis extract, volatile oil and ferulic acid content had reached the standard of Chinese Pharmacopoeia. Angelica sinensis sowed in late August at 2000 m altitude has the best yield and quality on root length, root diameter, plant height, leaf number, dry and fresh matter accumulation, followed by 1500 m altitude, and 2500 m worst. Therefore, altitude range of Angelica sinensis direct sowing cultivation area can be reduced to 1500-2000 m. Angelica sinensis sowed in late August, at 2000 m altitude, the indicators like root length,root diameter,plant height,leaf number,and dry and fresh matter accumulation showed the best, followed by 1500 m altitude, 2500 m worst. Therefore, altitude

  7. [Study on prediction model of Angelicae sinensis yield per unit in Min County].

    PubMed

    Yan, Hui; Duan, Jin-Ao; Wang, Rui-Qi; Guo, Zeng-Xiang; Qian, Da-Wei; Song, Bing-Sheng; He, Zi-Qing; Gao, Hong-Ying

    2012-04-01

    To establish a prediction model of Angelicae sinensis yield per unit in Min County, and the forecasting information can be provided to correlated department and organization. With the basis of the Angelicae sinensis yield statistics in Min County from 1995 to 2009 and weather data of development phase of each ten days, polynomial forecasting modeling was used and the stimulation forecasting of Angelicae sinensis yield in Min County was carried out. The results showed that the average accuracy of prediction model was reached to 97.2%, which basically met the demand for yield prediction. The prediction model of Angelicae sinensis yield per unit in Min County has good accuracy and relatively correct forecasting information about Angelicae sinensis yield, which provides methodology and important references for dynamic forecasting in the progress of Chinese medicinal materials production.

  8. [Study on quality standard of oil-processed Radix Angelica sinensis].

    PubMed

    Yang, Hong-Shen; Hua, Yong-Li; Guo, Yan-Sheng; Du, Tian-Xi; Yang, Xiao-Hu; Qinh, Liang; Qu, Ya-Ling; Wei, Yan-Ming

    2010-06-01

    To establish the quality standard of oil-processed Radix Angelica sinensis. Combined traditional identification, TLC and fingerprints of wine-processed Radix Angelica sinensis to control quality of oil-processed Radix Angelica sinensis. And referring to China Pharmacopoeia of 2005 edition, water, ash, and extract were also detected. The content of water, total ash, extract representatively was 7.30%, 8.70% and 50.9%. Eleven fingerprint peaks were defined, The eleven common peaks were appointed as fingerprint peaks by analyzing 14 representative samples, all the fingerprint peaks were quantified grounded on the peak of Ferulic acid. and quantified rested on the peak of ferulic acid. A multicomponent quantitative method for oil-processed Radix Angelica sinensis is established. The established method is feasible. The quality control standards of the oil-processed Radix Angelica sinensis is normative, systematic and accurate.

  9. 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.

  10. Spectroscopic Study of Green Tea (Camellia sinensis) Leaves Extraction

    NASA Astrophysics Data System (ADS)

    Marzuki, A.; Suryanti, V.; Virgynia, A.

    2017-04-01

    This paper reports the analysis of UV-VIS-NIR absorption spectra of different concentrations of green tea (Camellia sinensis) leaf extract in two different solvent systems (chloroform and ethyl acetate). In those solvents, two different peaks characterizing green tea are observed at different wavelengths, namely 296 nm and 329 nm (extracted in chloroform) and 391 nm and 534 nm (extracted in ethyl acetate). We then investigated the absorption spectra change as function of green tea concentration in both solvents. We found that light absorption increases linearly with the increase of green tea concentration. Different wavelengths, however, respond this change differently. However, the way it changes is wavelength dependence.

  11. 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.

  12. 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.

  13. On the reliability of fungal materials used in studies on Ophiocordyceps sinensis.

    PubMed

    Dong, C-H; Yao, Y-J

    2011-08-01

    Ophiocordyceps sinensis (≡Cordyceps sinensis) is one of the best known traditional Chinese medicines, with great benefits to human health and huge economic value. The reliability of fungal materials used in studies of the species is particularly important because contradictory results have been found in various studies in the past decades. Examination of fungal materials specified in reports on O. sinensis showed great variation in both sources and culture conditions of living strains. To test the reliability of the materials used, experiments were carried out to study the effect of culture conditions on the growth of living strains of O. sinensis by using six reliable strains representing the major production regions of the fungus on the Tibetan Plateau. The results showed that O. sinensis is a slow-growing fungus at comparatively low temperature, and that temperature and growth period are crucial factors which can be verified by experiment. Analyses of fungal materials used in 152 papers on O. sinensis from PubMed since 1998 showed that 41 papers lacked detailed information on the fungal materials; 26 used natural products, 11 used artificially cultivated fruit bodies, and 80 used fermentation products from living strains. Of the latter category (using fermentation products), 64 of the papers were found to use unreliable (45) or uncertain (19) strains for fermentation products based on the temperature and growth period for O. sinensis strains verified in this study. Apart from the natural products of O. sinensis, which require scientific identification, a total of at least 116 papers (over three-quarters) used unreliable, uncertain or unspecified materials, including so-called cultivated fruit bodies which were apparently from other species. The reliability of materials or living strains used in studies on O. sinensis is discussed in this paper, and suggestions are made for use of reliable fungal materials in further studies of this fungus.

  14. [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

  15. Transcriptomic analysis of flower development in tea (Camellia sinensis (L.)).

    PubMed

    Liu, Feng; Wang, Yu; Ding, Zhaotang; Zhao, Lei; Xiao, Jun; Wang, Linjun; Ding, Shibo

    2017-10-05

    Flowering is a critical and complicated process in plant development, involving interactions of numerous endogenous and environmental factors, but little is known about the complex network regulating flower development in tea plants. In this study, de novo transcriptome assembly and gene expression analysis using Illumina sequencing technology were performed. Transcriptomic analysis assembles gene-related information involved in reproductive growth of C. sinensis. Gene Ontology (GO) analysis of the annotated unigenes revealed that the majority of sequenced genes were associated with metabolic and cellular processes, cell and cell parts, catalytic activity and binding. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis indicated that metabolic pathways, biosynthesis of secondary metabolites, and plant hormone signal transduction were enriched among the DEGs. Furthermore, 207 flowering-associated unigenes were identified from our database. Some transcription factors, such as WRKY, ERF, bHLH, MYB and MADS-box were shown to be up-regulated in floral transition, which might play the role of progression of flowering. Furthermore, 14 genes were selected for confirmation of expression levels using quantitative real-time PCR (qRT-PCR). The comprehensive transcriptomic analysis presents fundamental information on the genes and pathways which are involved in flower development in C. sinensis. Our data also provided a useful database for further research of tea and other species of plants. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Fish tapeworm Khawia sinensis: an indicator of environmental microcystins?

    PubMed

    Palikova, Miroslava; Adamovsky, Ondrej; Blaha, Ludek; Mares, Jan; Kopp, Radovan; Navratil, Stanislav; Cutakova, Zdenka; Soukupova, Zdenka; Pikula, Jiri

    2013-01-01

    Parasites have recently been recognized as accumulation indicators that take up and bio-concentrate substances from environmental pollution. Interestingly, helminths of fish are known to accumulate metals from the ambient environment and to contain several orders of magnitude higher concentrations than hosts. While the majority of reports mention inorganic toxin accumulation in parasites, studies concerning effects of organic pollution are infrequent and little is known about the potential of parasites to bio-accumulate microcystins. The parasite-host system of tapeworm Khawia sinensis and common carp (Cyprinus carpio) was used to address this issue. Both the tapeworms and livers were dissected from experimental carps orally exposed to cyanobacterial biomass for 20 days. The total dose of microcystins amounted to 27 mg/kg of feed, i.e., 0.4 mg/kg of fish mass a day. Microcystin concentrations in tapeworms and carp liver tissues were measured using the LC-MS/MS method. Considering the three measured microcystin variants LR, YR and RR, only MC-RR was detected and its concentrations in tapeworms and carp liver tissue amounted to 5.78±3.78 ng/g and 2.11±0.74 ng/g fresh weight (p<0.05), respectively. Here we show accumulation of microcystin MC-RR in the tapeworm Khawia sinensis, a parasite of common carp (Cyprinus carpio). As this is the first report addressing this issue, further studies will be necessary to examine this specific parasite-host system.

  17. 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

  18. 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.

  19. Effects of Citrus sinensis juice on blood pressure.

    PubMed

    Asgary, Sedigheh; Keshvari, Mahtab

    2013-01-01

    Citrus sinensis juice (CSJ) is a rich source of dietary flavonoids which reduce the risk of adverse cardiovascular events. This study aimed to examine the effects of four-week intake of natural and commercial orange (Citrus sinensis) juice on blood pressure in healthy volunteers. In this single-blind randomized crossover study, 22 healthy subjects (age: 18-59 years old) were included and randomly divided into two groups of 11. Group A consumed commercial CSJ during the first four-week period. After a two-week washout period, they consumed natural CSJ for another four weeks. The procedure was reversed in group B. The participants were asked to drink 500 ml/day of either natural or commercial CSJ twice a day with breakfast and dinner. The effects of orange juice on blood pressure were evaluated. After drinking commercial CSJ, diastolic and systolic blood pressure were significantly decreased (5.13%; P = 0.03 and -5.91%; P = 0.003, respectively). However, consumption of natural CSJ did not have significant effects on either diastolic or systolic blood pressure. Commercial CSJ significantly decreased blood pressure. Higher flavonoid, pectin, and essential oils content of concentrated products compared to natural juice might have been responsible for this finding. Nevertheless, further studies to focus on dose-response effects are recommended.

  20. 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

  1. 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.

  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. 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

  4. 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

  5. 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

  6. 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

  7. 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.

  8. 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

  9. 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

  10. [Investigation of metabolic action of Cordyceps sinensis and its cultured mycelia on Escherichia coli by microcalorimetry].

    PubMed

    Zhou, Dan-Lei; Yan, Dan; Li, Bao-Cai; Wu, Yan-Shu; Xiao, Xiao-He

    2009-06-01

    This study is to investigate the effect of Cordyceps sinensis and its cultured mycelia on growth and metabolism of Escherichia coli, and microcalorimetric method was carried out to evaluate its biological activity. The study will provide the basis for the quality control of Cordyceps sinensis. Experimental result will show the effect of natural Cordyceps sinensis and its cultured mycelia on growth and metabolism of Escherichia coli, with index of P(1max) and effective rate (E) by microcalorimetry, the data of experiment were studied by cluster analysis. The results showed that Cordyceps sinensis and its cultured mycelia not only can promote growth and metabolism of Escherichia coli but also can regulate the balance of intestinal microecology efficiently. When the concentrations of samples > 6.0 mg mL(-1), natural Cordyceps sinensis can promote the growth and metabolism of Escherichia coli efficiently (P < 0.05) compared with the control group, and have better dose-effect relationship with concentration (r > 0.9), its cultured mycelia does not show conspicuous auxoaction (P > 0.05) and have not dose-effect relationship with concentration (r < 0.6); when the concentration of samples < 6.0 mg mL(-1), all samples does not show conspicuous auxoaction (P > 0.05). Natural Cordyceps sinensis and its cultured mycelia can be distinguished by cluster analysis. Microcalorimetry has a good prospect on the quality evaluation of the traditional Chinese medicine.

  11. 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

  12. [Evaluating the quality of Angelica sinensis with fuzzy matter-element model based on variation coefficient weight].

    PubMed

    Li, Yang; Xia, Peng-Fei; Ma, Xiao; Fan, Qin; Zhao, Lei; Wang, Ya-Li; Liu, Xiong

    2013-08-01

    To investigate the utilization of fuzzy matter-element model in evaluating the quality of Angelica sinensis. The quality of Angelica sinensis from different habitats was evaluated by determining six main compositions contained in the samples with fuzzy matter-element model based on variation coefficient weight. Angelica sinensis collected from 22 hatitats were divided into three ranges based on the values of approach degrees. Fuzzy matter-element model based on variation coefficient weight can judge the quality of Angelica sinensis objectively and feasibly.

  13. 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.

  14. Glucosinolate Diversity in Bretschneidera sinensis of Chinese Origin.

    PubMed

    Montaut, Sabine; Zhang, Wei-Dong; Nuzillard, Jean-Marc; De Nicola, Gina R; Rollin, Patrick

    2015-08-28

    The glucosinolate (GL) profile in several plant parts (leaf, branch, bark, root, and fruit) of Bretschneidera sinensis from three geographical regions of the People's Republic of China was established for the first time by HPLC. During this investigation, benzyl GL (1), 4-hydroxybenzyl GL (2), 2-hydroxy-2-methylpropyl GL (3), and 4-methoxybenzyl GL (4) were identified. In addition, one new GL, 3-hydroxy-4-methoxybenzyl GL (5), was isolated in a minor amount from the fruit and characterized by spectroscopic data interpretation. Furthermore, traces of 4-hydroxy-3-methoxyphenylacetonitrile were detected by GC-MS analysis in the fruits, thus confirming the presence of the regioisomeric 4-hydroxy-3-methoxybenzyl GL (6). GLs 1-5 were also quantified for the first time by HPLC in the various plant organs.

  15. 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.

  16. 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.

  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. 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

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

    PubMed

    Gjerde, Bjørn; Hilali, Mosaad

    2016-08-01

    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.

  1. Isolation and characterization of cDNAs encoding ribosome inactivating protein from Dianthus sinensis L.

    PubMed

    Cho, H J; Lee, S J; Kim, S; Kim, B D

    2000-04-30

    To isolate a ribosome inactivating protein (RIP) gene, six plant species were surveyed for antiviral activity. Crude proteins extracted from these plants were tested for the antiviral activity against tobacco mosaic virus (TMV) in Nicotiana glutinosa. All the plants, Spinacia oleracea, Amaranthus lividus, Dianthus superbus, Dianthus sinensis and Celosia cristata, with an exception of Oenanthe stolonifera, presented 70-90% inhibition of viral infectivity. In an attempt to search for the RIP gene from D. sinensis, partial cDNA was obtained by polymerase chain reaction (PCR) of the poly(A)+ RNA from D. sinensis leaves. DNA gel blot analysis showed that D. sinensis has multi-copy RIP genes. The expression of RIP gene was investigated in the flower, leaf, root and stem of D. sinensis, and was found to be most abundant in the leaf. Using the partial cDNA as a probe, seven full-length cDNAs were isolated from a library prepared from D. sinensis leaves. They were divided into three groups on the basis of their nucleotide sequence homology. The three representative clones, cDsRIP1, cDsRIP2 and cDsRIP3 were completely sequenced. They all had an open reading frame of 882 bp. The cDsRIP2 showed 79% homology with dianthin 30 and saporin genes; 59% with PAP and Mirabilis antiviral protein MAP genes. From the analysis of deduced amino acid sequences, it was predicted that D. sinensis RIP cDNAs might have a putative signal peptide of 23 amino acid residues at their N-terminus. When the cDNA was expressed in E. coli, the bacteria was unable to grow upon IPTG induction, suggesting that expression of the gene renders toxicity to E. coli cells.

  2. 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

  3. Angelica sinensis in China-A review of botanical profile, ethnopharmacology, phytochemistry and chemical analysis.

    PubMed

    Wei, Wen-Long; Zeng, Rui; Gu, Cai-Mei; Qu, Yan; Huang, Lin-Fang

    2016-08-22

    Angelica sinensis (Oliv.) Diels, known as Dang Gui (in Chinese), is a traditional medicinal and edible plant that has long been used for tonifying, replenishing, and invigorating blood as well as relieving pain, lubricating the intestines, and treating female irregular menstruation and amenorrhea. A. sinensis has also been used as a health product and become increasingly popular in China, Japan, and Korea. This paper aims to provide a systemic review of traditional uses of A. sinensis and its recent advances in the fields of phytochemistry, analytical methods and toxicology. In addition, possible trends, therapeutic potentials, and perspectives for future research of this plant are also briefly discussed. An extensive review of the literature was conducted, and electronic databases including China National Knowledge Infrastructure, PubMed, Google Scholar, Science Direct, and Reaxys were used to assemble the data. Ethnopharmacological literature and digitalised sources of academic libraries were also systematically searched. In addition, information was obtained from local books and The Plant List (TPL, www.theplantlist.org). This study reviews the progress in chemical analysis of A. sinensis and its preparations. Previously and newly established methods, including spectroscopy, thin-layer chromatography (TLC), gas chromatography (GC), high-performance liquid chromatography (HPLC), ultra-performance liquid chromatography(UPLC), and nuclear magnetic resonance analysis (NMR), are summarized. Moreover, identified bioactive components such as polysaccharides, ligustilide and ferulic acid were reviewed, along with analytical methods for quantitative and qualitative determination of target analytes, and fingerprinting authentication, quality evaluation of A. sinensis, and toxicology and pharmacodynamic studies. Scientific reports on crude extracts and pure compounds and formulations revealed a wide range of pharmacological activities, including anti-inflammatory activity

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

    PubMed Central

    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

    2015-01-01

    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. PMID:25618143

  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. 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.

  7. 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...

  8. 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...

  9. 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

  10. 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.

  11. 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

  12. [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.

