分离新型鸭源微RNA病毒进行全基因组测序并进行遗传进化分析。对本实验室2021年不同来源的种鸭和肉鸭病料进行PCR检测,初步确定存在一种未知分类的新型微RNA病毒感染。取病死鸭病料组织处理后接种SPF鸡胚分离病毒,设计引物对分离到的病...分离新型鸭源微RNA病毒进行全基因组测序并进行遗传进化分析。对本实验室2021年不同来源的种鸭和肉鸭病料进行PCR检测,初步确定存在一种未知分类的新型微RNA病毒感染。取病死鸭病料组织处理后接种SPF鸡胚分离病毒,设计引物对分离到的病毒进行PCR检测,通过重叠PCR方法进行全基因组扩增测序。将分离病毒各蛋白氨基酸序列两两比对,同时选取GenBank数据库中微RNA病毒代表毒株序列绘制系统进化树,并对主要蛋白P1、2C、3D序列比对分析。结果显示:共分离到三株微RNA病毒,分别命名为21101株、21016株和21075株(GenBank登录号:OQ927377~OQ927379)。基因组长度分别为7445、7445和7447 bp,均包含一个编码2141个氨基酸的开放阅读框(ORF),可划分为P1、P2、P3三个部分,符合微RNA病毒序列特征。基于全基因组序列遗传进化分析发现,三株分离病毒与本实验室前期分离的Duck/FC22/China/2017(GenBank登录号:MN102111)毒株及上海兽医研究所分离的Duck/AH15/CHN/2015(GenBank登录号:MT681985)位于同一分支,与鸭甲型肝炎病毒(Duck hepatitis A virus,DHAV)遗传距离最近。分离的三株鸭源微RNA病毒进行全基因组测序及遗传进化分析发现,与目前已知的两株微RNA毒株为同一类新型鸭源微RNA病毒。展开更多
采用荧光显微技术,对2006年长江口及近海水域20个站点的表层及10m层或潜水体冬、春两季的浮游病毒丰度进行了检测,对浮游病毒丰度在季节(冬、春两季)、水平分布和垂直分布上的变化进行了探讨。调查区浮游病毒丰度在冬、春季节上并无明...采用荧光显微技术,对2006年长江口及近海水域20个站点的表层及10m层或潜水体冬、春两季的浮游病毒丰度进行了检测,对浮游病毒丰度在季节(冬、春两季)、水平分布和垂直分布上的变化进行了探讨。调查区浮游病毒丰度在冬、春季节上并无明显差异,但在水平分布上存在很大差异,河口区浮游病毒直接检测量(Virus Direct Count,VDC)达到107个/ml,近海水域VDC为106个/ml,河口区的浮游病毒丰度都明显高于近海水域病毒丰度(P<0.01)。在垂直分布上,冬、春两季长江口水域水深小于10m的站位,表层浮游病毒丰度与底层病毒丰度无明显差别,水深大于10m的站位,表层水样的浮游病毒丰度都高于10m水层病毒丰度,说明长江口浮游病毒的垂直分布与站位总水深有关。还通过比较各站点VDC与叶绿素a含量的数据,分析了二者之间的相关性:冬季浮游病毒丰度与叶绿素a含量成正相关性;春季浮游病毒丰度与叶绿素a含量成负相关性,但病毒丰度受叶绿素a含量的影响仅为10%—11%。展开更多
In order to reveal the signaling pathways triggered by elicitor in plant-microbe interactions, the mechanisms of hypersensitive necrosis responses in Nicotiana tabacum L. cv. Gexin III induced by palmin were studied a...In order to reveal the signaling pathways triggered by elicitor in plant-microbe interactions, the mechanisms of hypersensitive necrosis responses in Nicotiana tabacum L. cv. Gexin III induced by palmin were studied at molecular and cellular level. The burst of superoxide, intercellular diffusion of hydrogen peroxide and process of cell death induced by palmin were investigated in tobacco plants by biochemical methods and Confocal microscopy. The results showed that a large amount of O-2(.-) was rapidly generated in tobacco cell elicited by palmin as a result of activation of NADPH oxidase, and the O-2(.-) was dismutated into H2O2 immediately by superoxide dismutase (SOD). Accumulation and intercellular diffusion of H2O2 were shown to be a trigger for hypersensitive cell death; and Ca2+ and some specific protein kinase were also shown to be involved in the activation of oxidative burst in tobacco cell induced by palmin.展开更多
MATLAB software and optimal complete subgraph algorithm were used to extract and reveal the microsatellite distribution features in the complete genomes of the tobacco vein clearing virus （NC-003 378.1） from the NCB...MATLAB software and optimal complete subgraph algorithm were used to extract and reveal the microsatellite distribution features in the complete genomes of the tobacco vein clearing virus （NC-003 378.1） from the NCBI database.The results showed that the repetitions number and their location of the N-base group has been extracted and displayed.The largest repetitions of N-base group in the complete genomes of the tobacco vein clearing virus was decreased as the exponential function with the increasing of N.The method used in this study could be applied to the extraction and revealing of the microsatellite distribution features in the complete genomes of other viruses,thereby provided a basis for the research of the structure and the law of function,inheritance and variation by the using of the microsatellite distribution features.展开更多
