Prohibitin 2基因参与猪圆环病毒2型PK-15细胞感染的研究
作者：
蒋一凡;罗旭英;吴锐;符世豪;张佳鑫;...
期刊：
中国兽医科学 ,2020年50(4):506-511 ISSN：1673-4696
作者机构：
湖南农业大学动物医学院,湖南长沙410128;兽用蛋白质工程疫苗湖南省重点实验室,湖南长沙410128;兽用疫苗逆向创制湖南省工程研究中心,湖南长沙410128;宜春学院数学与计算机科学学院,江西宜春336000;[蒋一凡; 王乃东; 张佳鑫; 吴锐; 罗旭英; 符世豪; 李周勉; 余婉婷] 湖南农业大学
关键词：
猪圆环病毒2型;猪肾上皮细胞;荧光定量PCR
摘要：
为探究Prohibitin2(PHB2)在PCV2感染细胞过程中发挥的作用,首先通过间接免疫荧光试验检测PCV2在PK-15细胞中的增殖变化,同时建立PHB2荧光定量PCR检测体系,然后以PCV2已知的宿主细胞互作蛋白gC1qR为对照,检测PCV2感染PK-15细胞后PHB2 mRNA的动态变化,最后鉴定PHB2 siRNA转染PK-15细胞中PCV2 Cap的表达。结果表明,PHB2在正常及PCV2感染的PK-15细胞中均有效表达。随着PCV2感染PK-15细胞,gC1qR转录水平在病毒感染3 h显著降低,感染6 h时升高,随后(12~ 48 hpi)转录水平下降;而PHB2转录水平在PCV2感染6 h内显著升高,随后(12~48 hpi)mRNA水平有所下降;RNA干扰PHB2对PCV2在PK-15细胞中复制起到显著的抑制作用。上述研究结果表明,PCV2可显著影响PK-15细胞中PHB2转录,而且PHB2对PCV2病毒复制具有调节作用。
语种：
中文
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猪圆环病毒3型Cap蛋白序列的生物信息学研究
作者：
李周勉;邹亚文;蒋一凡;胡意;秦艺文;...
期刊：
经济动物学报 ,2020年24(1):9-16 ISSN：1007-7448
作者机构：
湖南农业大学动物医学院,长沙410128;兽用蛋白质工程疫苗湖南省重点实验室,长沙410128;兽用疫苗逆向创制湖南省工程研究中心,长沙410128;[秦艺文; 蒋一凡; 湛洋; 李萌; 王乃东; 胡意; 李周勉; 王东亮; 邹亚文] 湖南农业大学
关键词：
猪圆环病毒3型;Cap蛋白;氨基酸突变位点;生物信息学
摘要：
猪圆环病毒3型（PCV3）是一种在世界范围内分布较广泛的新型猪病毒,其发病机制尚不清楚。为系统分析PCV3 Capsid（Cap）蛋白序列和结构特点,收集Gen Bank登录的来自不同国家和地区的210条PCV3全基因组和Cap蛋白基因序列,筛选获得46株PCV3代表序列;采用生物信息学（Bio Edit和MAGE 6.0等）技术分析PCV3 Cap序列同源性、结构特点、系统进化和热点突变位点情况。结果表明:46株PCV3 Cap序列同源性为97.66%～100%,PCV3 Cap预测的8个B细胞和10个T细胞表位中均存在序列突变,其中B细胞抗原表位E （109～146位氨基酸）和T细胞抗原表位D（72～81位氨基酸）存在的突变位点数最多。研究发现26株属于PCV3a,20株属于PCV3b。PCV3与PCV2 Cap序列比较分析发现,PCV3 Cap在NLS区域内有6处突变（R11K、R12K、H26M、R27K、V31A和R34K）; PCV3 Cap序列间比对分析发现,PCV3b Cap序列出现突变的几率（17/20,85%）高于PCV3a （19/26,73%）。而从临床表现有繁殖障碍和死胎的样品中检测出的PCV3序列均存在S77T和I150L突变位点;从伴有发热、肺炎的仔猪以及肺匀浆样本检测出的PCV3序列中均有R10K突变位点,这些特定的突变位点与临床症状的关系需后续深入研究。本研究结果为PCV3基因功能、进化、预防和控制等研究提供重要序列信息。
语种：
中文
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猪圆环病毒2型和3型双重PCR方法的建立与优化
作者：
张露华;李周勉;唐祎依;蔡云凤;田唯嘉;...
