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| 1 | 癌周及癌中心不同截短片段丙型肝炎病毒核心蛋白表达质粒的构建及表达显示文摘目的 研究不同截短片段丙型肝炎病毒(HCV)的核心蛋白(CORE)在HCV持续感染和肝癌发病机制中的作用,构建并表达不同HCV病毒株及不同截短片断CORE原核表达质粒。 方法 用PCR扩增基因型为HCV 1 b型的7个不同截短片段CORE基因:氨基酸(aa)长度分别为癌中心株(BT):B:1-172 aa、1-126 aa、1-58 aa、59-126 aa、127-172 aa;癌旁株(BNT):1-172 aa及C191(HCV-J6):1-172 aa,扩增产物用BamH Ⅰ及EcoR Ⅰ双酶切后将其插入到原核表达质粒pGEX-4T-1中,阳性克隆转染到BL21中,IPTG诱导表达GST-CORE融合蛋白,纯化定量,并用western blot加以验证。 结果 7个不同片段CORE在体外都得到相应表达,但表达量存在一定的差异,较长片段表达量相对较低,其中BT及BNT的1-172 aa截短片段CORE的表达高于同基因型等长度片段的C191,而BT以59-126 aa片段的表达量最高。BT、BNT及C191三株HCV基因1b型病毒株部分截短片段可形成二聚体。 结论 构建不同HCV病毒株及不同截短片段的CORE原核表达质粒获得成功,为研究HCV不同病毒株及不同区域CORE功能奠定基础;不同截短片段的CORE表达量不等,可能与不同片段具有不同的疏水基团、细胞毒性及在HCV结构和在HCV致病性中所起作用不同有关;HCV基因1b型的CORE二聚体的形成依耐于59-126 aa区域。 | 颜学兵 陈智 Delphine BOUCREUX Nicole PAVIO Christian BRECHOT | 2004 | 中华肝脏病杂志2004，12，11： | 5 |
| 2 | 丙型肝炎病毒核心蛋白结合蛋白激酶R的功能区域研究显示文摘目的构建并表达丙型肝炎病毒(HCV)不同病毒株:癌中心株(BT)、癌旁珠(BNT)、HCV-J及BT不同截短片段谷胱甘肽(GST)-核心融合蛋白;寻找核心蛋白(Core)与蛋白激酶R(PKR)相互作用区域,探讨它们在HCV持续感染及细胞癌(HCC)发病机制中的作用.方法用聚合酶链反应扩增不同片段HCV核心蛋白基因,并将7个不同的基因片段分别克隆到原核表达载体pGEX-4T-1,诱导表达并纯化表达蛋白,与两株细胞(HepG2和Huh-7)的PKR的进行相互作用试验.结果7个不同片段Core在体外都得到相应表达,不同片段结合PKR的能力存在一定差异.BT、BNT、C191的Core N端1~172氨基酸(aa)3个片段均能与PKP发生直接结合,BT与PKR结合的区域在Core N端的1~58aa.结论 不同片段Core在原核细胞中获得较好的表达;Core/PKP相互作,在HCV持续感染和HCC的发病机制中可能起重要作用. | 颜学兵 陈智 Delphine BOUCREUX Nicole PAVIO Christian BRECHOT | 2005 | 中华传染病杂志2005，23，1： | 4 |
| 3 | Selective accumulation of the X transcript of hepatitis B virus in patients negative for hepatitis B surface antigen with hepatocellular carcinoma显示文摘 | Patrizia Paterlini Karine Poussin Michael Kew Dominique Franco Christian Brechot | 1995 | Hepatology1995，，2： | 2 |
| 4 | Hepatitis C virus core proteins derived from different quasispecies of genotype 1b inhibit the growth of Chang liver cells显示文摘AIM: To investigate the influence of different quasispecies of hepatitis C virus (HCV) genotype 1b core protein on growth of Chang liver cells. METHODS: Three eukaryotic expression plasmids (pEGFP-N1/core) that contained different quasispecies truncated core proteins of HCV genotype 1b were constructed. These were derived from tumor (T) and non- tumor (NT) tissues of a patient infected with HCV and C191 (HCV-J6). The core protein expression plasmids were transiently transfected into Chang liver cells. At different times, the cell cycle and apoptosis was assayed by flow cytometry, and cell proliferation was assayed by methyl thiazolyl tetrazolium (MTT) assay. RESULTS: The proportion of S-phase Chang liver cells transfected with pEGFP-N1/core was significantly lower than that of cells transfected with blank plasmid at three different times after transfection (all P < 0.05). The proliferation ratio of cells transfected with pEGFP-N1/corewas significantly lower than that of cells transfected with blank plasmid. Among three different quasispecies, T, NT and C191 core expression cells, there was no significant difference in the proportion of S- and G0/G1-phase cells. The percentage of apoptotic cells was highest for T (T > NT > C191), and apoptosis was increased in cells transfected with pEGFP-N1/core as the transfection time increased (72 h > 48 h > 24 h). CONCLUSION: These results suggest that HCV genotype 1b core protein induces apoptosis, and inhibits cell- cycle progression and proliferation of Chang liver cells. Different quasispecies core proteins of HCV genotype 1b might have some differences in the pathogenesis of HCV persistent infection and hepatocellular carcinoma. | Xue-Bing Yan Lei Mei Xia Feng Mei-Rong Wan Zhi Chen Nicole Pavio Christian Brechot | 2008 | World Journal of Gastroenterology2008，14，18： | 2 |