  13. Development of Ophiocordyceps sinensis through Plant-Mediated Interkingdom Host Colonization

    PubMed Central

    Lei, Wei; Zhang, Guren; Peng, Qingyun; Liu, Xin

    2015-01-01

    Ophiocordyceps sinensis is a well-known entomogenous and medicinal fungus. After its anamorphs parasitize the larvae of the genus Thitarodes, fruit-bodies may form to be used as medicine. However, its developmental mechanisms remain unknown. The distribution of O. sinensis was determined in different tissues of the Thitarodes larvae and the dominant plant species using real-time quantitative polymerase chain reaction (qPCR) and fluorescence in situ hybridization (FISH) technique, respectively. We found that more fungal material was located in plants than in larvae, especially in Ranunculus tanguticus. A considerable amount was detected in larval intestinal-wall and plant roots. It is suggested that plants are the potential hosts of O. sinensis, which modifies our understanding of the life cycle of O. sinensis and indicates that the phytophagous larvae may become infected as they feed. Our research may contribute to the study of systematic evolution and population ecology of O. sinensis, elucidate its developmental mechanism and promote sustainable harvesting. PMID:26263972

  14. [Cordyceps sinensis enhances lymphocyte proliferation and CD markers expression in simulated microgravity environment].

    PubMed

    Hao, Tong; Li, Jun-Jie; Du, Zhi-Yan; Duan, Cui-Mi; Wang, Yan-Meng; Wang, Chang-Yong; Song, Jing-Ping; Wang, Lin-Jie; Li, Ying-Hui; Wang, Yan

    2012-10-01

    This study was aimed to explore the effect of cordyceps sinensis enhancing lymphocyte proliferation and surface CD marker expression in simulated microgravity environment. The splenic lymphocytes were separated from mice and cultured in the rotary cell culture system simulated microgravity environment. The cells were treated with different concentration of cordyceps sinensis solution (0, 6.25, 12.5, 25 and 50 µg/ml) for 24, 48 and 72 h respectively, then the cells were harvested, and analyzed for cell proliferation and the expression of cell surface markers (CD4 and CD8). The results showed that under simulated microgravity environment, the lymphocyte proliferation was inhibited. When the concentration of cordyceps sinensis was 25 or 50 µg/ml, the lymphocyte proliferation, CD4 and CD8 expressions all increased, but 50 µg/ml cordyceps sinensis could inhibit the proliferation ability with the time prolonging. It is concluded that the suitable concentration of cordyceps sinensis displayed the ability to enhance the lymphocyte proliferation and CD marker expression in simulated microgravity environment. These results may be valuable for screening drugs which can be potentially against immunosuppression under simulated microgravity.

  15. IDENTIFCATION AND VALIDATION OF NOVEL microRNA MOLECULE FROM THE PELODISCUS SINENSIS BY BIOINFORMATICS APPROACHES.

    PubMed

    Huang, Yong; Ren, Hong Tao; Wang, Zhan Bin; Sun, Xi Hong

    2015-01-01

    MicroRNAs (miRNAs) are a group of endogenous, short non-coding RNAs with the length of about 22 nt, which mediate gene expression at the post-transcriptional level through mRNA degradation or translational repression. Among them, some are highly evolutionally conserved in the animal kingdom; this provides a powerful strategy for identifying miRNAs in new species. The Chinese soft-shelled turtle (Pelodiscus sinensis) is one of the most important freshwater aquaculture reptilian species in China, but related miRNAs have not been identified up to now. In the present study, a total of 10 Pelodiscus sinensis miRNAs were identified according to Pelodiscus sinensis ESTs and GSSs information in NCBI database by bioinformatics approaches. The RT-PCR-based assays were performed and found that 10 Pelodiscus sinensis miRNAs were expressed. Using these miRNAs, 22 target genes were identified. These genes encode 22 proteins involved in metabolism, signal transduction, transcriptional regulation, and development. These miRNAs and their targets will serve as useful resources for their, functional analyses in miRNA-regulated processes in Pelodiscus sinensis breeding and genetic research.

  16. [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.

  17. 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.

  18. 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.

  19. 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

  20. Development of 12 genic microsatellite loci for a biofuel grass, Miscanthus sinensis (Poaceae).

    PubMed

    Ho, Chuan-Wen; Wu, Tai-Han; Hsu, Tsai-Wen; Huang, Jao-Ching; Huang, Chi-Chun; Chiang, Tzen-Yuh

    2011-08-01

    Miscanthus, a nonfood plant with high potential as a biofuel, has been used in Europe and the United States. The selection of a cultivar with high biomass, photosynthetic efficiency, and stress resistance from wild populations has become an important issue. New genic microsatellite markers will aid the assessment of genetic diversity for different strains. Twelve polymorphic microsatellite markers derived from the transcriptome of Miscanthus sinensis fo. glaber were identified and screened on 80 individuals of M. sinensis. The number of alleles per locus ranged from 6 to 12, and the mean expected heterozygosity was 0.75. Cross-taxa transferability revealed that all loci can be applied to all varieties of M. sinensis, as well as the closely related species M. floridulus. These new genic microsatellite markers are useful for characterizing different traits in breeding programs or to select genes useful for biofuel.

  1. Morphologic analysis of atretic follicles in the Chinese soft-shelled turtle, Pelodiscus sinensis.

    PubMed

    Wang, Wei; Li, Caiyan; Yu, Jianfeng; Jiang, Zhen; Si, Yuexiu; Yin, Shangjun; Qian, Guoying

    2013-12-01

    The Chinese soft-shelled turtle (Pelodiscus sinensis) and its eggs have been utilized as both a source of nutrition and medicine for thousands of years. In the present study, follicular atresia was investigated by morphological analysis as an important factor affecting clutch size. Results showed that follicular atresia can be divided into four stages: (1) development of follicular cell disorder; (2) massive cellular proliferation in the ooplasm with elimination of yolk granules; (3) large amounts of erythrocytes appearing in the ooplasm; (4) gradual degeneration of atretic follicles. For the first time, we have shown follicular atresia to be a dynamic process in P. sinensis and found that yolk granules are eliminated by both liquefaction as well as non-liquefaction processes. The present study may provide detailed morphologic analysis for further research into egg quality and clutch size in P. sinensis. Copyright © 2013 Elsevier Ltd. All rights reserved.

  2. 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

  3. Complete mitochondrial genome of Chinese bamboo rat, Rhizomys sinensis and species divergence comparison.

    PubMed

    Xu, Yu; Liu, Xiaohua; Tu, Feiyun

    2016-05-01

    The Chinese bamboo rat Rhizomys sinensis belongs to family Spalacidae, and is distributed in China, Myanmar, and Vietnam. In this study, the entire mitochondrial genome of R. sinensis was firstly determined. The genome is 16,564 bases in length, containing 13 protein coding genes, 22 transfer RNA genes, 2 ribosomal RNA genes and a putative control region. The composition and arrangement of its genes are identical to most other mammals. The whole base composition of the mitogenome is A 31.9%, G 12.0%, T 31.2% and C 24.9%, with an A+T rich pattern. The species divergence between R. sinensis and R. pruinosus is 0.113, in accordance with a "10× rule", as the intraspecific barcode variation averaged 1%. The mitogenome data of R. sumatrensis is required to better understand the phylogenetic relationships within Rhizomys.

  4. 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.

  5. Draft genome of the Chinese mitten crab, Eriocheir sinensis.

    PubMed

    Song, Linsheng; Bian, Chao; Luo, Yongju; Wang, Lingling; You, Xinxin; Li, Jia; Qiu, Ying; Ma, Xingyu; Zhu, Zhifei; Ma, Liang; Wang, Zhaogen; Lei, Ying; Qiang, Jun; Li, Hongxia; Yu, Juhua; Wong, Alex; Xu, Junmin; Shi, Qiong; Xu, Pao

    2016-01-01

    The Chinese mitten crab, Eriocheir sinensis, is one of the most studied and economically important crustaceans in China. Its transition from a swimming to a crawling method of movement during early development, anadromous migration during growth, and catadromous migration during breeding have been attractive features for research. However, knowledge of the underlying molecular mechanisms that regulate these processes is still very limited. A total of 258.8 gigabases (Gb) of raw reads from whole-genome sequencing of the crab were generated by the Illumina HiSeq2000 platform. The final genome assembly (1.12 Gb), about 67.5 % of the estimated genome size (1.66 Gb), is composed of 17,553 scaffolds (>2 kb) with an N50 of 224 kb. We identified 14,436 genes using AUGUSTUS, of which 7,549 were shown to have significant supporting evidence using the GLEAN pipeline. This gene number is much greater than that of the horseshoe crab, and the annotation completeness, as evaluated by CEGMA, reached 66.9 %. We report the first genome sequencing, assembly, and annotation of the Chinese mitten crab. The assembled draft genome will provide a valuable resource for the study of essential developmental processes and genetic determination of important traits of the Chinese mitten crab, and also for investigating crustacean evolution.

  6. 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).

  7. 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.

  8. Neuroprotective and Cytotoxic Phthalides from Angelicae Sinensis Radix.

    PubMed

    Gong, Wenxia; Zhou, Yuzhi; Li, Xiao; Gao, Xiaoxia; Tian, Junsheng; Qin, Xuemei; Du, Guanhua

    2016-04-26

    Seven phthalides, including a new dimeric one named tokinolide C (7), were isolated from Angelicae Sinensis Radix and characterized. The structures of these compounds were elucidated on the basis of comprehensive analysis of spectroscopic data and comparison with literature data. All of the compounds were evaluated for their cytotoxic activities against the A549, HCT-8, and HepG2 cancer cell lines. Riligustilide (4) showed cytotoxicity against three cancer cell lines, with IC50 values of 13.82, 6.79, and 7.92 μM, respectively. Tokinolide A (6) and tokinolide C (6) exerted low cytotoxicity in these cancer cell lines, while the remaining compounds were inactive. Flow cytometry analysis was employed to evaluate the possible mechanism of cytotoxic action of riligustilide (4). We observed that compound 4 was able to arrest the cell cycle in the G1, S phases and induce apoptosis in a time-dependent manner in HCT-8 cell lines. In addition, these compounds were evaluated for neuroprotective effect against SH-SY5Y cells injured by glutamate. The result showed that ligustilide (1), Z-butylidenephthalide (3) and tokinolide A (6) exhibited significant neuroprotective effects.

  9. 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.

  10. 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

  11. Epidemiological and Clinical Parameters Features of Patients with Clonorchiasis in the Geum River Basin, Republic of Korea

    PubMed Central

    Lee, Myoung-Ro; Ju, Jung-Won; Jeong, Byong-Suk; Park, Mi-Yeoun; Lee, Keoung-Sook

    2017-01-01

    This study was conducted to evaluate the infection rates of Clonorchis sinensis and laboratory findings in infected people. The 3,167 fecal samples, from nine villages in Okcheon-gun, were examined. PMID:28529523

  12. 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.

  13. Sequencing and analysis of the complete mitochondrial genome in Anopheles sinensis (Diptera: Culicidae).

    PubMed

    Chen, Kai; Wang, Yan; Li, Xiang-Yu; Peng, Heng; Ma, Ya-Jun

    2017-10-02

    Anopheles sinensis (Diptera: Culicidae) is a primary vector of Plasmodium vivax and Brugia malayi in most regions of China. In addition, its phylogenetic relationship with the cryptic species of the Hyrcanus Group is complex and remains unresolved. Mitochondrial genome sequences are widely used as molecular markers for phylogenetic studies of mosquito species complexes, of which mitochondrial genome data of An. sinensis is not available. An. sinensis samples was collected from Shandong, China, and identified by molecular marker. Genomic DNA was extracted, followed by the Illumina sequencing. Two complete mitochondrial genomes were assembled and annotated using the mitochondrial genome of An. gambiae as reference. The mitochondrial genomes sequences of the 28 known Anopheles species were aligned and reconstructed phylogenetic tree by Maximum Likelihood (ML) method. The length of complete mitochondrial genomes of An. sinensis was 15,076 bp and 15,138 bp, consisting of 13 protein-coding genes, 22 transfer RNA (tRNA) genes, 2 ribosomal RNA (rRNA) genes, and an AT-rich control region. As in other insects, most mitochondrial genes are encoded on the J strand, except for ND5, ND4, ND4L, ND1, two rRNA and eight tRNA genes, which are encoded on the N strand. The bootstrap value was set as 1000 in ML analyses. The topologies restored phylogenetic affinity within subfamily Anophelinae. The ML tree showed four major clades, corresponding to the subgenera Cellia, Anopheles, Nyssorhynchus and Kerteszia of the genus Anopheles. The complete mitochondrial genomes of An. sinensis were obtained. The number, order and transcription direction of An. sinensis mitochondrial genes were the same as in other species of family Culicidae.

  14. 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.

  15. 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

  16. 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).

  17. 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.

  18. Functional study of Cordyceps sinensis and cordycepin in male reproduction: A review.

    PubMed

    Chen, Yung-Chia; Chen, Ying-Hui; Pan, Bo-Syong; Chang, Ming-Min; Huang, Bu-Miin

    2017-01-01

    Cordyceps sinensis has various biological and pharmacological functions, and it has been claimed as a tonic supplement for sexual and reproductive dysfunctions for a long time in oriental society. In this article, the in vitro and in vivo effects of C. sinensis and cordycepin on mouse Leydig cell steroidogenesis are briefly described, the stimulatory mechanisms are summarized, and the recent findings related to the alternative substances regulating male reproductive functions are also discussed. Copyright © 2016. Published by Elsevier B.V.

  19. 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.

  20. 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

  1. Effect of Gleditsia sinensis Lam. Extract on Physico-Chemical Properties of Emulsion-Type Pork Sausages

    PubMed Central

    Jin, Sang-Keun; Yang, Han-Sul; Choi, Jung-Seok

    2017-01-01

    This study was performed to investigate the effect of Gleditsia sinensis Lam. extract on the physicochemical properties of emulsion-type pork sausages during storage at 10°C for 4 wk. Treatments were as follows: (C, control; T1, sodium ascorbate 0.05%; T2, Gleditsia sinensis Lam. 0.05%; T3, Gleditsia sinensis Lam. 0.1%; T4, Gleditsia sinensis Lam. 0.2%; T5, Gleditsia sinensis Lam. 0.1% + sodium ascorbate 0.05%). The values of pH, moisture content, lightness, redness, and sensory attributes were all significantly decreased, while the yellowness, chroma, hue angle, and texture properties were increased during storage with increase of the Gleditsia sinensis Lam. extract added. In addition, the antioxidant activity and antimicrobial activity in the sausages displayed significant increases (p<0.05). Therefore, although it was concluded that the addition of Gleditsia sinensis Lam. extract is not effective for improvement of the physical properties compared to chemical additives in sausages, it could be applied to meat products as a natural preservatives. PMID:28515651

  2. Immune-Modulating Activity of Extract Prepared from Mycelial Culture of Chinese Caterpillar Mushroom, Ophiocordyceps sinensis (Ascomycetes).

    PubMed

    Jang, Sun-Hee; Kim, Sae-Hae; Lee, Ha-Yan; Jang, Seung-Hwan; Jang, Hyonseok; Chae, Soo-Wan; Jung, Su-Jin; So, Byung-Ok; Ha, Ki-Chan; Sin, Hong-Sig; Jang, Yong-Suk

    2015-01-01

    Ophiocordyceps sinensis is a natural fungus that has been valued as a health food and traditional Chinese medicine for centuries. The fungus is parasitic and colonizes insect larva. Naturally occurring O. sinensis thrives at high altitude in cold and grassy alpine meadows on the Himalayan mountain ranges. Wild O. sinensis is becoming increasingly rare in its natural habitats, and its price is out of reach for clinical practice. For these reasons, development of a standardized alternative is a great focus of research to allow the use of O. sinensis as a medicine. To develop an alternative for wild O. sinensis, a refined standardized extract, CBG-CS-2, was produced by artificial fermentation and extraction of the mycelial strain Paecilomyces hepiali CBG-CS-1, which originated from wild O. sinensis. In this study, we analyzed the in vivo immune-modulating effect of CBG-CS-2 in mice. Oral administration of CBG-CS-2 supported splenocyte stimulation and enhanced Th1-type cytokine expression from the splenocytes. Importantly, the same treatment significantly enhanced the natural killer cell activity of the splenocytes. Finally, oral administration of CBG-CS-2 enhanced the potential for inflammatory responses. Together, these findings indicate that the mycelial culture extract prepared from O. sinensis exhibited immune-modulating activity and suggest its possible use in the treatment of diseases caused by abnormal immune function.

  3. Anti-diabetic properties of non-polar Toona sinensis Roem extract prepared by supercritical-CO2 fluid.

    PubMed

    Hsieh, Tusty-Jiuan; Tsai, Yi-Hong; Liao, Min-Chun; Du, Ying-Chi; Lien, Pei-Jung; Sun, Chuan-Chia; Chang, Fang-Rong; Wu, Yang-Chang

    2012-03-01

    Toona sinensis Roem (T. sinensis) leaves have been used as a nutritious vegetable and been suggested for medical applications; however, the reported bioactive compounds of T. sinensis leaves are, so far, from high to mid-high polar extracts. Our aims in this study were to reveal the non-polar constituents of the T. sinensis leave extract that were prepared by a method of using a supercritical-CO2 fluid and to investigate the anti-diabetic potential of this extract. Through a GC/MS analysis, we revealed 24 major components of the non-polar T. sinensis leave extract, the most abundant of which was phytol. The non-polar T. sinensis leave extract showed to prevent the progression of diabetes and hepatosteatosis, the rise of triglycerol levels and the decrease of adiponectin levels in the type 2 diabetic mice. Our results suggest that the non-polar extract of T. sinensis leaves prepared using the supercritical-CO2 fluid may contain effective constituents to prevent type 2 diabetes. Copyright © 2011 Elsevier Ltd. All rights reserved.

  4. 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.