Crimean-Congo Haemorrhagic Fever Virus (CCHFV) is a tick-born virus of the Nairovirus genus within the Bunyaviridae family,which is widespread and causes high fatality.The nucleocapsid of CCHFV is comprised of N prote...Crimean-Congo Haemorrhagic Fever Virus (CCHFV) is a tick-born virus of the Nairovirus genus within the Bunyaviridae family,which is widespread and causes high fatality.The nucleocapsid of CCHFV is comprised of N proteins that are encoded by the S segment.In this research,the N protein of CCHFV was expressed in insect cells using a recombinant baculovirus.Under an electron microscope,Virus-Like Particles (VLPs) with various size and morphology were observed in cytoplasmic vesicles in the infected cells.Sucrose-gradient purification of the cell lysate indicated that the VLPs were mainly located in the upper fraction after ultracentrifugation,which was confirmed by Western blot analysis and immuno-electron microscopy (IEM).展开更多
In recent years, increasingly evidences show that autophagy plays an important role in the pathogenesis and development of liver diseases, and the relationship between them has increasingly become a focus of concern. ...In recent years, increasingly evidences show that autophagy plays an important role in the pathogenesis and development of liver diseases, and the relationship between them has increasingly become a focus of concern. Autophagy refers to the process through which the impaired organelles, misfolded protein, and intruding microorganisms is degraded by lysosomes to maintain stability inside cells. This article states the effect of autophagy on liver diseases （hepatic fibrosis, fatty liver, viral hepatitis, and liver cancer）, which aims to provide a new direction for the treatment of liver diseases.展开更多
AIM: To investigate the reactivity of a panel of 8 mouse anti-hepatitis B surface antigen (HBsAg) monoclonal antibodies (mAbs) using a collection of 9 recombinant HBsAg mutants with a variety of amino acid substitutio...AIM: To investigate the reactivity of a panel of 8 mouse anti-hepatitis B surface antigen (HBsAg) monoclonal antibodies (mAbs) using a collection of 9 recombinant HBsAg mutants with a variety of amino acid substitutions mostly located within the “a” region.METHODS: The entire HBs genes previously cloned into a mammalian expression vector were transiently transfected into COS7 cells. Two standard unmutated sequences of the ayw and adw subtypes served as controls. Secreted mutant proteins were collected and measured by three commercial diagnostic immunoassays to assess transfection efficiency. Reactivity of anti-HBs mAbs with mutated HBsAgs was determined by sandwich enzyme-linked immunosorbent assay (ELISA).RESULTS: Reactivity of anti-HBs mAbs with mutated HBsAgs revealed different patterns. While three mutants reacted strongly with all mAbs, two mutants reacted weakly with only two mAbs and the remaining proteins displayed variable degrees of reactivity towards different mAbs. Accordingly, four groups of mAbs with different but overlapping reactivity patterns could be envisaged. One group consisting of two mAbs (37C5-S7 and 35C6-S11) was found to recognize stable linear epitopes conserved in all mutants. Mutations outside the “a” determinant at positions 120 (P→S), 123(T→N) and 161(M→T) were found to affect reactivity of these mAbs.CONCLUSION: Our findings could have important implications for biophysical studies, vaccination strategies and immunotherapy of hepatitis B virus (HBV) mutants.展开更多
