摘要:
Background: Sanguinarine (SA) is a quaternary benzo[c]phenanthridine alkaloid that is mainly present in the Papaveraceae family. SA has been extensively studied because of its antimicrobial, anti-inflammatory, antitumor, antihypertensive, antiproliferative and antiplatelet activities. Metabolic studies demonstrated that SA bioavailability is apparently low, and the main pathway of SA metabolism is iminium bond reduction resulting in dihydrosanguinarine (DHSA) formation. Nevertheless, the metabolic enzymes involved in SA reduction are still not known in detail. Thus, the aim of this study was to investigate the rat liver microsomes and cytosol-induced SA iminium bond reduction, and to examine the effects of cytosol reductase inhibitors on the reductive activity. Methods: DHSA formation was quantified by HPLC. The possible enzymes responsible for DHSA formation were examined using selective individual metabolic enzyme inhibitors. Results: When SA was incubated with liver microsomes and cytosol in the absence of NAD(P)H, DHSA, the iminium bond reductive metabolite was formed. The reductase activity of the liver microsomes and cytosol was also enhanced significantly in the presence of NADH. The amount of DHSA formed in the liver cytosol was 4.6-fold higher than in the liver microsomes in the presence of NADH. The reductase activity in the liver cytosol was inhibited by the addition of flavin mononucleotide and/or riboflavin. Inhibition studies indicated that menadione, dicoumarol, quercetin and 7-hydroxycoumarin inhibited rat liver cytosol-mediated DHSA formation in the absence of NADH. However, only menadione and quercetin inhibited rat liver cytosol-mediated DHSA formation in the presence of NADH. Conclusions: These results suggest that the SA iminium bond reduction proceeds via two routes in the liver cytosol. One route is direct non-enzymatic reduction by NAD(P)H, and the other is enzymatic reduction by possible carbonyl and/or quinone reductases in the liver cytosol.
摘要:
Capsid protein (Cap), the only structural protein of porcine circovirus 2 (PCV2), is involved in the host protective response and is a target for vaccine development. To find a rapid and easy way to fully map the antigenic epitopes of Cap, purified Cap-specific polyclonal antibodies were used to screen a random heptapeptide phage display library. After three rounds of screening, twenty phage clones that had binding activity to Cap-specific antibodies (tested by phage ELISA) were sequenced. When the inserted amino acid sequences were aligned with the Cap protein sequence, eight core regions in Cap ((50)SRTFGYT(56), (VRTPSW67)-V-62, (68)AVDMMR(73), (79)FLPPGG(84), (SNPRSVPF93)-S-86, (KVEFWP107)-K-102, (119)GSSXXXLDDN(128) and (PPLNP233)-P-229) were identified, three of which ((50)SRTFGYT(56), (SNPRSVPF93)-S-86 and (KVEFWP107)-K-102) for the first time. Nine phagetopes representing the eight regions were chosen to immunize Kunming mice. All except minotopes (50)SRTFGYT(56) and (PPLNP233)-P-229 induced antibodies against PCV2 when injected into Kunming mice. (C) 2013 Elsevier B.V. All rights reserved.
期刊:
Molecular and Cellular Probes,2013年27(3-4):145-148 ISSN:0890-8508
通讯作者:
Zhu, Xing-Quan
作者机构:
[Lin, Rui-Qing; Zhang, Yuan; Weng, Ya-Biao] South China Agr Univ, Coll Vet Med, Guangzhou 510642, Guangdong, Peoples R China.;[Song, Hui-Qun; Lin, Rui-Qing; Zhang, Yuan; Liu, Guo-Hua; Zhu, Xing-Quan] Chinese Acad Agr Sci, Lanzhou Vet Res Inst, Key Lab Vet Parasitol Gansu Prov, State Key Lab Vet Etiol Biol, Lanzhou 730046, Gansu, Peoples R China.;[Liu, Guo-Hua; Zhu, Xing-Quan] Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.;[D'Amelio, Stefano] Univ Roma La Sapienza, Dept Publ Hlth & Infect Dis, Parasitol Sect, I-00185 Rome, Italy.;[Zhu, Xing-Quan; Zou, Feng-Cai] Yunnan Agr Univ, Coll Anim Sci & Technol, Kunming 650201, Yunnan Province, Peoples R China.
通讯机构:
[Zhu, Xing-Quan] C;Chinese Acad Agr Sci, Lanzhou Vet Res Inst, Key Lab Vet Parasitol Gansu Prov, State Key Lab Vet Etiol Biol, Lanzhou 730046, Gansu, Peoples R China.
