通讯机构：
[Yu, Xinglong] H;Hunan Agr Univ, Coll Vet Med, Changsha, Hunan, Peoples R China.
摘要：
A double-antigen sandwich enzyme-linked immunosorbent assay (ELISA) is described for detection of porcine circovirus 2 (PCV2) antibodies using the well-characterized recombinant PCV2 capsid protein. In a comparative test of 394 pig sera against an indirect immunofluorescence (IIF) test and a commercial ELISA kit (also based on the recombinant PCV2 capsid protein), the results showed that the diagnostic sensitivity, specificity, and accuracy of the assay were, respectively, 90.61, 94.02, and 91.62% compared with IIF and 94.38, 95.28, and 94.67% compared with the commercial ELISA kit. Assay of 12 PCV-free pigs over a 5-week period produced only PCV2-negative titers by all 3 methods. These results and the seroprofiles of 4 pig farms obtained by both the commercial ELISA kit and the double-antigen sandwich ELISA indicate that the sandwich ELISA is a reliable method for detection of antibodies to PCV2. Additionally, the method described here permits the use of undiluted test serum samples simultaneously loaded with horseradish peroxidase (HRP)-conjugated antigen into the test well, and the complete test procedure can be performed in less than 90 min. This double-antigen sandwich ELISA should be a useful tool to aid swine industry professionals in deciding the intervention strategies for the control of PCV2-associated diseases.
作者：
Dai, R. S.*;Liu, G. H.;Song, H. Q.;Lin, R. Q.;Yuan, Z. G.;...
期刊：
JOURNAL OF HELMINTHOLOGY,2012年86(2):245-251 ISSN：0022-149X
通讯作者：
Dai, R. S.
作者机构：
[Liu, G. H.; Liu, W.; Dai, R. S.] Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.;[Liu, G. H.; Song, H. Q.; Liu, W.; Lin, R. Q.; Huang, S. Y.; Dai, R. S.; Zhu, X. Q.] CAAS, State Key Lab Vet Etiol Biol, Key Lab Vet Parasitol Gansu Prov, Lanzhou Vet Res Inst, Lanzhou 730046, Gansu, Peoples R China.;[Song, H. Q.; Lin, R. Q.; Yuan, Z. G.] S China Agr Univ, Coll Vet Med, Guangzhou 510642, Guangdong, Peoples R China.;[Li, M. W.] Guangdong Ocean Univ, Dept Vet Med, Zhanjiang 524088, Guangdong, Peoples R China.;[Zhu, X. Q.] Yunnan Agr Univ, Coll Anim Sci & Technol, Kunming 650201, Yunnan, Peoples R China.
通讯机构：
[Dai, R. S.] H;Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.
作者机构：
[Li, Zhong-Yuan; Chen, Jia; Liu, Guo-Hua; Zhu, Xing-Quan; Zhou, Dong-Hui] 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.;[Li, Zhong-Yuan; Chen, Jia] 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.
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：
Toxoplasma gondii is a highly prevalent protozoan parasite infecting a wide range of animals and humans. The epidemiological and biological diversity of T. gondii has resulted in a high genetic variation and unusual population structure in this parasite. This study examined sequence diversity in dense granule 5 (GRA5) gene among T. gondii isolates from different hosts and geographical regions. The entire genome region of the GRA5 gene was amplified and sequenced from 14 T. gondii isolates, and phylogenetic relationship among these T. gondii isolates was reconstructed using Bayesian inference (BI) and maximum parsimony (MP) based on the GRA5 sequences. The complete sequence of the GRA5 gene was 1614 bp in length for strains TgCatBr5 and MAS, but 1617 bp for the other 12 strains. Sequence analysis identified 41 (0–1.7%) variable nucleotide positions among all isolates, with 18 variations of these being in the coding region. Variable positions in the coding region resulted in 11 amino acid substitutions, and a deletion of 3 bp in the strains TgCatBr5 and MAS leading to the deletion of one amino acid. Sequence variations resulted in the existence of polymorphic restriction sites for endonucleases Aat II and Mlu I, allowing the differentiation of the three major clonal lineage types I, II and III by PCR-RFLP. Phylogenetic analyses using BI and MP supported the clear differentiation of the examined T. gondii strains into their respective genotypes. This study demonstrated the existence of sequence variability in the GRA5 gene sequence among T. gondii isolates from different hosts and geographical regions, which allowed the differentiation of the examined T. gondii strains into their respective genotypes, suggesting that this highly polymorphic GRA5 locus may provide a new genetic marker for population genetic studies of T. gondii isolates.