Patients who are chronically infected with the hepatitis C virus often develop chronic liver disease and assessment of the severity of liver injury is required prior to considering viral eradication therapy. This arti...Patients who are chronically infected with the hepatitis C virus often develop chronic liver disease and assessment of the severity of liver injury is required prior to considering viral eradication therapy. This article examines the various assessment methods currently available from gold standard liver biopsy to serological markers and imaging. Ultrasound is one of the most widely used imaging modalities in clinical practice and is already a first-line diagnostic tool for liver disease. Microbubble ultrasound contrast agents allow higher resolution images to be obtained and functional assessments of microvascular change to be carried out. The role of these agents in quantifying the state of hepatic injury is discussed as a viable method of determining the stage and grade of liver disease in patients with hepatitis C. Although currently confined to specialist centres, the availability of microbubble contrast-enhanced ultrasound will inevitably increase in the clinical setting.展开更多
Objective.To test whether intramuscular,intranasal,intrarectal and intravagina l administration of HPV6b L 1 virus-like particlescould induce immune response in mice and to as sess whether intra-muscular and mucosal v...Objective.To test whether intramuscular,intranasal,intrarectal and intravagina l administration of HPV6b L 1 virus-like particlescould induce immune response in mice and to as sess whether intra-muscular and mucosal vaccination against HPV is feasible.Me thods.HPV6b L1proteins self-assembled into VLPs in Sf-9cell in vitro.Mic e were immunized on day0and21with50ìg HPV6b L1VLPs intramuscularly,int ranasally,intrarectally and intravagi-nally respectively.Sera were collected for testing IgG titer after a further7days and3months respec-tively.Results .After immunizations,all mice developed significant anti-HPV6b L1antibody titers in serum by7days after the second immunization.The titer of the serum I gG antibody against HPV6b L1VLPs in the intramuscularly immunized group was h igher than that in the intranasally,intrarectally and intravaginally immunized groups respectively,indicating that both muscular and mucosal administration of HPV6b L1VLPs can stimulate a systemic HPV-specific antibody response.Sera of the mice in the in-tramuscularly immunized group still maintained a high tit er of the serum IgG antibody against HPV6b L1VLPs 3months after the immunizat ion.Conclusion.The results demonstrated that the HPV6b L1VLPs maintain stro ng antigenicity.Immu-nization with HPV6b L1VLPs via intramuscular and mucos al routes,without adjuvant ,can elicit spe-cific antibody in sera.These fin dings suggest that the VLPs are able to induce protective antibodies.展开更多
文摘分离新型鸭源微RNA病毒进行全基因组测序并进行遗传进化分析。对本实验室2021年不同来源的种鸭和肉鸭病料进行PCR检测,初步确定存在一种未知分类的新型微RNA病毒感染。取病死鸭病料组织处理后接种SPF鸡胚分离病毒,设计引物对分离到的病毒进行PCR检测,通过重叠PCR方法进行全基因组扩增测序。将分离病毒各蛋白氨基酸序列两两比对,同时选取GenBank数据库中微RNA病毒代表毒株序列绘制系统进化树,并对主要蛋白P1、2C、3D序列比对分析。结果显示:共分离到三株微RNA病毒,分别命名为21101株、21016株和21075株(GenBank登录号:OQ927377~OQ927379)。基因组长度分别为7445、7445和7447 bp,均包含一个编码2141个氨基酸的开放阅读框(ORF),可划分为P1、P2、P3三个部分,符合微RNA病毒序列特征。基于全基因组序列遗传进化分析发现,三株分离病毒与本实验室前期分离的Duck/FC22/China/2017(GenBank登录号:MN102111)毒株及上海兽医研究所分离的Duck/AH15/CHN/2015(GenBank登录号:MT681985)位于同一分支,与鸭甲型肝炎病毒(Duck hepatitis A virus,DHAV)遗传距离最近。分离的三株鸭源微RNA病毒进行全基因组测序及遗传进化分析发现,与目前已知的两株微RNA毒株为同一类新型鸭源微RNA病毒。