期刊：
黑龙江畜牧兽医 ,2020年(05):71-76 ISSN：1004-7034
作者机构：
湖南农业大学动物医学院兽用蛋白质工程疫苗湖南省重点实验室,长沙,410128;[田唯嘉; 唐祎依; 张露华; 湛洋; 蔡云凤; 王乃东; 黄坤; 李周勉; 王东亮] 湖南农业大学
关键词：
猪圆环病毒2型;猪圆环病毒3型;双重PCR;引物;退火温度;循环次数
摘要：
为了能够同时检测猪圆环病毒2型（Porcine circovirus type 2, PCV-2）和猪圆环病毒3型（Porcine circovirus type 3, PCV-3）,试验根据国内报道的PCV-2和PCV-3毒株全基因序列设计6对特异性引物（P3～P8）,并构建标准质粒进行引物特异性试验,然后通过优化扩增条件建立检测PCV-2和PCV-3混合感染的双重PCR方法。最后用PCV-2和PCV-3的感染性克隆同时转染PK-15细胞,盲传5代后用建立的双重PCR方法检测细胞培养液中的病毒核酸。结果表明:构建的PCV-2和PCV-3标准质粒浓度分别为290 ng/μL和380 ng/μL,双重PCR方法的最佳引物组合为P3/P6和P4/P8,最佳DNA聚合酶浓度为0.02 U/μL。使用引物组合P3/P6时,扩增得到的目的片段大小分别为308 bp和608 bp,最佳退火温度为58℃,最佳循环次数为28个;使用引物组合P4/P8时,扩增得到的目的片段大小分别为291 bp和645 bp,最佳退火温度为60℃,最佳循环次数为26个。运用建立好的双重PCR方法评估感染细胞模型,能有效鉴别PCV-2和PCV-3的单一感染和混合感染。说明优化得到的双重PCR检测方法可用于PCV-2和PCV-3的同时检测。
语种：
中文
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Inhibition of Abl or Src tyrosine kinase decreased porcine circovirus type 2 production in PK15 cells.
作者：
Zhang, Jiaxin;Lei, Xinnuo;Wang, Dongliang;Jiang, Yifan;Zhan, Yang;...
期刊：
Research in Veterinary Science ,2019年124:1-9 ISSN：0034-5288
通讯作者：
Wang, Naidong
作者机构：
[Zhan, Yang; Wang, Naidong; Qin, Yiwen; Wang, Dongliang; Zhou, Yawen; Jiang, Yifan; Wang, Aibing; Li, Meng; Zhang, Jiaxin; Lei, Xinnuo; Yang, Yi] Hunan Agr Univ, Coll Vet Med, Key Lab Anim Vaccine & Prot Engn, Changsha 410128, Hunan, Peoples R China.;[Liu, Jue] Beijing Acad Agr & Forestry Sci, Inst Anim Husb & Vet Med, Beijing Key Lab Prevent & Control Infect Dis Live, 9 Shuguang Garden Middle Rd, Beijing 100097, Peoples R China.
通讯机构：
[Wang, Naidong] H;Hunan Agr Univ, Coll Vet Med, Key Lab Anim Vaccine & Prot Engn, Changsha 410128, Hunan, Peoples R China.
关键词：
Antiviral activity;PK15 cells;Porcine circovirus 2;Tyrosine kinase inhibitor STI-571 and PP2;Virus like particles
摘要：
Porcine circovirus type 2 (PCV2) causes huge economic losses in the global swine industry and has a complex and poorly understood virus-host interaction mechanism. We reported that the C-terminal of the capsid protein of all PCV2 isolates shared a strictly conserved PXXP motif that may interact with SH3 domain-containing tyrosine kinases; however, its roles in PCV2 cell entry and replication remain unknown. In this study, we determined that mRNA levels of two SH3 domain-containing tyrosine kinases family (Abl and Src) had distinct profiles (wild-type and PXXP-mutated) during PCV2 infections of PK15 cells. Therefore, we hypothesized that activities of tyrosine kinases (Abl and Fyn) in PK15 cells may be hijacked by PCV2 via its PXXP motif of the Cap, to favor virus replication. Specific inhibitors PP2 of Lck/Fyn and STI-571 of Abl family kinases decreased viral production through suppression of DNA and Cap synthesis at the replication stage. However, based on indirect immunofluorescence assay (IFA), entry of PCV2 virus-like particles (VLPs) into PK15 cells was not altered. Elucidating mechanisms of PCV2-host interactions should provide new insights for development of new compounds to prevent or reduce PCV2 infections.
语种：
英文
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Development and application of a baculovirus-expressed capsid protein-based indirect ELISA for detection of porcine circovirus 3 IgG antibodies
作者：
Zhang, Sujiao;Wang, Dongliang;Jiang, Yifan;Li, Zhoumian;Zou, Yawen;...
期刊：
BMC Veterinary Research ,2019年15(1):1-10 ISSN：1746-6148
通讯作者：
Wang, Naidong
作者机构：
[Wang, Naidong; Yu, Haoyang; Wang, Dongliang; Jiang, Yifan; Zhang, Sujiao; Li, Meng; Li, Zhoumian; Zou, Yawen; Huang, Kun; Yang, Yi] Hunan Agr Univ, Coll Vet Med, Hunan Prov Key Lab Prot Engn Anim Vaccines, Lab Funct Prote,Res Ctr Reverse Vaccinol, Changsha 410128, Hunan, Peoples R China.
通讯机构：
[Wang, Naidong] H;Hunan Agr Univ, Coll Vet Med, Hunan Prov Key Lab Prot Engn Anim Vaccines, Lab Funct Prote,Res Ctr Reverse Vaccinol, Changsha 410128, Hunan, Peoples R China.
