鸭肝炎病毒感染和垂直传播的研究

用α-~(32)P-dATP-DHBV DNA探针做斑点杂交试验、检测了不同鸭种血清中携带DHBV情况。其中北京鸭与樱桃谷鸭的杂交种鸭杂交阳性率为5.1％;而不同鸭龄不同饲养条件的高邮麻鸭血清阳性率32.7—58.8％,经EM检测均在血清中找到病毒颗粒。将DHBV DNA不同的亲代配对所组成的四个组分群饲养,观察产卵后再孵化出的雏鸭DHBVDNA阳性率。第一组(♀-(?)-)为0％,第二组(♀ (?) )为100％,第三组(♀-(?) )28％,第四组(♀ (?)-)100％。然而亲代雌雄鸭均为DHBV DNA阴性的雌鸭所产卵在孵化到第9天时,经尿囊腔注射1×10~9病毒颗粒/m110μl的血清,孵出雏鸭时DHBV DNA阳性率达83.3％。     

不同种雏鸭建立鸭乙肝病毒感染模型及抗病毒效果的实验

目的探讨不同种雏鸭建立鸭乙肝病毒感染模型的影响因素,观察应用该模型抗病毒的效果。方法采集鸭血清,应用PCR方法定性检测鸭血清中病毒DNA;定量PCR方法检测鸭血清中病毒DNA载量变化;用抗病毒药物处理,观察其在鸭DHBV感染模型中的抗病毒效果。结果不同种鸭DHBV自然感染率不同,樱桃谷鸭为8.75%,湖北麻鸭两个批次分别为17.80%和10.68%;静脉注射和腹腔注射两途径均能致雏鸭感染DHBV,静脉注射感染率80%,腹腔注射感染率65%;鸭感染DHBV后,体内病毒载量维持在106～108copies/mL,可持续20 d以上;抗病毒药物处理后,在不同DHBV模型中其抗病毒效果变化趋势一致。结论鸭的种类和人工感染途径可影响DHBV感染率;雏鸭感染DHBV后其体内有持续性的病毒血症;DHBV感染模型是药物抗病毒研究较好模型。     

鸭乙型肝炎病毒实验感染后在外周血和肝脏中的动态

鸭乙型肝炎病毒(DHBV)与人乙型肝炎病毒(HBV)同属嗜肝病毒,两者的病毒大分子结构和复制过程有很多相似之处,了解DHBV实验感染规律和病毒在血液和肝脏内的动态,有助于研究病毒复制特点和抗肝炎药物的效果。 本文用DHBV-DNA杂交阳性和DHBV-DNA多聚酶阳性的上海鸭血清,静脉注射1～3     

“番北金”雏鸭的能量代谢和同化水平的测定

本文研究“番北金”雏鸭的能量代谢和同化率。其结果:代谢率与体重呈线性负相关(r=-0.6834,y=3.3801-0.0037x)。每日代谢能消耗随体重增长而下降。雏鸭的能量摄入、可代谢能量、代谢能消耗和生长能量均随体重增长而增多。单位体重的能量摄入变动在2.07—2.67千焦/克之间。能量同化率,1周龄时(70.84％)较低,2—5周龄时则保持在82.60％—84.20％之间,6周龄时(89.36％)有所提高。在可代谢能量中,代谢能消耗占58.75％,生长能量消耗占41.25％。“番北金”鸭是金定鸭、北京鸭和番鸭三元杂交的杂种鸭。该鸭具有生长快、产肉性能好和饲料报酬高的特点,颇受群众欢迎。本文就“番北金”雏鸭的能量代谢和同化水平进行研究,为拟定该鸭饲料的能量标准提供参考依据。现将测定结果报告如下。     