文摘采用荧光显微技术,对2006年长江口及近海水域20个站点的表层及10m层或潜水体冬、春两季的浮游病毒丰度进行了检测,对浮游病毒丰度在季节(冬、春两季)、水平分布和垂直分布上的变化进行了探讨。调查区浮游病毒丰度在冬、春季节上并无明显差异,但在水平分布上存在很大差异,河口区浮游病毒直接检测量(Virus Direct Count,VDC)达到107个/ml,近海水域VDC为106个/ml,河口区的浮游病毒丰度都明显高于近海水域病毒丰度(P<0.01)。在垂直分布上,冬、春两季长江口水域水深小于10m的站位,表层浮游病毒丰度与底层病毒丰度无明显差别,水深大于10m的站位,表层水样的浮游病毒丰度都高于10m水层病毒丰度,说明长江口浮游病毒的垂直分布与站位总水深有关。还通过比较各站点VDC与叶绿素a含量的数据,分析了二者之间的相关性:冬季浮游病毒丰度与叶绿素a含量成正相关性;春季浮游病毒丰度与叶绿素a含量成负相关性,但病毒丰度受叶绿素a含量的影响仅为10%—11%。
文摘In order to reveal the signaling pathways triggered by elicitor in plant-microbe interactions, the mechanisms of hypersensitive necrosis responses in Nicotiana tabacum L. cv. Gexin III induced by palmin were studied at molecular and cellular level. The burst of superoxide, intercellular diffusion of hydrogen peroxide and process of cell death induced by palmin were investigated in tobacco plants by biochemical methods and Confocal microscopy. The results showed that a large amount of O-2(.-) was rapidly generated in tobacco cell elicited by palmin as a result of activation of NADPH oxidase, and the O-2(.-) was dismutated into H2O2 immediately by superoxide dismutase (SOD). Accumulation and intercellular diffusion of H2O2 were shown to be a trigger for hypersensitive cell death; and Ca2+ and some specific protein kinase were also shown to be involved in the activation of oxidative burst in tobacco cell induced by palmin.
基金Supported by the Eleventh Five-year Development Planning Project for Instructional Science in Hubei Province (2006B131)~~
文摘MATLAB software and optimal complete subgraph algorithm were used to extract and reveal the microsatellite distribution features in the complete genomes of the tobacco vein clearing virus （NC-003 378.1） from the NCBI database.The results showed that the repetitions number and their location of the N-base group has been extracted and displayed.The largest repetitions of N-base group in the complete genomes of the tobacco vein clearing virus was decreased as the exponential function with the increasing of N.The method used in this study could be applied to the extraction and revealing of the microsatellite distribution features in the complete genomes of other viruses,thereby provided a basis for the research of the structure and the law of function,inheritance and variation by the using of the microsatellite distribution features.
基金The Knowledge Innovation Program of the Chinese Academy of Sciences,Grant No.KSCX2-YW-N-065the Knowledge Innovation Program of the Chinese Academy of Sciences,Grant No.KSCX2-EW-G-8+1 种基金National Key Basic Research Program(973Program),Grant No.2010CB530103National Basic Research Priorities Program of China,Grant No.2007FY210700
文摘Crimean-Congo Haemorrhagic Fever Virus (CCHFV) is a tick-born virus of the Nairovirus genus within the Bunyaviridae family,which is widespread and causes high fatality.The nucleocapsid of CCHFV is comprised of N proteins that are encoded by the S segment.In this research,the N protein of CCHFV was expressed in insect cells using a recombinant baculovirus.Under an electron microscope,Virus-Like Particles (VLPs) with various size and morphology were observed in cytoplasmic vesicles in the infected cells.Sucrose-gradient purification of the cell lysate indicated that the VLPs were mainly located in the upper fraction after ultracentrifugation,which was confirmed by Western blot analysis and immuno-electron microscopy (IEM).
