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| 1 | 生物信息学技术与新基因的研究显示文摘0引言
新基因的克隆化无异是生物医学领域创新知识源泉的重要组成部分.这一任务,不仅是人类基因组计划(HGP)的核心内容,同时也是后基因组计划(post-HGP)的重要内容. | 成军 刘妍 陆荫英 李克 王琳 | 2003 | 世界华人消化杂志2003，11，4： | 45 |
| 2 | 新基因结构与功能研究的策略显示文摘发现一种新的基因,同时把其编码的新蛋白的结构与生物学功能、与生物学和临床医学之间的相互关系、以及新基因表达调节的机制阐明,是目前基因的分子生物学研究领域中最具挑战性的工作。首先利用酵母双杂交(yeast twohybrid)技术、酵母单杂交技术(yeast one-hybrid)、抑制性消减杂交(SSH, suppression subtractive hybridization)技术、基因芯片(DNA chip)技术、噬菌体表面展示(phage display)技术等获得蛋白结合蛋白的编码基因、差异表达的基因、DNA/RNA结合的蛋白基因等,或利用生物信息学技术获得推测的编码基因。然后,利用分子生物学技术和生物信息学技术相结合的手段,阐明新基因的功能、基因表达启动子的结构和调节机制基础,最终阐明新基因的结构和新蛋白的生物学功能,以及这种基因研究的可能临床医学意义。生物信息学技术与分子生物学技术的结合,是目前基因的分子生物学研究领域中的重要、有效的研究技术和方法。 | 成军 | 2003 | 世界华人消化杂志2003，11，4： | 34 |
| 3 | Effects of heat shock on change of HSC70/HSP68,acid and alkaline phosphatases before and after rat partial hepatectomy显示文摘INTRODUCTIONOnly the liver has the great capability ofregeneration in mammal.Few hepatocytes are inthe phase of division in the normal liver of an adultmammal (including human beings),but theremaining hepatocytes can be induced to proliferatequickly by partial hepatectomy (PH),and,to somedegree,they stop dividing and re-differentiate intocells functioning as hepatocytes.This shows | Lu AL Xu CS | 2000 | World Journal of Gastroenterology2000，6，5： | 27 |
| 4 | TECA hybrid artificial liver support system in treatment of acute liver failure显示文摘AIM: To assess the efficacy and safety of TECA type hybrid artificial liver support system (TECA-HALSS) in providing liver function of detoxification, metabolism and physiology by treating the patients with acute liver failure (ALF).METHODS: The porcine liver cells (1 - 2 ) x 1010 were separated from the Chinese small swine and cultured in the bioreactor of TECA-BALSS at 37.0°C and circulated through the outer space of the hollow fiber tubes in BALSS. The six liver failure patients with various degree of hepatic coma were treated by TECA-HALSS and with conventional medicines. The venous plasma of the patients was separated by a plasma separator and treated by charcoal adsorbent or plasma exchange. The plasma circulated through the inner space of the hollow fiber tubes of BALSS and mixed with the patients' blood cells and flew back to their blood circulation. Some small molecular weight substances were exchanged between theplasma and porcine liver cells. Each treatment lasted 6.0-7.0 h.Physiological and biochemical parameters were measured before, during and after the treatment.RESULTS: The average of porcine liver cells was (1.0- 3.0)x 1010 obtained from each swine liver using our modified enzymatic digestion method. The survival rate of the cells was 85% - 93% by tnypan blue stain and AO/PI fluorescent stain. After cultured in TECA-BALSS bioreactor for 6 h, the survival rate of cells still remained 70% - 85%. At the end of TECA-HALSS treatment, the levels of