  5. Cordyceps sinensis (a traditional Chinese medicine) for kidney transplant recipients.

    PubMed

    Hong, Tao; Zhang, Minghua; Fan, Junming

    2015-10-12

    Kidney transplantation is the treatment of choice for patients with end-stage kidney disease (ESKD). Rising ESKD prevalence has substantially increased numbers of kidney transplants performed. Maintenance immunosuppression is long-term treatment to prevent acute rejection and deterioration of graft function. Although immunosuppressive treatment using drugs such as calcineurin inhibitors (CNIs, such as cyclosporin A (CsA) or tacrolimus) reduce acute rejection rates, long-term allograft survival rates are not significantly enhanced. CNI-related adverse effects contribute to reduced quality of life among kidney transplant recipients. Adjuvant immunosuppressive therapies that could offer a synergetic immunosuppressive effect, while minimising toxicity and reducing side effects, have been explored recently. Cordyceps sinensis, (Cordyceps) a traditional Chinese medicine, is used as an adjuvant immunosuppressive agent in maintenance treatment for kidney transplantation recipients in China, but there is no consensus about its use as an adjuvant immunosuppressive treatment for kidney transplantation recipients. This review aimed to evaluate the benefits and potential adverse effects of Cordyceps as an adjuvant immunosuppressive treatment for kidney transplant recipients. We searched the Cochrane Kidney and Transplant Specialised Register through contact with the Trials Search Co-ordinator to 7 September 2015 using search terms relevant to this review. We also searched Chinese language databases and other resources. We included all randomised controlled trials (RCTs) and quasi-RCTs evaluating the benefits and potential side effects of Cordyceps sinensis for kidney transplant recipients, irrespective of blinding or publication language. An inclusion criterion was that baseline immunosuppressive therapy must be the same in all study arms. Two authors extracted data. We derived risk ratios (RR) for dichotomous data and mean differences (MD) for continuous data with 95

  6. 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.

  7. Global transcriptome profiles of Camellia sinensis during cold acclimation.

    PubMed

    Wang, Xin-Chao; Zhao, Qiong-Yi; Ma, Chun-Lei; Zhang, Zong-Hong; Cao, Hong-Li; Kong, Yi-Meng; Yue, Chuan; Hao, Xin-Yuan; Chen, Liang; Ma, Jian-Qiang; Jin, Ji-Qiang; Li, Xuan; Yang, Ya-Jun

    2013-06-22

    Tea is the most popular non-alcoholic health beverage in the world. The tea plant (Camellia sinensis (L.) O. Kuntze) needs to undergo a cold acclimation process to enhance its freezing tolerance in winter. Changes that occur at the molecular level in response to low temperatures are poorly understood in tea plants. To elucidate the molecular mechanisms of cold acclimation, we employed RNA-Seq and digital gene expression (DGE) technologies to the study of genome-wide expression profiles during cold acclimation in tea plants. Using the Illumina sequencing platform, we obtained approximately 57.35 million RNA-Seq reads. These reads were assembled into 216,831 transcripts, with an average length of 356 bp and an N50 of 529 bp. In total, 1,770 differentially expressed transcripts were identified, of which 1,168 were up-regulated and 602 down-regulated. These include a group of cold sensor or signal transduction genes, cold-responsive transcription factor genes, plasma membrane stabilization related genes, osmosensing-responsive genes, and detoxification enzyme genes. DGE and quantitative RT-PCR analysis further confirmed the results from RNA-Seq analysis. Pathway analysis indicated that the "carbohydrate metabolism pathway" and the "calcium signaling pathway" might play a vital role in tea plants' responses to cold stress. Our study presents a global survey of transcriptome profiles of tea plants in response to low, non-freezing temperatures and yields insights into the molecular mechanisms of tea plants during the cold acclimation process. It could also serve as a valuable resource for relevant research on cold-tolerance and help to explore the cold-related genes in improving the understanding of low-temperature tolerance and plant-environment interactions.

  8. [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.

  9. 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.

  10. 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

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

    USDA-ARS?s Scientific Manuscript database

    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...

  12. Genetic and nongenetic factors influencing callus induction in Miscanthus sinensis (Anderss.) anther cultures.

    PubMed

    Glowacka, K; Jezowski, S

    2009-01-01

    Miscanthus sinensis is a promising species for biomass production. Influences of genetic and nongenetic factors on androgenesis induction efficiency were investigated. This is the first report on successful induction of pollen-derived callus in M. sinensis. The callus yield was strongly affected by genotype. A beneficial influence of cold pretreatment of spikes on androgenesis induction was observed. The highest yield of calli was obtained in cultures on a modified C17 medium. The results suggest that the high callus yield might be caused by the late culture initiation. The beginning of anther culture at the end of the flowering season caused a 17-fold increase in callus yield, in comparison to culture initiated at the height of the flowering season (August). It is likely, however, that the efficiency of androgenesis induction in the case of M. sinensis anther culture beginning in October could be related to a positive influence of growing donor plants in conditions of cooler and shorter day, i.e. 11-h day with temperature around 11xC and 13-h night with temperature around 5xC. Results of this study can significantly support the development of effective methods of M. sinensis haploidization, which could be used in crop improvement by breeding.

  13. 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.

  14. 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.

  15. 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. © 2011 The Authors. New Phytologist © 2011 New Phytologist Trust.

  16. 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.

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

    USDA-ARS?s Scientific Manuscript database

    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...

  18. Identification of miRNAs and their targets in tea (Camellia sinensis)#

    PubMed Central

    Zhu, Quan-wu; Luo, Yao-ping

    2013-01-01

    MicroRNAs (miRNAs) are endogenous small RNAs playing a crucial role in plant growth and development, as well as stress responses. Among them, some are highly evolutionally conserved in the plant kingdom, this provide a powerful strategy for identifying miRNAs in a new species. Tea (Camellia sinensis) is one of the most important commercial beverage crops in the world, but only a limited number of miRNAs have been identified. In the present study, a total of 14 new C. sinensis miRNAs were identified by expressed sequence tag (EST) analysis from 47 452 available C. sinensis ESTs. These miRNAs potentially target 51 mRNAs, which can act as transcription factors, and participate in stress response, transmembrane transport, and signal transduction. Analysis of gene ontology (GO), based on these targets, suggested that 37 biological processes were involved, such as oxidation-reduction process, stress response, and transport. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis inferred that the identified miRNAs took part in 13 metabolic networks. Our study will help further understanding of the essential roles of miRNAs in C. sinensis growth and development, and stress response. PMID:24101208

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

    PubMed

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

    2016-10-26

    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.

  20. Synergistic Antimicrobial Activity of Camellia sinensis and Juglans regia against Multidrug-Resistant Bacteria

    PubMed Central

    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 PMID:25719410

  1. 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

  2. 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

  3. 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

  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. Validation and characterization of Citrus sinensis microRNAs and their target genes.

    PubMed

    Song, Changnian; Yu, Mingliang; Han, Jian; Wang, Chen; Liu, Hong; Zhang, Yanping; Fang, Jinggui

    2012-05-15

    MicroRNAs play vital role in plant growth and development by changeable expression of their target genes with most plant microRNAs having perfect or near-perfect complementarities with their target genes but miRNAs in Citrus sinensis (csi-miRNAs) and their function have not been widely studied. In this study, 15 potential microRNAs in Citrus sinensis (csi-miRNAs) were identified and bioinformatically validated using miR-RACE, a newly developed method for determination of miRNAs prediction computationally. The expression of these fifteen C. sinensis miRNAs can be detected in leaves, stems, flowers and fruits of C. sinensis by QRT-PCR with some of them showed tissue-specific expression. Six potential target genes were identified for six csi-miRNAs and also experimentally verified by Poly (A) polymerase -mediated 3' rapid amplification of cDNA ends (PPM-RACE) and RNA ligase-mediated 5' rapid amplification of cDNA ends (RLM-RACE) which mapped the cleavage site of target mRNAs and detected expression patterns of cleaved fragments that indicate the regulatory function of the miRNAs on their target genes. Our results confirm that small RNA-mediated regulation whereby all csi-miRNAs regulate their target genes by degradation.

  6. 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.

  7. Niacin, thiamin, iron and protein status of humans as affected by the consumption of tea (Camellia sinensis) infusions.

    PubMed

    Wang, R S; Kies, C

    1991-10-01

    The objective of the project was to determine the effects of tea (Camellia sinensis) leaf beverage consumption on the apparent utilization of niacin, thiamin, and protein in human subjects. During two randomly arranged experimental periods of 14 days each, 10 adult female human subjects were fed a constant (same foods each day), measured, laboratory-controlled diet. Tea was fed during one period while no tea was allowed during the other period. A dehydrated black tea infusion product (instant tea, 8 g/subject/day/dry weight basis) was used which subjects were allowed to dilute in water. Subjects made complete collections of urine and stools. Fasting blood samples were drawn at the end of each experimental period. No effects on protein status were demonstrated. Urinary thiamin losses were depressed with the use of tea but niacin losses were increased. Blood serum concentrations of thiamin diphosphate also were depressed during the tea drinking period as compared to values during the non-tea period. No significant effect on blood serum levels of nicotinaminde or N'-methylnicotinamine were found but values tended to be lower during the tea than the non-tea period. These results suggest that tea consumption inhibits the utilization of thiamin. However, niacin availability was unaffected by tea drinking. Because of the decreased availability of thiamin, the need for niacin was depressed which caused a greater than expected urinary loss of this vitamin.

  8. 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

  9. 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

  10. [Potential Ecological Suitability Regionalization Analysis of Angelica sinensis Based on GIS and Fuzzy Matter Element Model].

    PubMed

    Shang, Zhong-hui; Wei, Hai-yan; Gu, Wei; Mao, Ya-juan; Zhu, Li-nan; Sang, Man-jie

    2015-07-01

    To study the potential ecological suitability regionalization of Angelica sinensis, for protecting wild resources and selecting cultivation location and designing rational production layout. Based on fuzzy matter element model, the relationship of fuzzy membership function between ferulic acid content and 14 ecological factors, including climate, topography and soil,were established. Then information entropy theory was used to determine the relative importance of each environmental factor, and thus to determine the most limiting habitat criteria. Finally, the probable spatial distribution of Angelica sinensis across ten provinces in Western China was determined based on GIS spatial analysis of habitat conditions. Meanwhile, the optimal index range of ecological factors was quantified. It was showed that the percentage of moderately and highly suitable habitats for Angelica sinensis in the study area was 9. 64%, its area was 306,768. 01 km2. The moderately and highly suitable habitats were mainly located in the southeast of Gansu ind Tibet,the north of Sichuan and the northwest of Yunnan. The results also showed that six dominant ecological factors controlling the distribution of Angelica sinensis. These six dominant features were as follows: (1) mean temperature of wettest quarter, (2) altitude, (3) precipitation of growth, (4) annual relative humidity, (5) average temperature of growth period, and (6) annual )recipitation. The habitat suitability assessment model based on GIS and fuzzy matter element model theory can accurately valuate the habitat suitability of Angelica sinensis, quantify the area of suitable habitat and analyze the spatial distribution. This informaion is of value to provide insight for choosing the most suitable cultivation sites,as well as the habitat protection zones.

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

    PubMed

    Yue, Huamei; Li, Chuangju; Du, Hao; Zhang, Shuhuan; Wei, Qiwei

    2015-01-01

    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. 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. 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 management of cultured fish.

  12. [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

  13. 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

  14. 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.

  15. 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

  16. 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. © The Author 2015. Published by Oxford University Press on behalf of the Society for Experimental Biology.

  17. 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

  18. Stable Carbon Isotope Composition of the Lipids in Natural Ophiocordyceps sinensis from Major Habitats in China and Its Substitutes.

    PubMed

    Guo, Lian-Xian; Xu, Xiao-Ming; Hong, Yue-Hui; Li, Yan; Wang, Jiang-Hai

    2017-09-18

    Ophiocordyceps sinensis is one rare medicinal fungus produced in the Qinghai-Tibetan Plateau. Its quality and price varies hugely with different habitat, and its numerous substitutes have sprung up in functional food markets. This paper aims to discriminate the geographic origin of wild O. sinensis and its substitutes via element analyzer-isotope ratio mass spectrometry and gas chromatography-isotope ratio mass spectrometry. The δ(13)C values of major fatty acids in the lipids of O. sinensis are characterized unanimously by the variation relation C18:0 < C18:2 ≈ C16:0 < C18:1, while their fluctuation intervals are notably different between those of neutral and polar lipids. The comparative analysis of the δ(13)C ratios of major fatty acids in lipids of O. sinensis suggests that the δ(13)C patterns may be sensitive potential indicators to discriminate its geographical origin. The δ(13)C values of individual major fatty acids of lipids from the cultivated stromata of Cordyceps militaris (SCM), the fermented mycelia of Hirsurella sinensis (FMH) and Paecilomyces epiali (FMP) range from -31.2‰ to -29.7‰, -16.9‰ to -14.3‰, and -26.5‰ to -23.9‰, respectively. Their δ(13)C pattern of individual major fatty acids may be used as a potential indicator to discriminate the products of natural O. sinensis and its substitutes.

  19. 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

  20. Biology, Bionomics and Molecular Biology of Anopheles sinensis Wiedemann 1828 (Diptera: Culicidae), Main Malaria Vector in China.

    PubMed

    Feng, Xinyu; Zhang, Shaosen; Huang, Fang; Zhang, Li; Feng, Jun; Xia, Zhigui; Zhou, Hejun; Hu, Wei; Zhou, Shuisen

    2017-01-01

    China has set a goal to eliminate all malaria in the country by 2020, but it is unclear if current understanding of malaria vectors and transmission is sufficient to achieve this objective. Anopheles sinensis is the most widespread malaria vector specie in China, which is also responsible for vivax malaria outbreak in central China. We reviewed literature from 1954 to 2016 on An. sinensis with emphasis on biology, bionomics, and molecular biology. A total of 538 references were relevant and included. An. sienesis occurs in 29 Chinese provinces. Temperature can affect most life-history parameters. Most An. sinensis are zoophilic, but sometimes they are facultatively anthropophilic. Sporozoite analysis demonstrated An. sinensis efficacy on Plasmodium vivax transmission. An. sinensis was not stringently refractory to P. falciparum under experimental conditions, however, sporozoite was not found in salivary glands of field collected An. sinensis. The literature on An. sienesis biology and bionomics was abundant, but molecular studies, such as gene functions and mechanisms, were limited. Only 12 molecules (genes, proteins or enzymes) have been studied. In addition, there were considerable untapped omics resources for potential vector control tools. Existing information on An. sienesis could serve as a baseline for advanced research on biology, bionomics and genetics relevant to vector control strategies.

  1. Studies on the Biochemical Formation Pathway of the Amino Acid l-Theanine in Tea (Camellia sinensis) and Other Plants.

    PubMed

    Cheng, Sihua; Fu, Xiumin; Wang, Xiaoqin; Liao, Yinyin; Zeng, Lanting; Dong, Fang; Yang, Ziyin

    2017-08-23

    Tea (Camellia sinensis) is the most widely consumed beverage aside from water. The flavor of tea is conferred by certain metabolites, especially l-theanine, in C. sinensis. To determine why more l-theanine accumulates in C. sinensis than in other plants, we compare l-theanine contents between C. sinensis and other plant species (Camellia nitidissima, Camellia japonica, Zea mays, Arabidopsis thaliana, and Solanum lycopersicum) and use a stable isotope labeling approach to elucidate its biosynthetic route. We quantify relevant intermediates and metabolites by mass spectrometry. l-Glutamic acid, a precursor of l-theanine, is present in most plants, while ethylamine, another precursor of l-theanine, specifically accumulates in Camellia species, especially C. sinensis. Most plants contain the enzyme/gene catalyzing the conversion of ethylamine and l-glutamic acid to l-theanine. After supplementation with [(2)H5]ethylamine, all the plants produce [(2)H5]l-theanine, which suggests that ethylamine availability is the reason for the difference in l-theanine accumulation between C. sinensis and other plants.

  2. Biology, Bionomics and Molecular Biology of Anopheles sinensis Wiedemann 1828 (Diptera: Culicidae), Main Malaria Vector in China

    PubMed Central

    Feng, Xinyu; Zhang, Shaosen; Huang, Fang; Zhang, Li; Feng, Jun; Xia, Zhigui; Zhou, Hejun; Hu, Wei; Zhou, Shuisen

    2017-01-01

    China has set a goal to eliminate all malaria in the country by 2020, but it is unclear if current understanding of malaria vectors and transmission is sufficient to achieve this objective. Anopheles sinensis is the most widespread malaria vector specie in China, which is also responsible for vivax malaria outbreak in central China. We reviewed literature from 1954 to 2016 on An. sinensis with emphasis on biology, bionomics, and molecular biology. A total of 538 references were relevant and included. An. sienesis occurs in 29 Chinese provinces. Temperature can affect most life-history parameters. Most An. sinensis are zoophilic, but sometimes they are facultatively anthropophilic. Sporozoite analysis demonstrated An. sinensis efficacy on Plasmodium vivax transmission. An. sinensis was not stringently refractory to P. falciparum under experimental conditions, however, sporozoite was not found in salivary glands of field collected An. sinensis. The literature on An. sienesis biology and bionomics was abundant, but molecular studies, such as gene functions and mechanisms, were limited. Only 12 molecules (genes, proteins or enzymes) have been studied. In addition, there were considerable untapped omics resources for potential vector control tools. Existing information on An. sienesis could serve as a baseline for advanced research on biology, bionomics and genetics relevant to vector control strategies. PMID:28848504

  3. [Effect of altitudes on the photosynthate accumulation and distribution pattern of Angelic sinensis].