关键词：
Porcine circovirus type 3;Diagnostic;Pig;ELISA
摘要：
Background: Porcine circovirus type 3 (PCV3), recently widely isolated from pigs with various clinical conditions, is likely globally epidemic. However, development of serological diagnosis for PCV3 in pigs is ongoing. Our objectives were to: 1) establish an indirect ELISA, using PCV3 capsid protein (Cap) prepared by Baculovirus Expression Vector System (BEVS) as a high-quality coating antigen for detection of PCV3-associated antibodies in serum samples; and 2) use this ELISA to conduct a serological survey for PCV3 in various regions of Hunan province, China. Results: The PCV3 positive rate to the ELISA assay (total of 190 serum samples) was higher in sows with reproductive failure compared to healthy sows (34/85, 40.0% versus 30/105, 28.6%), with similar results using qPCR assays. Further, in an additional 1038 serum samples collected from January 2016 to May 2018 in various regions of Hunan province and tested with this established ELISA, 20 to 84% were positive for PCV3 (according to region of sera collection), with high PCV3 seroprevalence (> 50%) in herds in Changde, Hengyang and Yueyang. Moreover, among serum samples from herds in Shaoyang and Changde, PCV3 seroprevalence was higher in sows than in other classes of pigs (i.e., suckling piglets, nursery pigs, gilts, growing-finishing pigs and boars). Conclusions: We developed a full-length PCV3 Cap-based ELISA using a eukaryotic expression system with excellent potential to elucidate PCV3 epidemiology. Based on this assay, PCV3 has been circulating in Hunan province. PCV3 prevalence was lower in healthy sows than in those with reproductive failure. Further studies are warranted to identify the PCV3 responsible for high seroprevalence in sows and determine pathogenesis of PCV3 in sows with reproductive failure. © 2019 The Author(s).
语种：
英文
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Protective humoral immunity in guinea pigs induced by PCV2 virus-like particles displaying the B cell linear epitope ((228)QQITDA(233)) of PPV1.
作者：
Wang, Naidong;Zhang, Sujiao;Wang, Dongliang;Li, Fuqiang;Liang, Lin;...
期刊：
Veterinary Microbiology ,2019年235:86-92 ISSN：0378-1135
通讯作者：
Tu, Di;Cui, Shangjin
作者机构：
[Wang, Dongliang; Yu, Wanting; Tu, Di; Zou, Yawen; Wang, Naidong; Zhan, Yang; Chen, Guanyu; Zhang, Sujiao; Deng, Zhibang] Hunan Agr Univ, Coll Vet Med, Res Ctr Reverse Vaccinol, Hunan Prov Key Lab Prot Engn Anim Vaccines,Lab Fu, Changsha 410128, Hunan, Peoples R China.;[Cui, Shangjin; Liang, Lin] Chinese Acad Agr Sci, IAS, Beijing 100193, Peoples R China.;[Li, Fuqiang; Li, Xiuli] Tianjin Anim Husb & Vet Res Inst, Tianjin 300381, Peoples R China.;[Cui, Shangjin; Liang, Lin] Minist Agr, Scientifc Observat & Expt Stn Vet Drugs & Diagnos, Beijing 100193, Peoples R China.
通讯机构：
[Tu, Di] H;[Cui, Shangjin] C;Hunan Agr Univ, Coll Vet Med, Res Ctr Reverse Vaccinol, Hunan Prov Key Lab Prot Engn Anim Vaccines,Lab Fu, Changsha 410128, Hunan, Peoples R China.;Chinese Acad Agr Sci, IAS, Beijing 100193, Peoples R China.
关键词：
Guinea pig;Porcine circovirus type 2;Porcine parvovirus;Subunit vaccine;Virus-like particles
摘要：
Although PCV2 infections generally cause mild disease in pigs, concurrent co-infections with other pathogens can damage the immune system and cause more severe diseases, collectively termed porcine circovirus associated diseases (PCVAD). Involvement of porcine parvovirus (PPV, a common cause of reproductive failure in naïve dams) in PCVAD caused by PCV2, has been reported. As this co-infection can be difficult to eliminate, there is a critical need to develop an effective vaccine to protect against PPV or synergistic effects of PCV2 and PPV under field conditions. In this study, we designed chimeric PCV2 virus-like particles (cVLPs) displaying a B-cell epitope derived from PPV1 structural protein around the surface of the 2-fold axes of PCV2 VLPs, based on 3D-structure analysis of the PCV2 capsid. The cVLPs were successfully prepared, verified by transmission electron microscopy and chromatography, with robust antibody titers against PCV2 and PPV1 produced in mice and guinea pigs. In addition, in guinea pigs challenged with 106 TCID50 PCV2, cVLPs conferred more effective immune protection (based on viral load) than a commercial PCV2 vaccine. Finally, antibody responses and immune protection against PPV were also evaluated. In guinea pigs vaccinated with cVLPs, although PPV antibodies detected by a hemagglutination inhibition (HI) assay appeared later after vaccination in the PCV2 cVLPs group than in the commercial PPV vaccine group, there were fewer PPV genomic DNA copies in the PCV2 cVLPs group than in a PBS group. In conclusion, guinea pigs vaccinated with cVLPs developed effective protective immunity against PCV2 challenge, with some protective immunity against PPV. This study provided valuable research data to pursue molecular design of chimeric epitopes PCV2 VLPs. © 2019
语种：
英文
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湖南地区猪圆环病毒2型的分离与鉴定
作者：
贺延峰;王东亮;邹亚文;董彦鹏;王乃东;...