plasma NH3, ALT, TB and DB were significantly decreased. The patients who were in the state of drowsiness or coma before the treatment improved their appetite significantly and regained consciousness, some patients resumed light physical work on a short period after the treatment. One to two days after the treatment, the ratio of PTA increased warkedly. During the treatment, the heart rates, blood pressure, respiration condition and serum electrolytes (K+, Na+ and Cl) were stable without thrombosis and bleeding in all the six patients.CONCLUSION: TECA-HALSS treatment could be a rapid,safe and efficacious method to provide temporary liver support for patients with ALF. | Yi-Long Xue~1 Shi-Feng Zhao~1 Yun-Luo~1 Xin-Jian Li~1 Zhong-Ping Duan~2 Xiao-Ping Chen~1 Wen-Ge Li~1 Xiao-Qiang Huang~1 Yan-Ling Li~1 Xin-Cui~1 Da-Guang Zhong~1 Zuo-Yun Zhang~1 Zhi-Qiang Huang~1 1 Chinese PLA General Hospital,Beijing 100853,China2 Beijing You-An Hospital,Beijing 100054,China | 2001 | World Journal of Gastroenterology2001，7，6： | 15 |
| 5 | Insights on augmenter of liver regeneration cloning and function显示文摘肝的激发器物质(HSS ) 是被指了一肝特定但是种类非特定的生长因素。HSS 蛋白质的坡度纯化和顺序分析显示它包含了肝新生(ALR ) 的 augmenter,也作为 hepatopoietin (HPO ) 知道。ALR,充当一个 hepatotrophic 生长因素,象肝细胞瘤细胞在试管内一样的有教养的 hepatocytes 的明确地刺激的增长,支持的肝新生和损坏 hepatocytes 和救的尖锐肝的失败在活体内的恢复。ALR 属于新 Erv1/Alr 蛋白质家庭,哪个从酵母在更低、更高的优核质被发现到人并且甚至在一些双 stranded DNA 病毒的成员。现在的评论文章集中于 ALR 的分子的生物学,检验从酵母的 ALR 基因和它到人和 ALR 蛋白质的生物功能的表示。ALR 蛋白质似乎是 non-liver-specific 作为,增加必要性扩大关于在处于正常、反常的细胞的生长的条件的不同纸巾,机关和发展阶段的哺乳动物的 ALR 蛋白质的研究是以前相信了。 | Elisavet Gatzidou Gregory Kouraklis Stamatios Theocharis | 2006 | World Journal of Gastroenterology2006，12，31： | 13 |
| 6 | 大鼠肝再生相关基因LRRP1的克隆化显示文摘目的:利用抑制性消减杂交技术构建大鼠肝脏部分切除术后差异表达基因的cDNA文库,并通过同源引物反转录PCR的方法克隆大鼠肝再生相关基因LRRP1的全长序列.方法:分别提取肝脏部分切除术后24h以及对照大鼠肝脏mRNA,以之为模板合成cDNA,称为测试和驱动.酶切消化后将测试分成两组,分别与不同的接头连接,分别与驱动进行杂交,然后混合两份杂交液,加入过量的驱动不经变性进行第二次杂交即消减杂交.之后进行两次PCR扩增,将产物克隆入质粒载体,构建差显文库,测序后以生物信息学技术进行序列同源性比较.通过同源引物反转录PCR的方法克隆基因全序列,并用软件分析其蛋白结构.结果:成功构建大鼠肝脏部分切除术后的cDNA差显文库,消减效果良好.将所得的部分基因进行测序分析,选择一个未知序列,通过同源引物反转录PCR的方法克隆到大鼠肝再生相关基因LRRP1的全长序列,并初步分析了其蛋白结构.结论:确定了大鼠肝再生相关蛋白LRRP1的基因序列,并证明其在大鼠肝再生过程中高度表达. | 王刚 刘妍 牟劲松 洪源 邵得志 张耀新 李莉 成军 | 2002 | 世界华人消化杂志2002，10，2： | 11 |
| 7 | 功能基因组学与肝脏疾病研究显示文摘功能基因组学是指应用整体的研究技术阐明基因和蛋白的生物学功能.因此,需要发展一些强大的分析技术,对基因和蛋白质的功能进行研究.这必将为肝脏病学的研究提供了一个前所未有的发展机遇.本文主要论述功能基因组学的先进技术在肝病领域的应用. | 成军 | 2004 | 世界华人消化杂志2004，12，1： | 9 |
| 8 | Cloning of cytochrome P-450 2C9 cDNA from human liver and its expression in CHL cells显示文摘AIM: Using bacterial, yeast, or mammalian cell expressing a human drug metabolism enzyme would seem good way to study drug metabolism-related problems. Human cytochrome P-450 2C9 ( CYP2 C9) is a polymorphic enzyme responsible for the metabolism of a large number of clinically important drugs. It ranks among the most important drug metabolizing enzymes in humans. In order to provide a sufficient amount of