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Indirect EliSA with Recombinant GP5 for Detecting Antibodies to Porcine Reproductive and Respiratory Syndrome Virus

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Virologica Sinica 2011年第1期26卷 61-66页

作者： yan Chen hong tian Jian-hui He jin-yin wu you-jun shang xiang-tao liu Lanzhou Veterinary Research Institute Chinese Academy of Agricultural Sciences State Key Laboratory of Veterinary Etiological Biology Key Laboratory of Animal Virology of Ministry of Agriculture Lanzhou 730046 China

Porcine reproductive and respiratory syndrome is caused by the PRRS virus (PRRSV),which has six structural proteins (GP2,GP3,GP4,GP5,M and N). GP5 and N protein are important targets for serological detection by enzyme-linked immunosorbent assay (EliSA) and other methods. Toward this goal,we developed an indirect EliSA with recombinant GP5 antigens and this method was validated by comparison to the LSI PRRSV-Ab EliSA kit. The results indicated that the optimal concentration of coated recombinant antigen was 0.2 μg/well for a serum dilution of 1:40. The rate of agreement with the LSI PRRSV-Ab kit was 88.7% (266/300). These results support the potential use of recombinant GP5 as an antigen for indirect EliSA to detect PRRSV antibodies in pigs.

关键词： Porcine reproductive and respiratory syndromevirus (PRRSV) GP5 protein Indirect EliSA

Genetic Variation Analyses of nsp2 Gene of PRRSV in Ningxia hui Autonomous Region of China

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Virologica Sinica 2009年第3期24卷 221-226页

作者： hong tian jing-yan wu shuang-hui yin you-jun shang Zi-ping MAN Na ZHAO Ye jin xiang-tao liu Key Laboratory of Animal Virology of Ministry of Agriculture State Key Laboratory of Veterinary Etiologic Biology Lanzhou Veterinary Research Institute Chinese Academy of Agricultural Sciences Lanzhou 730046 China

To gain a better understanding of the genetic diversity and evolution of PRRSV in the Ningxia hui Nationality Autonomous Region(Ningxia) of China,the nsp2 genes from a series of PRRSV strains collected from the region in 2007 were partially sequenced. These sequences were then analyzed along with the classical strain(ch-1a) and two other epidemic strains SD(3) and SD2006. Comparison of the nucleotide sequence with ch-1a indicated that nsp2 genes of seventeen Ningxia isolates(NX strain) have deletions of 87 nucleotides. Sequence analysis indicated that homology between the Ningxia strain and ch-1a was 60.3%-79.9% in the nucleotide sequence,and homology between the NX strains and SD strains was 80.3%-98.8% in the nucleotide sequence. The nsp2 genes of the seventeen isolates had 74.9%-100% nucleotide sequence identities with each other. This study was undertaken to assess the regional variation of prevalent PRRSV and to establish a sequence database for PRRSV molecular epidemiological studies.

关键词： PRRSV nsp2 gene Homology analysis PRRSV Ningxia isolates (NX strain)

Discrepancy of Classical Swine Fever Virus in Different Tissues by One-Step RT-PCR and Nested RT-PCR

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Animal Husbandry and Feed Science 2010年第3期2卷 18-20页

作者： yin shuang-hui tian hong shang you-jun yang shun-li wu jin-yan liu xiang-tao Lanzhou Veterinary Research Institute Chinese Academy of Agricultural SciencesState Key Laboratory of Veterinary Etiological Biologyand Key Laboratory of Animal Virology of the Ministry of AgricultureLanzhou 730046China

Reverse transcription polymerase chain reaction (RT-PCR) was used for the detection of classical swine fever virus (CSFV) in blood and tissue samples of field cases and experimentally inoculated pigs. The distribution of CSFV in different organ samples showed some discrepancies in infected pigs. Four weaner pigs were inoculated with C-strain vaccine virus, then samples of spleen, tonsil, lung, mesenteric lymph node, kidney and brain were collected after slaughter and tested for E2 and NS5B genes using one-step RT-PCR and nested RT-PCR. Using the same method, 12 field cases were simultaneously studied. A discrepancy of CSFV in different samples was found upon detecting the target gene. The most reliable diagnostic organs were spleen and tonsil, and the nested RT-PCR assay provided a highly sensitive and specific method with comparable performance to the one-step RT-PCR assay.

