INTRODUCTIONHepatocellular carcinoma(HCC)is one of the major causes of death in the word.The mechanism of carcinogenesis is unknown,although it is widely accepted that HBV and HCV are clsely related to liver cancer[1-...INTRODUCTIONHepatocellular carcinoma(HCC)is one of the major causes of death in the word.The mechanism of carcinogenesis is unknown,although it is widely accepted that HBV and HCV are clsely related to liver cancer[1-5[1-5].Previously,a variety of studies have described the differences in gene expression which distinguished tumor from nontumor[6-11].Cloning of the genes,especially the genes associated with HBV and HCV,is still very important to account for the development of liver cancer.展开更多
AIM To investigate the anti-HBV effect ofoxymatrine (oxy) in vivo.METHODS HBV transgenic mice were producedby micro-injection of a 4.2kb fragmentcontaining the complete HBV genomes.Expression level of HBsAg and HBcAg ...AIM To investigate the anti-HBV effect ofoxymatrine (oxy) in vivo.METHODS HBV transgenic mice were producedby micro-injection of a 4.2kb fragmentcontaining the complete HBV genomes.Expression level of HBsAg and HBcAg in thetransgenic mice liver was determined byimmunohistochemical assay.RESULTS Four groups (6 mice in each group)were injected intraperitoneally with oxy at thedosage of 100,200, and 300 mg/kg or with salineonce a day for 30 days. Both HBsAg and HBcAgwere positive in livers of all the six mice in thecontrol group (injected with saline), and werepositive in livers of two mice in 100 mg/kg groupand 300mg/kg group. In 200mg/kg group,HBsAg and HBcAg were negative in livers of allthe six mice. Based on the results, 200 mg/kg isthe ideal dosage to explore the effect of oxy atdifferent time points. According to the oxytreatment time, mice were divided into fourgroups: 10 d, 20 d, 30 d and 60 d (4 mice in eachgroup). Each mouse underwent liver biopsy twoweeks before the treatment of oxy. Down-regulation of HBsAg and HBcAg appeared aftertreatment of oxymatrine for 10 d and 20 d, Dane-like particles disappeared after the treatment ofoxy for 20d under electron microscopy,however, the expression level of HBsAg andHBcAg returned to normal 60 d later after oxytreatment.CONCLUSION oxymatrine can reduce thecontents of HBsAg and HBcAg in transgenic miceliver, longer treatment time and larger dosagedo not yield better effects.展开更多
INTRODUCTIONHepatitis B virus (HBV) belongs to the group ofhepatovirus, a major pathogen of human acute andchronic hepatitis B[1 4], which has a very closeassociation with human hepatocellular carcinoma(HCC)[5-8], For...INTRODUCTIONHepatitis B virus (HBV) belongs to the group ofhepatovirus, a major pathogen of human acute andchronic hepatitis B[1 4], which has a very closeassociation with human hepatocellular carcinoma(HCC)[5-8], For example, a statistical data from ahospital in Shanghai showed that 80% of HCCpatients were positive for HBsAg ( personalcommunication).展开更多