    PubMed

    Wang, Hui-Zhen; Jin, Ling; Zhang, En-He

    2012-08-01

    To study the effect of altitudes on the photosynthate accumulation and distribution pattern of Angelic sinensis in Gansu province and provide theontical for its expanding ecological planting region. Used field test to study the photosynthate accumulation and distribution pattern of Angelic sinensis on three different altitudinal gradients from 2 300 m to 2 800 m. Before September 25, total photosynthate accumulation were decreased with the increasing of elevation, which amaunt was 176 g/plant, 166 g/plant and 128 g/plant, respectively. The total photosynthate of low-altitude and middle-altitude were significantly higher than that of high-altitude (P < 0.05). After September 25, middle-altitude was significantly higher than the other two altitudes (P < 0.05), respectively, by 13.9% and 11.1%. The photosynthate accumulation rate existed the altitude effects, there was no significant difference between high-altitude (46.7%) and middle-altitude (43.7%), but they were even significantly higher than that of low-altitude (33.1%). The root distribution proportion (> 30%) existed the difference, that of high-altitude was 10 days earlier than the other two altitudes, and later that of high-altitude (about 54%) was higher than the other two altitudes (49.8% - 50.9%), it laid the foundation for yield formation. Yield of Angelic sinensis was as follows: middle-altitude (28.4 g/ plant), high-altitude (26.6 g/plant) and low-altitude (21.8 g/plant). Yield of Angelic sinensis middle-altitude and high-altitude were higher than that of low-altitude, respectively, by 30.2% and 22.2%, and it had a significant difference (P < 0.05), this result was consistent with the photosynthate accumulation rate. Altitudinal gradients affect yield formation of Angelic sinensis by changing the photosynthate distribution pattern and dry matter accumulation rate. So by appropriately increasing altitude, the root distribution proportion and yield are improved, this provides theoretical reference for

  4. Using Molecular Docking Analysis to Discovery Dregea sinensis Hemsl. Potential Mechanism of Anticancer, Antidepression, and Immunoregulation

    PubMed Central

    Liu, Xiujie; Shi, Yu; Deng, Yulin; Dai, Rongji

    2017-01-01

    Background: Dregea sinensis Hemsl. plant of the genus Dregea volubilis (Asclepiadaceae), plays a vital role in anticancer, antidepression, and immunoregulation. Steroidal glycosides are the main constituents of this herb, which were significant biological active ingredients. Objective: The objective of this study is to recognize the mechanism of anticancer, antidepression, and immunoregulation of D. sinensis Hemsl. Materials and Methods: Seventy-two steroidal glycosides of D. sinensis Hemsl. were evaluated on the docking behavior of tumor-associated proteins (PI3K, Akt, mTOR), depression-related proteins (MAO-A, MAO-B) and immune-related proteins (tumor necrosis factor-α [TNF-α], tumor necrosis factor receptor 2 [TNFR2], interleukin-2Rα [IL-2Rα]) using Discovery Studio version 3.1 (Accelrys, San Diego, USA). Results: The molecular docking analysis revealed that mostly steroidal glycosides of D. sinensis Hemsl. exhibited powerful interaction with the depression-related protein (MAO-A) and the immune-related proteins (TNFR2, IL-2Rα). Some ligands exhibited high binding energy for the tumor-associated proteins (PI3K, Akt, mTOR) and the immune-related protein (TNF-α), but MAO-B showed none interaction with the ligands. Conclusion: This study has paved better understanding of steroidal glycosides from D. sinensis Hemsl. as potential constituents to the prevention of associated cancer, depression and disorders of immunoregulation. SUMMARY The ligand database was consist of 72 steroidal glycosides from Dregea sinensis HemslSteroidal glycosides had the potential to dock with the tumor-associated proteins (PI3K, Akt, mTOR)Steroidal glycosides were bounded with MAO-A rather than MAO-B, accorded with the inhibitor selectivity of MAOs, can be considered as potent candidate inhibitors of MAO-A72 ligands got high interaction with TNFR2 and IL-2Rα, regard the steroidal glycoside as powerful candidate inhibitors of TNFR2 and IL-2Rα. Abbreviations used: PI3K: Phosphatidyl

  5. Using Molecular Docking Analysis to Discovery Dregea sinensis Hemsl. Potential Mechanism of Anticancer, Antidepression, and Immunoregulation.

    PubMed

    Liu, Xiujie; Shi, Yu; Deng, Yulin; Dai, Rongji

    2017-01-01

    Dregea sinensis Hemsl. plant of the genus Dregea volubilis (Asclepiadaceae), plays a vital role in anticancer, antidepression, and immunoregulation. Steroidal glycosides are the main constituents of this herb, which were significant biological active ingredients. The objective of this study is to recognize the mechanism of anticancer, antidepression, and immunoregulation of D. sinensis Hemsl. Seventy-two steroidal glycosides of D. sinensis Hemsl. were evaluated on the docking behavior of tumor-associated proteins (PI3K, Akt, mTOR), depression-related proteins (MAO-A, MAO-B) and immune-related proteins (tumor necrosis factor-α [TNF-α], tumor necrosis factor receptor 2 [TNFR2], interleukin-2Rα [IL-2Rα]) using Discovery Studio version 3.1 (Accelrys, San Diego, USA). The molecular docking analysis revealed that mostly steroidal glycosides of D. sinensis Hemsl. exhibited powerful interaction with the depression-related protein (MAO-A) and the immune-related proteins (TNFR2, IL-2Rα). Some ligands exhibited high binding energy for the tumor-associated proteins (PI3K, Akt, mTOR) and the immune-related protein (TNF-α), but MAO-B showed none interaction with the ligands. This study has paved better understanding of steroidal glycosides from D. sinensis Hemsl. as potential constituents to the prevention of associated cancer, depression and disorders of immunoregulation. The ligand database was consist of 72 steroidal glycosides from Dregea sinensis HemslSteroidal glycosides had the potential to dock with the tumor-associated proteins (PI3K, Akt, mTOR)Steroidal glycosides were bounded with MAO-A rather than MAO-B, accorded with the inhibitor selectivity of MAOs, can be considered as potent candidate inhibitors of MAO-A72 ligands got high interaction with TNFR2 and IL-2Rα, regard the steroidal glycoside as powerful candidate inhibitors of TNFR2 and IL-2Rα. Abbreviations used: PI3K: Phosphatidyl inositol 3-kinase; Akt: Protein kinase B; mTOR: Mammalian target of

  6. 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

  7. 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

  8. The protein-protein interaction network of eyestalk, Y-organ and hepatopancreas in Chinese mitten crab Eriocheir sinensis

    PubMed Central

    2014-01-01

    Background The protein-protein interaction network (PIN) is an effective information tool for understanding the complex biological processes inside the cell and solving many biological problems such as signaling pathway identification and prediction of protein functions. Eriocheir sinensis is a highly-commercial aquaculture species with an unclear proteome background which hinders the construction and development of PIN for E. sinensis. However, in recent years, the development of next-generation deep-sequencing techniques makes it possible to get high throughput data of E. sinensis tanscriptome and subsequently obtain a systematic overview of the protein-protein interaction system. Results In this work we sequenced the transcriptional RNA of eyestalk, Y-organ and hepatopancreas in E. sinensis and generated a PIN of E. sinensis which included 3,223 proteins and 35,787 interactions. Each protein-protein interaction in the network was scored according to the homology and genetic relationship. The signaling sub-network, representing the signal transduction pathways in E. sinensis, was extracted from the global network, which depicted a global view of the signaling systems in E. sinensis. Seven basic signal transduction pathways were identified in E. sinensis. By investigating the evolution paths of the seven pathways, we found that these pathways got mature in different evolutionary stages. Moreover, the functions of unclassified proteins and unigenes in the PIN of E. sinensis were predicted. Specifically, the functions of 549 unclassified proteins related to 864 unclassified unigenes were assigned, which respectively covered 76% and 73% of all the unclassified proteins and unigenes in the network. Conclusions The PIN generated in this work is the first large-scale PIN of aquatic crustacean, thereby providing a paradigmatic blueprint of the aquatic crustacean interactome. Signaling sub-network extracted from the global PIN depicts the interaction of different

  9. 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

  10. [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.

  11. 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/.

  12. 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.

  13. 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.

  14. 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.

  15. 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.

  16. 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

  17. 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

  18. 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.

  19. [Effects of integrated pest control techniques to growth of host larvae Cordyceps sinensis].

    PubMed

    Liu, Fei; Wu, Xiao-li; Zeng, Wei; Zhang, De-li; Chen, Shi-jiang; Yin, Ding-hua

    2008-12-01

    To study the effects of the integrated pest control techniques on growth of host larvae of Cordyceps sinensis. The integrated pest control techniques were compared with conventional techniques to evaluate the effects on growth of host larvae. The results showed that the techniques had broken the balance of the microbial living in the material, produced effective inhibition on the pests, raised the survival rate and promoted the growth of the host larvae at the same time.

  20. 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

  1. 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.

  2. 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

  3. 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

  4. 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. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. 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

  6. Production of secondary metabolites trimethyl xanthina by Camellia sinensis L suspension culture

    NASA Astrophysics Data System (ADS)

    Sutini, Sodiq, Mochamad; Muslihatin, Wirdhatul; Indra, Mochamad Rasjad

    2017-06-01

    Bioactive trimethyl xanthina can be obtained from the plant Camellia sinensis L. To obtain bioactive plant of which there are several hurdles for instance to wait up to five years to be harvested, also it needs land at a certain height from the sea level. Therefore, the production of secondary metabolites trimethyl xanthina need to be developed with suspense culture techniques. The purpose of this study obtained the production of bioactive trimethyl xanthina way culturally suspense in large scale with a relatively short time, potentially as anti-oxidants. Research methods include: (1) initiation of callus from pieces of leaves, shoots the youngest of the plant Camellia sinensis L in the media MS with the optimization of the addition of growth regulators, (2) the subculture of callus on media and plant growth regulator that is equal to the stage of initiation, (3) initiation of suspension culture using explants of callus Camellia sinensis L, (4) Analysis of secondary metabolites trimethyl xanthina growth in suspension culture, (5) the isolation and identification of trimethyl xanthina qualitatively and quantitatively using thin layer chromatography/high performance chromatography column. The results of the study suspension cultures containing bioactive trimethyl xanthina candidates that can be used as an antioxidant.

  7. 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

  8. 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

  9. 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.

  10. 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

  11. [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.

  12. 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

  13. 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.

  14. Conservation genetics of Annamocarya sinensis (Dode) Leroy, an endangered endemic species.

    PubMed

    Zhang, Z Y; Pang, X M; Han, J W; Wang, Y; Li, Y Y

    2013-03-11

    The endangered perennial plant Annamocarya sinensis (Dode) Leroy is a tertiary relict tree restricted to southeastern China and northern Vietnam. To explore endangerment mechanisms, develop protection strategies, and guide reintroduction efforts for this species, we investigated genetic diversity and population structure by surveying 70 individuals from three distinct populations using 12 polymorphic microsatellite markers. We found high genetic diversity for A. sinensis as indicated by high allelic diversity (allelic number = 4.667 ± 0.436, effective number of alleles = 2.913 ± 0.249), excess heterozygosity (observed heterozygosity = 0.586 ± 0.039, expected heterozygosity = 0.582 ± 0.029), and low fixation index (-0.028 ± 0.057). Our research revealed low genetic differentiation (FST = 0.066 ± 0.011) and no correlation between genetic distance and geographic distance. Analysis of molecular variance attributed 87% of the variance to differences within the population, whereas 13% was distributed among populations. The protection strategy should aim to protect as many populations as possible. Promoting sexual reproduction among various genotypes and establishing an outcrossing program are advisable for A. sinensis.

  15. Determination of hemicellulose, cellulose and lignin content using visible and near infrared spectroscopy in Miscanthus sinensis.

    PubMed

    Jin, Xiaoli; Chen, Xiaoling; Shi, Chunhai; Li, Mei; Guan, Yajing; Yu, Chang Yeon; Yamada, Toshihiko; Sacks, Erik J; Peng, Junhua

    2017-10-01

    Lignocellulosic components including hemicellulose, cellulose and lignin are the three major components of plant cell walls, and their proportions in biomass crops, such as Miscanthus sinensis, greatly impact feed stock conversion to liquid fuels or bio-products. In this study, the feasibility of using visible and near infrared (VIS/NIR) spectroscopy to rapidly quantify hemicellulose, cellulose and lignin in M. sinensis was investigated. Initially, prediction models were established using partial least squares (PLS), least squares support vector machine regression (LSSVR), and radial basis function neural network (RBF_NN) based on whole wavelengths. Subsequently, 23, 25 and 27 characteristic wavelengths for hemicellulose, cellulose and lignin, respectively, were found to show significant contribution to calibration models. Three determination models were eventually built by PLS, LS-SVM and ANN based on the characteristic wavelengths. Calibration models for lignocellulosic components were successfully developed, and can now be applied to assessment of lignocellulose contents in M. sinensis. Copyright © 2017 Elsevier Ltd. All rights reserved.

  16. 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.

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

    PubMed

    Guo, Jian-You; Han, Chun-Chao; Liu, Yong-Mei

    2010-09-01

    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.

  18. Phytochemical analysis and radical scavenging profile of juices of Citrus sinensis, Citrus anrantifolia, and Citrus limonum.

    PubMed

    Rauf, Abdur; Uddin, Ghias; Ali, Jawad

    2014-01-01

    The aim of the current investigation was to identify bioactive secondary metabolites including phenols, tannins, flavonoids, terpinedes, and steroids and compare the phytochemical analysis and antioxidant profile of the juice extracted from the fruits of Citrus sinensis, Citrus anrantifolia, and Citrus limonum. Phytochemical screening is important for the isolation of new, novel, and rare secondary metabolites before bulk extraction. Phytochemical analysis of the desired plant fruits of family Rutaceae revealed the presence of phenols, flavonoids, reducing sugars, steroids, terpinedes and tannins. The fruits of C. sinensis and C. anrantifolia exhibited the presence of phenols, flavonoids, reducing sugars, steroids, terpinedes and tannins, while the fruits of C. limonum indicated the presence of phenols, flavonoids, reducing sugars, terpinedes, and tannins. The fruits of selected plants were also subjected to antioxidant potential by 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay against ascorbic acid at various concentrations. Among the tested plants, C. sinensis showed promising antiradical effect (84.81%) which was followed by C. Anrantifolia (80.05%) at 100 μg/ml against ascorbic acid (96.36%). The C. limonum showed low antioxidant activity among the three selected plants of family Rutaceae. The current finding is baseline information in the use of the fruits of selected plants as food supplement which may be due to the presence of antioxidant molecules in the family Rutaceae. Further research is needed in this area to isolate the phenolic constituents which possess ideal antiradical potential.

  19. Phytochemical analysis and radical scavenging profile of juices of Citrus sinensis, Citrus anrantifolia, and Citrus limonum

    PubMed Central

    2014-01-01

    Background The aim of the current investigation was to identify bioactive secondary metabolites including phenols, tannins, flavonoids, terpinedes, and steroids and compare the phytochemical analysis and antioxidant profile of the juice extracted from the fruits of Citrus sinensis, Citrus anrantifolia, and Citrus limonum. Results Phytochemical screening is important for the isolation of new, novel, and rare secondary metabolites before bulk extraction. Phytochemical analysis of the desired plant fruits of family Rutaceae revealed the presence of phenols, flavonoids, reducing sugars, steroids, terpinedes and tannins. The fruits of C. sinensis and C. anrantifolia exhibited the presence of phenols, flavonoids, reducing sugars, steroids, terpinedes and tannins, while the fruits of C. limonum indicated the presence of phenols, flavonoids, reducing sugars, terpinedes, and tannins. The fruits of selected plants were also subjected to antioxidant potential by 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay against ascorbic acid at various concentrations. Among the tested plants, C. sinensis showed promising antiradical effect (84.81%) which was followed by C. Anrantifolia (80.05%) at 100 μg/ml against ascorbic acid (96.36%). The C. limonum showed low antioxidant activity among the three selected plants of family Rutaceae. Conclusions The current finding is baseline information in the use of the fruits of selected plants as food supplement which may be due to the presence of antioxidant molecules in the family Rutaceae. Further research is needed in this area to isolate the phenolic constituents which possess ideal antiradical potential. PMID:25024932

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

    PubMed

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

    2016-02-01

    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. 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. 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. 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. Agar well diffusion, antimicrobial activity, dental caries, Streptococcus mutans, Lactobacillus acidophilus.

  1. Antimutagenic and antioxidant activity of the essential oils of Citrus sinensis and Citrus latifolia.

    PubMed

    Toscano-Garibay, J D; Arriaga-Alba, M; Sánchez-Navarrete, J; Mendoza-García, M; Flores-Estrada, J J; Moreno-Eutimio, M A; Espinosa-Aguirre, J J; González-Ávila, M; Ruiz-Pérez, N J

    2017-09-13

    The essential oils of Citrus sinensis and Citrus latifolia showed antimycotic activity against Candida spp. isolated from the oral cavity; they are neither mutagenic on the Ames test nor cytotoxic. Their main components are R-(+)-limonene, β-thujene, α-myrcene and γ-terpinene. The aim of this work was to evaluate their antimutagenic and antioxidant capacities. Antimutagenic properties were evaluated against MNNG and ENNG on S. typhimurium TA100; against 2AA on strain TA98 and in front of 4NQO and NOR on strain TA102. Both were antimutagenic against MNNG (p < 0.001) but only C. latifolia was antimutagenic against ENNG (p < 0.001). Both presented antimutagenic activity against 2AA (p < 0.001). They were antioxidant against the ROS-generating compound 4NQO (p < 0.001) and the antibiotic NOR (p < 0.001). In the antioxidant evaluation, the activity in DPPH assay was in a range of 6-23% for C. sinensis and of 22-71% for C. latifolia. Both were antioxidant compared with BHT in β-carotene bleaching assay and were able to decreased apoptosis in HaCat cells stimulated with H2O2. The levels of intracellular superoxide ion were lower in the presence of both oils. In conclusion, the essential oils of C. sinensis and C. latifolia are antimutagenic against at least three types of mutagens and have antioxidants properties.