鸭肝炎病毒新血清型基因组序列分析

[目的]为了揭示鸭肝炎病毒新血清型(DHV-N)G株的演化及其与DHV-1的基因序列差异.[方法]运用RT-PCR结合5'-/3'-RACE策略对DHV-N G株基因组进行扩增,克隆测序后进行序列分析.[结果]DHV-NG株全基因组序列除12 nt poly(A)尾外全长共7774 nt,含有一个大的开放阅读框,编码2251 aa的蛋白前体,其基因组结构:5'UTR-L-VP0-VP3-VP1-2A1-2A2-2B-2C-3A-3B-3C-3D-3'UTR;3'UTR长为366 nt,比1型鸭肝炎病毒C80株多52 nt;VP1蛋白与DHV-1相比发生较大的变异,特别在其第140～221位;其基因组与DHV-1、韩国DHV-N和台湾DHV-N的同源率分别为72.8%～73.4%、96.3%～96.5%和78.3%.[结论]DHV-N G株基因组结构与DHV-1存在明显差异,在DHV-N成员DHV-N G株与韩国DHV-N关系更为密切.     

异源副粘病毒感染对雏鸭血清抗氧化酶活性的影响

以雏鸭为试验对象,研究不同来源的副粘病毒感染对雏鸭血清中抗氧化酶活性的影响。选择20日龄健康雏鸭100只,随机分成Ⅰ、Ⅱ、Ⅲ、Ⅳ组,Ⅰ组为对照组,皮下注射0.5 mL/只生理盐水,Ⅱ、Ⅲ、Ⅳ组分别皮下注射0.5 mL/只稀释的鸡源、鹅源及鸭源副粘病毒SPF胚液。分别在攻毒后7、14、21、28 d时,随机抽取Ⅰ、Ⅱ、Ⅲ、Ⅳ组雏鸭各5只进行血液采集与血清分离,测定雏鸭血清中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-Px)及丙二醛(MDA)的活性与含量,研究异源副粘病毒感染对雏鸭血清中抗氧化酶活性的影响,以及脂质过氧化程度与抗氧化酶活性的变化趋势。结果表明,在攻毒后不同时间内,感染异源副粘病毒的雏鸭血清中的SOD、CAT、GSH-Px活性和MDA活性与含量变化显著不同,揭示异源副粘病毒对雏鸭血清自由基代谢产生了不同程度影响,脂质过氧化程度与SOD、GSH-Px活性密切相关。     

1型鸭肝炎病毒CL株感染性分子克隆的构建

摘要：【目的】构建1型鸭肝炎病毒（DHV）的感染性克隆,用于研究其基因组的结构与功能。【方法】用RT-PCR方法扩增出覆盖整个1型鸭肝炎病毒CL株基因组3个忠实性片段,并按顺序组装进载体pBR322中,获得全长cDNA克隆（BR-CL）。将BR-CL在体外转录出的RNA转染鸭胚肾细胞,并传至第6代,利用RT-PCR方法和间接免疫荧光试验进行鉴定。将获得的子代病毒（CL-R）在SPF鸡胚上传代,观察鸡胚死亡及胚体病变情况。通过胶体金免疫电镜观察子代病毒粒子的形态。【结果】RT-PCR、间接免疫荧光和胶体金免     

绍兴鸭血液生化指标与产蛋性状间相关的通径分析

本研究估测了125羽60日龄的绍兴鸭血液中的总蛋白（TP）、胰岛素样生长因子-I（IGF-I）的含量和谷草转氨酶（GOT）、谷丙转氨酶（GPT）的活性与3个产蛋性状（300日总蛋数、300日总蛋重、平均蛋重）的表型相关,再分别以3个产蛋性状为依变量对60日龄的性状相关进行了通径分析,为绍兴鸭的早期选育提供科学依据.相关分析结果表明:60日龄绍兴鸭血液中GPT与300日总蛋数、300日总蛋重的表型相关为中等程度相关（0.5274、0.6234）,与平均蛋重的表型相关为强相关（0.6733）,都达极显著水平（P <0.01）.TP与3个产蛋性状的表型相关为弱相关（0.2067、0.2629、0.3026）,但都达显著水平（P<0.05）.通径分析结果为:GPT对3个产蛋性状的正直接影响均较大（0.5275、0.6109、0.6395）.     

