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| 1 | 慢性病毒性肝炎发病机制的分子生物学研究显示文摘由乙型肝炎病毒(HBV)和丙型肝炎病毒(HCV)的感染引起的急性和慢性肝病目前还没有满意的治疗方法.新型治疗方法和治疗药物的研究开发,依赖于肝炎病毒致病的分子生物学机制的研究进展.关于HBV感染个体准种概念的引入,使我们对于HBV基因变异的研究,从单一病毒的基因变异上升到群体病毒的基因变异?从静态的基因突变上升到动态的基因变异,从而对HBV存在状态的看法发生了根本的改变.HBV感染肝细胞的相关受体蛋白虽然进行了多年的研究,但还没有最终确定.利用新型技术对于HBV表面抗原蛋白的结合蛋白进行筛选是一个重要的方向.HBV和HCV感染与肝细胞癌之间的关系已经得到确定,但是具体的分子生物学机制还有许多工作要做.研究这2种肝炎病毒反式激活作用的把基因是阐明其引起肝细胞癌分子生物学机制的重要途径.在肝细胞中表达的肝炎病毒蛋白不是孤立存在的,或者与其自身结合形成同二聚体,或者与病毒的其他蛋白?肝细胞蛋白结合形成异二聚体,从而对于肝细胞的生长?代谢?甚至是恶性转化产生重要影响. | 成军 | 2002 | 世界华人消化杂志2002，10，2： | 141 |
| 2 | 截短型乙型肝炎病毒表面抗原中蛋白上调c-myc基因表达的研究显示文摘目的:应用抑制性消减杂交方法构建截短型乙型肝炎病毒表面抗原中蛋白(MHBst)反式调节基因的消减文库,用免疫印迹方法验证截短型乙型肝炎病毒表面抗原中蛋白对c-myc基因的上调表达.方法:构建羧基末端截短的乙型肝炎病毒表面抗原中蛋白的真核表达载体pcDNA3.1(-)-Mt,分别以重组表达质粒pcDNA3.1(-)-Mt和空载体pcDNA3.1(-)瞬时转染HepG2细胞,提取细胞mRNA并逆转录为cDNA,用抑制性消减杂交技术,将实验组与对照组cDNA进行两次消减杂交及两次抑制性多聚酶链反应(PCR),构建cDNA消减文库,并转染大肠杆菌进行文库扩增,随机挑选克隆PCR扩增后进行测序及同源性分析.生物信息学分析结果显示部分与肿瘤发生密切相关的基因,如癌基因c-myc.结果:显示截短型乙肝病毒表面抗原中蛋白可以反式激活c-myc基因,并上调其表达.结论:成功构建截短型乙肝病毒表面抗原中蛋白反式调节基因的消减文库,细胞原癌基因c-myc是MHBst反式激活作用的靶基因,为进一步阐明乙型肝炎病毒致癌的分子生物学机制提供理论基础. | 韩萍 刘妍 成军 王刚 陆荫英 李克 李莉 | 2002 | 世界华人消化杂志2002，10，2： | 66 |
| 3 | 乙型肝炎病毒X基因酵母表达载体构建及表达显示文摘目的:为进一步探讨乙型肝炎病毒(HBV)X蛋白(HBxAg)的功能,在真核生物酵母细胞中表达X基因。 方法:用多聚酶链反应(PCR)的方法以HBV ayw亚型全序质粒pCP10为模板扩增X基因,克隆到pGEM-T载体中扩增并测序鉴定,EcoRI和PstI双酶切后回收连接到酵母表达载体pGBKT-7中并转化酵母AH109,色氨酸缺陷型培养基(SD/-Trp)上筛选阳性菌落,提取酵母蛋白质,进行十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和Western免疫印迹分析,以明确HaxAg是否在其中表达。 结果:成功地扩增出HBx基因,测序鉴定符合Gene Bank报告序列;酶切回收的HBx基因成功地克隆入酵母表达载体pGBKT-7并转化入酵母细胞AH109中,Western免疫印迹显示X基因在酵母细胞中表达,表达产物在胞内存在,相对M_r为35000左右。 结论:成功地构建了HBxAg在酵母表达载体,并在酵母细胞中表达。 | 陆荫英 李克 成军 王琳 刘妍 段惠娟 张玲霞 | 2002 | 世界华人消化杂志2002，10，1： | 44 |
| 4 | 清除乙型肝炎病毒的非细胞裂解机制显示文摘乙型肝炎病毒(HBV)的感染过程,也是机体清除HBV的过程,两者密不可分。近年来的研究表明,除了细胞裂解并清除HBV感染的机制以外,还存在清除HBV的非细胞裂解机制,而后者更具有实际应用价值,因为在达到清除HBV的目的同时,还能不损伤HBV感染的肝细胞,或者只有很轻程度的肝细胞损害。研究中发现干扰素、白介素、肿瘤坏死因子、一氧化氮以及一些未知蛋白质因子的参与,是清除HBV的非细胞裂解机制的重要因素。研究清除HBV的非细胞裂解机制,具有十分重要的实际应用前景。 | 成军 李莉 | 2002 | 世界华人消化杂志2002，10，1： | 49 |
| 5 | Hepatoprotective role of ganoderma lucidum polysaccharide against BCG-induced immune liver injury in mice显示文摘AIM: To examine the effect of ganoderma lucidumpolysaccharide (GLP) on the immune liver injuryinduced by BCG infection, and investigate therelationship between degrees of hepatic damage andNO production in mice.METHODS: Immune hepatic injury was markedlyinduced by BCG-pretreatment (125 mg.kg-1, 2-week, iv)or by BCG-pretreatment plus lipopolysaccharide (LPS,125 μg.kg-1, 12-hour, iv) in mice in vivo.Hepatocellulardamage induced by BCG-pretreated plus inflammatorycytokines mixture (CM), which was included TNF-α, IL1β, IFN-γ and LPS in culture medium in vitro.Administration of GLP was performed by oral orincubating with culture medium at immune stimulisimultaneity. Liver damage was determined by activityof alanine aminotransferase (ALT) in serum and inhepatocytes cultured supernatant, by liver weightchanges and histopathological examination. NOproduction in the cultured supematant was determinedby the Griess reaction. Moreover, inducible nitric oxidesynthase (iNOS) protein expression was alsoexaminated by immunohistochemi1cal method.RESULTS: Immune hepatic injury was markedly inducedby BCG or BCG plus inflammatory