  2. 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

  3. Antimicrobial effects of citrus sinensis peel extracts against periodontopathic bacteria: an in vitro study.

    PubMed

    Hussain, Khaja Amjad; Tarakji, Bassel; Kandy, Binu Purushothaman Panar; John, Jacob; Mathews, Jacob; Ramphul, Vandana; Divakar, Darshan Devang

    2015-01-01

    Use of plant extracts and phytochemicals with known antimicrobial properties may have great significance in therapeutic treatments. 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 periodontal pathogens. Aqueous and ethanol (cold and hot) extracts prepared from peel of Citrus sinensis were screened for in vitro antimicrobial activity against Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis and Prevotella intermedia, 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. Confidence level and level of significance were set at 95% and 5% respectively. Prevotella intermedia and Porphyromonas gingivalis were resistant to aqueous extracts while Aggregatibacter actinomycetemcomitans was inhibited at very high cncentrations. Hot ethanolic extracts showed significantly higher zone of inhibition than cold ethanolic extract. Minimum inhibitory concentration of hot and cold ethanolic extracts of Citrus sinensis peel ranged between 12-15 mg/ml against all three periodontal pathogens. Both extracts were found sensitive and contain compounds with therapeutic potential. Nevertheless, clinical trials on the effect of these plants are essential before advocating large-scale therapy.

  4. 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.

  5. Gene expression profile indicates involvement of NO in Camellia sinensis pollen tube growth at low temperature.

    PubMed

    Pan, Junting; Wang, Weidong; Li, Dongqin; Shu, Zaifa; Ye, Xiaoli; Chang, Pinpin; Wang, Yuhua

    2016-10-18

    Nitric oxide (NO) functions as a critical signaling molecule in the low-temperature stress responses in plants, including polarized pollen tube growth in Camellia sinensis. Despite this, the potential mechanisms underlying the participation of NO in pollen tube responses to low temperature remain unclear. Here, we investigate alterations to gene expression in C. sinensis pollen tubes exposed to low-temperature stress and NO using RNA-Seq technology, in order to find the potential candidate genes related to the regulation of pollen tube elongation by NO under low-temperature stress. Three libraries were generated from C. sinensis cv. 'Longjingchangye' pollen tubes cultured at 25 °C (CsPT-CK) and 4 °C (CsPT-LT) or with 25 μM DEA NONOate (CsPT-NO). The number of unigenes found for the three biological replications were 39,726, 40,440 and 41,626 for CsPT-CK; 36,993, 39,070 and 39,439 for CsPT-LT; and 39,514, 38,298 and 39,061 for CsPT-NO. A total of 36,097 unique assembled and annotated sequences from C. sinensis pollen tube reads were found in a BLAST search of the following databases: NCBI non-redundant nucleotide, Swiss-prot protein, Kyoto Encyclopedia of Genes and Genomes, Cluster of Orthologous Groups of proteins, and Gene Ontology. The absolute values of log2Ratio > 1 and probability > 0.7 were used as the thresholds for significantly differential gene expression, and 766, 497 and 929 differentially expressed genes (DEGs) were found from the comparison analyses of the CK-VS-LT, CK-VS-NO and LT-VS-NO libraries, respectively. Genes related to metabolism and signaling pathways of plant hormones, transcription factors (TFs), vesicle polarized trafficking, cell wall biosynthesis, the ubiquitination machinery of the ubiquitin system and species-specific secondary metabolite pathways were mainly observed in the CK-VS-LT and CK-VS-NO libraries. Differentially expressed unigenes related to the inhibition of C. sinensis pollen tube growth under low

  6. [Effects of NaCl stress on Hovenia dulcis and Gleditsia sinensis seedlings growth, chlorophyll fluorescence, and active oxygen metabolism].

    PubMed

    Feng, Lei; Bai, Zhi-ying; Lu, Bing-she; Cai, Sheng-wen; Feng, Li-na

    2008-11-01

    With potted Hovenia dulcis and Gleditsia sinensis seedlings as test materials, their plant growth, chlorophyll fluorescence characteristics, and active oxygen metabolism under stress of different concentration (0, 0.15%, 0.30%, 0.45%, and 0.60%) NaCl were studied. The results showed that with increasing concentration of NaCl, the plant growth, leaf chlorophyll content, photochemical efficiency of PS II (Fv/Fm), quantum yield of PS II (phi(PS II)), and photochemical quenching (q(P)) decreased gradually, while the non-photochemical quenching of fluorescence (q(N)) was in adverse. After 10 days of 0. 15% NaCl stress, the leaf chlorophyll content, Fv/Fm, phi(PS II), and q(P) of H. dulcis seedlings decreased by 19.77%, 2.94%, 29.03%, and 8.16%, respectively, with significant differences (P<0.05) to the control, while no significant differences were observed for G. sinensis seedlings. Compared with the control, the Fv/Fm and phi(PS II), of G. sinensis seedlings in treatment 0.30% NaCl decreased significantly by 1.91% and 14.66%, and the chlorophyll content and q(P) of the seedling in treatment 0.45% NaCl decreased significantly by 29.28% and 11.36%, respectively (P<0.05). With increasing concentration of NaCl, the SOD activity of G. sinensis seedlings showed a consistent increasing trend, and that of H. dulcis seedlings increased first and decreased then. The POD and CAT activities of G. sinensis and H. dulci seedlings tended to increase first and decrease then, with the increment being higher for G. sinensis than for H. dulci, while the MDA content of the seedlings had an increasing trend, with the increment being higher for H. dulcis than for G. sinensis, suggesting that the cell membrane lipid peroxidation of H. dulcis was more serious than that of G. sinensis. It was concluded that G. sinensis had greater salt tolerance than H. dulcis, which was related toits higher anti-oxidation enzyme activities.

  7. 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

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

    USDA-ARS?s Scientific Manuscript database

    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...

  9. Immunoglobulin and cytokine production from mesenteric lymph node lymphocytes is regulated by extracts of Cordyceps sinensis in C57Bl/6N mice.

    PubMed

    Park, Dong Ki; Choi, Wahn Soo; Park, Pyo-Jam; Kim, Eun Kyung; Jeong, Yong Jun; Choi, Se Young; Yamada, Koji; Kim, Jong Dai; Lim, Beong Ou

    2008-12-01

    Cordyceps sinensis, one of the well-known fungi used in traditional Chinese medicine, is recognized to play a role in the metabolic process of inflammation and immunity. The purpose of this study was to evaluate the effects of water extracts of C. sinensis on the immune function of mesenteric lymph node (MLN) lymphocytes in C57Bl/6N mice. C. sinensis-treated mice were administered the respective extract by oral gavage for 4 weeks. Immunoglobulin E concentrations in serum and MLN lymphocytes were significantly lower in C. sinensis-treated mice than in control mice. In contrast, the immunoglobulin A concentration from the C. sinensis group was higher than that in control mice. C. sinensis increased the proportion of CD4(+) and CD8(+) T cells in MLN lymphocytes. C. sinensis significantly decreased interleukin-4 and interleukin-10 cytokine concentrations. Therefore, water extracts of C. sinensis modulate immune parameters through regulation of immunoglobulin production resulting from decreased T-lymphocyte helper 2 cytokine secretion and reduce cytokine secretion in MLN lymphocytes.

  10. 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

  11. [Correlation Analysis Between Common Peaks of Angelica sinensis HPLC Fingerprint and Mineral Elements in Its Growing Soil].

    PubMed

    Ding, Jun-xia; Gu, Zhi-rong; Wang, Ya-li; Zhang, Ya-ya; Wang, Yao-peng; Sun, Yu-jing

    2015-08-01

    To investigate the correlation between common peaks of Angelica sinensis HPLC fingerprint and mineral elements in its growing soil. The fingerprints of 120 batches of Angelica sinensis from 12 habitats were determined by HPLC. The contents of Pb, As, Cr, Sb, Hg, Cu, Cd, Ni, Zn, Mg, Mn, Ca, Fe, Na and K in corresponding soil were determined by ICP-MS and AAS. Bivariate and multiple linear regression were used to analyze the correlation. There were significant ( P < 0. 01 or P < 0.05) positive and negative correlation between many common peaks in HPLC fingerprint of Angelica sinensis and mineral elements in its growing soil. The contribution of mineral elements in soil on peak 1 were Zn > K > Sb > Fe > Na; on peak 6 (3-butylphthalide) were Mn > Mg > Ca; on peak 7 were Cr > Zn; on peak 8 were Mn > Na; on peak 11 were As > K > Fe > Cd; on peak 12 were Zn > Mn > K; on peak 13 (Z-butylidenephthalide) were Mn > Zn > Cd; on peak 15 were Zn > K; on peak 16 were Fe > Ni; on peak 17 were Zn > Mn > Ni > Fe > K; on peak 18 were Zn > Na; peaks 2,3 (ferulic acid), 4 and 14 (Z-ligustilide) was mainly affected by As, Zn, Sb and Cu, respectively. The relationship between HPLC fingerprint peak of Angelica sinensis and mineral elements in its growing soil shows complexity, multiplicity and interactivity, which should be selectively examined during manuring micronutrient fertilizer and Angelica sinensis cultivation.

  12. 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.

  13. Mycelia extracts of fungal strains isolated from Cordyceps sinensis differently enhance the function of RAW 264.7 macrophages.

    PubMed

    Meng, Lan-Zhen; Lin, Bao-Qin; Wang, Bo; Feng, Kun; Hu, De-Jun; Wang, Lan-Ying; Cheong, Kit-Leong; Zhao, Jing; Li, Shao-Ping

    2013-07-30

    Cordyceps sinensis, an entomogenous fungus used in traditional Chinese medicine with multiple pharmacological activities. However, its usage has been limited due to the high price and short supply. Isolate of fungi strains from natural Cordyceps sinensis to achieve a large-scale production by fermentation is an alternative choice. The aim of this study was to investigate and compare the effects of mycelia extracts of different fungal stains isolated from natural Cordyceps sinensis on macrophage functions in vitro. Macrophages' proliferation, phagocytosis, nitric oxide (NO) production, cytokines secretion, iNOS, NF-κB p65 activation and translocation were investigated by the MTT assay, flow cytometry assay, Griess reagent method, ELISA, western blot and immunostaining assay, respectively. The results showed that the effects of cultured Cordyceps mycelia of different fungal strains isolated from natural Cordyceps sinensis on macrophages greatly variant. Among 17 Cordyceps aqueous extracts, only five extracts (UM01, QH11, BNQM, GNCC and DCXC) could significantly increase cell proliferation and NO production of RAW 264.7 mouse macrophages. Moreover, the five extracts, especially UM01 and QH11, significantly enhanced phagocytosis and promoted cytokines release of macrophages. Polysaccharides in cultured UM01 mycelia were found to be the main immune stimulating compounds. The variation of biological effects of fermented mycelia of different fungal strains from natural Cordyceps sinensis may be derived from their chemical diversity, especially polysaccharides, which need further study in future. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

  14. Characterization of the complete mitochondrial genome of Ortleppascaris sinensis (Nematoda: Heterocheilidae) and comparative mitogenomic analysis of eighteen Ascaridida nematodes.

    PubMed

    Zhao, J H; Tu, G J; Wu, X B; Li, C P

    2017-06-22

    Ortleppascaris sinensis (Nematoda: Ascaridida) is a dominant intestinal nematode of the captive Chinese alligator. However, the epidemiology, molecular ecology and population genetics of this parasite remain largely unexplored. In this study, the complete mitochondrial (mt) genome sequence of O. sinensis was first determined using a polymerase chain reaction (PCR)-based primer-walking strategy, and this is also the first sequencing of the complete mitochondrial genome of a member of the genus Ortleppascaris. The circular mitochondrial genome (13,828 bp) of O. sinensis contained 12 protein-coding, 22 transfer RNA and 2 ribosomal RNA genes, but lacked the ATP synthetase subunit 8 gene. Finally, phylogenetic analysis of mtDNAs indicated that the genus Ortleppascaris should be attributed to the family Heterocheilidae. It is necessary to sequence more mtNDAs of Ortleppascaris nematodes in the future to test and confirm our conclusion. The complete mitochondrial genome sequence of O. sinensis reported here should contribute to molecular diagnosis, epidemiological investigations and ecological studies of O. sinensis and other related Ascaridida nematodes.

  15. 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.

  16. Fungus-larva relation in the formation of Cordyceps sinensis as revealed by stable carbon isotope analysis.

    PubMed

    Guo, Lian-Xian; Hong, Yue-Hui; Zhou, Qian-Zhi; Zhu, Qing; Xu, Xiao-Ming; Wang, Jiang-Hai

    2017-08-10

    For more than one thousand years, Cordyceps sinensis has been revered as a unique halidom in the Qinghai-Tibetan Plateau for its mysterious life history and predominant medicinal values. This mysterious fungus-larva symbiote also attracted the over-exploitation, while several problems on the initial colonization of Ophiocordyceps sinensis in the host larva have constrained artificial cultivation. In this work, stable carbon isotope analysis was employed to analyse the subsamples of C. sinensis from 5 representative habitats. The results demonstrated that these samples possessed similar δ(13)C profiles, i.e., a steady ascending trend from the top to the bottom of stroma, occurrence of the δ(13)C maximum at the head, a slight decrease from the head to the end of thorax, a sharply descent trend from the end of thorax to the forepart of abdomen, and maintenance of lower δ(13)C values in the rest parts of abdomen. Based on the data, we consider that the site near the head of the host larva may be the initial target attacked by O. sinensis, and the fungus growth is closely related to the digestive tract of its host larva. The growth stages of O. sinensis are accordingly speculated as the symptom-free, symptom-appearing, and stroma-germinating stages.

  17. Cardioprotective effect of the Hibiscus rosa sinensis flowers in an oxidative stress model of myocardial ischemic reperfusion injury in rat

    PubMed Central

    Gauthaman, Karunakaran K; Saleem, Mohamed TS; Thanislas, Peter T; Prabhu, Vinoth V; Krishnamoorthy, Karthikeyan K; Devaraj, Niranjali S; Somasundaram, Jayaprakash S

    2006-01-01

    Background The present study investigates the cardioprotective effects of Hibiscus rosa sinensis in myocardial ischemic reperfusion injury, particularly in terms of its antioxidant effects. Methods The medicinal values of the flowers of Hibiscus rosa sinensis (Chinese rose) have been mentioned in ancient literature as useful in disorders of the heart. Dried pulverized flower of Hibiscus rosa sinensis was administered orally to Wistar albino rats (150–200 gms) in three different doses [125, 250 and 500 mg/kg in 2% carboxy methyl cellulose (CMC)], 6 days per week for 4 weeks. Thereafter, rats were sacrificed; either for the determination of baseline changes in cardiac endogenous antioxidants [superoxide dismutase, reduced glutathione and catalase] or the hearts were subjected to isoproterenol induced myocardial necrosis. Results There was significant increase in the baseline contents of thiobarbituric acid reactive substances (TBARS) [a measure of lipid per oxidation] with both doses of Hibiscus Rosa sinensis. In the 250 mg/kg treated group, there was significant increase in superoxide dismutase, reduced glutathione, and catalase levels but not in the 125 and 500 mg/kg treated groups. Significant rise in myocardial thiobarbituric acid reactive substances and loss of superoxide dismutase, catalase and reduced glutathione (suggestive of increased oxidative stress) occurred in the vehicle treated hearts subjected to in vivo myocardial ischemic reperfusion injury. Conclusion It may be concluded that flower of Hibiscus rosa sinensis (250 mg/kg) augments endogenous antioxidant compounds of rat heart and also prevents the myocardium from isoproterenol induced myocardial injury. PMID:16987414

  18. 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

  19. 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.

  20. Comparison of two DNA extraction protocols from leave samples of Cotinus coggygria, Citrus sinensis and Genus juglans.

    PubMed

    Fallah, F; Minaei Chenar, H; Amiri, H; Omodipour, S; Shirbande Ghods, F; Kahrizi, D; Sohrabi, M; Ghorbani, T; Kazemi, E

    2017-02-28

    High quality DNA is essential for molecular research. Secondary metabolites can affect the quantity and quality DNA. In current research two DNA isolation methods including CTAB and Delaporta (protocols 1 & 2 respectively) were applied in three leave samples from Cotinus coggygria, Citrus sinensis and Genus juglans that their leaves are rich of secondary metabolites. We successfully isolated DNA from C. coggygria, C. sinensis and Genus Juglans using the two protocols described above. Good quality DNA was isolated from C. coggygria, C. sinensis and Genus Juglans using protocol 1, while protocol 2 failed to produce usable DNA from these sources. The highest amount of DNA (1.3-1.6) was obtained from them using protocol 1. As we discovered, procedure 1 may work better for plants with secondary metabolites.

  1. Application of microscopy in authentication of valuable Chinese medicine I--Cordyceps sinensis, its counterfeits, and related products.

