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Control of hepatitis B virus replication by interferons and Toll-like receptor signaling pathways 被引量:21
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作者 Rong-Juan Pei Xin-Wen Chen Meng-Ji Lu 《World Journal of Gastroenterology》 SCIE CAS 2014年第33期11618-11629,共12页
Hepatitis B virus (HBV) infection is one of the major causes of liver diseases, affecting more than 350 million people worldwide. The interferon (IFN)-mediated innate immune responses could restrict HBV replication at... Hepatitis B virus (HBV) infection is one of the major causes of liver diseases, affecting more than 350 million people worldwide. The interferon (IFN)-mediated innate immune responses could restrict HBV replication at the different steps of viral life cycle. Indeed, IFN-&#x003b1; has been successfully used for treatment of patients with chronic hepatitis B. However, the role of the innate immune response in HBV replication and the mechanism of the anti-HBV effect of IFN-&#x003b1; are not completely explored. In this review, we summarized the currently available knowledge about the IFN-mediated anti-HBV effect in the HBV life cycle and the possible effectors downstream the IFN signaling pathway. The antiviral effect of Toll-like receptors (TLRs) in HBV replication is briefly discussed. The strategies exploited by HBV to evade the IFN- and TLR-mediated antiviral actions are summarized. 展开更多
关键词 Hepatitis B virus INTERFERON Toll-like receptor Interferon stimulated genes Innate immune response
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Detection of hepatitis C virus NS5 protein and genome in Chinese carcinoma of the extrahepatic bile duct and its significance 被引量:19
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作者 Ming Yi Chen Zhi Qiang Huang +3 位作者 Le Zhen Chen Ya Bing Gao Rui Yun Peng De Wen Wang 《World Journal of Gastroenterology》 SCIE CAS CSCD 2000年第6期800-804,共5页
AIM To investigate the hepatitis C virus(HCV)infection in the tissues of carcinoma ofextrahepatic bile duct and study theircorrelation.METHODS HCV NS5 protein and HCV RNA weredetected by labeled streptavidin biotin(LS... AIM To investigate the hepatitis C virus(HCV)infection in the tissues of carcinoma ofextrahepatic bile duct and study theircorrelation.METHODS HCV NS5 protein and HCV RNA weredetected by labeled streptavidin biotin(LSAB)method and in situ reverse transcriptionpolymerase chain reaction(IS-RT-PCR)insections of 51 cases of carcinoma ofextrahepatic bile duct and 34 cases of controlgroup(without malignant biliary disease).RESULTS In 51 cases of carcinoma ofextrahepatic bile duct,HCV NS5 protein wasdetected in 14(27.5%),which was clearlystained in the cytoplasm of cancer cell but not inthe nucleus or cell membrane.HCV RNA wasdetected in 18(35.4%),which was located inthe nucleus of cancer cell in 12 cases and in thecytoplasm in 6 cases.HCV NS5 protein and RNAcoexistence was found in 2 cases.In 34 cases ofcontrol group,HCV RNA was detected in 2(5.9%).HCV