cytokines in BALB/cmice in vivoand in vitro. Under BCG-stimulated condition,augment of the liver weight and increase of the serum/supernatant ALT level were observed, as well asgranuloma forming and inflammatory cells soakage wereobserved by microscopic analysis within liver tissues.Moreover, NO production was also increased by BCG or/and CH stimuli in the culture supernatant, and a lot ofiNOS positive staining was observed in BCG-prestimulated hepatic sections. Application of GLPsignificantly mitigated hepatic tumefaction, decreasedALT enzyme release and NO production in serum/supernatant, improved the pathological changes ofchronic and acute inflammation induced by BCG-stimuliin mice. Moreover, the immunohistochemical resultshowed that GLP inhibited iNOS protein expression inBCG-immune hepatic damage model.CONCLUSION: The present study indicates that NOparticipates in immune liver injury induced byMycobacterium bovis BCG infection. The mechanismsof protective roles by GLP for BCG-induced immune liverinjury may be due to influence NO production in mice. | Guo-Liang Zhang Ye-Hong Wang Wei Ni Hui-Ling Teng Zhi-Bin Lin Department of Pharmacology,School of Basic Medical Sciences,Beijing University,Beijing 100083,China | 2002 | World Journal of Gastroenterology2002，8，4： | 47 |
| 6 | Overexpression of p28/gankyrin in human hepatocellular carcinoma and its clinical significance显示文摘AIM: To investigate the expression of p28/gankyringene and its role in the carcinogenetic process of humanhepatocellular carcinoma (HCC).METHODS: 64 specimens of HCC and para-carcinomatissues, 22 specimens of non-tumor liver tissues (7normal, 15 cirrhosis), 10 specimens of normal humantissues and 5 hepatoma cell lines were studied for theexpression of p28/gankyrin by Northern blot. Theexpression of p28/gankyrin protein was detectedimmunohistochemically by using the specificpolyclonal antibody. RESULTS: Northern blot analysis indicated that theexpression of p28/gankyrin mRNA was intensivelydistributed in brain and heart, weakly in lung, spleenand muscle, undetectable in digestive system includingliver, pancreas, stomach, small and large intestines.p28/gankyrin mRNA was absent in normal liver, weaklydetected in liver cirrhosis and in 18 of 64 para-carcinoma liver tissues. In contrast, the expression ofp28/gankyrin mRNA was intensively detected in ali 5hepatoma cell lines tested, markedly increased in 57of 64 and moderately increased in 5 of 64 HCC samples.In comparison with liver cirrhosis and para-carcinomaliver tissues, the average expression of p28/gankyrin mRNA in HCC was increased 3.6- (2.901+0.507 vs 0.805 + 0.252, P<0.05) and 5.2-fold (2.901 +0.507 vs 0.557+0.203, p<0.01), respectively, Inaddition, p28/gankyrin mRNA expression level washigher in HCC with portal vein tumor thrombus andmicroscopic hepatic vein involvement (P--0.021 andP=-0.047, respectively). The overexpression of p28/gankyrin protein in HCC was targeted in hepatic tumorcells, not in bile duct cells and other interstitial cells.CONCLUSION: Overexpression of p28/gankyrin in HCCplays an important role and contributes to themetastasis potential in the process of carcinogenesis.p28/gankyrin may become a specific biological tissuemarker for the pathological diagnosis of HCC. | Xiao-YongFu Hong-YangWang | 2002 | World Journal of Gastroenterology2002，8，4： | 38 |