the enzyme for drug metabolic research, the CYP2 C9 eDNA was cloned and expressed stably in CHL cellsMETHODS: After extraction of total RNA from human livertissue, the human CYP2C9 eDNA was amplified withreverse transcription-polymerase chain reaction (RT-PCR),and cloned into cloning vector pGEM-T. The cDNA fragmentwas identified by DNA sequencing and subcloned into amammalian expression vector pREP9. A transgenic cell linewas established by transfecting the recombinant vector ofpREP9-CYP2C9 into CHL cells. The enzyme activity ofCYP2C9 catalyzing oxidation of tolbutamide to hydroxytolbutamide in S9 fraction of the cell was determined by highperformance liquid chromatography(HPLC).RESULTS: The amino acid sequence predicted from theeDNA segment was identical to that of CYP2 C9 * 1, the wildtype CYP2 C9. However, there were two base differences, i.e. 21T > C, 1146C > T, but the encoding amino acidsequence was the same, L7, P382. The S9 fraction of theestablished cell line metabolizes tolbutamide to hydroxytolbutamide; tolbutamide hydroxylass activity was found to be0.465 ± 0.109 μmol@ min-1 . g1 S9 protein or 8.62 ± 2.02 mol@ min 1 ~mol-1 CYP, but was undetectable in parental CHL cell.CONCLUSION: The cDNA of human CYP2C9 was successfullycloned and a cell line of CHL- CYP2C9, efficiently expressingthe protein of CYP2C9, was established. | Ge-Jian Zhu Ying-Nian Yu,Department of Pathophysiology and Laboratory of Medical Molecular Biology,Zhejiang University School of Medicine,Hangzhou 310031,Zhejiang Province,China Xin Li,Department of pharmaceutical analysis & drug metabolism,College of Pharmacology Science,Zhejiang University,Hangzhou 310031,Zhejiang Province,China Yu-Li Qian, Present address:Center of laboratory,Women’s hospital,School of Medicine,Zhejiang University,Hangzhou 310031,Zhejiang Province,China | 2002 | World Journal of Gastroenterology2002，8，2： | 9 |
| 9 | Establishment of a transgenic cell line stably expressing human cytochrome P450 2C18 and identification of a CYP2C18 clone with exon 5 missing显示文摘AIM: The human cytochrome P-450 2C18(CYP2C18) hasbeen characterized. However, the protein has not beenpurified from liver and very little is known regarding thespecific substrate of CYP2C18. In order to study its enzymaticactivity for drug metabolism, the CYP2C18cDNA was clonedand a stable CHL cell line expressing recombinant CYP 2C18was established.METHODS: The human CYP2C18cDNA was amplified withreverse transcription-polymerase chain reaction (RT-PCR)from total RNAs extracted from human liver and cloned intopGEM-T vector. The cDNA segment was identified by DNAsequencing and subcloned into a mammalian expressionvector pREP9. A transgenic cell line was established bytransfecting the recombinant plasmid of pREPg-CYP2C18toChinese hamster lung (CHL) cell. The enzyme activity ofCYP2C18 catalyzing oxidation of tolbutamide