NS5 protein and RNA positive cellswere found either scattered or in clusters.CONCLUSION The prevalence of hepatitis C viral infection in the tissues of carcinoma ofextrahepatic bile duct was significantly higherthan in control group(X^2=9.808,P=0.002).The findings suggest a correlation between HCVinfection and carcinoma of extrahepatic bileduct,which is different from the traditionalviewpoint.HCV infection might be involved inthe development of carcinoma of extrahepaticbile duct. 展开更多
关键词 hepatitis C virus bile duct neoplasm polymerase chain reaction immunohistochemistry risk factors genes suppressor tumor TRANSFECTION genome
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抗呼吸道合胞病毒M2-1基因pshRNA载体质粒的构建及效应研究 被引量:7
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作者 崔玉霞 周娟 +3 位作者 方平 蒋利萍 王莉佳 杨锡强 《中华儿科杂志》 CAS CSCD 北大核心 2005年第11期858-862,共5页
目的构建针对人类呼吸道合胞病毒(RSV)M2-1基因mRNA的pshRNA表达载体,并在培养细胞内观察其对RSV复制的影响,为从基因水平抑制RSV感染的研究奠定基础。方法将构建成功的表达载体pshRNA7816转染Hep2细胞,然后利用光镜观察pshRNA7816对RSV... 目的构建针对人类呼吸道合胞病毒(RSV)M2-1基因mRNA的pshRNA表达载体,并在培养细胞内观察其对RSV复制的影响,为从基因水平抑制RSV感染的研究奠定基础。方法将构建成功的表达载体pshRNA7816转染Hep2细胞,然后利用光镜观察pshRNA7816对RSV致Hep2细胞病变效应的影响,空斑实验检测RSV滴度变化以及四甲基偶氮唑盐比色实验检测pshRNA7816对被RSV感染后细胞的保护作用。结果成功构建了针对RSV M2-1基因mRNA的pshRNA7816载体质粒,研究发现pshRNA7816能明显改善RSV所致的病变效应,降低RSV在细胞内复制的病毒滴度,并且能明显提高被RSV感染后细胞的存活率(P<0.001),非特异性质粒无抗RSV效应。结论针对RSV M2-1基因的pshRNA7816的载体质粒具有明显的和特异性的抗RSV效应的作用。 展开更多
关键词 呼吸道合胞病毒 基因 病毒 质粒 病毒复制
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Transactivating effect of complete S protein of hepatitis B virus and cloning of genes transactivated by complete S protein using suppression subtractive hybridization technique 被引量:6
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作者 Gui-QinBai YanLiu +4 位作者 JunCheng Shu-LinZhang Ya-FeiYue Yan-PingHuang Li-YingZhang 《World Journal of Gastroenterology》 SCIE CAS CSCD 2005年第25期3893-3898,共6页
AIM: To investigate the transactivating effect of complete S protein of hepatitis B virus (HBV) and to construct a subtractive cDNA library of genes transactivated by complete S protein of HBV by suppression subtracti... AIM: To investigate the transactivating effect of complete S protein of hepatitis B virus (HBV) and to construct a subtractive cDNA library of genes transactivated by complete S protein of HBV by suppression subtractive hybridization (SSH) technique and to clone genes associated with its transactivation activity, and to pave the way for elucidating the pathogenesis of hepatitis B virus infection. METHODS: pcDNA3.1(-)-complete S containing full-length HBV S gene was constructed by insertion of HBV complete S gene into BamH I/Kpn I sites. HepG2 cells were cotransfected with pcDNA3.1(-)-complete S and pSV-lacZ. After 48 h, cells were collected and detected for the expression of β-galactosidase (β-gal). Suppression subtractive hybridization and bioinformatics techniques were used. The mRNA of HepG2 cells transfected with pcDNA3.Incomplete S and pcDNA3.1(-) empty vector was isolated, and detected for the expression of complete S