期刊：
中国兽医科学 ,2019年49(2):213-219 ISSN：1673-4696
作者机构：
湖南农业大学动物医学院;兽用蛋白质工程疫苗湖南省重点实验室;[董彦鹏] 江苏南农高科技股份有限公司;[湛洋; 王乃东; 贺延峰; 王东亮; 邹亚文] 湖南农业大学
关键词：
猪圆环病毒2型;分离;鉴定
摘要：
为分离鉴定湖南地区2个猪场的猪圆环病毒2型(PCV2),从临床表现为断奶仔猪多系统衰竭综合征(PMWS)猪体采集组织病料,验证为PCV2感染的病料,用无污染猪肾细胞系(PK15)细胞分离培养。采用间接免疫荧光试验(IFA)、分子克隆及生物信息学等方法对分离病毒进行分析。结果显示,分别获得了2株PCV2:1株为PCV2d(命名为ChenZ-2-1,GenBank号MH718995),另1株为PCV2b(命名为YiY-3-27, GenBank号KU317482),且两分离株的病毒效价TCID_(50)分别为1×10~(5.66)/ mL和1×10~5.5/ mL。本研究有利于提高PCV2的诊断水平,对深入研究PCV2的发病机理及开发PCV2新型疫苗奠定基础。
语种：
中文
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猪圆环病毒3型检测和致病性的研究进展
作者：
蒋一凡;张南乡子;李周勉;张佳鑫;张素姣;...
期刊：
经济动物学报 ,2019年23(03):181-186 ISSN：1007-7448
作者机构：
湖南农业大学动物医学院,长沙410128;兽用蛋白质工程疫苗湖南省重点实验室及兽用疫苗逆向创制湖南省工程研究中心,长沙410128;[张南乡子; 蒋一凡; 王乃东; 张佳鑫; 李周勉; 张素姣] 湖南农业大学
关键词：
猪圆环病毒3型;繁殖障碍;致病性;混合感染
摘要：
猪圆环病毒3型（Porcine circovirus 3,PCV3）是2016年首次在美国报道的一种新的猪圆环病毒。之后,中国、波兰、韩国、丹麦、意大利和西班牙等国家陆续报道PCV3单独或混合感染的检测,引起研究人员的广泛关注。目前认为PCV3与母猪繁殖障碍、发热、呼吸系统疾病以及多系统炎症等有一定的相关性,且检测报道预示PCV3会引起新生仔猪先天性震颤和心肌炎等临床疾病。但PCV3确切致病机理仍需进一步调查研究。文中主要对PCV3基因组序列的特点、感染检测和致病性相关报道进行综述,为PCV3流行病学调查和致病机理研究提供参考。
语种：
中文
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高校创新创业教育的突出问题与发展策略研究
作者：
杨成林;邬静;王乃东;陈宇科;陈美伊
期刊：
大学教育 ,2019年(10):180-182 ISSN：2095-3437
作者机构：
湖南农业大学动物医学院,湖南 长沙,410128;[杨成林; 王乃东; 陈宇科; 邬静; 陈美伊] 湖南农业大学
关键词：
高校;创新创业教育;发展与现状;突出问题;发展策略
摘要：
近年来,随着国家"大众创业,万众创新"口号的积极号召,社会各界特别是高校对创新创业教育工作的重视提到了新高度,它是推动我国经济、科技快速发展的迫切需要.高校作为创新创业人才培养的主阵地,其创新创业教育工作的扎实推进是决定培养成效的关键.要解决当前我国高校创新创业教育存在的突出问题,必须思想引领,端正学生认知;专兼结合,强化师资建设;层级递进,构建课程体系;搭建平台,推进成果转化.
语种：
中文
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非洲猪瘟影响下的猪场生物安全设计
作者：
戴永强;闫之春;王乃东;李修松;周均青;...
期刊：
中国猪业 ,2019年14(1):35-37 ISSN：1673-4645
作者机构：
湖南农业大学动物医学院;[周均青; 李修松; 冯晓龙] 青岛大牧人机械股份有限公司;[闫之春] 新希望六和股份有限公司;浙江大学生物系统工程与食品科学学院;[王乃东; 戴永强] 湖南农业大学
关键词：
猪场;非洲猪瘟;生物安全;流向
摘要：
国内的养猪场单场母猪规模从早先的几十头一直到目前的上万头,并且存栏母猪规模还有继续增大的趋势。在当前非洲猪瘟防控形势极其严峻的背景下,猪场生物安全显得迫切而重要。在猪场规划设计之初,必须制定好猪场的生物安全措施,规划好猪场的各个流向,给猪场的后期运营提供强有力的生物安全保障。
语种：
中文
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湖南省部分地区猪流产胎儿组织中猪圆环病毒3型的检测及其Cap序列分析
作者：
李周勉,,;邹亚文,,;蒋一凡,,;谭磊,,;王教顺,,;...