关键词： Classical swine fever virus E2 gene NS5B gene Tissues RT-PCR

Progressive liver injury and increased mortality risk in COVID-19 patients:A retrospective cohort study in China

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World Journal of Gastroenterology 2021年第9期27卷 835-853页

作者： Shui-Sheng Zhang li Dong Gao-Ming Wang Yuan tian Xiao-Fang Ye Yue Zhao Zheng-yin liu Jia-Yu Zhai Zhi-ling Zhao jun-hong Wang hui-Min Zhang Xiao-Long li Chang-Xin wu cai-Ting yang li-Juan yang Hai-Xia Du hui Wang Qing-Gang Ge Dian-Rong Xiu Ning Shen Department of General Surgery Peking University Third HospitalBeijing 100191China Institutes of Biomedical Sciences Shanxi UniversityTaiyuan 030006Shanxi ProvinceChina Department of Radiotherapy Oncology Shandong Provincial Qianfoshan HospitalThe First Hospital Affiliated with Shandong First Medical UniversityJinan 250014Shandong ProvinceChina Department of Respiratory and Critical Medicine Beijing Anzhen HospitalCapital Medical UniversityBeijing 100029China Department of Infectious Diseases Peking Union Medical College HospitalChinese Academy of Medical SciencesBeijing 100005China Department of Rheumatology Peking University Third HospitalBeijing 100191China Department of Critical Care Medicine Peking University Third HospitalBeijing 100191China Department of Emergency Medicine Peking University Third HospitalBeijing 100191China Department of Cardiac Surgery Peking University Third HospitalBeijing 100191China Department of Urology Peking University Third HospitalBeijing 100191China Department of Gynecology Peking University Third HospitalBeijing 100191China Department of Rehabilitation Peking University Third HospitalBeijing 100191China Department of Gastroenterology Peking University Third HospitalBeijing 100191China Department of Respiratory and Critical Medicine Peking University Third HospitalBeijing 100191China

BACKGROUND liver injury is common and also can be fatal,particularly in severe or critical patients with coronavirus disease 2019(COVID-19).AIM To conduct an in-depth investigation into the risk factors for liver injury and into the effective measures to prevent subsequent mortality *** A retrospective cohort study was performed on 440 consecutive patients with relatively severe COVID-19 between January 28 and March 9,2020 at Tongji Hospital,wuhan,*** on clinical features,laboratory parameters,medications,and prognosis were *** COVID-19-associated liver injury more frequently occurred in patients aged≥65 years,female patients,or those with other comorbidities,decreased lymphocyte count,or elevated D-dimer or serum ferritin(Pliver injury(severe patients:Odds ratio[OR]=2.86,95%confidence interval[CI]:1.78-4.59;critical patients:OR=13.44,95%CI:7.21-25.97).The elevated levels of on-admission aspartate aminotransferase and total bilirubin indicated an increased mortality risk(Pliver *** drugs tended to be of assistance to treat the liver injury and improve the prognosis of patients with COVID-19-associated liver *** More intensive monitoring of aspartate aminotransferase or total bilirubin is recommended for COVID-19 patients,especially patients aged≥65 years,female patients,or those with other *** hepatotoxicity of antibiotics and intravenous nutrition should be alert for COVID-19 patients.

关键词： COVID-19 liver injury Prognosis Risk factors Drugs Alanine aminotransferase

The first data release(DR1) of the LAMOST regular survey

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Research in Astronomy and Astrophysics 2015年第8期15卷 1095-1124页