| 7 | Activating mechanism of transcriptor NF-kappaB regulated by hepatitis B virus X protein in hepatocellular carcinoma显示文摘AIM:To investigate the mechanism and significance of NF-κB activation regulated by hepatitis B virus X protein (HBx) in hepatitis B virus (HBV)-associated hepatocellular carcinoma (HCC).METHODS:The expression levels of HBx, p65,IκB-α and ubiquitin were detected by immunohistochemistry in HCC tissue microarrays (TMA) respectively, and IκB-α was detected by Western blot in HCC and corresponding liver tissues.RESULTS: The percentage of informative TMA samples was 98.8% in 186 cases with a total of 367 samples. Compared with corresponding liver tissues (60.0%),the HBx expression was obviously decreased in HBV-associated HCC (47.9%,u=2.24,P<0.05).On the contrary, the expressions of p65 (20.6% vs45.3%, u=4.85, P<0.01) and ubiquitin (8.9% vs 59.0%,u=9.68,P<0.01) were notably elevated in HCC.In addition, IκB-α had a tendency to go up. Importantly, positive relativity was observed between HBx and p65 (X^2=10.26,P<0.01), p65 and IκB-α (x^2=16.86,P<0.01), IκB-α and ubiquitin (x^2=8.90, P<0.01) in HCC, respectively.CONCLUSION:Both active and non-active forms of NF-κB are increased in HBV-associated HCC. Variant HBx is the major cause of the enhancement of NF-κB activity. The activation always proceeds in nucleus and the proteasome complexes play an important role in the activation. | TaoWang YiWang Meng-ChaoWu Xin-YuanGuan Zheng-FengYin | 2004 | World Journal of Gastroenterology2004，10，3： | 36 |
| 8 | Expression of NF-кB and human telomerase reverse transcriptase in gastric cancer and precancerous lesions显示文摘AIM:To investigate the expression of NF-κBp65 protein and human telomerase reverse transcriptase (hTERT) and their correlation in gastric cancer and precancerous lesions.METHODS: Forty-one patients with primary gastric cancer,15 with dysplasia, 23 intestinal metaplasia and 10 with normal gastric mucosa were included in this study.Expression of NF-κBp65 protein,hTERT mRNA and protein were determined by immunohistochemistry and in situ hybridization.RESULTS:The rate of p65 expression in normal gastric mucosa, intestinal metaplasia,dysplasia and carcinoma was 0%, 34.78%, 53.33% and 60.98%,respectively,while the rate of hTERT mRNA expression was 10.00%,39.13%,66.67% and 85.37% and the rate of hTERT protein expression was 0%,30.43%,60.00% and 78.05%,respectively.All the three parameters were significantly increased in dysplasia and carcinoma compared to normal mucosa, while the expression levels were also significantly higher in carcinoma than in intestinal metaplasia (P<0.05).In gastric cancer tissues, nuclear staining rates of p65 and hTERT protein were both significantly associated with the degree of differentiation, lymph node metastasis,clinical stage and invasion depth (P<0.05).However,hTERT mRNA expression was only significantly associated with clinical stage.There was a positive correlation between p65 and hTERT mRNA(rs=0.661-0.752, P<0.01),and between hTERT protein and hTERT mRNA (rs=0.609-0.750, P<0.01).CONCLUSION: NF-κBp65 and hTERT expressions are upregulated at the early stage of gastric carcinogenesis.NF-κB activation may contribute to hTERT expression and thereby enhance telomerase activity, which represents an important step in carcinogenesis progress. | WeiWang He-ShengLuo Bao-PingYu | 2004 | World Journal of Gastroenterology2004，10，2： | 30 |