    PubMed

    Au, Dawn; Wang, Lijing; Yang, Dajian; Mok, Daniel K W; Chan, Albert S C; Xu, Hongxi

    2012-01-01

    Light and polarized microscope was applied to authenticate 32 Cordyceps and 6 artificial counterfeits and 8 fermented Cordyceps as well as 7 Cordyceps capsules available in Hong Kong markets. Results showed that transverse sections of stroma and powder of larvae can be used to differentiate C. sinensis from its counterfeits. The fermented Cordyceps are in powder form. Among the eight fermented Cordyceps collected, half of them were pure; three were a mixture of fermented Cordyceps and soya beans; one was a mixture of unknown plant tissues and soya beans. For the seven Cordyceps capsules, the powders of five samples were a mixture of fermented Cordyceps and soya beans; the powders of other two were a mixture of C. sinensis stroma powder and fermented Cordyceps. The study indicated that the microscopy is an unambiguous method that requests fewer sample for the authentication of valuable Chinese medicine-C. sinensis and its related products. Copyright © 2011 Wiley Periodicals, Inc.

  2. PCR-RFLP identification of four Chinese soft-shelled turtle Pelodiscus sinensis strains using mitochondrial genes.

    PubMed

    Zhang, Chao; Xu, Xiao-Jun; Zhang, Hai-Qi; Mu, Chang-Kao; He, Zhong-Yang; Wang, Chun-Lin

    2015-08-01

    A PCR-RFLP method to confirm the identification of four Pelodiscus sinensis strains (Taihu Lake strain, Taiwan strain, Yellow River strain and Japanese strain) was developed and evaluated. In this study, we sequenced and analyzed the partial sequences of mitochondrial NADH4, COX I and NADH5-NADH6 genes of 60 individuals from the four P. sinensis strains. Bgl I, Hpa II and Cla I were selected for cutting NADH4, COX I and NADH5-NADH6 PCR products, respectively, and each strain has its unique restriction band patterns. The result showed that all the 140 samples tested can be correctly identified based on the combination of the three digested fragments pattern. This study provides an effective method to distinguish the four main strains of P. sinensis.

  3. Supplementation with different teas from Camellia sinensis prevents memory deficits and hippocampus oxidative stress in ischemia-reperfusion.

    PubMed

    Martins, Alexandre; Schimidt, Helen L; Garcia, Alexandre; Colletta Altermann, Caroline Dalla; Santos, Francielli W; Carpes, Felipe P; da Silva, Weber Cláudio; Mello-Carpes, Pâmela B

    2017-09-01

    Memory and cognition impairments resultant of ischemic stroke could be minimized or avoided by antioxidant supplementation. In this regard, the neuroprotective potential of Green tea from Camellia sinensis has been investigated. However, there is a lack of information regarding the neuroprotective potential of others teas processed from the Camellia sinensis. Here we investigate the neuroprotective role of green, red, white and black tea on memory deficits and brain oxidative stress in a model of ischemic stroke in rats. Our findings show that green and red teas prevent deficits in object and social recognition memories, but only green tea protects against deficits in spatial memory and avoids hippocampal oxidative status and intense necrosis and others alterations in the brain tissue. In summary, green tea shows better neuroprotection in ischemic stroke than the others teas from Camellia sinensis. Copyright © 2017 Elsevier Ltd. All rights reserved.

  4. Identification and characterization of larval and adult anopheline mosquito habitats in the Republic of Korea: potential use of remotely sensed data to estimate mosquito distributions.

    PubMed

    Sithiprasasna, Ratana; Lee, Won Ja; Ugsang, Donald M; Linthicum, Kenneth J

    2005-07-13

    Plasmodium vivax malaria reemerged in the Republic of Korea in 1993, with more than 2,000 cases reported in the northwestern part of the country over the last 10 years. To better assess the risk of malaria transmission we conducted a surveillance study to identify and characterize the habitats that produce potential Anopheles vector mosquitoes. Immature and adult mosquito collection data were incorporated into a Geographic Information System (GIS) along with remotely sensed satellite imagery, and imagery classified to land use to determine if remote sensing data could be used to estimate mosquito habitats. More than 2,100 anopheline larvae were collected and mapped from 186 immature habitats, which were categorized into 9 types. Anopheles sinensis was the most commonly collected species, representing more than 97% of the specimens, followed by Anopheles pullus 1.0%, Anopheles lesteri 1.0%, and Anopheles sineroides 0.8%. Anopheles sinensis, pullus, and lesteri were found most frequently in rice paddies followed by: ditches, flooded areas, ground pools, wheel tracks, swamps, irrigation canals, and stream margins. Anopheles sineroides was found most commonly in flooded areas. Supervised classification was conducted on a LANDSAT 7 ETM+ image to identify 5-6 land-use classes that were considered to be of interest. The spatial distribution of the larval mosquito collections was overlaid on the land-use image derived from the LANDSAT image, and the number of mosquitoes habitats in each class calculated. On Ganghwa Island 67% of the immature habitats containing Anopheles sinensis were in the Paddy land-use class, although the class only represented 17% of the land area. In Paju District 53% of the immature habitats containing Anopheles sinensis were in the Paddy land-use class which represented only 9 % of the area of the district. There was significant (p < 0.05) correlation between habitats containing all four species and land use on Ganghwa Island; however, only

  5. Identification and characterization of larval and adult anopheline mosquito habitats in the Republic of Korea: potential use of remotely sensed data to estimate mosquito distributions

    PubMed Central

    Sithiprasasna, Ratana; Ja Lee, Won; Ugsang, Donald M; Linthicum, Kenneth J

    2005-01-01

    Background Plasmodium vivax malaria reemerged in the Republic of Korea in 1993, with more than 2,000 cases reported in the northwestern part of the country over the last 10 years. To better assess the risk of malaria transmission we conducted a surveillance study to identify and characterize the habitats that produce potential Anopheles vector mosquitoes. Immature and adult mosquito collection data were incorporated into a Geographic Information System (GIS) along with remotely sensed satellite imagery, and imagery classified to land use to determine if remote sensing data could be used to estimate mosquito habitats. Results More than 2,100 anopheline larvae were collected and mapped from 186 immature habitats, which were categorized into 9 types. Anopheles sinensis was the most commonly collected species, representing more than 97% of the specimens, followed by Anopheles pullus 1.0%, Anopheles lesteri 1.0%, and Anopheles sineroides 0.8%. Anopheles sinensis, pullus, and lesteri were found most frequently in rice paddies followed by: ditches, flooded areas, ground pools, wheel tracks, swamps, irrigation canals, and stream margins. Anopheles sineroides was found most commonly in flooded areas. Supervised classification was conducted on a LANDSAT 7 ETM+ image to identify 5–6 land-use classes that were considered to be of interest. The spatial distribution of the larval mosquito collections was overlaid on the land-use image derived from the LANDSAT image, and the number of mosquitoes habitats in each class calculated. On Ganghwa Island 67% of the immature habitats containing Anopheles sinensis were in the Paddy land-use class, although the class only represented 17% of the land area. In Paju District 53% of the immature habitats containing Anopheles sinensis were in the Paddy land-use class which represented only 9 % of the area of the district. There was significant (p < 0.05) correlation between habitats containing all four species and land use on Ganghwa Island

  6. 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

  7. Genetic Diversity and Distribution Patterns of Host Insects of Caterpillar Fungus Ophiocordyceps sinensis in the Qinghai-Tibet Plateau

    PubMed Central

    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

  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. Larvicidal, oviposition, and ovicidal effects of Artemisia annua (Asterales: Asteraceae) against Aedes aegypti, Anopheles sinensis, and Culex quinquefasciatus (Diptera: Culicidae).

    PubMed

    Cheah, Shao-Xiong; Tay, Jia-Wei; Chan, Lai-Keng; Jaal, Zairi

    2013-09-01

    This study focuses on the larvicidal, oviposition, and ovicidal effects of a crude extract of Artemisia annua against Aedes aegypti, Anopheles sinensis, and Culex quinquefasciatus. Dried cells of Artemisia annua from cell suspension cultures were extracted using hexane. The extract showed moderate larvicidal effects against mosquitoes. At 24-h post treatment, the LC50 values for Anopheles sinensis, Aedes aegypti, and Culex quinquefasciatus were recorded as 244.55, 276.14, and 374.99 ppm, respectively. The percentage mortality of larvae was directly proportional to the tested concentration. Anopheles sinensis was found to be the most susceptible species, whereas Culex quinquefasciatus was the most tolerant to the Artemisia annua extract. The results indicated that the Artemisia annua extract showed concentration-dependent oviposition deterrent activity and had a strong deterrent effect. At 500 ppm, the percentage effective repellency was more than 85% compared with the control group for all the species, with oviposition activity index values of -0.94, -0.95, and -0.78 for Aedes aegypti, Anopheles sinensis, and Culex quinquefasciatus, respectively. In the ovicidal assay, the percentage hatchability of eggs after treatment with 500 ppm of Artemisia annua extract was significantly lower than the control, with values of 48.84 ± 4.08, 38.42 ± 3.67, and 79.35 ± 2.09% for Aedes aegypti, Anopheles sinensis, and Culex quinquefasciatus, respectively. Artemisia annua was found to be more effective against Aedes aegypti and Anopheles sinensis compared with Culex quinquefasciatus. This study indicated that crude extract of A. annua could be a potential alternative for use in vector management programs.

  10. Anti-hyperglycemic activity of natural and fermented Cordyceps sinensis in rats with diabetes induced by nicotinamide and streptozotocin.

    PubMed

    Lo, Hui-Chen; Hsu, Tai-Hao; Tu, Shih-Te; Lin, Kwo-Chuan

    2006-01-01

    Our previous study demonstrated that the fruiting bodies of Cordyceps sinensis, a traditional Chinese medicine, attenuated diabetes-induced weight loss, polydipsia, and hyperglycemia in rats. In the present study, we further compared the anti-hyperglycemic activity of the fermented mycelia and broth of Cordyceps sinensis with that of the fruiting bodies. Male Wistar rats orally administered a placebo (STZ group), fruiting bodies (FB group, 1 g/day), fermented mycelia (MCS group, 1 g/day), fermented broth (BCS group, 1 g/day), or fermented mycelia plus broth (XCS group, 0.5 g/day of each) of Cordyceps sinensis (d1 to d28) were injected with nicotinamide (200 mg/kg) and streptozotocin (65 mg/kg) on d15. Rats fed with a placebo and injected with saline served as the control (CON) group. The amount of water and food consumption (d15 to d29), the 2-hour-postprandial blood glucose concentrations (d21 and d28), and the serum concentrations of fructosamine (d29) were significantly lower in the FB, MCS, BCS, and XCS groups than in the STZ group (one-way ANOVA, p < 0.05). The diabetic rats had significantly higher blood glucose concentrations as measured by the oral glucose tolerance test than the control rats; moreover, these changes were significantly reduced by ingesting the fruiting bodies, fermented mycelia and/or broth of Cordyceps sinensis. Our results revealed that the fermented mycelia and broth of Cordyceps sinensis have anti-hyperglycemic activities similar to those of the fruiting bodies. Therefore, the fermented products of Cordyceps sinensis could be developed as potential anti-diabetic agents or functional foods for persons with a high risk of diabetes mellitus.

  11. 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.

  12. [Therapeutic effects of ethanol extracts of Angelica sinensis and Astragalus mongholicus in diabetic gastroparesis rats].

    PubMed

    Jing, Qi; Wu, Guo-Tai; Du, Li-Dong; Liu, Wu-Zhou; Li, Ying-Dong; Ren, Yuan

    2014-08-01

    To observe therapeutic effects of the ethanol extract of Angelica sinensis and Astragalus mongholicus (1:2) in diabetic gastroparesis (DG) rats. Diabetic rats were induced by intraperitoneal injection of alloxan (200 mg/kg). DG was based on gastrointestinal motility index and the character of stool in diabetic rats. The metformin and cisapride mixed solution (containing met-formin 175 mg/kg and cisapride 3.5 mg/kg)was intragastric administrated in the positive control group, the ethanol extract of Angelica sinensis and Astragalus mongholicus was intragastric treated in the high dose (10. 5 g/kg) and low dose(5. 2 g/kg) group rats, 1/d for 42 consecutive days. The body weight, fasting blood glucose (FBG) value and 24 h intake and drinking amount were detected at interval of 14 days. The gastrointestinal propulsion index, motilin (MOT), glucagon (GLG) and gastrin (GAS) contents in the blood were detected after the last administration. And then the gastric antrum smooth muscle cells and interstitial cells of Cajal were observed in microscope. The general state and stool had been improved,and the body weight and 24 h intake were significantly increased, and 24 h drinking in the high and low dose groups at 28 d of administration were reduced. At the 42th day,the body weight and 24 h in- take were increased, 24 h drinking water and FBG were reduced, and the MOT, GAS and GLG contents were decreased in the high dose group. Gastric mucosa and gastric smooth muscle tissue morphology were significantly improved. The therapeutic effect of ethanol extracts of Angelica sinensis and Astragalus mongholicus was obvious in DG rats. Its mechanism of action was associated with FBG, MOT and GAS levels decreasing.

  13. 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

  14. 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.

  15. 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.

  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. 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

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

    PubMed

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

    2014-05-01

    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. To compare the effect of Citrus sinensis and carbimazole (CARB) on blood levels of thyroid hormones (T4 and T3) and TSH. 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. 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. 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.

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

    PubMed

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

    2015-10-01

    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. 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). 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. 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 conservation policies and management strategies for

  20. Danggui to Angelica sinensis root: are potential benefits to European women lost in translation? A review.

    PubMed

    Hook, Ingrid L I

    2014-02-27

    Danggui (Chinese Angelica root; Dong quai; Angelica sinensis (Oliv.) Diels.) is a traditional Chinese herbal remedy with a long history of use in China, Korea and Japan. Even today it is still one of the herbs most commonly used by Traditional Chinese Medicine (TCM) practitioners in China, as well as Europe. It is mainly used for the treatment of women's reproductive problems, such as dysmenorrhea, amenorrhoea, menopause, among others. Using Angelica sinensis (Oliv.) Diels. root as the example, this Review examines the ease with which the use of a Traditional Chinese Herbal Remedy can be transposed from one culture to another. By examining the more recent literature, a number of aspects are considered by the author to be potentially lost in translation: (i) identity and quality (phytochemistry); (ii) tradition of use and processing (smoke-drying, stir-frying, with and without wine); (iii) method of use and traditional types of Chinese herbal medicines; (iv) ethnic differences (Caucasian vs. Asian); (v) efficacy, safety and potential for western drug-herb interactions. This review is based on evaluation of the literature available in scientific journals, textbooks, electronic sources such as ScienceDirect, PubMed, Scopus, etc., as well as other web-sites. A vast amount of information concerning the use of Angelica sinensis exists in the public domain. Many aspects associated with the use of the root are deemed problematical, such as identity, processing, amount and types of constituents, tradition of use in combination with other Chinese herbs, ethnicity of users, etc. Numerous constituents have been isolated with phthalides, ferulic acid and polysaccharides showing biological activities. In spite of the potential activities associated with the traditional use of danggui, and the many trials using the Chinese system of 'Zheng differentiation', well-designed western-style clinical trials carried out using the authenticated, chemically standardized crude drug material

  1. 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.

  2. 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.

  3. 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

  4. 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.

  5. Complete mitochondrial genomes of three mitten crabs, Eriocheir sinensis, E. hepuensis, and E. japonica.

    PubMed

    Wang, Jun; Huang, Lei; Cheng, Qixuan; Lu, Guoqing; Wang, Chenghui

    2016-01-01

    Taxonomic classification of three mitten crabs (Eriocheir sinensis, E. hepuensis, and E. japonica) has long been controversial. In this study, the complete mitogenomes of the three crabs were reported. The three mitogenomes were conserved in the organization of genes with 13 protein-coding genes, 2 rRNA genes, 22 tRNA genes, and 1 control region. Nucleotide variations among the crabs were identified in both coding and non-coding regions. In addition, variable numbers of tandem repeats in control region were identified in the mitten crabs. The mitogenome sequences provide a valuable resource to elucidate taxonomic relationship and evolutionary history of the three mitten crabs.

  6. Cytotoxicity and anti-Leishmania amazonensis activity of Citrus sinensis leaf extracts.

    PubMed

    Garcia, Andreza R; Amaral, Ana Claudia F; Azevedo, Mariana M B; Corte-Real, Suzana; Lopes, Rosana C; Alviano, Celuta S; Pinheiro, Anderson S; Vermelho, Alane B; Rodrigues, Igor A

    2017-12-01

    Leishmania amazonensis is the main agent of diffuse cutaneous leishmaniasis, a disease characterized by lesional polymorphism and the commitment of skin surface. Previous reports demonstrated that the Citrus genus possess antimicrobial activity. This study evaluated the anti-L. amazonensis activity of Citrus sinensis (L.) Osbeck (Rutaceae) extracts. Citrus sinensis dried leaves were subjected to maceration with hexane (CH), ethyl acetate (CEA), dichloromethane/ethanol (CD/Et - 1:1) or ethanol/water (CEt/W - 7:3). Leishmania amazonensis promastigotes were treated with C. sinensis extracts (1-525 μg/mL) for 120 h at 27 °C. Ultrastructure alterations of treated parasites were evaluated by transmission electron microscopy. Cytotoxicity of the extracts was assessed on RAW 264.7 and J774.G8 macrophages after 48-h treatment at 37 °C using the tetrazolium assay. In addition, Leishmania-infected macrophages were treated with CH and CD/Et (10-80 μg/mL). CH, CD/Et and CEA displayed antileishmanial activity with 50% inhibitory activity (IC50) of 25.91 ± 4.87, 54.23 ± 3.78 and 62.74 ± 5.04 μg/mL, respectively. Parasites treated with CD/Et (131.2 μg/mL) presented severe alterations including mitochondrial swelling, lipid body formation and intense cytoplasmic vacuolization. CH and CD/Et demonstrated cytotoxic effects similar to that of amphotericin B in the anti-amastigote assays (SI of 2.16, 1.98 and 1.35, respectively). Triterpene amyrins were the main substances in CH and CD/Et extracts. In addition, 80 μg/mL of CD/Et reduced the number of intracellular amastigotes and the percentage of infected macrophages in 63% and 36%, respectively. The results presented here highlight C. sinensis as a promising source of antileishmanial agents.