摘要：
Zearalenone is a mycotoxin produced mainly by Fusarium. There are numerous incidences of mycotoxicosis in laboratory and domestic animals, especially in pigs. However, little is known about molecular mechanisms of zearalenone toxicity. Granulosa cells are the maximal cell population in follicles, and they play an essential role in the development and maturation of follicles. The objective of this study was to explore the effect of zearalenone at high concentrations on proliferation and apoptosis of porcine granulosa cells and uncover signaling pathway underlying the cytotoxicity of zearalenone. We found that zearalenone reduced the proliferation of porcine granulosa cells in a dose-dependent manner as shown by the MTT assay and zearalenone resulted in an obvious apoptosis and necrosis in porcine granulosa cells as determined by the TUNEL analysis and flow cytometry. In addition, TUNEL assay with caspase inhibitors showed that zearalenone triggered a caspase-3- and caspase-9-dependent apoptotic process in porcine granulosa cells. Fluorescence spectrophotometer displayed that zearalenone led to a loss of mitochondrial transmembrane potential of porcine granulosa cells but enhanced reactive oxygen species (ROS) levels of the cells. Notably, Western blots revealed that caspase-3 and caspase-9 were activated by zearalenone in porcine granulosa cells. Collectively, our results suggest that zearalenone induces apoptosis and necrosis of porcine granulosa cells in a dose-dependent manner via a caspase-3- and caspase-9-dependent mitochondrial pathway. This study thus offers a novel insight into molecular mechanisms by which zearalenone has adverse cytotoxicity on reproduction. J. Cell. Physiol. 227: 18141820, 2012. (C) 2011 Wiley Periodicals, Inc.
期刊：
International Journal of Mass Spectrometry,2012年309:133-140 ISSN：1387-3806
通讯作者：
Sun, Zhi-Liang
作者机构：
[Sun, Zhi-Liang; Yu, Chun-Hong; Liu, Zhao-Ying] Hunan Agr Univ, Coll Vet Med, Natl Res Ctr Engn & Technol Utilizat Bot Funct In, Biol Vet Drugs Branch, Changsha 410128, Hunan, Peoples R China.;[Wan, Leren] Shimadzu Int Trading Shanghai Co Ltd, Shanghai 200052, Peoples R China.
通讯机构：
[Sun, Zhi-Liang] H;Hunan Agr Univ, Coll Vet Med, Natl Res Ctr Engn & Technol Utilizat Bot Funct In, Biol Vet Drugs Branch, Changsha 410128, Hunan, Peoples R China.
关键词：
Trichothecenes;Ion trap/time-of-flight mass spectrometry;Accurate mass;Fragmentation
摘要：
Five trichothecenes, T-2 toxin, HT-2 toxin, deoxynivalenol (DON), 3-acetyl-DON (3-AcDON) and nivalenol (NIV), major class of mycotoxins produced by Fusarium, were investigated by electrospray hybrid ion trap/time-of-flight mass spectrometry (IT–TOF-MS). Fragmentation mechanisms are proposed based on the MS2 and MS3 experiments and accurate mass measurements. The sodium adduct ions [M+Na]+ of type-A trichothecenes, such as T-2 toxin and HT-2 toxin, were observed in the MS spectra. However, the [M−H]− ions of type-B trichothecenes, such as NIV, and 3-AcDON, were observed in the MS spectra. Eliminations of isovaleryloxy group and acetic acid were the common fragmentation pathways of T-2 toxin and HT-2 toxin. However, the cleavage of epoxy was the common fragmentation pathways of DON, 3-AcDON and NIV. The other fragmentation pathways were found to be dependent on the substitutent group. The data reported here may help to identify analogues of the trichothecenes that are commonly co-produced by Fusarium in cultures or naturally contaminated samples.
期刊：
INTERNATIONAL JOURNAL OF BIOLOGICAL SCIENCES,2012年8(5):640-649 ISSN：1449-2288
通讯作者：
Zhu, Xing-Quan
作者机构：
[Li, Chun; Li, Jia-Yuan; Liu, Guo-Hua; Zhu, Xing-Quan; Zhou, Dong-Hui; Zou, Feng-Cai] 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.;[Liu, Guo-Hua; Zhu, Xing-Quan] Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.;[Li, Chun] Sichuan Ctr Anim Dis Control & Prevent, Chengdu 610041, Sichuan Provinc, Peoples R China.;[Li, Jia-Yuan; Lin, Rui-Qing] S China Agr Univ, Coll Vet Med, Guangzhou 510642, Guangdong, Peoples R China.;[Xiong, Rong-Chuan] Chinese Acad Sci, Chengdu Inst Biol, Chengdu 610041, 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.