作者： A-li Luo Yong-Heng Zhao Gang Zhao li-cai Deng Xiao-Wei liu Yi-peng jing Gang Wang Hao-Tong Zhang Jian-Rong Shi xiang-Qun Cui Yao-Quan Chu Guo-Ping li Zhong-Rui Bai Yue wu yan cai Shu-Yun Cao Zi-Huang Cao Jeffrey L.Carlin Hai-Yuan Chen Jian-jun Chen Kun-Xin Chen li Chen xue-Lei Chen Xiao-yan Chen ying Chen Norbert Christlieb Jia-Ru Chu Chen-Zhou Cui Yi-Qiao Dong Bing Du Dong-Wei Fan Lei Feng Jian-Ning Fu peng Gao xue-Fei Gong Bo-Zhong Gu yan-Xin Guo Zhan-Wen Han Bo-liang He jin-liang Hou Yong-hui Hou Wen Hou hong-Zhuan Hu Ning-Sheng Hu Zhong-Wen Hu Zhi-ying Huo Lei Jia Fang-Hua Jiang xiang Jiang Zhi-Bo Jiang Ge jin Xiao Kong Xu Kong Ya-Juan Lei Ai-Hua li Chang-Hua li Guang-Wei li Hai-Ning li Jian li Qi li shuang li Sha-Sha li Xin-Nan li yan li yin-Bi li Ye-Ping li Yuan liang Chien-Cheng lin Chao liu Gen-Rong liu Guan-Qun liu Zhi-Gang liu Wen-Zhi Lu Yu Luo yin-Dun Mao Heidi Newberg Ji-jun Ni Zhao-xiang Qi Yong-jun Qi Shi-yin Shen Huo-Ming Shi jing Song Yi-Han Song Ding-Qiang Su hong-jun Su Zheng-hong Tang Qing-Sheng tao Yuan tian Dan Wang Da-Qi Wang Feng-Fei Wang Guo-Min Wang Hai Wang hong-Chi Wang Jian Wang Jia-Ning Wang Jian-ling Wang Jian-Ping Wang jun-Xian Wang Lei Wang Meng-Xin Wang Shou-Guan Wang Shu-Qing Wang Xia Wang Ya-Nan Wang you Wang Yue-Fei Wang you-Fen Wang peng Wei Ming-Zhi Wei hong wu Ke-Fei wu xue-Bing wu Yu-Zhong wu Xiao-Zheng Xing ling-Zhe Xu Xin-Qi Xu yan Xu Tai-Sheng yan De-Hua yang Hai-Feng yang hui-Qin yang Ming yang Zheng-Qiu Yao Yong Yu hui Yuan Hai-Bo Yuan Hai-Long Yuan Wei-Min Yuan Chao Zhai En-peng Zhang Hua-Wei Zhang Jian-Nan Zhang li-Pin Zhang Wei Zhang Yong Zhang yan-Xia Zhang Zheng-Chao Zhang Ming Zhao Fang Zhou Xu Zhou Jie Zhu Yong-tian Zhu Si-Cheng Zou Fang Zuo National Astronomical Observatories Chinese Academy of Sciences Department of Astronomy Peking University shanghai Astronomical Observatory Chinese Academy of Sciences Nanjing Institute of Astronomical Optics & Technology National Astronomical ObservatoriesChinese Academy of Sciences University of Science and Technology of China Department of Physics Applied Physics and AstronomyRensselaer Polytechnic InstituteTroyNY 12180USA Department of Physics and Astronomy Earlham CollegeRichmondIN 47374USA Universitt Heidelberg Zentrum für AstronomieLandessternwarteKnigstuhl 1269117 HeidelbergGermany Department of Astronomy Beijing Normal University Yunnan Observatories Chinese Academy of Sciences Purple Mountain Observatory Chinese Academy of Sciences

The Large sky Area Multi-Object Fiber Spectroscopic Telescope（LAMOST） general survey is a spectroscopic survey that will eventually cover approximately half of the celestial sphere and collect 10 million spectra of stars, galaxies and QSOs. Objects in both the pilot survey and the first year regular survey are included in the LAMOST DR1. The pilot survey started in October 2011 and ended in june 2012, and the data have been released to the public as the LAMOST Pilot Data Release in August 2012. The regular survey started in September 2012, and completed its first year of operation in june 2013. The LAMOST DR1 includes a total of 1202 plates containing 2 955 336 spectra, of which 1 790 879 spectra have observed signalto-noise ratio（SNR） ≥ 10. All data with SNR ≥ 2 are formally released as LAMOST DR1 under the LAMOST data policy. This data release contains a total of 2 204 696 spectra, of which 1 944 329 are stellar spectra, 12 082 are galaxy spectra and 5017 are quasars. The DR1 not only includes spectra, but also three stellar catalogs with measured parameters： late A,FGK-type stars with high quality spectra（1 061 918 entries）, A-type stars（100 073 entries）, and M-type stars（121 522 entries）. This paper introduces the survey design, the observational and instrumental limitations, data reduction and analysis, and some caveats. A description of the FITS structure of spectral files and parameter catalogs is also provided.