作者机构：
湖南农业大学动物医学院;兽用蛋白质工程疫苗湖南省重点实验室;兽用疫苗逆向创制湖南省工程研究中心
会议名称：
第三届中国猪业科技大会暨中国畜牧兽医学会2019年学术年会
会议时间：
2019-09-19
会议地点：
中国山东青岛
会议论文集名称：
第三届中国猪业科技大会暨中国畜牧兽医学会2019年学术年会论文集
摘要：
<正>引言猪圆环病毒2型（PCV2）是猪圆环病毒相关病（PCVAD）的主要病原,临床表现为断奶仔猪多系统衰竭综合征（PMWS）,猪皮炎肾病综合征（PDNS）等,给全球养猪业带来了巨大的经济损失。猪圆环病毒3型（PCV3）是一种新的圆环病毒,其致病性及致病机制仍不清楚。本研究对2015-2017年湖南省部分地区的猪流产胎儿样品进行了PCR检测,采用生物信息学技术分析了PCV3 Cap序列同源性、结构特点、系统进化和突变位点情况,旨在为PCV3基因功能、进化和防控等研究提供重要的序列信息。材料与方法
语种：
中文
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Src和Abl酪氨酸蛋白激酶家族参与病原微生物感染的研究进展
作者：
张佳鑫;蒋一凡;雷昕诺;秦艺文;湛洋;...
期刊：
微生物学通报 ,2019年46(10):2781-2786 ISSN：0253-2654
作者机构：
湖南农业大学动物医学院 湖南长沙410128;宜春学院数学与计算机科学学院 江西宜春336000;兽用蛋白质工程疫苗湖南省重点实验室 湖南长沙410128;兽用疫苗逆向创制湖南省工程研究中心 湖南长沙410128;[秦艺文; 蒋一凡; 湛洋; 王乃东; 雷昕诺; 张佳鑫] 湖南农业大学
关键词：
非受体酪氨酸激酶;微生物感染
摘要：
Src和Abl家族激酶属于非受体型酪氨酸激酶(Nonreceptor tyrosine kinase,NRTK)家族重要成员,广泛存在于各种细胞中,参与细胞内信号传递并调节细胞生理过程,它们在维持细胞、组织和器官稳态功能中发挥着至关重要的作用。研究表明,Src和Abl家族激酶通过多种机制参与病原微生物的感染(如与病原微生物的脯氨酸基序-PXXP互作)。因此,从Src和Abl家族激酶角度出发探究病原微生物感染机制逐渐成为一个热点。本文就Src和Abl家族激酶的结构特点以及参与病原微生物感染的研究报道进行综述,以期为病原微生物感染的致病机制、防控和药物研发提供参考。
语种：
中文
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Autophagy and Apoptosis Interact to Modulate T-2 Toxin-Induced Toxicity in Liver Cells
作者：
Wu, Jing;Zhou, Yu;Yuan, Zhihang;Yi, Jine;Chen, Jingshu;...
期刊：
TOXINS ,2019年11(1):45- ISSN：2072-6651
通讯作者：
Wang, Naidong;Tian, Yanan
作者机构：
[Wu, Jing; Wang, Naidong; Wang, ND; Tian, Yanan; Zhou, Yu; Yi, Jine; Yuan, Zhihang] Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.;[Wu, Jing; Yuan, Zhihang] Hunan Agr Univ, Hunan Collaborat Innovat Utilizat Bot Funct Ingre, Changsha 410128, Hunan, Peoples R China.;[Wu, Jing; Yuan, Zhihang] Hunan Agr Univ, Hunan Engn Res Ctr Vet Drug, Changsha 410128, Hunan, Peoples R China.;[Chen, Jingshu; Tian, Yanan] Texas A&M Univ, Coll Vet Med, Dept Vet Physiol & Pharmacol, College Stn, TX 77843 USA.
通讯机构：
[Wang, ND; Tian, YA] H;[Tian, Yanan] T;Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.;Texas A&M Univ, Coll Vet Med, Dept Vet Physiol & Pharmacol, College Stn, TX 77843 USA.
关键词：
T-2 toxin;toxicity;autophagy;apoptosis
摘要：
T-2 toxin is a mycotoxin generated by Fusarium species which has been shown to be highly toxic to human and animals. T-2 toxin induces apoptosis in various tissues/organs. Apoptosis and autophagy are two closely interconnected processes, which are important for maintaining physiological homeostasis as well as pathogenesis. Here, for the first time, we demonstrated that T-2 toxins induce autophagy in human liver cells (L02). We demonstrated that T-2 toxin induce acidic vesicular organelles formation, concomitant with the alterations in p62/SQSTM1 and LC3-phosphatidylethanolamine conjugate (LC3-II) and the enhancement of the autophagic flux. Using mRFP-GFP-LC3 by lentiviral transduction, we showed T-2 toxin-mediated lysosomal fusion and the formation of autophagosomes in L02 cells. The formation of autophagosomes was further confirmed by transmission electron microcopy. While T-2 toxin induced both autophagy and apoptosis, autophagy appears to be a leading event in the response to T-2 toxin treatment, reflecting its protective role in cells against cellular damage. Activating autophagy by rapamycin (RAPA) inhibited apoptosis, while suppressing autophagy by chloroquine greatly enhanced the T-2 toxin-induced apoptosis, suggesting the crosstalk between autophagy and apoptosis. Taken together, these results indicate that autophagy plays a role in protecting cells from T-2 toxin-induced apoptosis suggesting that autophagy may be manipulated for the alleviation of toxic responses induced by T-2 toxin.