关键词:
Contracaecum;Cytochrome c oxidase subunit (cox1);Genetic variation;Mitochondrial DNA;NADH dehydrogenase subunits 1 and 4 (nad1 and nad4);Phylogenetic analysis;Sibling species;Small subunit of rRNA (rrnS)
摘要:
The present study investigated sequence variability in four mitochondrial DNA (mtDNA) regions, namely cytochrome c oxidase subunit (cox1), NADH dehydrogenase subunits 1 and 4 (nad1 and nad4), and small subunit of rRNA (rrnS), among Contracaecum rudolphii A, C. rudolphii B, C. rudolphii C and Contracaecum septentrionale from different hosts and geographical origins in China, Italy, Spain and the USA. Regions in the cox1, nad1, nad4 and rrnS genes (designated pcox1, pnad1, pnad4 and prrnS, respectively) were amplified separately from individual nematodes by PCR, sequenced and compared to estimate sequence variability. While sequence variation within each of the Contracaecum species was 0-2.6% for pcox1, 0.3-2.5% for pnad1, 0-1.9% for pnad4 and 0-2.9% for prrnS, differences between species was significantly higher, being 3.3-12%, 9.8-15.2%, 9.6-18.3% and 3.5-11.12% for these regions, respectively. Phylogenetic analyses of pcox1, pnad1, pnad4 and prrnS sequence data using maximum likelihood (ML), maximum parsimony (MP) and neighbour joining (NJ) showed that the specimens of each Contracaecum species clustered together. These results provide additional genetic evidence for the existence of sibling species within C. rudolphii sensu lato.
作者机构:
[Dong, Bo; Ren, Xiaofeng; Zhu, Weijuan] Northeast Agr Univ, Dept Vet Prevent Med, Coll Vet Med, Harbin 150030, Peoples R China.;[Lin, Hai; Tu, Di; Feng, Jing; Su, Dingding; Dong, Bo; Yang, Qing; Li, Lanxiang] Hunan Agr Univ, Dept Vet Clin Sci, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.;[Su, Dingding] Hunan Agr Univ, Hunan Prov Key Lab Germplasm Innovat & Utilizat C, Changsha 410128, Hunan, Peoples R China.;[Ren, Xiaofeng] Northeast Agr Univ, Dept Vet Prevent Med, Coll Vet Med, 59 Mucai St, Harbin 150030, Peoples R China.
通讯机构:
[Ren, Xiaofeng] N;Northeast Agr Univ, Dept Vet Prevent Med, Coll Vet Med, 59 Mucai St, Harbin 150030, Peoples R China.
关键词:
DNA vaccine;ORF2 gene;PCV2;Porcine interluekin-15
摘要:
Porcine circovirus type 2 (PCV2) is associated with many kinds of diseases including postweaning multisystemic wasting syndrome (PMWS). It affects the immune system of swine and causes huge epidemic losses every year. In our previous study, we provided evidence that DNA plasmid bearing porcine IL-15 (pVAX-pIL-15) might serve as an immune enhancer for DNA plasmid encoding porcine reproductive and respiratory syndrome virus GP5 gene. In this study, PCV2 open reading frame (ORF)2 gene was cloned into the eukaryotic expression vector pVAX, resulting in the plasmid pVAX-PCV2-ORF2. Transient expression of the plasmid in BHK-21 cells could be detected using immunofluorescence assay. Experimental mice were divided into 5 groups and immunized with PBS, pVAX, pVAX-pIL-15, pVAX-PCV2-ORF2 or pVAX-pIL-15 plus pVAX-PCV2-ORF2. The results showed that the mice co-inoculated with pVAX-PCV2-ORF2 plus pVAX-pIL-15 had higher humoral and cellular immune responses than the others. In addition, DNA plasmid bearing PCV2 ORF2 gene had a protective effect against challenge with PCV2 in mice which could be promoted with the utilization of pIL-15.
作者:
Xiao, Hong-Bo*;Lu, Xiang-Yang;Zhang, Heng-Bo;Sun, Zhi-Liang;Fang, Jun
期刊:
Journal of Physiology and Biochemistry,2013年69(4):719-725 ISSN:1138-7548
通讯作者:
Xiao, Hong-Bo
作者机构:
[Xiao, Hong-Bo; Sun, Zhi-Liang] Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.;[Lu, Xiang-Yang] Hunan Agr Bioengn Res Inst, Changsha 410128, Hunan, Peoples R China.;[Zhang, Heng-Bo] Furong Dist Red Cross Hosp, Changsha 410126, Hunan, Peoples R China.;[Sun, Zhi-Liang] Hunan Agr Univ, Natl Res Ctr Engn & Technol Utilizat Bot Funct In, Biol Vet Drugs Branch, Changsha 410128, Hunan, Peoples R China.;[Fang, Jun] Hunan Agr Univ, Coll Biosci & Biotechnol, Changsha 410128, Hunan, Peoples R China.