关键词： techniques: spectroscopic survey—data release—catalog

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猪瘟重组蛋白E2的兔体免疫保护试验

华北农学报 2009年第B12期24卷 225-228页

作者：田宏吴锦艳尹双辉尚佑军郑海学刘湘涛中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室农业部畜禽病毒学重点实验室甘肃兰州730046

鉴于猪瘟病毒不能使家兔发病,但能使之产生免疫原性,而且猪瘟兔化弱度苗能在兔体的定型热反应。本研究选用家兔为试验模型,将自制的亚单位疫苗用弗氏佐剂乳化后免疫家兔,持续一定的免疫周期后,用猪瘟活疫苗(II)攻击免疫及对照试验家兔... 详细信息

鉴于猪瘟病毒不能使家兔发病,但能使之产生免疫原性,而且猪瘟兔化弱度苗能在兔体的定型热反应。本研究选用家兔为试验模型,将自制的亚单位疫苗用弗氏佐剂乳化后免疫家兔,持续一定的免疫周期后,用猪瘟活疫苗(II)攻击免疫及对照试验家兔。根据T淋巴细胞增殖试验、阻断EliSA及攻毒保护试验对所制备的抗原进行了全面的评价。结果表明,该免疫原具有良好的免疫原性,而且能保护免疫家兔为4/4。可以进行下一步的开发和利用价值。以期该疫苗能为猪瘟的防治做出贡献。

关键词：猪瘟病毒亚单位疫苗 E2蛋白免疫试验

猪圆环病毒2型和猪生殖与呼吸综合征病毒混合感染的检测

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中国兽医科学 2009年第8期39卷 702-706页

作者：蔡承如尚佑军蔺芳田宏尹双辉吴锦艳杨顺利刘湘涛甘肃农业大学动物医学院甘肃兰州730070 中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室甘肃兰州730046

对采自甘肃省天水市某猪场的24份病猪组织器官样品(肺、脾、肾和淋巴结)进行了病毒分离,同时设计特异性引物,建立了分别检测猪圆环病毒2型(PCV-2)和猪生殖与呼吸综合征病毒(PRRSV)的PCR和RT-PCR方法,应用EliSA方法检测了血清中PCV-2及PR... 详细信息

对采自甘肃省天水市某猪场的24份病猪组织器官样品(肺、脾、肾和淋巴结)进行了病毒分离,同时设计特异性引物,建立了分别检测猪圆环病毒2型(PCV-2)和猪生殖与呼吸综合征病毒(PRRSV)的PCR和RT-PCR方法,应用EliSA方法检测了血清中PCV-2及PRRSV抗体,以调查该猪场PCV-2和PRRSV混合感染的情况。结果显示,有14份样品均扩增出了PCV-2和PRRSV的目的基因片段,且PCV-2及PRRSV抗体均呈阳性。证实,该猪场出现了PCV-2和PRRSV的混合感染,阳性率为58.33%。

关键词：猪圆环病毒2型猪生殖与呼吸综合征病毒混合感染检测

Timing analysis of the black hole candidate EXO 1846–031 with Insight-HXMT monitoring

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Research in Astronomy and Astrophysics 2021年第3期21卷 193-204页