目的:研究乙型肝炎病毒(Hepatitis B virus,HBV)对microRNA形成相关因子Pasha表达的调控,并对其作用机制进行初步探讨。方法:用RT-PCR,Western blot的方法检测HepG2细胞及稳定表达HBV的HepG2-H7细胞中Pasha的表达差异。构建Pasha启动子...目的:研究乙型肝炎病毒(Hepatitis B virus,HBV)对microRNA形成相关因子Pasha表达的调控,并对其作用机制进行初步探讨。方法:用RT-PCR,Western blot的方法检测HepG2细胞及稳定表达HBV的HepG2-H7细胞中Pasha的表达差异。构建Pasha启动子的萤火虫荧光素酶报告质粒,分别转染HepG2细胞及HepG2-H7细胞,检测HBV对Pasha启动子的影响。将Pasha启动子质粒与HBV 4种蛋白的真核表达质粒共转染HepG2细胞,寻找对启动子影响较大的HBV蛋白。结果:RT-PCR和Western blot的结果显示Pasha在HepG2-H7细胞中的表达较HepG2细胞明显下降。萤火虫荧光素酶活性分析显示HBV能抑制Pasha启动子的活性,且HBx和HBs蛋白对其影响较大。结论:HBV蛋白可以通过抑制Pasha启动子活性调节其在HepG2-H7细胞中的表达。展开更多
目的:探讨乙型肝炎病毒X蛋白(hepatitis B virus X protein,HBx)参与人肝癌细胞hsp90α基因表达的调控机制。方法:采用半定量RT-PCR、蛋白质印迹法及荧光素酶活性检测技术,检测不同剂量HBx表达载体转染的HepG2细胞中hsp90α基因mRNA和...目的:探讨乙型肝炎病毒X蛋白(hepatitis B virus X protein,HBx)参与人肝癌细胞hsp90α基因表达的调控机制。方法:采用半定量RT-PCR、蛋白质印迹法及荧光素酶活性检测技术,检测不同剂量HBx表达载体转染的HepG2细胞中hsp90α基因mRNA和蛋白表达及启动子活性的变化,并观察转录因子c-Myc阻断剂10058-F4对上述效应的影响。采用启动子实验分析c-Myc对hsp90α基因启动子的激活作用。RT-PCR和蛋白质印迹法检测表达HBx的HepG2细胞中c-Myc mRNA和蛋白表达情况;蛋白质印迹法及ELISA分析细胞外信号调节激酶1/2(ERK1/2)和磷酸化ERK1/2(p-ERK1/2)表达水平及内源性ERK的活性。结果:HBx基因转染HepG2细胞中hsp90α基因的mRNA和蛋白表达水平以及启动子活性呈剂量-依赖性增强,P=0.006 1;c-Myc阻断剂抑制了HBx对hsp90α基因表达的上调效应。c-Myc通过与c-Myc位点(-1094/-1088)结合转录激活hsp90α基因启动子活性。HBx使HepG2细胞中c-Myc mRNA和蛋白表达增强,并伴有ERK1/2蛋白磷酸化水平增加,ERK活性明显提高,P=0.032。结论:HBx通过ERK信号通路反式激活肝癌细胞c-Myc介导的hsp90α基因启动子活性,从而上调hsp90α基因表达。展开更多
目的探讨贵州省世居侗族和汉族人群趋化因子RANTES基因（regulated upon activation normal T cell expressed and secreted）-403G／A和-28C／G位点多态性与乙型肝炎病毒（HBV）感染后结局的相关性。方法采取病例对照方法，募集侗族和...目的探讨贵州省世居侗族和汉族人群趋化因子RANTES基因（regulated upon activation normal T cell expressed and secreted）-403G／A和-28C／G位点多态性与乙型肝炎病毒（HBV）感染后结局的相关性。方法采取病例对照方法，募集侗族和汉族共计229例HBV持续感染者、161例HBV清除者作为研究对象，同时选取200例正常者作为对照组。应用TaqMan—MGB探针实时荧光聚合酶链反应SNP分型技术对趋化因子RANTES基因-403G／A和-28C／G位点进行分型测定。结果趋化因子RANTES基因-403G／A和-28C／G基因型频率在侗族汉族HBV持续感染组、HBV清除组和正常对照组间差异无统计学意义（P〉0．05），单因素分析发现-403AG和-28GG增加HBV持续感染的危险度（OR=1．292，OR=1．151）。趋化因子RANTES-28C／G基因型频率在侗族和汉族HBV持续感染组分布差异有统计学意义（P〈0．05），而RANTES-403G／A各基因型在两族人群中分布的差异无统计学意义（P〉0．05）。结论携带趋化因子RANTES-403AG和-28GG基因型HBV持续感染风险性高。RANTES-403A有助于HBV清除。展开更多
文摘INTRODUCTIONHepatocellular carcinoma(HCC)is one of the major causes of death in the word.The mechanism of carcinogenesis is unknown,although it is widely accepted that HBV and HCV are clsely related to liver cancer[1-5[1-5].Previously,a variety of studies have described the differences in gene expression which distinguished tumor from nontumor[6-11].Cloning of the genes,especially the genes associated with HBV and HCV,is still very important to account for the development of liver cancer.