tohydroxytolbutamide in postmitochondrial supernant(Sg)fraction of the cell was determined by high performanceliquid chromatography(HPLC).RESULTS: The amino acid sequence predicted from thecloned cDNA segment was identical to that of reported byRomkes et al(GenBank accession number: M61856,J05326).The S9 fraction of the established cell line metabolizestolbutamide to hydroxytolbutamide. Tolbutamide hydroxylaseactivity was found to be 0.509±0.052 μmol.min-1.g-1 S9protein or 8.82±0.90 mol.min-1.mol-1 CYP, but wasundetectable in parental CHL cell. In addition, we haveidentified a CYP2C18cDNA clone with exon 5 missing.CONCLUSION: The cDNA of human CYP2C18 wassuccessfully cloned and a cell line, CHL-CYP2C18, efficientlyexpressing the protein of CYP2C18, was established. Aspliced variant of CYP2C18 with exon 5 missing was identifiedin the cloning process. | JianZhu-GE Ying-NianYu | 2002 | World Journal of Gastroenterology2002，8，5： | 6 |
| 10 | 猪丙型肝炎病毒核心蛋白结合蛋白6同源基因的克隆化研究显示文摘利用不同种属动物之间重要基因序列高度同源的理论 ,应用分子生物学与生物信息学技术和方法 ,克隆猪丙型肝炎病毒 (HCV)核心蛋白结合蛋白 6(HCBP6)的同源基因。首先应用酵母双杂交技术 ,以表达HCV核心蛋白的表达载体作为诱饵 ,对于百万级的肝细胞cDNA文库酵母进行配合、筛选 ,首先获得人HCBP6的全长编码基因 ,然后应用美国国立生物工程中心 (NCBI)建立的核苷酸序列数据库 (GenBank)的同源基因的检索 ,搜索与之同源的来源于猪的表达序列标签 (EST)。然后根据基因同源性的原则 ,确定猪HCBP6的同源基因。获得了与HCV核心蛋白结合蛋白的 3 6个基因片段 ,其中之一命名为HCBP6。根据基因同源性搜索 ,获得了来源于猪的EST基因序列片段 ,最终确立了猪HCBP6的同源基因。利用不同物种之间基因同源性的原理、NCBI数据库GenBank同源基因的搜索 ,获得了猪HCBP6同源基因。 | 成军 李克 王琳 陆荫英 刘妍 王刚 张玲霞 | 2003 | 生物学杂志2003，20，4： | 5 |
| 11 | 人肝再生增强因子上调细胞周期素B2基因的表达显示文摘目的研究人肝再生增强因子(ALR)转基因表达对细胞周期素B2启动子(cyclin B2p)转录的调节作用,从而进一步探讨ALR的生物学作用及机制。方法根据文献确定细胞周期素B2p区域,聚合酶链反应(PCR)扩增细胞周期素B2p,克隆至报告基因表达载体pCAT3-Basic中,构建pCAT3-cyclin B2p报告载体;以该质粒转染肝癌细胞系HepG2细胞系,同时与ALR的真核表达载体pcDNA3.1(-)-ALR共转染HepG2细胞系,用酶联免疫吸附法(ELISA)检测报告基因氯霉素乙酰转移酶(CAT)的表达活性。结果成功构建pCAT3- cyclin B2p报告基因表达载体;pCAT3-cyclin B2p与pcDNA3.1(-)-ALR共转染HepG2细胞系的CAT表达活性是pCAT3-Basic空载体的25.13倍,是pCAT3-cyclin B2p单转染的2.60倍。结论人ALR对细胞周期素B2p基因的转录表达具有反式激活作用。 | 邵凤娟 成军 马英骥 郭江 刘妍 王巧侠 | 2006 | 中国地方病学杂志2006，25，6： | 3 |
| 12 | 构象敏感凝胶电泳快速检测HBV准种序列异质性的方法显示文摘目的:建立一种快速检测乙型肝炎病毒(HBV)准种遗传异质性的方法并验证其敏感性和特异性。以满足研究HBV准种需要检测大量病毒序列的要求。 方法:在构象敏感凝胶电泳(CSGE)检测方法的基础上,针对HBV核酸序列组成的特殊性,调整凝胶中使用的甲酰胺和乙二醇浓度并加入一定量的尿素,另外通过一次预电泳优选驱动序列。最后用改进地CSGE检测已知序列的HBV C-ORF区片段克隆,并把检测结果与DNA序列分析结果进行比对,验证该方法的敏感性和特异性。 结果:用改进地CSGE检测克隆的HVC C-ORF区片段获得了清楚地电泳图谱。对已知序列的HBC C-ORF区克隆片段进行检测:在34个克隆中发现了27种不同的克隆型,准种内病毒株(克隆)频率介于2.9~17.6%,克隆型之间的遗传距离介于0.2~2%,CSGE所得结果与DNA序列分析结果高度一致。 结论:我们建立的CSGE具有高度的敏感性和特异性,完全可以用于检测HBV准种序列的异质性及其他相关研究。 | 兰林 王宇明 | 2002 | 世界华人消化杂志2002，10，4： | 3 |
| 13 | 肝细胞生成素快速诱导肝再生相关基因mRNA的表达显示文摘目的:观察人重组肝细胞生成素(rhHPO)迅速诱导瞬时早期反应基因表达情况.方法:利用PT-PCR方法观察HPO刺激原代培养大鼠肝细胞24h内选择性肝再生相关基因mRNA表达情况.结果:HPO在原代培养大鼠肝细胞体系中可迅速诱导瞬时早期反应基因c-fos,LRF-1d的表达,对PC3和Tec基因也有诱导表达的作用,高峰出现在刺激后的0.5-2h,表达量增加3-5倍.结论:HPO可迅速诱导瞬时早期反应基因表达,可能是HPO促肝细胞增生的分子作用机制,首次报道PC3和Tec基因是一种与肝再生调控密切相关的早期反应基因. | 王阁 冷恩仁 胡辂 王军 房殿春 杨晓明 张咏 贺福初 | 2002 | 世界华人消化杂志2002，10，2： | 0 |
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