作者： He-Xin liu Yue Huang Guang-Cheng Xiao Qing-Cui Bu jin-Lu Qu Shu Zhang shuang-Nan Zhang Shu-Mei Jia Fang-jun Lu xiang Ma lian tao Wei Zhang li Chen li-Ming Song Ti-Pei li Yu-peng Xu xue-Lei Cao Yong Chen Cong-Zhan liu Ce cai Zhi Chang Gang Chen tian-xiang Chen Yi-Bao Chen Yu-peng Chen Wei Cui Wei-Wei Cui jing-Kang Deng Yong-Wei Dong Yuan-Yuan Du Min-xue Fu Guan-Hua Gao He Gao Min Gao Ming-Yu Ge Yu-Dong Gu Ju Guan Cheng-Cheng Guo Da-Wei Han Jia Huo Lu-Hua Jiang Wei-Chun Jiang jing jin Yong-Jie jin ling-Da Kong Bing li Cheng-Kui li Gang li Mao-shun li Wei li Xian li Xiao-Bo li Xu-Fang li yang-Guo li Zheng-Wei li Xiao-Hua liang jing-Yuan liao Bai-Sheng liu Guo-Qing liu hong-Wei liu Xiao-jing liu Yi-Nong liu Bo Lu xue-Feng Lu Qi Luo tao Luo Bin Meng Yi Nang Jian-yin Nie Ge Ou Na Sai Ren-Cheng shang Xin-ying Song liang Sun ying Tan you-li Tuo Chen Wang Guo-Feng Wang Juan Wang ling-jun Wang Weng-Shuai Wang Yu-Sa Wang xiang-yang Wen Bai-yang wu Bo-Bing wu Mei wu Shuo Xiao Shao-lin Xiong He Xu Jia-Wei yang Sheng yang yan-Ji yang Yi-jung yang Qi-Bin Yi Qian-Qing yin Yuan you Ai-Mei Zhang Cheng-Mo Zhang Fan Zhang hong-Mei Zhang Juan Zhang Tong Zhang Wan-Chang Zhang Wen-Zhao Zhang Yi Zhang Yi-Fei Zhang Yong-Jie Zhang Yuan-Hang Zhang Yue Zhang Zhao Zhang Zhi Zhang Zi-liang Zhang Hai-Sheng Zhao Xiao-Fan Zhao Shi-Jie Zheng Yao-Guang Zheng Deng-Ke Zhou Jian-Feng Zhou Yu-Xuan Zhu Ren-lin Zhuang Yue Zhu Key Laboratory of Particle Astrophysics Institute of High Energy PhysicsChinese Academy of SciencesBeijing 100049China University of Chinese Academy of Sciences Chinese Academy of SciencesBeijing 100049China Institut fur Astronomie und Astrophysik Kepler Center for Astro and Particle PhysicsEberhard Karls Universität72076 TubingenGermany Department of Astronomy Beijing Normal UniversityBeijing 100088China Department of Astronomy Tsinghua UniversityBeijing 100084China Department of Physics Tsinghua UniversityBeijing 100084China Department of Engineering Physics Tsinghua UniversityBeijing 100084China Key Laboratory of Space Astronomy and Technology National Astronomical ObservatoriesChinese Academy of SciencesBeijing 100101China College of Physics Sciences&Technology Hebei UniversityBaoding 071002Hebei ProvinceChina

We present the observational results from a detailed timing analysis of the black hole candidate EXO 1846-031 during its outburst in 2019 with the observations of Insight-HXMT,NICER and *** outburst can be classified roughly into four different ***-C quasi-periodic oscillations(QPOs)observed by NICER(about 0.1-6 Hz)and Insight-HXMT(about 0.7-8 Hz)are also reported in this ***,we study various physical quantities related to QPO *** QPO rms-frequency relationship in the energy band 1-10 keV indicates that there is a turning pointing in frequency around2 Hz,which is similar to that of GRS 1915+105.A possible hypothesis for the relationship above may be related to the inclination of the source,which may require a high inclination to explain *** relationships between QPO frequency and QPO rms,hardness,total fractional rms and count rate have also been found in other transient sources,which can indicate that the origin of type-C QPOs is non-thermal.

关键词： X-ray binaries black hole candidate QPO EXO 1846–031

口蹄疫病毒OH株结构蛋白基因VP1的克隆与表达

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中国兽医科学 2006年第7期36卷 515-518页

作者：龚真莉刘湘涛尚佑军田宏吴锦艳尹双辉陈国栋中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室农业部畜禽病毒学重点开放实验室

采用RT-PCR方法扩增获得了O型口蹄疫病毒的主要免疫原VP1基因,将其插入pMD18-T载体进行序列分析,结果表明,所获得的基因片段含有完整的FMDV结构蛋白VP1编码区。根据表达载体pQE-Trisystem的克隆位点序列和该VPl基因片段的末端序列设计了... 详细信息

采用RT-PCR方法扩增获得了O型口蹄疫病毒的主要免疫原VP1基因,将其插入pMD18-T载体进行序列分析,结果表明,所获得的基因片段含有完整的FMDV结构蛋白VP1编码区。根据表达载体pQE-Trisystem的克隆位点序列和该VPl基因片段的末端序列设计了1对表达引物,以重组pMD-T-VP1阳性质粒为模板,扩增获得了VP1基因,通过酶切将其克隆至表达载体pQE-Trisystem上。经测序证实,重组表达质粒所含的外源基因VP1编码框正确无误。将重组表达质粒pQE-VP1转化至大肠埃希氏菌M15,通过IPTG诱导促使VP1基因高效表达,SDS- PAGE和Western-blot分析表明,表达产物大小与预期的结果(26 ku)一致,且具有良好的反应原性。以2 mmol／L IPTG诱导表达5 h时表达量最高,其中70％-80％的目的蛋白存在于菌体裂解后的上清中,表明外源基因VP1主要以可溶性方式表达。

关键词：口蹄疫病毒 VP1基因基因克隆表达