通讯机构:
[Xiao, Hong-Bo] H;Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.
关键词:
Undaria pinnatifida;Soluble fiber;Angiopoietin-like protein 3;Hyperlipidemia;Mice
摘要:
Angiopoietin-like protein 3 (Angptl3)-lipoprotein lipase (LPL) pathway may be a useful pharmacologic target for hyperlipidemia. The present study was conducted to test the effect of soluble fiber extracted from Undaria pinnatifida (UP), on hyperlipidemia in apolipoprotein E-deficient (ApoE(-/-)) mice. Forty mice were divided into four groups (n = 10): control group (C57BL/6J mice), ApoE(-/-) mice group, and two groups of ApoE(-/-) mice treated with UP fiber (5 or 10 % per day). UP soluble fiber treatment significantly decreased plasma and hepatic total cholesterol, triglycerides levels, plasma low-density lipoprotein cholesterol, and malondialdehyde concentrations and increased plasma high-density lipoprotein cholesterol level and downregulated protein expression of Angptl3 concomitantly with upregulated protein expression of LPL. In addition, T0901317 caused elevated expression of hepatic Angptl3 protein, and the effect of T0901317 was also abrogated by UP soluble fiber in C57BL/6J mice. The present results suggest that the UP soluble fiber regulates Angptl3-LPL pathway to lessen hyperlipidemia in mice.
作者机构:
[Song, Hui-Qun; Lin, Rui-Qing; Zhang, Yuan; Liu, Guo-Hua; Zhu, Xing-Quan] Chinese Acad Agr Sci, Lanzhou Vet Res Inst, Key Lab Vet Parasitol Gansu Prov, State Key Lab Vet Etiol Biol, Lanzhou 730046, Gansu, Peoples R China.;[Yuan, Zi-Guo; Lin, Rui-Qing; Zhang, Yuan; Weng, Ya-Biao] S China Agr Univ, Coll Vet Med, Guangzhou 510642, Guangdong, Peoples R China.;[Liu, Guo-Hua] Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.;[Li, Ming-Wei] Guangdong Ocean Univ, Coll Agr, Zhanjiang 524088, Guangdong, Peoples R China.;[Zhu, Xing-Quan; Zou, Feng-Cai] Yunnan Agr Univ, Coll Anim Sci & Technol, Kunming 650201, Yunnan Province, Peoples R China.
通讯机构:
[Zhu, Xing-Quan] C;Chinese Acad Agr Sci, Lanzhou Vet Res Inst, Key Lab Vet Parasitol Gansu Prov, State Key Lab Vet Etiol Biol, Lanzhou 730046, Gansu, Peoples R China.
摘要:
In this study, sequence variation in three mitochondrial DNA regions, namely cytochrome c oxidase subunit (cox1) and NADH dehydrogenase subunits 1 and 4 (nad1 and nad4), between Oesophagostomum dentatum and O. quadrispinulatum isolated from pigs in different geographical origins in Mainland China was examined, and their phylogenetic relationships were reconstructed. A partial of the cox1 (pcox1), nad1, and nad4 genes (pnad1 and pnad4) were amplified separately from individual nodule worms by PCR and were subjected to direct sequencing in order to define sequence variations. While the intraspecific sequence variations within each of the two species were 0.3-5.2% for pcox1, 0-4.9% for pnad1, and 0-7.1% for pnad4, the interspecific sequence differences were significantly higher, being 10.7-13.4% for pcox1, 11-14.6% for pnad1, and 14.9-18% for pnad4, respectively. There were a number of nucleotide positions in the pcox1, pnad1, and pnad4 sequences with no apparent intraspecific variation but distinct interspecific differences among those samples of Oesophagostomum spp. examined, which may be used as genetic makers for the identification and differentiation of the Oesophagostomum spp. Phylogenetic analyses using three inference methods, namely Bayesian inference, maximum likelihood, and maximum parsimony based on the combined sequences of pcox1, pnad1, and pnad4 revealed that the O. dentatum and O. quadrispinulatum form monophyletic groups, respectively. These findings demonstrated clearly the usefulness of the three mitochondrial sequences for studying systematics, population genetic structures, and the molecular ecology of Oesophagostomum spp.