protein by reverse transcription polymerase chain reaction (RT-PCR) method, and cDNA was synthesized. After digestion with restriction enzyme RsaI, cDNA fragments were obtained. Tester cDNA was then divided into two groups and ligated to the specific adaptors 1 and 2, respectively. After tester cDNA had been hybridized with driver cDNA twice and underwent nested PCR twice, amplified cDNA fragments were subcloned into pGEM-Teasy vectors to set up the subtractive library. Amplification of the library was carried out within E. coli strain DH5α. The cDNA was sequenced and analyzed in GenBank with BLAST search after polymerase chain reaction (PCR) amplification. RESULTS: The complete S mRNA could be detected by RT-PCR in HepG2 cells transfected with the pcDNA3.1(-)-complete S. The activity of β-gal in HepG2 cells transfected with the pcDNA3.1(-)-complete s was 6.9 times higher than that of control plasmid. The subtractive library of genes transactivated by HBV complete S protein was constructed successfully. The amplified library contains 86 positive clones. Colony PCR showed that 86 clones con 展开更多
关键词 Complete S protein Transactivated genes Hepatitis virus B
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重型乙型肝炎患者血清乙型肝炎病毒全基因克隆及测序 被引量:3
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作者 吴炜 李兰娟 +3 位作者 陈瑜 李君 钱香 程东庆 《中华肝脏病杂志》 CAS CSCD 北大核心 2005年第10期734-737,共4页
目的建立重型乙型肝炎患者血清乙型肝炎病毒(HBV)DNA克隆并测序,从全基因水平分析HBV基因变异与重型乙型肝炎发病的关系。方法10例重型乙型肝炎患者血清提取HBV DNA,聚合酶链反应(PCR)扩增HBV全基因。PCR产物构建到pUCm-T载体上,转化至... 目的建立重型乙型肝炎患者血清乙型肝炎病毒(HBV)DNA克隆并测序,从全基因水平分析HBV基因变异与重型乙型肝炎发病的关系。方法10例重型乙型肝炎患者血清提取HBV DNA,聚合酶链反应(PCR)扩增HBV全基因。PCR产物构建到pUCm-T载体上,转化至大肠杆菌感受态DH-5α细胞, 经酶切鉴定,获得含3.2 Kb HBV DNA的重组克隆菌,全基因测序, 分析各读码框核苷酸和氨基酸变化。结果4例成功构建HBV DNA克隆,并完成全基因测序。其中3例在前C区发生G1896A变异,产生一个终止密码子,导致HBeAg缺失; 1例在C启动子区1762、1764双位点出现突变;有多处点突变及缺失变异分布于PreS2区及C区已知细胞毒T淋巴细胞、B淋巴细胞和T淋巴细胞的细胞表位。结论该法可用于临床研究HBV病毒基因结构与重型乙型肝炎发病的关系,并为进一步研究其HBV基因功能奠定基础。 展开更多
关键词 肝炎病毒 乙型 基因 病毒 肝炎 重型 血清乙型肝炎病毒 重型乙型肝炎 乙型肝炎患者 全基因测序 全基因克隆 乙型肝炎病毒(HBV) 聚合酶链反应(PCR)
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ISG20 inhibits bluetongue virus replication 被引量:3
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作者 Di Kang Shandian Gao +6 位作者 Zhancheng Tian Guorui Zhang Guiquan Guan Guangyuan Liu Jianxun Luo Junzheng Du Hong Yin 《Virologica Sinica》 SCIE CAS CSCD 2022年第4期521-530,共10页
ISG20 is an interferon-inducible exonuclease that inhibits virus replication.Although ISG20 is thought to degrade viral RNA,the antiviral mechanism and specificity of ISG20 remain unclear.In this study,the antiviral r... ISG20 is an interferon-inducible exonuclease that inhibits virus replication.Although ISG20 is thought to degrade viral RNA,the antiviral mechanism and specificity of ISG20 remain unclear.In this study,the antiviral role of ovine ISG20(o ISG20)in bluetongue virus(BTV)infection was investigated.It was found that BTV infection upregulated the transcription of ovine ISG20(o ISG20)in a time-and BTV multiplicity of infection(MOI)-dependent manner.Overexpression of o ISG20 suppressed