基金Supported by the National Natural Science Foundation of China(81373465)
文摘In recent years, increasingly evidences show that autophagy plays an important role in the pathogenesis and development of liver diseases, and the relationship between them has increasingly become a focus of concern. Autophagy refers to the process through which the impaired organelles, misfolded protein, and intruding microorganisms is degraded by lysosomes to maintain stability inside cells. This article states the effect of autophagy on liver diseases （hepatic fibrosis, fatty liver, viral hepatitis, and liver cancer）, which aims to provide a new direction for the treatment of liver diseases.
基金Supported by Tehran University of Medical Sciences
文摘AIM: To investigate the reactivity of a panel of 8 mouse anti-hepatitis B surface antigen (HBsAg) monoclonal antibodies (mAbs) using a collection of 9 recombinant HBsAg mutants with a variety of amino acid substitutions mostly located within the “a” region.METHODS: The entire HBs genes previously cloned into a mammalian expression vector were transiently transfected into COS7 cells. Two standard unmutated sequences of the ayw and adw subtypes served as controls. Secreted mutant proteins were collected and measured by three commercial diagnostic immunoassays to assess transfection efficiency. Reactivity of anti-HBs mAbs with mutated HBsAgs was determined by sandwich enzyme-linked immunosorbent assay (ELISA).RESULTS: Reactivity of anti-HBs mAbs with mutated HBsAgs revealed different patterns. While three mutants reacted strongly with all mAbs, two mutants reacted weakly with only two mAbs and the remaining proteins displayed variable degrees of reactivity towards different mAbs. Accordingly, four groups of mAbs with different but overlapping reactivity patterns could be envisaged. One group consisting of two mAbs (37C5-S7 and 35C6-S11) was found to recognize stable linear epitopes conserved in all mutants. Mutations outside the “a” determinant at positions 120 (P→S), 123(T→N) and 161(M→T) were found to affect reactivity of these mAbs.CONCLUSION: Our findings could have important implications for biophysical studies, vaccination strategies and immunotherapy of hepatitis B virus (HBV) mutants.
基金the United Kingdom Department of Health, British Medical Research Council, Grant No. G99000178 and the United Kingdom National Health Service Research and Development Initiative
文摘Patients who are chronically infected with the hepatitis C virus often develop chronic liver disease and assessment of the severity of liver injury is required prior to considering viral eradication therapy. This article examines the various assessment methods currently available from gold standard liver biopsy to serological markers and imaging. Ultrasound is one of the most widely used imaging modalities in clinical practice and is already a first-line diagnostic tool for liver disease. Microbubble ultrasound contrast agents allow higher resolution images to be obtained and functional assessments of microvascular change to be carried out. The role of these agents in quantifying the state of hepatic injury is discussed as a viable method of determining the stage and grade of liver disease in patients with hepatitis C. Although currently confined to specialist centres, the availability of microbubble contrast-enhanced ultrasound will inevitably increase in the clinical setting.
文摘Objective.To test whether intramuscular,intranasal,intrarectal and intravagina l administration of HPV6b L 1 virus-like particlescould induce immune response in mice and to as sess whether intra-muscular and mucosal vaccination against HPV is feasible.Me thods.HPV6b L1proteins self-assembled into VLPs in Sf-9cell in vitro.Mic e were immunized on day0and21with50ìg HPV6b L1VLPs intramuscularly,int ranasally,intrarectally and intravagi-nally respectively.Sera were collected for testing IgG titer after a further7days and3months respec-tively.Results .After immunizations,all mice developed significant anti-HPV6b L1antibody titers in serum by7days after the second immunization.The titer of the serum I gG antibody against HPV6b L1VLPs in the intramuscularly immunized group was h igher than that in the intranasally,intrarectally and intravaginally immunized groups respectively,indicating that both muscular and mucosal administration of HPV6b L1VLPs can stimulate a systemic HPV-specific antibody response.Sera of the mice in the in-tramuscularly immunized group still maintained a high tit er of the serum IgG antibody against HPV6b L1VLPs 3months after the immunizat ion.Conclusion.The results demonstrated that the HPV6b L1VLPs maintain stro ng antigenicity.Immu-nization with HPV6b L1VLPs via intramuscular and mucos al routes,without adjuvant ,can elicit spe-cific antibody in sera.These fin dings suggest that the VLPs are able to induce protective antibodies.