| 5 | Hepatocellular Carcinoma Is Associated With an Increased Risk of Hepatitis B Virus Recurrence After Liver Transplantation显示文摘 | Luciana C. Faria Michelle Gigou Anne M. Roque–Afonso Mylene Sebagh Bruno Roche Guillaume Fallot Teresa C.A. Ferrari Catherine Guettier Elisabeth Dussaix Denis Castaing Christian Brechot Didier Samuel | 2008 | Gastroenterology2008，，7： | 2 |
| 6 | Hepatitis C virus: molecular biology and genetic variability 显示文摘 | Brechot C | 1996 | Dig Dis Sci1996，41，12： | 1 |
| 7 | A second heterozygous MDR3 nonsense mutation associated with intrahepatic cholestasis of pregnancy显示文摘 | Gendrot C Bacq Y Brechot MC | 2003 | J Med Genet2003，40，： | 1 |
| 8 | A second heterozygous MDR3 nonsense mutation associated with intrahepatic cholestasis of pregnancy显示文摘 | Gendrot C Bacq Y Brechot M C | 2003 | J Med Genet2003，40，3： | 1 |
| 9 | Pathogenesis of hepatitis B virus-related hepatocellular carcinoma:old and new paradigms显示文摘 | Brechot C | 2004 | Gastreentemlogy2004，127，： | 1 |
| 10 | The detection of hepatitis B virus (HBV) in HBsAg negative individuals with primary liver cancer 显示文摘 | Paterlini P Brechot C | 1991 | Dig Dis Sci1991，36，8： | 1 |
| 11 | Immunotherapy of chronic hepatitis B by anti HBV vaccine: from present to future显示文摘 | Michel ML Pol S Brechot C | 2001 | Vac cine2001，19，： | 1 |
| 12 | What role do viruses play in nosoco- mial pneumonia显示文摘 | Luyt C E Brechot N Chastre J | 2014 | Curr Opin Infect Dis2014，27，2： | 1 |
| 13 | What is the niche for extracorporeal membrane oxygenation in severe acute respiratory distress syndrome? 显示文摘 | Combes A Brechot N Luyt CE | 2012 | Curr Opin Crit Care2012，18，5： | 1 |
| 14 | Pulmonary metastasis from endometrial stromal sarcoma may benefit from hormone therapy显示文摘 | Brechot J M Kamboucher M Brauner M | 2007 | Rev Mal Respir2007，24，1： | 1 |
| 15 | Gene therapy of hepatocarcinoma:a long way from the concept to the therapeutical impact显示文摘 | GEROLAMI R UCH R BRECHOT C | 2003 | Can Gene Ther2003，10，9： | 1 |
| 16 | A second heterozygous MDR3 nonsense mutation associated with intrahepatic eholestasis of pregnancy显示文摘 | Gendrot C Bacq Y Brechot MC | | 0，，： | 1 |
| 17 | Intrahepatic cholestasis of pregnancy:a French prospective study显示文摘 | Bacq Y Sapey T Brechot MC | 1997 | Hepatology1997，26，： | 1 |
| 18 | Persistent hepatitis B virus infection in subjects without hepatitis B surface antigen:clinically significant or purely 'occult'? 显示文摘 | BRECHOT C THIER V KREMSDORF D | 2001 | Hepatology2001，34，1： | 1 |
| 19 | Interaction between alcohol and hepatitis virus in the liver显示文摘 | Brechot C Nalpas B Feitelson M A | | 0，，： | 1 |
| 20 | Persistent hepatitis B virus infection in subjects without hepatitis B surface antigen clinically significant or purely occult 显示文摘 | BRECHOT C THIERS V KREMSDORF D | 2001 | Hepatology2001，34，： | 1 |
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