| 9 | Applying a highly specific and reproducible cDNA RDA method to clone garlic up-regulated genes in human gastric cancer cells显示文摘AIM: To develop and optimize cDNA representationaldifference analysis (cDNA RDA) method and to identify andclone garlic up-regulated genes in human gastric cancer(HGC) cells.METHODS: We performed cDNA RDA method by usingabundant double-stranded cDNA messages provided by twoself-constructed cDNA libraries (Allitridi-trested and paternalHGC cell line BGC823 cells cDNA libraries respectively).BamH Ⅰ and Xho I restriction sites harbored in the libraryvector were used to select representations. Northern andSlot blots analyses were employed to identify the obtaineddifference products.RESJLTS: Fragments released from the cDNA library vectorafter restriction endonuclease digestion acted as goodmarker indicating the appropriate digestion degree for libraryDNA. Two novel expressed sequence tags (ESTs) and arecombinant gene were obtained. Slot blots result showed a8-fold increase of gila-derived nexin/protease nexin 1 (GDN/PN1 ) gene expression level and 4-fold increase of hepatitis Bvirus x-interacting protein (XIP) mRNA level in BGC823 cellsafter Allitridi treatment for 72 h.CONCLUSION: Elevated levels of GDN/PN1 and XIP mRNAsinduced by Allitridi provide valuable molecular evidence forelucidating the garlic' s efficacies against neurodegenerativeand inflammatory diseases. Isolation of a recombinant geneand two novel ESTs further show cDNA RDA based on cDNAlibraries to be a powerful method with high specificity andreproducibility in cloning differentially expressed genes. | Yong Li You-Yong Lu,Beijing Institute for Cancer Research,Beijing Laboratory of Molecular Oncology,School of Oncology,Peking University,Beijing 100034,China | 2002 | World Journal of Gastroenterology2002，8，2： | 24 |
| 10 | 病毒性肝炎发病机制中的反式调节机制显示文摘病毒性肝炎的发病机制中涉及到复杂的反式调节(transregulation)机制.肝炎病毒的反式调节机制至少包括三方面的含义:一方面是肝炎病毒蛋白对于肝细胞基因组表达调节的反式调节,即肝炎病毒蛋白与肝细胞基因组启动子DNA结合,对于肝细胞基因表达谱产生影响;第二方面是肝细胞蛋白对于肝炎病毒基因表达的反式调节.肝炎病毒属于简单的生物类型,要完成其生活周期,必须借助于肝细胞中的蛋白成分,肝细胞中特有的一些蛋白成分,与肝炎病毒基因组DNA/RNA结合,对于肝炎病毒基因表达进行调节,这也是决定肝炎病毒嗜肝特性的重要的分子生物学机制;第三方面是肝炎病毒蛋白对于肝炎病毒基因组表达的反式调节.这是所有病毒复制和表达调节的一般机制.关于肝炎病毒反式调节机制的研究,主要是阐明肝炎病毒为何在肝细胞中的复制和表达处于最佳状态,阐明肝炎病毒的致病机制,有助于据此设计新型的抗肝炎病毒的治疗技术和治疗方法. | 成军 | 2003 | 世界华人消化杂志2003，11，7： | 23 |
| 11 | 扶正抑瘤颗粒对食管癌、胃癌组织细胞周期及核转录因子的影响显示文摘目的 :观察扶正抑瘤颗粒对食管癌、胃癌患者肿瘤组织细胞周期及核转录因子 (NF κB)的影响。方法 :76例食管癌、胃癌患者随机分为 2组 ,服药组服用扶正抑瘤颗粒 15日后 ,与对照组分别采取手术切除肿瘤组织 ,用流式细胞仪检测肿瘤组织细胞周期、凋亡率和NF κB水平。结果 :服药组较对照组NF κB水平明显升高 (P <0 0 5 ) ;肿瘤细胞G0 /G1期比率明显升高 (P <0 0 5 ) ,S期比率明显降低 (P <0 0 5 )。同时 ,服药组癌细胞有明显的凋亡现象 ,其凋亡率与对照组比较差异有显著性 (P <0 0 1)。结论 :扶正抑瘤颗粒可提高肿瘤细胞NF κB的表达 ,阻滞肿瘤细胞增殖 ,促进肿瘤细胞凋亡。 | 赵健雄 曲勇 陈学忠 沈世林 | 2003 | 中国中西医结合杂志2003，23，12： | 21 |