the production of BTV genome,proteins,and virus titer,whereas the knockdown of o ISG20 increased viral replication.o ISG20 was found to co-localize with BTV proteins VP4,VP5,VP6,and NS2,but only directly interacted with VP4.Exonuclease defective o ISG20 significantly decreased the inhibitory effect on BTV replication.In addition,the interaction of mutant o ISG20 and VP4 was weakened,suggesting that binding to VP4 was associated with the inhibition of BTV replication.The present data characterized the anti-BTV effect of o ISG20,and provides a novel clue for further exploring the inhibition mechanism of double-stranded RNA virus by ISG20. 展开更多
关键词 Bluetongue virus(BTV) Interferon-stimulated genes(ISGs) Ovine ISG20 virus replication Antiviral immunity
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Distribution of antibiotic resistance genes and their association with bacteria and viruses in decentralized sewage treatment facilities 被引量:3
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作者 Jiaheng Zhao Bing Li +3 位作者 Pin Lv Jiahui Hou Yong Qiu Xia Huang 《Frontiers of Environmental Science & Engineering》 SCIE EI CSCD 2022年第3期91-104,共14页
The distribution of antibiotic resistance genes(ARGs)has been intensively studied in large-scale wastewater treatment plants and livestock sources.However,small-scale decentralized sewage treatment facilities must als... The distribution of antibiotic resistance genes(ARGs)has been intensively studied in large-scale wastewater treatment plants and livestock sources.However,small-scale decentralized sewage treatment facilities must also be explored due to their possible direct exposure to residents.In this study,six wastewater treatment facilities in developed rural areas in eastern China were investigated to understand their risks of spreading ARGs.Using metagenomics and network analysis tools,ARGs and bacterial and viral communities were identified in the influent(INF)and effluent(EFF)samples.The dominant ARGs belonged to the bacitracin class,which are different from most of municipal wastewater treatment plants(WWTPs).The dominant hosts of ARGs are Acidovorax in bacterial communities and Prymnesiovirus in viral communities.Furthermore,a positive relationship was found between ARGs and phages.The ARGs significantly correlated with phages were all hosted by specific genera of bacteria,indicating that phages had contributed to the ARG’s proliferation in sewage treatment facilities.Paying significant concern on the possible enhanced risks caused by bacteria,viruses and their related ARGs in decentralized sewage treatment facilities is necessary. 展开更多
关键词 Decentralized sewage treatment facilities Antibiotic resistance genes virus METAGENOMICS Network analysis
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人原发性肝内胆管细胞癌中HCV基因及其抗原表达的意义 被引量:4
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作者 王文亮 王春杰 王冰枫 《世界华人消化杂志》 CAS 2001年第5期541-545,共5页