摘要：
Sparganosis, caused by the plerocercoid larvae of members of the genus Spirometra, can cause significant public health problem and considerable economic losses. In the present study, the complete mitochondrial DNA (mtDNA) sequence of Spirometra erinaceieuropaei from China was determined, characterized and compared with that of S. erinaceieuropaei from Japan. The gene arrangement in the mt genome sequences of S. erinaceieuropaei from China and Japan is identical. The identity of the mt genomes was 99.1% between S. erinaceieuropaei from China and Japan, and the complete mtDNA sequence of S. erinaceieuropaei from China is slightly shorter (2 bp) than that from Japan. Phylogenetic analysis of S. erinaceieuropaei with other representative cestodes using two different computational algorithms [Bayesian inference (BI) and maximum likelihood (ML)] based on concatenated amino acid sequences of 12 protein-coding genes, revealed that S. erinaceieuropaei is closely related to Diphyllobothrium spp., supporting classification based on morphological features. The present study determined the complete mtDNA sequences of S. erinaceieuropaei from China that provides novel genetic markers for studying the population genetics and molecular epidemiology of S. erinaceieuropaei in humans and animals.
摘要：
The objective was to investigate the effects of a novel DNA vaccine (pcISI) harboring two copies of inhibin alpha (1-32) fragments on immune response, hormone concentrations and reproductive performance in rats. Female Wistar rats (n = 18 per group) were immunized (twice, 4 wk apart) with 10, 50, or 100 mu g (T-1, T-2 and T-3, respectively), of the pcISI plasmid. At 4 wk after the second immunization, plasma antibody titers were higher (P < 0.05) in T-3 than in either T-1 or T-2 (0.341 +/- 0.123, 0.236 +/- 0.068, and 0.251 +/- 0.077, respectively, mean +/- SD). Concurrrently, plasma concentrations of FSH and estradiol were highest (P < 0.05) in T-3, and were higher (P < 0.05) in T-1 and T-2 than in control groups. For antibody-positive rats, there was a correlation (P < 0.01) between antibody titer and FSH concentrations after two pcISI immunizations. The number of mature follicles in the T-3 group (46.00 +/- 4.65) was higher (P < 0.05) than in two control groups (29.25 +/- 3.72 and 27.92 +/- 3.48), and also higher (P < 0.05) than in T-1 and T-2 (37.17 +/- 4.99 and 38.75 +/- 7.09). Antibody-positive rats had more mature ovarian follicles than negative rats (46.75 +/- 4.23 vs. 35.60 +/- 3.38, P < 0.05). Moreover, litter size and number of placentas were increased (P < 0.05) in the pcISI immunization groups, except for the T-1 group, compared to the control groups. In conclusion, the pcISI DNA vaccine successfully induced a humoral immune response, improved reproductive hormone concentrations, stimulated follicular development, and increased number of placentas and litter size. Furthermore, 100 mu g yielded the best immune response. (c) 2012 Elsevier Inc. All rights reserved.
摘要：
Until recently, there was no cell line that could produce continuously high-titer duck hepatitis virus type 1 (DHV-1). In this study, a duck embryo fibroblast (DEF) cell line was established, and the susceptibility of this cell line to DHV-1 was determined. The primary culture of DEF cells was from a duck embryo that was partially digested with trypsin. Digested tissue pieces were cultured at 37 degrees C in Dulbecco's Modified Eagle Medium supplemented with 10% fetal bovine serum. The cultured DEF cells, which had the morphology of fibroblast, proliferated to 100% confluence four days later. An immortalized DEF cell line, named DEF-TA, was established and subcultured to passage 33, and the susceptibility of that cell line to DHV-1 was determined. In the DHV-1 susceptibility tests, cytopathic effects and the propagation of virus were observed in DEF-TA cells after DHV-1 infection. This continuous DHV-1-susceptible DEF cell line may serve as a valuable cell line for studies of cell-virus interactions and the pathogenesis of DHV-1 and may be useful for the development of an inactivated vaccine. Crown Copyright (C) 2012 Published by Elsevier B.V. All rights reserved.
作者机构：
[Li, Guangxing; Ren, Xiaofeng; Zhu, Weijuan] NE Agr Univ, Coll Vet Med, Harbin 150030, Peoples R China.;[Ren, Yudong] NE Agr Univ, Coll Engn, Harbin 150030, Peoples R China.;[Su, Dingding] Hunan Agr Univ, Hunan Prov Key Lab Germplasm Innovat & Utilizat G, Changsha 410128, Hunan, Peoples R China.;[Yang, Qing] Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.;[Ren, Xiaofeng] NE Agr Univ, Coll Vet Med, 59 Mucai St, Harbin 150030, Peoples R China.