| 12 | Overexpression of HBxAg in hepatocellular carcinoma and its relationship with Fas/FasL system显示文摘AIM: To study the expression and serum level of HBxAg,Fas and FasL in tissues of HCC patients, and to assess the relationship between HBxAg and Fas/FasL system.METHODS: Tissues from 50 patients with HCC were tested for the expression of HBxAg, Fas and FasL by S-P immunohistochemistry. Serum levels of sFas/sFasL and HBsAg/HBeAg were measured by ELISA assay. HBV X gene was detected by PCR in serum and confirmed by automatic sequencing. Fifty cases of liver cirrhosis and 30 normal controls were involved in serum analysis.RESULTS: The expression of HBxAg, Fas and FasL in carcinoma tissues was 96 %, 84 % and 98 %, respectively.Staining of HBxAg, Fas and FasL was observed predominately in cytoplasms, no significant difference was found in intensity between HBxAg, Fas and FasL (P>0.05). HBxAg, Fas and FasL might express in the same area of carcinoma tissues and this co-expression could be found in most patients with HCC. The mean levels of sFas in serum from HCC, cirrhosis and normal controls were 762.29±391.56 μg@ L-1 835.36±407.33 μg@L-1 and 238.27±135.29 μg@L-1. The mean levels of sFasL in serum from HCC, cirrhosis and normal controls were 156.36±9.61iμg@ L-1, 173.63±18.74 μg@L-1 and 121.96±7.83 μg@ L-1.Statistical analysis showed that both sFas and sFasL in HCC and cirrhosis patients were significantly higher than those in normal controls (P<0.01). Serum HBV X gene was found in 32 % of HCC patients and ,46 % of cirrhotic patients.There was no significant relationship between serum level of sFas/sFasL and serum X gene detection (P>0.05). Eight percent of HCC patients with negative HBsAg and HBeAg in serum might have X gene in serum and HBxAg expression in carcinoma tissues.CONCLUSION: Our data suggest that HBxAg and Fas/FasL system plays an important role in the development of human HCC. Expression of HBxAg can leads to expression of Fas/FasL system which and reverse apoptosis of hepatocellular carcinoma induced by FasL. | Xiao-ZhongWang Xiao-ChunChen Yun-XinChen Li-JuanZhang DanLi Feng-LinChen Zhi-XinChen Hong-YingChen Qi-MingTao | 2003 | World Journal of Gastroenterology2003，9，12： | 20 |
| 13 | 核因子-κB与大肠肿瘤细胞凋亡及增生的关系显示文摘目的:通过观察大肠腺瘤、腺癌和正常大肠组织中的细胞凋亡、细胞增生及其调控基因Bcl-2,Bcl-Xl,C-myc和核因子-κB的表达状态,探讨核因子-κB在大肠癌发病过程中的作用。 方法:利用DNA缺口末端标记技术和NF-κB,Bcl-2,Bcl-Xl,C-myc,PCNA蛋白免疫组化染色,原位检测30例大肠腺瘤和30例大肠腺癌中凋亡细胞、细胞增生和NF-κB,Bcl-2,Bcl-Xl,C-myc阳性表达细胞的密度和分布,以10例正常大肠组织作对照。 结果:NF-κB,Bcl-2,Bcl-Xl,C-myc蛋白的表达率和表达强度腺癌和腺瘤高于正常组织(70.9±7.2 vs 54.0±4.5 vs 30.6±2.4;78.2±8.3 vs 55.6±6.5 vs 11.0±1.9;77.3±6.5 vs 56.4±4.7vs 9.3±1.1;70.3±6.7 vs 50.1±4.2 vs 14.2±1.7,P<0.01),腺癌高于腺瘤(70.9±7.2 vs 54.0±4.5;78.2±8.3 vs 55.6±6.5;77.3±6.5 vs 56.4±4.7;70.3±6.7 vs 50.1±4.2,P<0.01)。腺瘤和腺癌中的凋亡细胞指数均高于正常组织(105.9±6.1 vs31.5±3.7 vs 10.0±1.5,P<0.01),腺瘤高于腺癌(105.9±6.1vs 31.5±3.7,P<0.01)。腺癌和腺瘤中的细胞增生高于正常组织(75.4±5.6 vs 50.7±4.9 vs 29.6±3.0,P<0.01);腺癌高于腺瘤(75.4±5.6 vs 50.7±4.9,P<0.01)。 结论:NF-κB在大肠肿瘤中促进细胞增生、抑制细胞凋亡。 | 於亮亮 于皆平 冉宗学 于红刚 | 2002 | 世界华人消化杂志2002，10，3： | 18 |