目的检测HCV基因及其抗原在人原发性肝内胆管细胞癌中的表达,探讨肝内胆管细胞癌的病因及发病机制。方法用免疫组化技术检测HCV抗原,用原位分子杂交技术检测HCV基因(HCV RNA)。结果在40例肝内胆管细胞癌中,HCVC33c抗原、核心抗原及HCV ... 目的检测HCV基因及其抗原在人原发性肝内胆管细胞癌中的表达,探讨肝内胆管细胞癌的病因及发病机制。方法用免疫组化技术检测HCV抗原,用原位分子杂交技术检测HCV基因(HCV RNA)。结果在40例肝内胆管细胞癌中,HCVC33c抗原、核心抗原及HCV RNA阳性率分别为52.5%,72.5%和57.5%;在21例癌旁肝组织中,其阳性率分别为63.6%,45.5%和48.5%。结论人原发性肝内胆管细胞癌的发生除与HBV有密切关系外,与HCV慢性感染也有密切关系。 展开更多
关键词 原发性肝内胆管细胞癌 HCV基因 HCV抗原 发病机理
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H5N1禽流感病毒headless HA基因在杆状病毒中的表达 被引量:3
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作者 张立霞 于在江 +3 位作者 周剑芳 秦堃 许应天 舒跃龙 《中华实验和临床病毒学杂志》 CAS CSCD 2012年第3期179-181,共3页
目的建立能有效表达禽流感病毒A/Hubei/1/2010(H5N1)无头HA(headlessHA)的杆状病毒系统。方法利用RT—PCR扩增获得headlessHA基因,克隆至杆状病毒转移载体pFastBacl,命名为pFastBaclheadlessHA。将pFastBaclheadlessHA转化到DH... 目的建立能有效表达禽流感病毒A/Hubei/1/2010(H5N1)无头HA(headlessHA)的杆状病毒系统。方法利用RT—PCR扩增获得headlessHA基因,克隆至杆状病毒转移载体pFastBacl,命名为pFastBaclheadlessHA。将pFastBaclheadlessHA转化到DH10Bae感受态细胞,获得重组穿梭质粒HeadlessHA—Bac。利用Bac—to—Bac杆状病毒表达系统,转染Sf9昆虫细胞,获得重组杆状病毒,并进行WesternBlot和红细胞凝集试验检测。结果HeadlessHA在St9细胞中能有效表达,不能使红细胞发生凝集。结论本研究为利用headlessHA表达蛋白研制广谱流感疫苗奠定了基础。 展开更多
关键词 基因 流感病毒A型 杆状病毒科 表达的序列标记
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上海地区2010年冬季甲型H1N1流行性感冒病毒株变异分析 被引量:2
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作者 高颖阳 居丽雯 +4 位作者 汪千力 蒋露芳 熊海燕 朱雯 姜庆五 《中华传染病杂志》 CAS CSCD 北大核心 2012年第1期10-15,共6页
目的了解上海地区2010年冬季人群甲型H1N1流行性感冒(流感)病毒流行株基因及抗原的变异。方法采集2010年12月至2011年1月间上海地区哨点医院流感样患者咽拭子标本137份,接种犬。爵细胞(MDCK),分离流感病毒,直接免疫荧光法(DIF... 目的了解上海地区2010年冬季人群甲型H1N1流行性感冒(流感)病毒流行株基因及抗原的变异。方法采集2010年12月至2011年1月间上海地区哨点医院流感样患者咽拭子标本137份,接种犬。爵细胞(MDCK),分离流感病毒,直接免疫荧光法(DIF)鉴定流感病毒型,RT—PCR鉴定甲型H1N1,对部分甲型H1N1流感病毒株进行血凝素(HA)、神经氨酸酶(NA)、病毒聚合酶(PB2)片段全基因测序,分析基因及氨基酸位点变异。结果共分离到53株人流感病毒,48株为甲型H1N1流感病毒,按简单随机抽样法抽取19株测序。HA进化树分析发现,与2010年6月前分离的甲型H1N1毒株比较,绝大部分不位于同一主干上;HA蛋白的氨基酸位点分析显示,部分毒株在抗原决定位点上发生变异。NA蛋白酶活性中心及周围相关位点氨基酸组成保守,未检测到耐奥司他韦和扎那米韦的变异位点。PB2蛋白第627位和701位点分别是谷氨酸和天冬氨酸,仍是禽源流感病毒特征,但第677位点出现E677G突变。结论2010年冬季上海地区人群甲型H1N1流感病毒流行株与之前春夏季分离株比较已经有一定变异,出现了一些抗原漂移和在哺乳动物宿主内的适应性进化。 展开更多
关键词 流感病毒A型 H1N1亚型 基因 病毒 抗原
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用RT-PCR方法检测登革病毒E基因核酸 被引量:2
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作者 郝牧 左丽 +1 位作者 舒莉萍 李永念 《贵阳医学院学报》 CAS 2003年第5期380-382,共3页
目的:应用逆转录聚合酶链反应(RT-PCR)技术建立检测登革2型病毒E基因的方法。方法:分别抽提登革2型病毒NGC株和从登革出血热病人血清中分离得到的2型登革热病毒(DV)核酸(RNA),合成cDNA第一链,经过RT-PCR得到cDNA产物,用HinfⅠ限制性内... 目的:应用逆转录聚合酶链反应(RT-PCR)技术建立检测登革2型病毒E基因的方法。方法:分别抽提登革2型病毒NGC株和从登革出血热病人血清中分离得到的2型登革热病毒(DV)核酸(RNA),合成cDNA第一链,经过RT-PCR得到cDNA产物,用HinfⅠ限制性内切酶酶切鉴定。结果:通过RT-PCR检测登革2型病毒核酸。结论:登革2型病毒E基因核酸的RT-PCR检测方法的建立为快速诊断登革病毒感染提供了可靠的方法。 展开更多
关键词 登革热病毒 基因 结构 病毒 逆转录聚合酶链反应
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2例TTV感染者中TTV DNA的基因变异分析 被引量:1
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作者 侯周华 谭德明 +2 位作者 谢玉桃 刘水平 李聪智 《中国医师杂志》 CAS 2006年第9期1169-1171,共3页
目的探讨TTV的基因变异情况。方法PCR扩增BD与CSH血清中的TTV DNA后进行分子克隆,每例挑选10个不同TTV DNA克隆进行测序。不同克隆之间及其与G1 a亚型TA278之间进行序列比较并进行进化树分析。结果BD存在2种不同的TTV病毒株,都为G1 a亚... 目的探讨TTV的基因变异情况。方法PCR扩增BD与CSH血清中的TTV DNA后进行分子克隆,每例挑选10个不同TTV DNA克隆进行测序。不同克隆之间及其与G1 a亚型TA278之间进行序列比较并进行进化树分析。结果BD存在2种不同的TTV病毒株,都为G1 a亚型。CSH中存在7种不同的TTV病毒株,分别属于G1 a和G1b亚型。结论CSH中TTV的基因变异比BD中的复杂得多。 展开更多