通讯机构：
[Ren, Xiaofeng] N;NE Agr Univ, Coll Vet Med, 59 Mucai St, Harbin 150030, Peoples R China.
关键词：
Antibody;ELISA;Porcine parvovirus;VP2
摘要：
Viral protein 2 (VP2) of porcine parvovirus (PPV) is the major viral structural protein and responsible for eliciting neutralizing antibodies in immunized animals. In this study, the gene encoding VP2 of PPV was amplified by PCR. The VP2 gene was then cloned into the prokaryotic expression vector, pET-32a followed by expression in Escherichia coli Rosetta. The VP2 protein expression was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Rabbit polyclonal antiserum was generated using the recombinant VP2 protein. The optimal titer of the anti-VP2 antibody was determined by ELISA. The anti-VP2 antibody was able to distinguish PPV from other viruses in ELISA. (C) 2012 PVJ. All rights reserved
作者机构：
[Guo, Chengzhi; Yuan, Hui] Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.;[He, Zuping; Guo, Chengzhi] Shanghai Jiao Tong Univ, Sch Med, Renji Hosp, Clin Stem Cell Res Ctr, Shanghai 200127, Peoples R China.;[He, Zuping] Georgetown Univ, Med Ctr, Dept Biochem & Mol & Cellular Biol, Washington, DC 20057 USA.;[Yuan, Hui] Hunan Agr Univ, Coll Vet Med, 19 E Lake Rd, Changsha 410128, Hunan, Peoples R China.
通讯机构：
[Yuan, Hui] H;Hunan Agr Univ, Coll Vet Med, 19 E Lake Rd, Changsha 410128, Hunan, Peoples R China.
关键词：
Apoptosis;Melamine;Normal rat kidney (NRK)-52e cells;p38 mitogen-activated protein kinase (MAPK) pathway;reactive oxygen species (ROS)
摘要：
There was an outbreak of urinary stones associated with consumption of melamine-tainted milk products in 2008 in China, leading to serious illness of many infants and even death. We have recently demonstrated that melamine causes oxidative damage on the NRK (normal rat kidney)-52e cells. The objective of this study was to explore the cellular signalling pathway that mediates the cell apoptosis induced by melamine in the NRK-52e cells. Fluorescence microscope showed that melamine enhanced intracellular ROS (reactive oxygen species) levels of the NRK-52e cells. AO/EB (acridine orange/ ethidium bromide) staining and flow cytometry revealed that melamine increased apoptotic and necrotic percentages of the NRK-52e cells in a dose-dependent manner. Notably, MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide] assays and flow cytometry displayed that SB203580, an inhibitor for p38 MAPK (mitogen-activated protein kinase) pathway, increased the proliferation of the NRK-52e cells and reduced the apoptotic and necrotic percentages of the NRK-52e cells. Western blots further demonstrated that p38 phosphorylation was activated by melamine in the NRK-52e cells and inhibitor SB203580 blocked the increase of p38 phosphorylation induced by melamine. Together, these results suggested that melamine causes apoptosis of the NRK-52e cells via excessive intracellular ROS and the activation of p38 MAPK pathway. This study thus offers a novel insight into molecular mechanisms by which melamine has adverse cytotoxicity on renal tubular epithelial cells.
摘要：
In this study, the stress degradation of olaquindox under conditions of hydrolysis (neutral, acidic and basic), oxidation and photolytic stress was investigated. In order to characterize each degradation product, we developed a rapid, sensitive and reliable high-performance liquid chromatography combined with hybrid ion trap/time-of-flight mass spectrometry (LC/MS-IT-TOF) method. The degradation products formed under different forced conditions were separated using an ODS-C18 column with gradient elution. Multiple scans of degradation products in MS and MS/MS modes and accurate mass measurements were performed through data-dependent acquisition. The structural elucidations of degradation products were performed by comparing the changes in the accurate molecular masses and fragment ions generated from precursor ions with those of parent drug. The present results showed that maximum degradation was observed in hydrolysis, especially in the acidic condition. The drug was also degraded significantly under photolytic conditions. A total of 12 degradation products of olaquindox were detected and characterized using the developed method. The main degradation product was formed by the complete cleavage of side chain to form 3-methyl-2-hydroxylquinoxaline-4-oxide. A degradation pathway of olaquindox was also tentatively proposed for the first time based on these characterized structures. (C) 2011 Elsevier B.V. All rights reserved.