  7. 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

  8. Cordyceps sinensis increases hypoxia tolerance by inducing heme oxygenase-1 and metallothionein via Nrf2 activation in human lung epithelial cells.

    PubMed

    Singh, Mrinalini; Tulsawani, Rajkumar; Koganti, Praveen; Chauhan, Amitabh; 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 NF κ B (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, NF κ B 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 NF κ B levels. These findings provide a basis for possible use of Cordyceps sinensis in tolerating hypoxia.

  9. 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.

  10. Phylogenomic and MALDI-TOF MS Analysis of Streptococcus sinensis HKU4T Reveals a Distinct Phylogenetic Clade in the Genus Streptococcus

    PubMed Central

    Tse, Herman; Chen, Jonathan H.K.; Tang, Ying; Lau, Susanna K.P.; Woo, Patrick C.Y.

    2014-01-01

    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. PMID:25331233

  11. The abundance and host-seeking behavior of culicine species (Diptera: Culicidae) and Anopheles sinensis in Yongcheng city, people's Republic of China

    PubMed Central

    2011-01-01

    Background The knowledge of mosquito species diversity and the level of anthropophily exhibited by each species in a region are of great importance to the integrated vector control. Culicine species are the primary vectors of Japanese encephalitis (JE) virus and filariasis in China. Anopheles sinensis plays a major role in the maintenance of Plasmodium vivax malaria transmission in China. The goal of this study was to compare the abundance and host-seeking behavior of culicine species and An. sinensis in Yongcheng city, a representative region of P. vivax malaria. Specifically, we wished to determine the relative attractiveness of different animal baits versus human bait to culicine species and An. sinensis. Results Culex tritaeniorhynchus was the most prevalent mosquito species and An. sinensis was the sole potential vector of P. vivax malaria in Yongcheng city. There were significant differences (P < 0.01) in the abundance of both An. sinensis and Cx. tritaeniorhynchus collected in distinct baited traps. The relative attractiveness of animal versus human bait was similar towards both An. sinensis and Cx. tritaeniorhynchus. The ranking derived from the mean number of mosquitoes per bait indicated that pigs, goats and calves frequently attracted more mosquitoes than the other hosts tested (dogs, humans, and chickens). These trends were similar across all capture nights at three distinct villages. The human blood index (HBI) of female An. sinensis was 2.94% when computed with mixed meals while 3.70% computed with only the single meal. 19:00~21:00 was the primary peak of host-seeking female An. sinensis while 4:00~5:00 was the smaller peak at night. There was significant correlation between the density of female An. sinensis and the average relative humidity (P < 0.05) in Wangshanzhuang village. Conclusions Pigs, goats and calves were more attractive to An. sinensis and Cx. tritaeniorhynchus than dogs, humans, and chickens. Female An. sinensis host-seeking activity

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

    USDA-ARS?s Scientific Manuscript database

    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...

  13. 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 ...

  14. 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

  15. 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

  16. 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.

  17. The ethanolic extract of Juglans sinensis leaves and twigs attenuates CCl4-induced hepatic oxidative stress in rats

    PubMed Central

    Yang, Heejung; Sung, Sang Hyun; Kim, Young Choong

    2015-01-01

    Background: The nuts of Juglans sinensis Dode, walnut tree, are rich in unsaturated fatty acids and bioactive compounds with antioxidant activity on liver damages. However, hepatoprotective activity of the leaves and twigs of J. sinensis have not intensively studied yet. Objective: Hepatoprotective activity of the refined ethanolic extract of J. sinensis (JSE3) was evaluated using carbon tetrachloride (CCl4)-intoxicated rats. Materials and Methods: Hepatotoxicity was induced in Sprague Dawley rats by intraperitoneal injection of CCl4 for 6 weeks in the presence or absence of JSE3 (100 and 200 mg/kg body weight). The hepatoprotective activity of JSE3 was assessed by biochemical parameters including plasma aspartate aminotransferase (AST), alanine aminotransferase (ALT), and antioxidant enzymes, such as superoxide dismutase (SOD), glutathione reductase, glutathione peroxide, reduced glutathione and oxidized glutathione, along with histopathological studies on hepatic tissue. Results: JSE3 significantly decreased the elevated levels of AST and ALT and restored the reduced levels of antioxidant enzymes. JSE3 also decreased the amounts of collagen content accumulated by CCl4 intoxication. Conclusion: These results suggested that the refined extract of J. sinensis may have a potential to be developed as a therapeutic agent to treat hepatic diseases, such as fatty liver and hepatic fibrosis. PMID:26246728

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

    PubMed

    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

    2014-05-01

    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. 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 (1)H-NMR-based metabolomic analysis. 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. Oral administration of Cordyceps sinensis significantly attenuates the liver and heart injuries in CKD rats. The (1)H 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.

  19. Structural characterization of the thermally-tolerant pectin methylesterase purified from Citrus sinensis fruit and its gene sequence

    USDA-ARS?s Scientific Manuscript database

    Despite the longstanding importance for the thermally-tolerant pectin methylesterase (TT-PME) activity in citrus juice processing and product quality, unequivocal identification of the protein and its corresponding gene has remained elusive. We purified TT-PME from sweet orange [Citrus sinensis (L.)...

  20. 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.

  1. 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

  2. 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. Copyright © 2011 Elsevier Masson SAS. All rights reserved.

  3. 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.

  4. 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 ...

  5. 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. Copyright © 2016 Elsevier Ltd. All rights reserved.

  6. Biotransformation of isofraxetin-6-O-β-d-glucopyranoside by Angelica sinensis (Oliv.) Diels callus.

    PubMed

    Zhou, Di; Zhang, Yuhua; Jiang, Zhe; Hou, Yue; Jiao, Kun; Yan, Chunyan; Li, Ning

    2017-01-15

    Isofraxetin-6-O-β-d-glucopyranoside, identified from traditional medicinal herbal Xanthoceras sorbifolia Bunge, has been demonstrated to be a natural neuroinflammatory inhibitor. In order to obtain more derivatives with potential anti-neuroinflammatory effects, biotransformation was carried out. According to the characteristics of coumarin skeleton, suspension cultures of Angelica sinensis (Oliv.) Diels callus (A. sinensis callus) were employed because of the presence of diverse phenylpropanoids biosynthetic enzymes. As a result, 15 products were yielded from the suspension cultures, including a new coumarin: 8'-dehydroxymethyl cleomiscosin A (1), together with 14 known compounds. Their structures were elucidated by extensive spectroscopic analysis. Furthermore, the biotransformed pathways were discussed. Among them, compound 13 was transformed from isofraxetin-6-O-β-d-glucopyranoside, while compounds 1-6, 10-12, 14-15 were derived from the culture medium stimulated by the substrate. The biotransformation processes include hydroxylation, oxidation and esterification. Furthermore, their inhibitory effects on lipopolysaccharide (LPS)-activated nitric oxide (NO) production were evaluated in BV2 microglial cells. It is worth noting that, 1, 1'-methanediylbis(4-methoxybenzene) (3), obtucarbamates A (5), 2-nonyl-4-hydroxyquinoline N-oxide (10) and 1H-indole-3-carbaldehyde (11) exhibited significant inhibitory effect against neuroinflammation with IC50 values at 1.22, 10.57, 1.02 and 0.76μM respectively, much stronger than that of the positive control minocycline (IC50 35.82μM). Copyright © 2016 Elsevier Ltd. All rights reserved.

  7. 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

  8. 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.

  9. 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

  10. Characterization of Citrus sinensis type 1 mitochondrial alternative oxidase and expression analysis in biotic stress.

    PubMed

    Daurelio, Lucas Damián; Checa, Susana Karina; Barrio, Jorgelina Morán; Ottado, Jorgelina; Orellano, Elena Graciela

    2009-10-09

    The higher plant mitochondrial electron transport chain contains an alternative pathway that ends with the AOX (alternative oxidase). The AOX proteins are encoded by a small gene family composed of two discrete gene subfamilies. Aox1 is present in both monocot and eudicot plants, whereas Aox2 is only present in eudicot plants. We isolated a genomic clone from Citrus sinensis containing the Aox1a gene. The orange Aox1a consists of four exons interrupted by three introns and its promoter harbours diverse putative stress-specific regulatory motifs including pathogen response elements. The role of the Aox1a gene was evaluated during the compatible interaction between C. sinensis and Xanthomonas axonopodis pv. citri and no induction of the Aox1a at the transcriptional level was observed. On the other hand, Aox1a was studied in orange plants during non-host interactions with Pseudomonas syringae pv. tomato and Xanthomonas campestris pv. vesicatoria, which result in hypersensitive response. Both phytopathogens produced a strong induction of Aox1a, reaching a maximum at 8 h post-infiltration. Exogenous application of salicylic acid produced a slight increase in the steady-state level of Aox1a, whereas the application of fungi elicitors showed the highest induction. These results suggest that AOX1a plays a role during biotic stress in non-host plant pathogen interaction.

  11. 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.

  12. 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.

  13. Effects of temperature and salinity on Ruppia sinensis seed germination, seedling establishment, and seedling growth.

    PubMed

    Gu, Ruiting; Zhou, Yi; Song, Xiaoyue; Xu, Shaochun; Zhang, Xiaomei; Lin, Haiying; Xu, Shuai; Zhu, Shuyu

    2017-08-16

    As typical submerged aquatic vegetation, Ruppia species are facing population reductions due to anthropogenic impacts. In this study, we investigated the effects of temperature and salinity on seed germination and seedling establishment of Ruppia sinensis seeds collected from northern China. The effects of seven salinities (0-50) and six water temperatures (0-30°C) on seed germination were investigated to identify the environmental conditions that could potentially limit survival and growth. We found that: 1) optimum seed germination was salinity 5 at 30°C; 2) high salinity (salinity 40-50) and low temperature (0°C) significantly inhibited seed germination; 3) seed germination with increasing temperature showed a bimodal pattern at suitable salinities (5-10); 4) storing seeds at high salinities (40-50) or low temperature (0°C) promoted germination after transferal to optimal germination conditions. These findings may serve as useful information for R. sinensis habitat establishment and restoration programs. Copyright © 2017 Elsevier Ltd. All rights reserved.

  14. 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

  15. 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

  16. 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

  17. [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.

  18. Global warming and the regional persistence of a temperate-zone insect (Tenodera sinensis)

    SciTech Connect

    Rooney, T.P.; Smith, A.T.; Hurd, L.E.

    1996-07-01

    Models based on the paleoecological record predict that animals in temperate regions will respond to global warming by migrating poleward to remain within their temperature tolerance ranges. The effect of global warming on invertebrates is of great concern because of their critical role in ecosystem structure and function. Migration poses a problem for many species because of their limited dispersal abilities. The life cycle of a typical temperature zone univoltine insect. Tenodera sinensis (Mantodea: Mantidae) is constrained by degree-days per season: too few prevent maturation before the killing frost in the autumn; too many allow egg hatch before a killing frost. We used field and laboratory observation on the life history and ecology of this species to predict the effect of global warming on the regional distribution of this insect by the end of the next century. Based on the simplified, best-case, biological assumptions of our model, the geographical range of T. sinensis in eastern North America would be compressed toward the northern part of its present contiguous regional distribution. This and other univoltine temperate species with long maturation periods and low vagility could face regional extinction if global warming predictions are accurate. 61 refs., 3 figs.

  19. 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

  20. 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.

  1. 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.

  2. 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.

  3. 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

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

    PubMed

    Prokop, Jakub; Pecharová, Martina; Ren, Dong

    2016-06-20

    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.

  5. 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. Copyright © 2011 Elsevier Ltd. All rights reserved.

  6. Augmentation of various immune reactivities of tumor-bearing hosts with an extract of Cordyceps sinensis.

    PubMed

    Yamaguchi, N; Yoshida, J; Ren, L J; Chen, H; Miyazawa, Y; Fujii, Y; Huang, Y X; Takamura, S; Suzuki, S; Koshimura, S

    1990-01-01

    In order to enhance general reactivity of immune system in the tumor-bearing host, we employed extract of Cordyceps sinensis (CSE) as a biological response modifier. Cordyceps sinensis is an interesting material produced by a kind of mushroom parasitic to larval moths and was used to hasten recovery from exhaustion in ancient China. In this experiment, C57BL/6 mice implanted subcutaneously with syngeneic EL-4 lymphoma cells were employed as the host. Oral administration of the extract leads to a reduction of tumor size and prolongation of the host survival time. As judged by plaque-forming cells against T-dependent (sheep erythrocytes) and T-independent (bacterial lipopolysaccharide) antigens, CSE showed to augment the antibody responses. As for the activities of peritoneal macrophages, chemotaxis was dramatically depressed within a few days after EL-4 transplantation up to the end of life, but treatment with CSE at -14, -7, -4, +4, +7 and +10 days after the tumor transplantation augmented the activity about four times stronger than that of control. Phagocytic activity of macrophages was also decreased in tumor-bearing mice treated with cyclophosphamide (100 mg/kg) 3 and 5 days after tumor transplantation. But administration of CSE restored the activity to more than the normal level. The overall efficacy of CSE was tested with protective activity against systemic infection by Salmonella enteritides. The tumor-bearing mice receiving this medicine lived significantly longer than any other groups without CSE.

  7. The sulfur-fumigation reduces chemical composition and biological properties of Angelicae Sinensis Radix.

    PubMed

    Zhan, Janis Ya-Xian; Yao, Ping; Bi, Cathy Wen-Chuan; Zheng, Ken Yu-Zhong; Zhang, Wendy Li; Chen, Jian-Ping; Dong, Tina Ting-Xia; Su, Zi-Ren; Tsim, Karl Wah-Keung

    2014-09-25

    Angelica Sinensis Radix (roots of Angelica sinensis; ASR) is a popular herbal supplement in China for promoting blood circulation. Today, sulfur-fumigation is commonly used to treat ASR as a means of pest control; however, the studies of sulfur-fumigation on the safety and efficacy of ASR are very limited. Here, we elucidated the destructive roles of sulfur-fumigation on ASR by chemical and biological assessments. After sulfur-fumigation, the chemicals in ASR were significantly lost. The biological activities of anti-platelet aggregation, induction of NO production and estrogenic properties were compared between the water extracts of non-fumigated and sulfur-fumigated ASR. In all cases, the sulfur-fumigation significantly reduced the biological properties of ASR. In addition, application of water extract deriving from sulfur-fumigated ASR showed toxicity to cultured MCF-7 cells. In order to ensure the safety and to achieve the best therapeutic effect, it is recommended that sulfur-fumigation is an unacceptable approach for processing herbal materials. Copyright © 2014 Elsevier GmbH. All rights reserved.

  8. Establishment and characterization of a cell line from the Chinese soft-shelled turtle Pelodiscus sinensis.

    PubMed

    Guo, Haijie; Xia, Zhaonan; Tang, Wei; Mao, Zhijuan; Qian, Guoying; Wang, Caisheng

    2016-06-01

    The establishment and partial characterization of Pelodiscus sinensis continuous cell line is described here. A novel P. sinensis fibroblast cell line, designated PSF, was established from heart tissue by the semi-digestion explant culture technique. Since its initiation in July 2013, the cell line has been subcultured at 30°C in minimal essential medium (MEM) containing 15% (v/v) fetal bovine serum for more than 50 passages. The growth curve of the cell line revealed the population doubling time was 51.1 h. Karyotyping analysis indicated the modal chromosome number was 66, and no microbial contamination was detected. The PSF cell line produced significant fluorescent signals after transfection with plasmid pEGFP-C3. Analysis of mitochondrial cytochrome D-loop sequences revealed 96% identity among other Chinese turtle subspecies. Several cell line characterizations included morphological analysis and immunocytochemistry, which revealed the origin of the PSF cell line was fibroblast-like cells. Measurement of the isoenzymes lactic dehydrogenase and malic dehydrogenase showed no cross-contamination of this cell line with other species. This newly established cell line will be a valuable tool for transgenic and genetic manipulation studies and will act as an efficient instrument for studies of the viral diseases of the soft-shelled turtle.

  9. [Use of Phaseolus vulgaris and Vigna sinensis in a fermented dairy drink].

    PubMed

    Granito, Marisela; Trujillo, Lesma; Guerra, Marisa

    2004-06-01

    The objective of this work was to develop a new kind fermented dairy drink, partially substituted with clear varieties of Phaseolus vulgaris (caraota) and Vigna sinensis (frijol). The formulation of fermented dairy drinks included sterile extracts of caraota and frijol, as partial substitutes which replaced milk: 10, 20 and 30%. The mixtures were inoculated with 2% of a mixture of Lactobacillus acidophillus, Streptococcus thermophilus and Bifidobacterium sp. and were incubated at 42 degrees C for 7 hours. Mango and guava jams were used as flavorings at 20%. On the basis of the sensorial evaluation the mixtures 10% frijol-mango, 10% frijol-guava, 30% caraota-mango and 20% caraota-guava were selected. In the selected fermented dairy drinks, the levels of protein, soluble and insoluble fiber, available and resistant starches were increased and the protein digestibility was 81%. The technical feasibility of partial substitution of milk with extracts of Phaseolus vulgaris or Vigna sinensis. For the elaboration of a fermented dairy drink similar to the liquid yogurt kind was demonstrated.