关键词 输血传播病毒 DNA病毒感染 变异(遗传学) 基因 病毒
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Genome Annotation and Comparative Genomics of ORF Virus 被引量:1
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作者 A. K. M. Firoj Mahmud K. M. Zillur Rahman +2 位作者 Shuvra Kanti Dey Tahsina Islam Ali Azam Talukder 《Advances in Microbiology》 2014年第15期1117-1131,共15页
ORF virus (ORFV), the etiological agent of contagious pustular dermatitis in small ruminants, belongs to members of the genus Parapoxvirus of the Poxviridae. The genome of the ORFV is dsDNA of 139,962 bp which has abo... ORF virus (ORFV), the etiological agent of contagious pustular dermatitis in small ruminants, belongs to members of the genus Parapoxvirus of the Poxviridae. The genome of the ORFV is dsDNA of 139,962 bp which has about 89% coding region, 63% GC content and codes 130 proteins. There are four unique genes within the genome revealed by homology search of them two posses’ strong regulatory region and transmembrane helices. One of the ORF-039 contains signal peptide indicating the possibilities to be secretory protein coding gene. Comparative genomic analysis reveals significant differences in Bovine Papular Stomatitis Virus (BPSV) strain BV-AR02 and ORFV strain OV-SA00, and these may account for differences in host range. Interspecies sequence variability is observed in all functional classes of genes but is the highest in putative virulence/host range genes. Notably, ORFV contains genes which are homologous of Vaccinia virus. Phylogenetic analysis reveals that although divergent, ORFV virus is distinct from other known mammalian cowpox virus. An improved understanding of Parapoxvirus (PPV) biology will permit the engineering of novel vaccine viruses and expression vectors with enhanced efficacy and greater versatility. The novel vaccine will have a significant role in the economy of a country through the control of disease in an economically important and small ruminant caused by ORFV. 展开更多
关键词 ORF virus Contagious PUSTULAR DERMATITIS Comparative GENOMICS GLIMMER ACT Uniue genes Novel VACCINE
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Hepatitis B and circadian rhythm of the liver 被引量:1
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作者 Ivana Skrlec Jasminka Talapko 《World Journal of Gastroenterology》 SCIE CAS 2022年第27期3282-3296,共15页
The circadian rhythm in humans is determined by the central clock located in the hypothalamus’s suprachiasmatic nucleus,and it synchronizes the peripheral clocks in other tissues.Circadian clock genes and clock-contr... The circadian rhythm in humans is determined by the central clock located in the hypothalamus’s suprachiasmatic nucleus,and it synchronizes the peripheral clocks in other tissues.Circadian clock genes and clock-controlled genes exist in almost all cell types.They have an essential role in many physiological processes,including lipid metabolism in the liver,regulation of the immune system,and the severity of infections.In addition,circadian