语种：
英文
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Truncated Rep protein of porcine circovirus 2 (PCV2) caused by a naturally occurring mutation reduced virus replication in PK15 cells
作者：
Hu, Yi;Cai, Xiong;Zhan, Yang;Yuan, Xiaomin;Liu, Tanbin;...
期刊：
BMC Veterinary Research ,2019年15(1):1-10 ISSN：1746-6148
通讯作者：
Wang, Aibing;Guo, Shiyin
作者机构：
[Yang, Lingchen; Liu, Wei; Wang, Aibing; Tan, Lei; Li, Yalan; Yuan, Xiaomin; Hu, Yi; Liu, Tanbin] Hunan Agr Univ HUNAU, Coll Vet Med, Key Lab Anim Vaccine & Prot Engn, LAMFG, Changsha 410128, Hunan, Peoples R China.;[Cai, Xiong] Hunan Univ Chinese Med, Inst Innovat & Appl Res Chinese Med, Changsha 410208, Hunan, Peoples R China.;[Zhan, Yang; Wang, Naidong; Deng, Zhibang; Yang, Yi] HUNAU, Coll Vet Med, Key Lab Anim Vaccine & Prot Engn, LFP, Changsha 410128, Hunan, Peoples R China.;[Zhang, Lijie] Nankai Univ, State Key Lab Med Chem Biol, Tianjin 300071, Peoples R China.;[Zhang, Lijie] Nankai Univ, Coll Pharm, Tianjin 300071, Peoples R China.
通讯机构：
[Wang, Aibing; Guo, Shiyin] H;Hunan Agr Univ HUNAU, Coll Vet Med, Key Lab Anim Vaccine & Prot Engn, LAMFG, Changsha 410128, Hunan, Peoples R China.;HUNAU, Coll Food Sci & Technol, Changsha 410128, Hunan, Peoples R China.
关键词：
Porcine circovirus 2(PCV2);Truncated Rep protein;Infectious clone;Rescued viruses;Virus replication;PCV2 evolution
摘要：
Background: Porcine circovirus 2 (PCV2) is the causative agent of porcine circovirus-associated diseases (PCVADs). The infection of PCV2 is widespread and has serious consequence, thereby causing significant economic losses in the swine industry worldwide. Previously, we found that a strain named YiY-3-2-3 has a naturally occurring point mutation (G710 to A710) in ORF1 region, which leads to a shorten product of the rep gene (945 to 660 base pair). Importantly, the Rep protein is responsible for genome replication of PCV2. To explore the effects of this mutation on the PCV2 replication, in the current study we constructed infectious clone of this IF-YiY-3-2-3, as well as those of its two parental strains of IF-YiY-3-2-1 and IF-YiY-3-2-10. Subsequently, these infectious clones which have 1.1 copy of PCV2 genome of their corresponding strains were transfected into PK15 cells to obtain rescued viruses, respectively. Results: Though all of the three infectious clones could be rescued, the copy number and infectivity of these rescued viruses were significantly different, as analyzed by fluorescence quantitative PCR, Tissue culture infectious dose 50 (TCID50), and indirect immunofluorescence assay (IFA). Notably, whether the PCV2 copy number, viral titer or the infectivity of rescued viruses from infectious clone IF-YiY-3-2-3 was significantly less than those of its parental clones. Meanwhile, the spatial structure of the Rep protein from the IF-YiY-3-2-3 displayed an apparent truncation at the C-terminal. Conclusions: These findings therefore suggest that the Rep protein with truncated C-terminal would reduce virus replication and infectivity, and there might also exist both favorable and unfavorable mutations in the ORF1 of PCV2 in the process of its evolution. © 2019 The Author(s).
语种：
英文
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单颗粒示踪技术及其在动物病毒入侵机制研究中的应用
作者：
余婉婷;王乃东
期刊：
经济动物学报 ,2019年23(01):57-62 ISSN：1007-7448
作者机构：
湖南农业大学动物医学院,长沙410128;兽用蛋白质工程疫苗湖南省重点实验室,长沙410128;[王乃东; 余婉婷] 湖南农业大学
关键词：
单颗粒示踪技术;病毒;细胞亚结构;内吞;荧光标记
摘要：
病毒入侵宿主细胞的分子机制十分复杂,是病毒学界的研究热点。单颗粒示踪技术能够直接观察单个病毒入侵细胞的动态行为,有助于解析病毒入侵宿主细胞的分子机制。综述单颗粒示踪技术用于病毒与宿主细胞相互作用的不同过程,有助于动物病毒入侵机制研究和新型抗病毒药物与疫苗的研发。文中主要论述单颗粒示踪技术的发展历程,介绍该技术用于病毒和细胞亚结构的标记方法和进展,以及单颗粒示踪技术在动物病毒入侵机制研究中的应用进展,并总结现阶段单颗粒示踪技术在病毒学发展中的优势和局限性。
语种：
中文
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猪圆环病毒2d型的分子生物学特征和流行病学研究进展
作者：
黄坤;王东亮;邹亚文;王乃东;董彦鹏;...