关键词：
Latex agglutination test;antibody against human T-cell
摘要：
Based on human T-cell as a source of antigen, a simple and rapid latex agglutination test (LAT) was developed to detect pig antibodies against human T-cell instead of the time-consuming erythrocyte rosette inhibition test (ERIT). A total of 380 pig serum samples were subjected to the LAT and the sensitivity and the specificity were assessed in comparison with ERIT. The corresponding sensitivity and specificity were 92.05% and 98.61%.
期刊：
Toxicology and Applied Pharmacology,2011年257(3):405-411 ISSN：0041-008X
通讯作者：
Xiao, Hong-Bo
作者机构：
[Xiao, Hong-Bo] Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.;[Zhang, Heng-Bo] Furong Dist Red Cross Hosp, Changsha 410126, 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：
Apolipoprotein e-deficient mouse;Cluster of differentiation 44;Inflammation;Kaempferol;Osteopontin
摘要：
Recent studies show that osteopontin (OPN) and its receptor cluster of differentiation 44 (CD44) are two pro-inflammatory cytokines contributing to the development of atherosclerosis. The objective of this study was to explore the inhibitory effect of kaempferol, a naturally occurring flavonoid compound, on atherogenesis and the mechanisms involved. The experiments were performed in aorta and plasma from C57BL/6J control and apolipoprotein E-deficient (ApoE(-/-)) mice treated or not with kaempferol (50 or 100 mg/kg, intragastrically) for 4 weeks. Kaempferol treatment decreased atherosclerotic lesion area, improved endothelium-dependent vasorelaxation, and increased the maximal relaxation value concomitantly with decrease in the half-maximum effective concentration, plasma OPN level, aortic OPN expression, and aortic CD44 expression in ApoE(-/-) mice. In addition, treatment with kaempferol also significantly decreased reactive oxygen species production in mice aorta. The present results suggest that kaempferol regulates OPN-CD44 pathway to inhibit the atherogenesis of ApoE(-/-) mice. (C) 2011 Elsevier Inc. All rights reserved.
作者机构：
[Song, Hui-Qun; Liu, Guo-Hua; Zhu, Xing-Quan] CAAS, 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] S China Agr Univ, Coll Vet Med, Parasitol Lab, Guangzhou 510642, Guangdong, Peoples R China.;[Zhang, Kou-Xing] Sun Yat Sen Univ, Affiliated Hosp 3, Dept Infect Dis, Guangzhou 510630, Guangdong, Peoples R China.;[Li, Ming-Wei] Guangdong Ocean Univ, Coll Agr, Dept Vet Med, Zhanjiang 524088, Guangdong, Peoples R China.;[Liu, Wei; Liu, Yi; Liu, Guo-Hua] Hunan Agr Univ, Coll Vet Med, Parasitol Lab, Changsha 410128, Hunan, Peoples R China.
通讯机构：
[Zhu, Xing-Quan] C;CAAS, Lanzhou Vet Res Inst, Key Lab Vet Parasitol Gansu Prov, State Key Lab Vet Etiol Biol, Lanzhou 730046, Gansu, Peoples R China.
摘要：
Mitochondrial (mt) genome sequences provide useful markers for investigating population genetic structures, systematics and phylogenetics of organisms. Although Taenia multiceps, T. hydatigena, and T. taeniaeformis are common taeniid tapeworms of ruminants, pigs, dogs, or cats, causing significant economic losses, no published study on their mt genomes is available. The complete mt genomes of T. multiceps, T. hydatigena, and T. taeniaeformis were amplified in two overlapping fragments and then sequenced. The sizes of the entire mt genome were 13700 bp for T. multiceps, 13489 bp for T. hydatigena, and 13647 bp for T. taeniaeformis. Each of the three genomes contains 36 genes, consisting of 12 genes for proteins, 2 genes for rRNA, and 22 genes for tRNA, which are the same as the mt genomes of all other cestode species studied to date. All genes are transcribed in the same direction and have a nucleotide composition high in A and T. The contents of A+T of the complete genomes are 71.3% for T. multiceps, 70.8% for T. hydatigena, and 73.0% for T. taeniaeformis. The AT bias had a significant effect on both the codon usage pattern and amino acid composition of proteins. T. multiceps and T. hydatigena had two noncoding regions, but T. taeniaeformis had only one. Phylogenetic analyses based on concatenated amino acid sequences of 12 protein-coding genes revealed that T. multiceps, T. hydatigena, and T. taeniaeformis were more closely related to the other members of the Taenia genus, consistent with results of previous morphological and molecular studies. The present study determined the complete mt genome sequences for three Taenia species of animal and human health significance, providing useful markers for studying the systematics, population genetics, and molecular epidemiology of these cestode parasites of animals and humans.