  10. 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.

  11. 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

  12. 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.

  13. 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.

  14. 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.

  15. 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.

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

    PubMed

    Vinholes, Juliana; Vizzotto, Márcia

    2017-01-01

    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. 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. 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. 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. 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. Alfa-glucosidase and antioxidant Interaction between Camellia sinensis L. Kuntze and Eugenia uniflora L. ethanolic extracts was investigated.Extracts showed synergistic effect over alpha-glucosidase and peroxyl radicals

  18. Potential of Moringa oleifera root and Citrus sinensis fruit rind extracts in the treatment of ulcerative colitis in mice.

    PubMed

    Gholap, Prashant A; Nirmal, Sunil A; Pattan, Shashikant R; Pal, Subodh C; Mandal, Subhash C

    2012-10-01

    The plant Moringa oleifera Lam (Moringaceae), commonly known as the drumstick tree, is an indigenous species in India. This species has been of interest to researchers because traditionally its roots are reported in the treatment of ulcerative colitis (UC). Traditionally it is reported that Citrus sinensis Linn (Rutaceae) fruit rind when combined with M. oleifera will increase the efficacy of the plant in the treatment of UC. The present work was undertaken to determine the effectiveness of M. oleifera root alone and in combination with C. sinensis fruit rind in the treatment of UC. Ethanol and aqueous extracts of M. oleifera roots (100 and 200 mg/kg, body weight) were screened alone and in equal combination with ethanol extract of C. sinensis fruit rind, i.e., 50 mg/kg each of C. sinensis and M. oleifera for their activity on acetic acid-induced UC in mice. Treatment with combination of extracts of M. oleifera root and C. sinensis fruit rind (50 mg/kg, each) showed less ulceration and hyperemia than individual extract (200 mg/kg) in histopathological observation. Acetic acid increased myeloperoxidase (MPO) level in blood and colon tissue to 342 U/mL and 384 U/mg, respectively. Combination of ethanol extract of M. oleifera root with C. sinensis fruit rind extract significantly (p<0.05) decreased MPO in blood and tissue to 278 U/mL and 291 U/mg, respectively. MPO in blood and tissue in control group was 85 ± 1.2 U/mL and 96 ± 1.3 U/mg, respectively. Similarly this combination significantly reduced malondialdehyde (MDA) level in blood and tissue to 7.11 nmol/mL and 8.19 nmol/mg, from 11.20 nmol/mL and 13.20 nmol/mg, respectively. MDA in blood and tissue in control group was 2.76 ± 1.2 nmol/mL and 3.76 ± 1.2 nmol/mg, respectively. Results show that a combination of M. oleifera root extracts with C. sinensis fruit rind extract is effective in the treatment of UC and results are comparable with the standard drug prednisolone.

  19. Vector capacity of Anopheles sinensis in malaria outbreak areas of central China.

    PubMed

    Pan, Jia-Yun; Zhou, Shui-Sen; Zheng, Xiang; Huang, Fang; Wang, Duo-Quan; Shen, Yu-Zu; Su, Yun-Pu; Zhou, Guang-Chao; Liu, Feng; Jiang, Jing-Jing

    2012-07-09

    Both falciparum and vivax malaria were historically prevalent in China with high incidence. With the control efforts, the annual incidence in the whole country has reduced to 0.0001% except in some areas in the southern borders after 2000. Despite this, the re-emergence or outbreak of malaria was unavoidable in central China during 2005-2007. In order to understand the role of the vector in the transmission of malaria during the outbreak period, the vector capacity of An. sinensis in Huanghuai valley of central China was investigated. The study was undertaken in two sites, namely Huaiyuan county of Anhui province and Yongcheng county of Henan province. In each county, malaria cases were recorded for recent years, and transmission risk factors for each study village including anti-mosquito facilities and total number of livestock were recorded by visiting each household in the study sites. The specimens of mosquitoes were collected in two villages, and population density and species in each study site were recorded after the identification of different species, and the blood-fed mosquitoes were tested by ring precipitation test. Finally, various indicators were calculated to estimate vector capacity or dynamics, including mosquito biting rate (MBR), human blood index (HBI), and the parous rates (M). Finally, the vector capacity, as an important indicator of malaria transmission to predict the potential recurrence of malaria, was estimated and compared in each study site.About 93.0% of 80 households in Huaiyuan and 89.3% of 192 households in Yongcheng had anti-mosquito facilities. No cattle or pigs were found, only less than 10 sheep were found in each study village. A total of 94 and 107 Anopheles spp. mosquitos were captured in two study sites, respectively, and all of An. sinensis were morphologically identified. It was found that mosquito blood-feeding peak was between 9:00 pm and 12:00 pm. Man biting rate of An. sinensis was 6.0957 and 5.8621 (mosquitoes

  20. Vector capacity of Anopheles sinensis in malaria outbreak areas of central China

    PubMed Central

    2012-01-01

    Background Both falciparum and vivax malaria were historically prevalent in China with high incidence. With the control efforts, the annual incidence in the whole country has reduced to 0.0001% except in some areas in the southern borders after 2000. Despite this, the re-emergence or outbreak of malaria was unavoidable in central China during 2005–2007. In order to understand the role of the vector in the transmission of malaria during the outbreak period, the vector capacity of An. sinensis in Huanghuai valley of central China was investigated. Findings The study was undertaken in two sites, namely Huaiyuan county of Anhui province and Yongcheng county of Henan province. In each county, malaria cases were recorded for recent years, and transmission risk factors for each study village including anti-mosquito facilities and total number of livestock were recorded by visiting each household in the study sites. The specimens of mosquitoes were collected in two villages, and population density and species in each study site were recorded after the identification of different species, and the blood-fed mosquitoes were tested by ring precipitation test. Finally, various indicators were calculated to estimate vector capacity or dynamics, including mosquito biting rate (MBR), human blood index (HBI), and the parous rates (M). Finally, the vector capacity, as an important indicator of malaria transmission to predict the potential recurrence of malaria, was estimated and compared in each study site. About 93.0% of 80 households in Huaiyuan and 89.3% of 192 households in Yongcheng had anti-mosquito facilities. No cattle or pigs were found, only less than 10 sheep were found in each study village. A total of 94 and 107 Anopheles spp. mosquitos were captured in two study sites, respectively, and all of An. sinensis were morphologically identified. It was found that mosquito blood-feeding peak was between 9:00 pm and 12:00 pm. Man biting rate of An. sinensis was 6.0957 and

  1. 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.

  2. 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

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

    PubMed

    Bhaskar, Anusha; Nithya, V

    2012-01-01

    To investigate the wound-healing potency of the ethanolic extract of the flowers of Hibiscus rosa sinensis. 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. 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. The ethanolic extract of H. rosa sinensis had greater wound-healing activity than the nitrofurazone ointment.

  4. Comparative transcriptome analysis of the accessory sex gland and testis from the Chinese mitten crab (Eriocheir sinensis).

    PubMed

    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.

  5. 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

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

    PubMed

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

    2016-01-01

    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. 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. 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. 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.

  7. 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

  8. Assessment of Angelica sinensis (Oliv.) Diels as a repellent for personal protection against mosquitoes under laboratory and field conditions in northern Thailand.

    PubMed

    Champakaew, Danita; Junkum, Anuluck; Chaithong, Udom; Jitpakdi, Atchariya; Riyong, Doungrat; Wannasan, Anchalee; Intirach, Jitrawadee; Muangmoon, Roongtawan; Chansang, Arpaporn; Tuetun, Benjawan; Pitasawat, Benjawan

    2016-06-29

    revealed strong repellency from both 25 % AHEv and 25 % DEETv, with complete protection (100 %) against a wide range of local mosquito populations. A total of 5,718 adult female mosquitoes, with the most predominant being Culex quinquefasciatus (41.47 %), Armigeres subalbatus (41.13 %), and Culex vishnui (10.53 %), was collected during field applications. No local skin reaction or other allergic responses was observed during both laboratory and field study periods. Angelica sinensis proved to have not only impressive repellency against both laboratory Ae. aegypti and a wide range of natural mosquito populations, but also relative stability in physical and biological performance.

  9. Protective effects of orange (Citrus sinensis L.) peel aqueous extract and hesperidin on oxidative stress and peptic ulcer induced by alcohol in rat.

    PubMed

    Selmi, Slimen; Rtibi, Kais; Grami, Dhekra; Sebai, Hichem; Marzouki, Lamjed

    2017-08-14

    Massive alcohol drinking can lead to gastric ulcer. In the present study we investigated the gastroprotective effect of Citrus sinensis peel aqueous extract (CSPE) and Hesperidin (H) in ethanol (EtOH) induced oxidative stress and peptic ulcer in rats. Seventy adult male Wistar rats were divided into seven groups of 10 each: control, EtOH (4 g/kg b.w.), EtOH + various doses of CSPE (100, 200 and 400 mg/kg, b.w.), EtOH + Hesperidin (50 mg/kg, p.o.) and EtOH + Omeprazole (OM, 20 mg/kg, p.o.). Animals were perorally (p.o.) pre-treated with CSPE during 15 days and intoxicated with a single oral administration of EtOH (4 g/kg b.w.) during 2 h. Gastric ulcer was induced in rats with a single dose of ethanol (EtOH). Ulcer index, gene expression of gastric cyclooxygenase-2 (COX-2), tumor necrosis factor alpha (TNF-α), malondialdhyde (MDA), hydrogen peroxide H2O2 and Thiol groups (-SH) content in stomach and antioxidant enzymes superoxide dismutase (SOD), catalase (CAT) and gluthation peroxidise (GPx) were measured. Furthermore, histopathological examinations were performed. The results showed that ethanol induced gastric damage, improving oxidative stress markers level such as MDA (121 ± 4.45 nmol/mg proteins) and H2O2 (24.62 ± 1.04 μmol/mg proteins), increased pro-inflammatory cytokine (TNF-α level), as well as the expression of COX-2 in the ethanol group. However, a significant depletion of enzymatic and non-enzymatic antioxidants were observed, such as, GPx (72%), SOD (57.5%), CAT (41.6%) and -SH (50%). The lesions were associated with severe histopathological damage. The both Citrus sinensis peel aqueous extract (CSPE) and hesperidin significantly protect against all gastric damages caused by ethanol administration in rats. We propose that CSPE and hesperidin exhibit protective effects in EtOH-induced peptic ulcer in rat. This protection might be related in to part its antioxidant properties as well as its opposite effects on some studied intracellular

  10. 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.

  11. 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

  12. 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. © 2015 Wiley Periodicals, Inc.

  13. 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.

  14. Notes on the Distribution of Anopheles (Anopheles) sinensis Wiedemann (Diptera: Culicidae) in China and the Status of Some Anopheles Hyrcanus Group Type Specimens from China

    DTIC Science & Technology

    2005-01-10

    ANSI Std Z39-18 236 PROCEEDINGS OF THE ENTOMOLOGICAL SOCIETY OF WASHINGTON Table 1. Collections of Anopheles (Anopheles) sinensis in the Institute of... PROCEEDINGS OF THE ENTOMOLOGICAL SOCIETY OF WASHINGTON differences of rDNA-ITS2 and species-diagnostic PCR assay of Anopheles sinensis and Anopheles...Harrison and Scanlon 1975), and Nepal ( Darsie and Prad- han 1990). Several sources were used to document the distribution record of An. si- hensis in

  15. 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

  16. 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

  17. SOME ENGINEERING PROPERTIES OF SHELLED AND KERNEL TEA (Camellia sinensis) SEEDS.

    PubMed

    Altuntas, Ebubekir; Yildiz, Merve

    2017-01-01

    Camellia sinensis is the source of tea leaves and it is an economic crop now grown around the World. Tea seed oil has been used for cooking in China and other Asian countries for more than a thousand years. Tea is the most widely consumed beverages after water in the world. It is mainly produced in Asia, central Africa, and exported throughout the World. Some engineering properties (size dimensions, sphericity, volume, bulk and true densities, friction coefficient, colour characteristics and mechanical behaviour as rupture force of shelled and kernel tea (Camellia sinensis) seeds were determined in this study. This research was carried out for shelled and kernel tea seeds. The shelled tea seeds used in this study were obtained from East-Black Sea Tea Cooperative Institution in Rize city of Turkey. Shelled and kernel tea seeds were characterized as large and small sizes. The average geometric mean diameter and seed mass of the shelled tea seeds were 15.8 mm, 10.7 mm (large size); 1.47 g, 0.49 g (small size); while the average geometric mean diameter and seed mass of the kernel tea seeds were 11.8 mm, 8 mm for large size; 0.97 g, 0.31 g for small size, respectively. The sphericity, surface area and volume values were found to be higher in a larger size than small size for the shelled and kernel tea samples. The shelled tea seed's colour intensity (Chroma) were found between 59.31 and 64.22 for large size, while the kernel tea seed's chroma values were found between 56.04 68.34 for large size, respectively. The rupture force values of kernel tea seeds were higher than shelled tea seeds for the large size along X axis; whereas, the rupture force values of along X axis were higher than Y axis for large size of shelled tea seeds. The static coefficients of friction of shelled and kernel tea seeds for the large and small sizes higher values for rubber than the other friction surfaces. Some engineering properties, such as geometric mean diameter, sphericity, volume, bulk

  18. SOME ENGINEERING PROPERTIES OF SHELLED AND KERNEL TEA (Camellia sinensis) SEEDS

    PubMed Central

    Altuntas, Ebubekir; Yildiz, Merve

    2017-01-01

    Background: Camellia sinensis is the source of tea leaves and it is an economic crop now grown around the World. Tea seed oil has been used for cooking in China and other Asian countries for more than a thousand years. Tea is the most widely consumed beverages after water in the world. It is mainly produced in Asia, central Africa, and exported throughout the World. Some engineering properties (size dimensions, sphericity, volume, bulk and true densities, friction coefficient, colour characteristics and mechanical behaviour as rupture force of shelled and kernel tea (Camellia sinensis) seeds were determined in this study. Materials and Methods: This research was carried out for shelled and kernel tea seeds. The shelled tea seeds used in this study were obtained from East-Black Sea Tea Cooperative Institution in Rize city of Turkey. Shelled and kernel tea seeds were characterized as large and small sizes. Results: The average geometric mean diameter and seed mass of the shelled tea seeds were 15.8 mm, 10.7 mm (large size); 1.47 g, 0.49 g (small size); while the average geometric mean diameter and seed mass of the kernel tea seeds were 11.8 mm, 8 mm for large size; 0.97 g, 0.31 g for small size, respectively. The sphericity, surface area and volume values were found to be higher in a larger size than small size for the shelled and kernel tea samples. The shelled tea seed’s colour intensity (Chroma) were found between 59.31 and 64.22 for large size, while the kernel tea seed’s chroma values were found between 56.04 68.34 for large size, respectively. The rupture force values of kernel tea seeds were higher than shelled tea seeds for the large size along X axis; whereas, the rupture force values of along X axis were higher than Y axis for large size of shelled tea seeds. The static coefficients of friction of shelled and kernel tea seeds for the large and small sizes higher values for rubber than the other friction surfaces. Conclusion: Some engineering properties

  19. Adult Learning, Adult Teaching.

    ERIC Educational Resources Information Center

    Daines, John; And Others

    This book is designed to provide specific and practical suggestions for teachers and tutors of adult students. It is divided into two major parts. Part 1, which is devoted to adult learning, includes information on the characteristics of adult learners, their expectations and motivation, and issues related to equal opportunities and access. Part 2…

  20. 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.

  1. 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.

  2. 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

  3. The complete mitochondrial genome of Chinese green hydra, Hydra sinensis (Hydroida: Hydridae).

    PubMed

    Pan, Hong-Chun; Qian, Xiao-Cheng; Li, Ping; Li, Xiao-Fei; Wang, An-Tai

    2014-02-01

    The complete mitochondrial genome of Chinese green hydra, Hydra sinensis (Hydroida: Hydridae) is a linear molecule of 16,189 bp in length, containing 13 protein-coding genes, small and large subunit ribosomal RNAs, methionine and tryptophan transfer RNAs, a pseudogene consisting of a partial copy of COI and terminal sequences at two ends of the linear mitochondrial DNA. The A + T content of the overall base composition of H-strand is 77.2% (T: 41.7%; C: 10.9%; A: 35.5%; and G: 11.9%). COI and ND1 genes begin with GTG as start codon, while other 11 protein-coding genes start with a typical ATG initiation codon. COII, ATP8, ATP6, COIII, ND5, ND6, ND3, ND1, ND4 and COI genes are terminated with TAA as stop codon, ND4L ends with TAG, ND2 ends with TA and Cyt b ends with T.

  4. 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.

  5. The use of green tea (Camellia sinensis) as a phytogenic substance in poultry diets.

    PubMed

    Khan, Sohail H

    2014-04-15

    This review examined the use of green tea (Camellia sinensis) in the diets of poultry. Research findings were obtained from various recent studies, where much attention was focused on the role of green tea in the promotion of both animal and human health. The review involved some of the currently available information about green tea, pertaining to its chemical composition, anticoccidial and antimicrobial effect, effect on broiler and layer performance and on blood and egg yolk constituents. To the author's knowledge this is the first review paper on this topic. It will be helpful for poultry nutritionists and the poultry industry, although more detailed studies are still needed to elucidate the effects of green tea in poultry nutrition under various circumstances.

  6. 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

  7. 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.

  8. 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.

  9. 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.

  10. 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.

  11. 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.

  12. 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 a