rhythm genes can stimulate the immune response of host cells to virus infection.Hepatitis B virus(HBV)infection is the leading cause of liver disease and liver cancer globally.HBV infection depends on the host cell,and hepatocyte circadian rhythm genes are associated with HBV replication,survival,and spread.The core circadian rhythm proteins,REV-ERB and brain and muscle ARNTL-like protein 1,have a crucial role in HBV replication in hepatocytes.In addition to influencing the virus’s life cycle,the circadian rhythm also affects the pharmacokinetics and efficacy of antiviral vaccines.Therefore,it is vital to apply antiviral therapy at the appropriate time of day to reduce toxicity and improve the effectiveness of antiviral treatment.For these reasons,understanding the role of the circadian rhythm in the regulation of HBV infection and host responses to the virus provides us with a new perspective of the interplay of the circadian rhythm and anti-HBV therapy.Therefore,this review emphasizes the importance of the circadian rhythm in HBV infection and the optimization of antiviral treatment based on the circadian rhythm-dependent immune response. 展开更多
关键词 Circadian rhythm Clock genes Hepatitis B virus Immune system LIVER
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RNA干扰对大鲵蛙病毒主要功能基因表达及其增殖的影响 被引量:2
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作者 牟维豪 周燕 +6 位作者 耿毅 汪开毓 余泽辉 李亚军 黄小丽 欧阳萍 陈德芳 《南方水产科学》 CAS CSCD 北大核心 2017年第4期80-86,共7页
采用q PCR与病毒滴度测定观察RNA干扰(RNA interference,RNAi)对大鲵蛙病毒(Chinese giant salamander ranavirus,CGSRV)主要衣壳蛋白(major capsid protein,MCP)、甲基转移酶(methyltransferases,MTases)、DNA多聚酶(DNA polymerase)... 采用q PCR与病毒滴度测定观察RNA干扰(RNA interference,RNAi)对大鲵蛙病毒(Chinese giant salamander ranavirus,CGSRV)主要衣壳蛋白(major capsid protein,MCP)、甲基转移酶(methyltransferases,MTases)、DNA多聚酶(DNA polymerase)基因表达与病毒增殖的影响。结果表明,小干扰RNA(small interfering RNA,siRNA)能推迟鲤鱼上皮瘤细胞(epithelioma papillosum cyprini,EPC)出现病变,且病变程度也较对照组轻。在各功能基因表达量上,NC-FAM对照组与阴性对照组差异不显著(P>0.05);而干扰组的干扰率极显著高于阴性对照组,其中siR-DP-1组siRNA对MCP基因的干扰率为79%,极显著高于其余组(P<0.01);siR-MT-1、siR-MT-2组siRNA对MTases基因的干扰率为77%,极显著高于其余组(P<0.01);siR-DP-1组siRNA对DNA polymerase基因的干扰率为79%,极显著高于其余组(P<0.01)。干扰实验组病毒滴度与NC-FAM对照组和阴性对照组相比也有不同程度的降低。阴性对照组lg TCID50为8.362,NC-FAM对照组lg TCID50为7.848,siR-MCP、siR-MT、siR-DP实验组最低lg TCID50分别为5.764、5.317、5.362。证实RNAi能够抑制CGSRV主要功能基因的表达并影响病毒的复制。 展开更多
关键词 大鲵蛙病毒 RNA干扰 基因表达 病毒滴度
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急性丙型肝炎患者病毒株部分基因克隆及其序列分析 被引量:1
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作者 蒋道荣 姚登福 +2 位作者 董志珍 邱历伟 陆建新 《中华肝脏病杂志》 CAS CSCD 2000年第3期153-155,共3页
目的探讨HCV感染在肝炎发生中的病原学作用。方法对89例急性肝炎进行HCV标志物分析,并从HCV RNA阳性的5例非甲非乙型急性肝炎患者血清中提取HCV RNA,经随机引物逆转录合成cDNA,先以型别特异的PCR 法分... 目的探讨HCV感染在肝炎发生中的病原学作用。方法对89例急性肝炎进行HCV标志物分析,并从HCV RNA阳性的5例非甲非乙型急性肝炎患者血清中提取HCV RNA,经随机引物逆转录合成cDNA,先以型别特异的PCR 法分型,再用巢式PCR扩增部分核心基因区序列,将产物连接P-GEM-T载体,在大肠杆菌中表达后测序分析。结果非甲非乙型急性肝炎中HAV、HBV和HCV阳性分别占47.2%、28.1%和15.7%,HAV和HBV双重感染占 14.6%,非甲、非乙和非丙肝炎占9%; HCV分型显示Ⅱ型、 Ⅲ型和Ⅱ/Ⅲ混合型分别占858%、71%和71%; Ⅱ型血清用于序列分析,扩增序列424hP与原设计完全一致。急性肝炎株间核苷酸同源性在981%~995%。氨基酸同源性在97.6%~99.2%;前者的同源性在1型为91.90/0,在Ⅱ型为943%~95.6%;后者与Ⅰ型、Ⅱ型的同源性在92.3%~958%。结论HCV为引起非甲非乙型急性肝炎的主要病毒之一,以Ⅱ型为主,应引起临床重视。 展开更多