| 14 | Hepatitis B virus X gene induces human telomerase reverse transcriptase mRNA expression in cultured normal human cholangiocytes显示文摘AIM: To study the transcriptional regulation of human telomerase reverse transcriptase (hTERT) mRNA in normal human cholangiocytes (HBECs) after hepatitis B virus X (HBx) gene transfection and to elucidate the possible mechanism of HBV infection underlying cholangiocarcinoma.METHODS: HBECs were cultured in vitroand co-transfected with a eukaryotic expression vector containing the HBx coding region and a doning vector containing coding sequences of enhanced green fluorescent protein (EGFP) using lipidmediated gene transfer. The transfection efficiency was determined by the expression of EGFP. The expressions of hTERT mRNA and HBx protein in HBECs were detected by RT-PCR and immunocytochemical stain, respectively.RESULTS: The transfection efficiencies were about 15% for both HBx gene expression plasmid and empty vector.No hTERT mRNA was expressed in HBECs when transfected with OPTI-NEN medium and empty vector, but a dramatic increase was observed for hTERT mRNA expression in HBECs when transfected with HBx expression vector. HBx proyein was only expressed in HBECs when transfected with HBx expression vector.CONCLUSION: HBx transfection can activate thet ranscriptional expression of hTERT mRNA. Cis-activation of hTERT mRNA by HBx gene is the primary mechanism underlying the proliferation, differentiation and tumorigenesis of biliary epithelia. | Sheng-QuanZou Zhen-LiangQu Zhan-FeiLi XinWang | 2004 | World Journal of Gastroenterology2004，10，15： | 15 |
| 15 | Anti-intlammatory effect of cholecystokinin ana its signal transduction mechanism in endotoxic shock rat显示文摘AIM: To study the anti-inflammatory effects ofcholecystokinin-octapeptide (CCK-8) onlipopolysaccharide (LPS)-induced endotoxic shock (ES)and further investigate its signal transduction pathwaysinvolving p38 mitogen-activated protein kinase (MAPK)METHODS: Eighty-four rats were divided randomly intofour groups: LPS (8 mg @ kg-1, iv) induced ES; CCK-8(40 μg @ kg-1, iv) pretreatment 10 min before LPS (8mg @ kg-1); CCK-8(40 μ g @ kg-1, iv) or normal saline(control) groups.The inflammatory changes of lung andspleen, phagocytic function of alveolar macrophage,quantification of inflammatory calls in bronchoalveolarlavage (BAL) were investigated in rats by usinghematoxylin and eosin (HE) staining, phagocytosis ofCandida albicans and differential cell counting. Nitricoxide (NO) production in serum, lung and spleen wasmeasured with the Griess reaction. The mechanisminvestigated by Western blot.RESULTS: Inflammatory changes of lung and spleeninduced by LPS were alleviated by CCK-8, the increaseof NO induced by LPS in serum, lung and spleen wassignificantly inhibited and the neutrophil infiltration inBAL was significantly reduced by CCK-8. The numberof neutrophils was (52±10)× 106 cells. L-1 in LPS group,while it decreased to (18±4)×106 cells@ L-1 in CCK-8+LPS (P<0.01). The phagocytic rate of CCK-8 groupincreased to (62.49±9.49) %, compared with controlgroup (48.16±14.20) %, P<0.05. The phagocytosisrate was (85.14±4.64) % in LPS group, which reducedto (59.33±3.14) % in CCK-8+LPS group (P<0.01). Theresults of phagocytosis indexes showed similar changes.CCK-8 may play an important role in increasing theexpression of p38 MAPK and decreasing the degradation of IκB-αin Iung and spleen of ES rats.CONCLUSION: CCK-8 can result in anti-inflammatoryeffects, which may be related to activation of p38 MAPK and inhibition on the degradation of IκB-α. | Ai-Hong Meng Yi-Ling Ling Jun-Lan Zhang Department of Pathophysiology,Hebei Medical University,Shijiazhuang 050017,Hebei Province,China Xiao-Peng Zhang Department of Cardiothoracic Surgery,Hebei Provincial People’s Hospital,Shijiazhuang 050071,Hebei Province,China | 2002 | World Journal of Gastroenterology2002，8，4： | 15 |