基金Supported by Projects of the Science Development Foundation of Shanghai, No. 994919033Tackling Key Problems in Science+1 种基金 Technology from the State ScienceTechnology Ministry, TJ99-LA01
文摘AIM To investigate the anti-HBV effect ofoxymatrine (oxy) in vivo.METHODS HBV transgenic mice were producedby micro-injection of a 4.2kb fragmentcontaining the complete HBV genomes.Expression level of HBsAg and HBcAg in thetransgenic mice liver was determined byimmunohistochemical assay.RESULTS Four groups (6 mice in each group)were injected intraperitoneally with oxy at thedosage of 100,200, and 300 mg/kg or with salineonce a day for 30 days. Both HBsAg and HBcAgwere positive in livers of all the six mice in thecontrol group (injected with saline), and werepositive in livers of two mice in 100 mg/kg groupand 300mg/kg group. In 200mg/kg group,HBsAg and HBcAg were negative in livers of allthe six mice. Based on the results, 200 mg/kg isthe ideal dosage to explore the effect of oxy atdifferent time points. According to the oxytreatment time, mice were divided into fourgroups: 10 d, 20 d, 30 d and 60 d (4 mice in eachgroup). Each mouse underwent liver biopsy twoweeks before the treatment of oxy. Down-regulation of HBsAg and HBcAg appeared aftertreatment of oxymatrine for 10 d and 20 d, Dane-like particles disappeared after the treatment ofoxy for 20d under electron microscopy,however, the expression level of HBsAg andHBcAg returned to normal 60 d later after oxytreatment.CONCLUSION oxymatrine can reduce thecontents of HBsAg and HBcAg in transgenic miceliver, longer treatment time and larger dosagedo not yield better effects.
基金This work was supported by Projects of Tackling Key Problems in ScienceTechnology from the State Science+2 种基金Technology Ministry (TJ99-LA01) Shanghai ScienceTechnology Commission (994919033 )
文摘INTRODUCTIONHepatitis B virus (HBV) belongs to the group ofhepatovirus, a major pathogen of human acute andchronic hepatitis B[1 4], which has a very closeassociation with human hepatocellular carcinoma(HCC)[5-8], For example, a statistical data from ahospital in Shanghai showed that 80% of HCCpatients were positive for HBsAg ( personalcommunication).
文摘目的:探讨乙型肝炎病毒X蛋白(hepatitis B virus X protein,HBx)参与人肝癌细胞hsp90α基因表达的调控机制。方法:采用半定量RT-PCR、蛋白质印迹法及荧光素酶活性检测技术,检测不同剂量HBx表达载体转染的HepG2细胞中hsp90α基因mRNA和蛋白表达及启动子活性的变化,并观察转录因子c-Myc阻断剂10058-F4对上述效应的影响。采用启动子实验分析c-Myc对hsp90α基因启动子的激活作用。RT-PCR和蛋白质印迹法检测表达HBx的HepG2细胞中c-Myc mRNA和蛋白表达情况;蛋白质印迹法及ELISA分析细胞外信号调节激酶1/2(ERK1/2)和磷酸化ERK1/2(p-ERK1/2)表达水平及内源性ERK的活性。结果:HBx基因转染HepG2细胞中hsp90α基因的mRNA和蛋白表达水平以及启动子活性呈剂量-依赖性增强,P=0.006 1;c-Myc阻断剂抑制了HBx对hsp90α基因表达的上调效应。c-Myc通过与c-Myc位点(-1094/-1088)结合转录激活hsp90α基因启动子活性。HBx使HepG2细胞中c-Myc mRNA和蛋白表达增强,并伴有ERK1/2蛋白磷酸化水平增加,ERK活性明显提高,P=0.032。结论:HBx通过ERK信号通路反式激活肝癌细胞c-Myc介导的hsp90α基因启动子活性,从而上调hsp90α基因表达。
文摘目的探讨贵州省世居侗族和汉族人群趋化因子RANTES基因（regulated upon activation normal T cell expressed and secreted）-403G／A和-28C／G位点多态性与乙型肝炎病毒（HBV）感染后结局的相关性。方法采取病例对照方法，募集侗族和汉族共计229例HBV持续感染者、161例HBV清除者作为研究对象，同时选取200例正常者作为对照组。应用TaqMan—MGB探针实时荧光聚合酶链反应SNP分型技术对趋化因子RANTES基因-403G／A和-28C／G位点进行分型测定。结果趋化因子RANTES基因-403G／A和-28C／G基因型频率在侗族汉族HBV持续感染组、HBV清除组和正常对照组间差异无统计学意义（P〉0．05），单因素分析发现-403AG和-28GG增加HBV持续感染的危险度（OR=1．292，OR=1．151）。趋化因子RANTES-28C／G基因型频率在侗族和汉族HBV持续感染组分布差异有统计学意义（P〈0．05），而RANTES-403G／A各基因型在两族人群中分布的差异无统计学意义（P〉0．05）。结论携带趋化因子RANTES-403AG和-28GG基因型HBV持续感染风险性高。RANTES-403A有助于HBV清除。