婴儿围产期感染乙型肝炎病毒后各项血清学指标的动态变化

本文观察了102名新生儿出生后至18月龄的HBV血清学指标的动态变化,婴儿分成乙型肝炎疫苗按种组(63人)和对照组(39人), 观察期间HBsAg始终阴性的70名婴儿,出生后6、12和18月龄的抗-HBc阳性率依次为90％、30％和4.3％;而HBsAg阳转的27名婴儿,18月龄时抗-HBc全都阳性,但仅有6名婴儿在6月龄时测出IgM抗-HBc,疫苗接种组婴儿出生后1、3、6、12和18月龄的抗-HBs阳性率,依次为28.6％、76.2％、77。8％、82.5％和82.5％;对照组婴儿18月龄时抗-HBs阳性率仅为12.8％。     

鸭乙型肝炎病毒基因组结构及其编码蛋白的研究进展

鸭乙型肝炎病毒(DHBV)属于禽嗜肝病毒属、嗜肝DNA病毒科病毒,与人乙型肝炎病毒(HBV)具有相同的复制方式及相似的基因结构、抗原结构。DHBV基因组是闭合环状不完全双链DNA,3个主要ORF-PreS/S、ORF-PreC/C、ORF-P位于负链,编码L蛋白、S蛋白、C蛋白和P蛋白。本文通过对DHBV基因组结构特征及其编码病毒重要蛋白的基本特性、结构特点与功能进行全面阐释,旨在对DHBV的深入研究提供重要参考和以DHBV人工感染建立的动物模型研究HBV提供重要理论依据。     

鸭甲型病毒性肝炎的研究进展

以下综述了鸭甲型肝炎病毒的命名与溯源、分子遗传与进化,分析了鸭甲型病毒性肝炎的流行病学、临床特征及监测手段,总结了国内外鸭甲型病毒性肝炎的研究现状,进一步阐述了研究该病的必要性和紧迫性,旨在为从事该领域的科研人员提供参考依据。     

Complete genomic sequence of a Chinese isolate of Duck hepatitis virus

The complete genomic sequence of Duck hepatitis virus 1 (DHV-1) ZJ-V isolate was sequenced and determined to be 7 691 nucleotides (nt) in length with a 5'-terminal un-translated region (UTR) of 626 nt and a 3'-terminal UTR of 315 nt (not including the poly(A) tail). One large open reading frame (ORF) was found within the genome (nt 627 to 7 373) coding for a polypeptide of 2 249amino acids. Our data also showed that the poly (A) tail of DHV-1 has at least 22 A's. Sequence comparison revealed significant homology (from 91.9% to 95.7%) between the protein sequences of the virus in the Picornaviridae family, its genome showed some unique characteristics. DHV-1 contains 3copies of the 2A gene and only 1 copy of the 3B gene, and its 3'-NCR is longer than those of other picornaviruses. Phylogenetic analysis to do sequence homology based on the VP1 protein sequences showed that the ZJ-V isolate shares high sequence homology with the reported DHV-1 isolates (from 92.9% to 99.2%), indicating that DHV-1 is genetically stable.     

鸭病毒性肝炎二价灭活疫苗 (DHAV-SH和DHAV-FS株) 的制备和评价

我国鸭甲型肝炎病毒(DHAV)的快速变异及广泛流行给养鸭业造成了较大的经济损失,为研究鸭病毒性肝炎二价灭活疫苗(DHAV-SH和DHAV-FS株)的制备和评价方法,首先对我国多个鸭场进行血清型流行病学分析,从201株DHAV-1、38株DHAV-3中筛选出6株毒力较强的DHAV,验证了DHAV-1和DHAV-3为我国流行优势株,并对6株DHAV进行ELD50和LD50测定,筛选疫苗候选株后经传代确定疫苗毒种,选取F5代鸭胚胚体研磨液作为疫苗毒种,经甲醛灭活后制成3批水包油包水(W/O/W)型乳剂二价灭活疫苗实验室制品。通过安全性试验、抗体中和试验、攻毒保护及免疫交叉保护试验发现:疫苗安全性良好,雏鸭免疫后第7天可以检测到DHAV中和抗体,第14-21天的攻毒保护率为90%-100%,免疫持续期可达5周以上,DHAV-SH和DHAV-FS之间的交叉保护为20%-30%。研究表明本试验研制的鸭病毒性肝炎二价灭活疫苗实验室制品安全且高效,为我国DHAV预防和控制提供了一种新方法和新产品。     