期刊：
经济动物学报 ,2019年23(01):51-56 ISSN：1007-7448
作者机构：
湖南农业大学动物医学院;兽用蛋白质工程疫苗湖南省重点实验室;[董彦鹏] 江苏南农高科技股份有限公司;[湛洋; 王乃东; 黄坤; 王东亮; 邹亚文] 湖南农业大学
关键词：
猪圆环病毒2型;感染;核衣壳蛋白;流行病学
摘要：
猪圆环病毒2型(Porcine circovirus Type 2,PCV2)被认为是猪圆环病毒相关疾病(PCVAD)的主要病原体,分为多个基因型和亚型,具有很高的突变率,给猪场的疫病防控带来了巨大的挑战。虽然PCV2不同基因型间存在一定的交叉免疫保护作用,但是,在国内外多个接种了PCV2疫苗的猪群中,报道分离得到了突变型PCV2d毒株(又称PCV2b-1C,PCV2 mutant或m PCV2b)。为更好地防控PCV2d毒株的流行,文中就PCV2d的分子生物学特征和流行病学情况分别进行了综述。
语种：
中文
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Loop EF区嵌合猪细小病毒B细胞表位对猪圆环病毒2型病毒样颗粒组装的影响
作者：
张素姣;王东亮;李萌;蒋一凡;湛洋;...
期刊：
中国兽医科学 ,2019年49(2):176-182 ISSN：1673-4696
作者机构：
湖南农业大学动物医学院兽用蛋白质工程疫苗湖南省重点实验室兽用疫苗逆向创制湖南省工程研究中心,湖南长沙,410128;[蒋一凡; 湛洋; 李萌; 王乃东; 张素姣; 王东亮] 湖南农业大学
关键词：
猪圆环病毒2型;病毒样颗粒;外源表位;突变体蛋白
摘要：
Loop EF是猪圆环病毒2型(PCV2) Cap蛋白的重要Loop结构,然而它在PCV2组装过程中的作用仍不清楚.本研究通过模拟PCV2病毒样颗粒(VLPs) 3D结构,比较分析PCV2和PCV1、PCV3的氨基酸序列以及预测Loop EF的突变位点,筛选出PCV2 Loop EF中可供外源表位替换或插入的capsid表面位点,即第133位丙氨酸.探索性地针对该位点设计3个突变体,分别为猪细小病毒1型(PPV1)B细胞线性表位(228QQITDA233)插入PCV2 Cap第132和133位氨基酸、第133和134位氨基酸之间及替换第133位氨基酸.利用over-lap PCR等技术构建上述3个突变体的重组质粒,在大肠杆菌(BL21/DE3)中表达,并利用镍柱亲和层析纯化3个突变体蛋白,最后在体外进行VLPs组装,电镜结果显示3个突变体蛋白均可体外组装成完整的VLPs.结果 表明,Loop EF中第133位丙氨酸的替换或插入外源表位突变不影响VLPs组装,提示PCV2 VLPs Loop EF展示外源抗原表位具有可行性.
语种：
中文
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The protective effects of DL-Selenomethionine against T-2/HT-2 toxins-induced cytotoxicity and oxidative stress in broiler hepatocytes.
作者：
Yang, Lingchen;Tu, Di;Wang, Naidong;Deng, Zhibang;Zhan, Yang;...
期刊：
Toxicology in Vitro ,2019年54:137-146 ISSN：0887-2333
通讯作者：
Guo, Shiyin;Wang, Aibing
作者机构：
[Yang, Lingchen; Liu, Wei; Wang, Aibing; Tu, Di; Tan, Lei; Li, Yalan; Hu, Yi; Liu, Tanbin] Hunan Agr Univ HUNAU, LAMFG, Key Lab Anim Vaccine & Prot Engn, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.;[Wang, Naidong; Zhan, Yang; Deng, Zhibang] Hunan Agr Univ HUNAU, LFP, Key Lab Anim Vaccine & Prot Engn, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.;[Guo, Shiyin] HUNAU, Coll Food Sci & Technol, Changsha 410128, Hunan, Peoples R China.;[Guo, Shiyin; Wang, AB] Hunan Agr Univ, 1 Nongda Rd, Changsha 410128, Hunan, Peoples R China.
通讯机构：
[Guo, SY; Wang, AB] H;Hunan Agr Univ, 1 Nongda Rd, Changsha 410128, Hunan, Peoples R China.