| 16 | Anti-HBV effect of TAT-HBV targeted ribonuclease显示文摘AIM: To prepare and purify TAT-HBV targeted ribonuclease fusion protein, evaluate its transduction activity and investigate its effect on HBV replication in 2.2.15 cells.METHODS: The prokaryotic expression vector pTAT containing TR gene was used in transforming E. coli BL21(DE3) LysS and TR was expressed with the induction of IPTG. The TAT-TR fusion protein was purified using Ni-NTA-agrose and PD-10 desalting columns, and analyzed by SDSPAGE. Transduction efficiency of TAT-TR was detected with immunofluorescence assay and the concentration of HBeAg in the supernatant of the 2.2.15 cells was determined via solid-phase radioimmunoassay (spRIA). MTT assay was used to detect the cytotoxicity of TAT-TR.RESULTS: The SDS-PAGE showed that the TAT-TR fusion protein was purified successfully, and the purity of TAT-TR was 90 %. The visualization of TAT-TR by immunofiuorescence assay indicated its high efficiency in transducing 2.2.15 cells.RIA result suggests that TAT-TR could inhibit the replication of HBV effectively, it didn′t affect cell growth and had no cytotoxicity.CONCLUSION: TAT-TR possesses a significant anti-HBV activity and the preparation of TAT-TR fusion protein has laid the foundation for the use of TR in the therapeutic trial of HBV infection. | Jin Ding Jun Liu Cai-Fang Xue Wei-Dong Gong Ying-Hui Li Ya Zhao Department of Etiology,Fourth Military Medical University,Xi’an 710032,Shaanxi Province,China | 2003 | World Journal of Gastroenterology2003，9，7： | 12 |
| 17 | Transfection and expression of hepatitis B virus x gene and its effect on apoptosis in HL-7702 cells显示文摘AIM: To investigate the effects of hepatitis B virus x gene and its protein product HBxAg on apoptosis in hepatocyte line HL-7702. METHODS: The reconstituted plasmid pcDNA3-x was established through recombination DNA technique; pcDNA3X was transfected into HL-7702 cells by lipid-mediated trasfection. Positive clones were screened by G418, and HL7702/HI3x cells were analysed by the RT-PCR to confirm the steady expression of X gene in HL-7702 cells. The apoptosis rate in HL-7702 cells was determined by flow cytometry, TUNEL technology, electronic microscope. At the mean time, pcDNA3-X was transfected transiently into HL-7702 cells, and total RNA from HL-7702 cells was extracted 24, 48, 72, 96 and 120 h after the transient transfection, and semiquantitative analysis was performed by RT-PCR to detect the expression of HI3V X gene. Furthermore, apoptosis rate in HL-7702 cells was determined by flow cytometry analysis at the different times. RESULTS: RT-PCR analysis showed that HI3V X gene could be expressed stably in HL-7702 cells. Both flow cytometry and TUNEL technology revealed that the