Antisense therapy of hepatitis B virus infection

Chronic infection with the hepatitis B virus (HBV) is a major health problem worldwide. The only established therapy is interferon-a with an efficacy of only 30–40% in highly selected patients. The discovery of animal viruses closely related to the HBV has contributed to active research on antiviral therapy of chronic hepatitis B. The animal model tested and described in this article are Peking ducks infected with the duck hepatitis B virus (DHBV). Molecular therapeutic strategies aimed at blocking gene expression include antisense DNA. An antisense oligodeoxynucleotide directed against the 5′-region of the preS gene of DHBV inhibited viral replication and gene expression in vitro in primary duck hepatocytes and in vivo in Peking ducks. These results demonstrate the potential clinical use of antisense DNA as antiviral therapeutics.     

Expression and characterization of hepatitis C virus core protein fused to hepatitis B virus core antigen

Recombinant plasmids were constructed by fusing the gene fragments encoding the full-length (1-191aa) and the truncated (1-40aa and 1-69aa) HCV core proteins (HCc) respectively to the core gene of HBV at the position of amino acid 144 and expressed in E. coli. The products were analyzed by ELISA, Western blotting as well as the immunization of the mice. The results showed that those fusion proteins (B144C191, B144C69, B144C40) possessed the dual antigenicity and immunogenicity of both hepatitis B virus core antigen (HBcAg) and hepatitis C virus core protein (HCc). Analysis by electron microscopy and CsCl density gradient ultra-centrifugation revealed that similar to the HBcAg itself, all fusion proteins were able to form particles. Comparison of the antigenicity and immunogenicity of those fusion proteins showed that the length of HCc gene fused to HBeAg had no much effect on the antigenicity and immunogenicity of HBcAg, however, B144C69 and B144C40 induced higher titres antibodies against HCc than B14d     

The birth and life of lipid droplets: learning from the hepatitis C virus

LDs (lipid droplets) are probably the least well-characterized cellular organelles. Having long been considered simple lipid storage depots, they are now considered to be dynamic organelles involved in many biological processes. However, most of the mechanisms driving LDs biogenesis, growth and intracellular movement remain largely unknown. As for other cellular mechanisms deciphered through the study of viral models, HCV (hepatitis C virus) is an original and relevant model for investigations of the birth and life of these organelles. Recent studies in this model have raised the hypothesis that the HCV core protein induces the redistribution of LDs through the regression and regeneration of these organelles in specific intracellular domains.     

Role of lipid metabolism in hepatitis C virus assembly and entry

HCV (hepatitis C virus) represents a major global health problem. A consistent body of evidence has been accumulating, suggesting a peculiar overlap between the HCV life cycle and lipid metabolism. This association becomes evident both for the clinical symptoms of HCV infection and the molecular mechanisms underlying the morphogenesis and entry process of this virus. The HCV core–lipid droplets association seems to be central to the HCV morphogenesis process. Moreover, the biogenesis pathway of very‐low‐density lipoproteins has been shown to be involved in HCV morphogenesis with MTP (microsomal triacylglycerol transfer protein), ApoB (apolipoprotein B) and ApoE (apolipoprotein E) as essential elements in the production of infectious HCV particles. HCV infectivity also correlates with the lipidation status of the particles. Furthermore, some HCV cellular receptors and the regulation of the entry process are also connected to lipoproteins and lipid metabolism. Specifically, lipoproteins modulate the entry process and the cholesterol transporter SR‐BI (scavenger receptor class B type I) is a cellular entry factor for HCV. The present review aims to summarize the advances in our understanding of the HCV–lipid metabolism association, which may open new therapeutic avenues.