关键词：
Cytotoxicity;DL-Selenomethionine;Hepatocytes;Oxidative stress;Protective effects;T-2/HT-2 toxins
摘要：
T-2 and HT-2 toxins can cause cytotoxicity and oxidative stress in animals, while DL-Selenomethionine plays an important role in preventing oxidative stress and improving cell viability. However, the role of DL-Selenomethionine in T-2/HT-2 toxins-induced cell damage is still unknown. In this study, we investigated whether DL-Selenomethionine plays a protective role against T-2/HT-2-induced cytotoxicity and oxidative stress in primary hepatocytes. Our results demonstrated that T-2/HT-2 toxins-exposed broiler hepatocytes exhibited significantly decreased cell viability and intracellular glutathione (GSH) concentration while increased Lacate dehydrogenase (LDH) leakage, intracellular reactive oxygen species (ROS), glutathione peroxidase (GSH-PX), malondialdehyde (MDA) and catalase (CAT) levels, as well as elevated expression levels of genes related to oxidative stress, in a toxin dose-dependent manner (P < 0.05). However, the application of DL-Selenomethionine into T-2/HT-2 treated hepatocytes effectively alleviated the adverse effects of T-2/HT-2, as demonstrated by increased cell viability, decreased LDH leakage, declined intracellular ROS and MDA levels, increased expression of oxidative stress-related genes, as well as accordingly enhanced activities of GSH, GSH-PX, SOD and CAT as compared to the control groups (P < 0.05). Therefore, our in vitro data demonstrate that DL-Selenomethionine can function as an effectively protective agent against T-2/HT-2-induced cytotoxicity and oxidative stress.
语种：
英文
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First molecular detection of porcine circovirus type 3 in dogs in China
作者：
Zhang, Jiaxin;Liu, Zhenguo;Zou, Yawen;Zhang, Nanxiangzi;Wang, Dongliang;...
期刊：
Virus Genes ,2018年54(1):140-144 ISSN：0920-8569
通讯作者：
Wang, Naidong;Jiang, Ping
作者机构：
[Yang, Linchen; Wang, Naidong; Wang, Dongliang; Tu, Di; Deng, Zhibang; Zhang, Jiaxin; Zou, Yawen; Zhang, Nanxiangzi; Yang, Yi] Hunan Agr Univ, Coll Vet Med, RCRV, Changsha 410128, Hunan, Peoples R China.;[Yang, Linchen; Wang, Naidong; Wang, Dongliang; Tu, Di; Deng, Zhibang; Zhang, Jiaxin; Zou, Yawen; Zhang, Nanxiangzi; Yang, Yi] Hunan Agr Univ, Coll Vet Med, LFP, Changsha 410128, Hunan, Peoples R China.;[Liu, Zhenguo] Cent S Univ, Dept Infect Dis, Xiangya Hosp 3, Changsha 410013, Hunan, Peoples R China.;[Jiang, Ping] Nanjing Agr Univ, Coll Vet Med, Key Lab Anim Dis Diagnost & Immunol, Minist Agr, Nanjing 210095, Jiangsu, Peoples R China.
通讯机构：
[Wang, Naidong] H;[Jiang, Ping] N;Hunan Agr Univ, Coll Vet Med, RCRV, Changsha 410128, Hunan, Peoples R China.;Hunan Agr Univ, Coll Vet Med, LFP, Changsha 410128, Hunan, Peoples R China.;Nanjing Agr Univ, Coll Vet Med, Key Lab Anim Dis Diagnost & Immunol, Minist Agr, Nanjing 210095, Jiangsu, Peoples R China.
关键词：
Porcine circovirus 3;Molecular detection;Genetic diversity;Canine
摘要：
Porcine circovirus type 3 (PCV3) has recently been isolated from diseased pigs within the USA. The objective was to detect the presence of PCV3 in dogs. Nested polymerase chain reactions (PCR) with PCV3-specific primers for the capsid gene were used to detect PCV3 genomic DNA in serum samples from dogs (n = 44) in China. There was PCV3 DNA detected in 4 of 44 dogs [all were negative for PCV2 and canine circovirus (CanineCV)]. Based on sequence analysis, positive sequences were grouped into PCV3 genotypes. However, these isolates had close evolutionary relationships with FoxCV (KP941114) and CanineCV (JQ821392). Further investigations of the epidemiology, evolutionary biology, and pathobiology of PCV3 to dogs are warranted.
语种：
英文
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犬圆环病毒检测和致病性的研究进展
作者：
张南乡子;张佳鑫;邹亚文;杨凌宸;屠迪;...
期刊：
经济动物学报 ,2018年22(01):53-56 ISSN：1007-7448
作者机构：
湖南农业大学动物医学院,长沙,410128;[张南乡子; 王乃东; 张佳鑫; 杨凌宸; 屠迪; 邹亚文] 湖南农业大学
关键词：
犬圆环病毒;出血性腹泻;脉管炎;致病性
摘要：
犬圆环病毒（Canine circovirus,Canine CV）是2012年首次在美国报道的一种新型哺乳动物圆环病毒。之后,意大利、德国和中国陆续报道检测到该病毒。目前已证实Canine CV可引起犬发生坏死性血管炎和淋巴结肉芽肿,临床表现为出血性腹泻和呕吐等,但该病毒的致病机理尚不清楚。文中主要对Canine CV的检测和致病性研究进展进行综述,为Canine CV的流行病学调查和致病机理研究等提供参考。
语种：
中文
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