apoptosis rates of HL-7702/HI3x cells were much higher than those of HL-7702/ pcDNA3 and HL-7702 cells. Furthermore, the apoptotic phenomena and apoptotic body were observed in HL-7702/ HI3x cells under electronic microscope, but not in HL-7702/ pcDNA3 and HL-7702 cells. In the experiment of transient transfection, RT-PCR reveals that X gene was expressed most at 72 h after transfection; and the apoptosis rate reached the highest at the same time. After that, the apoptosis rate was reduced with the decrease of the X gene expression. CONCLUSION: HBV X gene and X protein can promote the apoptosis in hepatocyte. And there exist a quantity-effect relationship between the X gene expression and apoptosis rate in hepatocyte. | Hong-YingChen Nan-HongTang Xiu-JinLi Sheng-JunZhang Zhi-XinChen Xiao-ZhongWang | 2004 | World Journal of Gastroenterology2004，10，7： | 12 |
| 18 | Nuclear and mitochondrial DNA microsatellite instability in hepatocellular carcinoma in Chinese显示文摘AIM:To study the nuclear microsatellite instability (nMSI) at BAT26 and mitochondral microsalellite instability (mtMSI) in the occurrence and development of hepatocellular carcinoma and the relationship between nMSI and mtMSI.METHODS: nMSI was observed with PCR and mtMSI with PCR-SSCP in 52 cases of hepatocellular carcinoma.RESULTS:mtMSI was detected in 11 out of the 52 cases of hepatocellular carcinoma (21.2%). Among the 11 cases of hepatocellular carcinoma with mtMSI, 7 occured in one locus and 4 in 2 loci. The frequency of mtMSI in the 52 cases of hepatocellular carcinoma showed no correlation to sex, age,infection of hepatitis B, liver cirrhosis as well as positive AFP of the patients (P>0.05). In addition, nMSI was detected in 3 out of 52 cases of hepatocellular carcinoma (5.8%) and there was no correlation of the incidence of mtMSI to that of nMSI (P>0.05).CONCLUSION:mtMSI may be involved in the coccurrence and development of hepatocellular carcinoma and it is independent of nMSI. | Dian-ChunFang LiFang Rong-QuanWang Shi-MingYang | 2004 | World Journal of Gastroenterology2004，10，3： | 10 |
| 19 | 乙型肝炎病毒X蛋白反式激活基因10的克隆化研究显示文摘目的:乙型肝炎病毒(HBV)X蛋白(HBxAg)是一种具有反式激活作用的病毒蛋白质.为了探索HBxAg蛋白反式激活作用新的靶基因,利用基因芯片技术(DNA chips)对于表达和不表达HBxAg的hepG2细胞的基因表达谱进行比较,以期发现HBxAg蛋白反式激活作用的新靶点,为阐明HBxAg的反式激活作用分子生物学机制,以及HBV相关的肝细胞癌(HCC)形成的分子生物学机制开辟新的研究方向.方法:以含有2拷贝头尾连接的HBV DNA的质粒pCP10作为模板,根据ayw亚型的HBV DNA序列设计、合成引物,PCR扩增产物克隆到真核表达载体pcDNA3.1(-)中,转染肝母细胞瘤细胞系hepG2,提取RNA并进行逆转录.进行基因芯片技术分析.以生物信息学技术,克隆、鉴定新基因.结果:在16种HBxAg上调的靶基因中,其中包括未知功能基因,利用生物信息学技术,获得HBxAg蛋白反式激活作用的新型靶基因,开放读码框架(ORF)长度为1 206个核苷酸(nt),编码产物由401个氨基酸残基(aa)组成,命名为XTP10.结论HBxAg是一种具有反式激活作用的蛋白.基因芯片技术是进行基因表达谱分析的可靠、有效技术.分子克隆技术结合生物信息学技术,是目前鉴定、克隆未知功能新基因的有效技术途径. | 成军 刘妍 洪源 王琳 钟彦伟 董菁 王刚 | 2003 | 世界华人消化杂志2003，11，7： | 9 |
| 20 | 乙肝病毒X基因真核表达载体的构建及人肝细胞株HL-7702转染显示文摘目的:构建表达乙肝病毒(HBV)X基因的人肝细胞株. 方法:用PCR法扩增HBVX基因序列,将其添A后连至PUCmT载体上,用EcolⅠ和Hind Ⅲ双酶切PUCmT-X和PcDNA3载体,连接酶切片段PcDNA3及X片段以构建重组质粒PcDNA3-X.用脂质体转染法将PcDNA3-X及空质粒PcDNA3导入肝细胞HL-7702中,G418选择培养,RT- PCR鉴定其稳定表达. 结果:已构建的PcDNA3-X经序列测定含有完整的HBV X 基因片段,转入HL-7702细胞后经RT-PCR证实该细胞有稳定表达X蛋白. 结论:成功构建了表达HBV X基因的肝细胞株,为进一步探讨HBV X基因在肝炎与肝癌发生中的作用提供了理想的实验模型. | 陈红英 唐南洪 张生君 陈治新 王小众 | 2004 | 世界华人消化杂志2004，12，3： | 9 |
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