关键词 序残分析 丙型肝炎 基因克隆 HCV
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原发性肝细胞癌病人PBMCs中HBV X、P基因的整合 被引量:1
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作者 王淑霞 闫志勇 +3 位作者 王力秋 赵海燕 刘成玉 王斌 《青岛大学医学院学报》 CAS 2006年第3期225-227,共3页
目的探讨外周血单个核细胞(PBMCs)中乙型肝炎病毒(HBV)X、P基因整合与原发性肝细胞癌(HCC)的关系。方法分离HBsAg阳性的40例原发性肝癌病人PBMCs,提取基因组DNA;分别设计HBV X基因、P基因PCR引物,检测HCC病人PBMCs基因组中HBV的X基因和... 目的探讨外周血单个核细胞(PBMCs)中乙型肝炎病毒(HBV)X、P基因整合与原发性肝细胞癌(HCC)的关系。方法分离HBsAg阳性的40例原发性肝癌病人PBMCs,提取基因组DNA;分别设计HBV X基因、P基因PCR引物,检测HCC病人PBMCs基因组中HBV的X基因和P基因的整合情况。结果HCC病人X基因在PBMCs中的整合率为77.5%(31/40),P基因整合率为15.0%(6/40),二者差异有显著性(χ2=31.43,P<0.001)。结论PBMCs中存在HBV X、P基因的整合,X基因片段的整合及表达是HCC发生的主要因素。 展开更多
关键词 肝细胞 肝炎病毒 乙型 基因 病毒整合
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萤光素酶报告基因在病毒学研究中的应用 被引量:1
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作者 赵海卫 韩佩君 +1 位作者 吕欣 尹文 《生物技术通讯》 CAS 2015年第1期131-134,共4页
萤光素酶是一类在氧气存在下能催化其底物特异性发光的酶,检出灵敏度高、特异性强。萤光素酶催化底物发光因无须外源光的激发,避免了非特异性干扰,具有其他报告基因不可替代的优势。萤光素酶基因作为报告基因已成为医学科学研究领域的... 萤光素酶是一类在氧气存在下能催化其底物特异性发光的酶,检出灵敏度高、特异性强。萤光素酶催化底物发光因无须外源光的激发,避免了非特异性干扰,具有其他报告基因不可替代的优势。萤光素酶基因作为报告基因已成为医学科学研究领域的重要标记工具,具有良好的应用前景。我们简要综述了萤光素酶报告基因在病毒学研究中的应用进展。 展开更多
关键词 萤光素酶 报告基因 病毒
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基因-病毒治疗系统CNHK300-小鼠内皮抑素对裸鼠胃癌的抗肿瘤作用 被引量:1
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作者 聂明明 方国恩 +2 位作者 王星华 苏长青 钱其军 《中华胃肠外科杂志》 CAS 2007年第6期565-569,共5页
目的探讨新构建的基因-病毒治疗系统CNHK300-小鼠内皮抑素murineendostatin(CNHK300-mE)对胃癌的抑制作用。方法通过胃癌SGC-7901细胞裸鼠皮下移植瘤模型观察该病毒治疗系统对胃癌生长和肿瘤血管生成的抑制作用。用电镜观察该病毒在... 目的探讨新构建的基因-病毒治疗系统CNHK300-小鼠内皮抑素murineendostatin(CNHK300-mE)对胃癌的抑制作用。方法通过胃癌SGC-7901细胞裸鼠皮下移植瘤模型观察该病毒治疗系统对胃癌生长和肿瘤血管生成的抑制作用。用电镜观察该病毒在肿瘤细胞中的复制情况及肿瘤细胞的超微结构改变。用酶联免疫吸附(ELISA)法检测mE基因在体内的表达。用免疫组织化学的方法检测腺病毒外壳蛋白六邻体(hexon)、增殖细胞核抗原(PCNA)及vWF因子相关抗原的表达情况。采用TUNEL法检测细胞凋亡。结果CNHK300-mE能在肿瘤细胞内复制,高表达mE,在第7天时,达到(2115±770)ng/ml(范围1745~3000ng/ml);明显抑制胃癌皮下移植瘤的生长及瘤内的血管生成,并可引起胃癌细胞的凋亡[治疗组小鼠肿瘤细胞凋亡率(78.4±9.1)%,对照组仅(15.2±0.5)%,P〈0.01],抑制肿瘤细胞增殖[治疗组小鼠PCNA指数(55.0±1.4)%,对照组为(74.1±0.4)%,P〈0.05]。结论CNHK300-mE能在小鼠胃癌内增殖复制,高效表达mE基因,抑制胃癌生长。 展开更多
关键词 胃肿瘤 基因治疗 病毒 端粒酶 内皮抑素
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金黄色葡萄球菌肠毒素A基因在C57BL/6小鼠淋巴细胞中的表达 被引量:1
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作者 汪宇 陆洪光 +2 位作者 程波 麦跃 余德厚 《中华皮肤科杂志》 CAS CSCD 北大核心 2005年第5期297-299,共3页
目的通过复制缺陷型腺病毒载体的介导,将金黄色葡萄球菌肠毒素A(SEA)基因转染入C57BL/6小鼠淋巴细胞中,观察基因表达产物对B16细胞的作用。方法通过MTT法直接测定重组腺病毒颗粒感染的淋巴细胞的增殖情况,采用双抗体夹心ABC-ELISA法,测... 目的通过复制缺陷型腺病毒载体的介导,将金黄色葡萄球菌肠毒素A(SEA)基因转染入C57BL/6小鼠淋巴细胞中,观察基因表达产物对B16细胞的作用。方法通过MTT法直接测定重组腺病毒颗粒感染的淋巴细胞的增殖情况,采用双抗体夹心ABC-ELISA法,测定淋巴细胞培养上清液中白介素2的水平。MTT法测定活细胞数目,以了解活化的C57BL/6小鼠淋巴细胞对B16细胞的杀伤效应。结果加入不同滴度的重组腺病毒颗粒后,C57BL/6小鼠淋巴细胞明显增殖;同时淋巴细胞培养上清液中白介素2水平也明显增加。重组腺病毒颗粒转染的C57BL/6小鼠淋巴细胞可明显杀伤B16细胞,效/靶比越高,杀伤效应越明显。结论SEA基因可以在C57BL/6小鼠淋巴细胞中表达,并且表达产物能够活化C57BL/6小鼠淋巴细胞,进而杀伤B16细胞。 展开更多
关键词 C57BL/6小鼠 金黄色葡萄球菌肠毒素 淋巴细胞 细胞培养上清液 C57BL/6小鼠 B16细胞 缺陷型腺病毒载体 基因表达产物 病毒颗粒 MTT法 白介素2 杀伤效应 双抗体夹心 SEA基因 基因转染 直接测定 细胞数目 重组 ABC 增殖 水平
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