作者机构:
[Song, Hui-Qun; Lin, Rui-Qing; Liu, Guo-Hua; Zhu, Xing-Quan] Chinese Acad Agr Sci, State Key Lab Vet Etiol Biol, Lanzhou Vet Res Inst, Key Lab Vet Parasitol Gansu Prov, Lanzhou 730046, Gansu, Peoples R China.;[Lin, Rui-Qing; Zhang, Yuan] S China Agr Univ, Coll Vet Med, Guangzhou 510642, Guangdong, Peoples R China.;[Liu, Guo-Hua; Zhu, Xing-Quan] Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.;[Hu, Min] Huazhong Agr Univ, Coll Vet Med, State Key Lab Agr Microbiol, Wuhan 430070, Peoples R China.;[Wu, Xiang-Yun] S China Sea Inst Oceanol, Lab Marine Bioresources Sustainable Utilizat, Guangzhou 510301, Guangdong, Peoples R China.
通讯机构:
[Zhu, Xing-Quan] C;Chinese Acad Agr Sci, State Key Lab Vet Etiol Biol, Lanzhou Vet Res Inst, Key Lab Vet Parasitol Gansu Prov, Lanzhou 730046, Gansu, Peoples R China.
作者机构:
[Xu, Min-Jun; Su, Ang; Liu, Guo-Hua; Zhu, Xing-Quan] Chinese Acad Agr Sci, State Key Lab Vet Etiol Biol, Key Lab Vet Parasitol Gansu Prov, Lanzhou Vet Res Inst, Lanzhou, Gansu, Peoples R China.;[Liu, Guo-Hua; Zhu, Xing-Quan] Hunan Agr Univ, Coll Vet Med, Changsha, Hunan, Peoples R China.;[Gasser, Robin B.] Univ Melbourne, Dept Vet Sci, Werribee, Vic 3030, Australia.;[Su, Ang; Lin, Rui-Qing] S China Agr Univ, Coll Vet Med, Guangzhou, Guangdong, Peoples R China.;[Nejsum, Peter] Univ Copenhagen, Dept Vet Dis Biol, Frederiksberg, Denmark.
通讯机构:
[Liu, Guo-Hua] C;Chinese Acad Agr Sci, State Key Lab Vet Etiol Biol, Key Lab Vet Parasitol Gansu Prov, Lanzhou Vet Res Inst, Lanzhou, Gansu, Peoples R China.
摘要:
Author Summary Trichuriasis is a neglected tropical disease (NTD) caused by parasitic nematodes of the genus Trichuris (Nematoda), causing significant human and animal health problems as well as considerable socio-economic consequences world-wide. Although Trichuris species are considered to be relatively host specific, there has been significant controversy as to whether Trichuris infecting humans (recognized as T. trichiura) is a distinct species from that found in pigs (recognized as T. suis), or not. In the present study, we sequenced, annotated and compared the complete mitochondrial genomes of Trichuris from these two hosts and undertook a phylogenetic analysis of the mitochondrial datasets. This analysis showed clear genetic distinctiveness and strong statistical support for the hypothesis that T. trichiura and T. suis are separate species, consistent with previous studies using nuclear ribosomal DNA sequence data. Future studies could explore, using mitochondrial genetic markers defined in the present study, cross-transmission of Trichuris between pigs and humans in endemic regions, and the population genetics of T. trichiura and T. suis.
摘要:
For many years, whipworms (Trichuris spp.) have been described with a relatively narrow range of both morphological and biometrical features. Moreover, there has been insufficient discrimination between congeners (or closely related species). In the present study, we determined the complete mitochondrial (mt) genomes of two whipworms Trichuris ovis and Trichuris discolor, compared them and then tested the hypothesis that T. ovis and T. discolor are distinct species by phylogenetic analyses using Bayesian inference, maximum likelihood and maximum parsimony) based on the deduced amino acid sequences of the mt protein-coding genes. The complete mt genomes of T. ovis and T. discolor were 13,946 bp and 13,904 bp in size, respectively. Both mt genomes are circular, and consist of 37 genes, including 13 genes coding for proteins, 2 genes for rRNA, and 22 genes for tRNA. The gene content and arrangement are identical to that of human and pig whipworms Trichuris trichiura and Trichuris suis. Taken together, these analyses showed genetic distinctiveness and strongly supported the recent proposal that T. ovis and T. discolor are distinct species using nuclear ribosomal DNA and a portion of the mtDNA sequence dataset. The availability of the complete mtDNA sequences of T. ovis and T. discolor provides novel genetic markers for studying the population genetics, diagnostics and molecular epidemiology of T. ovis and T. discolor. (C) 2012 Elsevier B.V. All rights reserved.