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| 1 | Detection of micrometastasis of gastric carcinoma in peripheral blood circulation显示文摘AIM:To detect the micrometastasis of gastric carcinoma in peripheral blood circulation using immunomagnetic beads sorting technique and RT-PCR technique,and to discuss its significance and the difference between the two methods.METHODS:Density gradient centrifugation was used to isolate mononuclear cells from peripheral blood,immunomagnetic beads sorting technique and RT-PCR technique were used to detect the disseminated carcinoma cells.HE,immunocytochemical and immunofluorescence staining were also used to identify the characteristics of the cells separated with immunomagnetic beads sorting technique.RESULTS:Cells expressing cytokeratin were separated and enriched from the peripheral blood specimens of patients suffering from gastric carcinoma or chronic gastritis. After HE staining, two kinds of ceils with little cytoplasm were found.Majority of these cells had small and round nuclei, even chromatins and the thickness of nuclear membrane was normal. Immunohistochemical staining indicated that there were CD34 and CD45 expression on the cell membrane ofthis kind of cells and these cells also showed expressed human telomerase reverse transcriptase by immunofluorescence staining, but the expression of carcinoembryonic antigen was absent.So,these cells might hematopoiesis precursors.Another kind of cells had larger and abnormal nuclei with thicker nuclear membranes. Massed chromatins and polynudeoli were found in the nudei. These cells expressed human telomerase reverse transcriptase and carcinoembryonic antigen,but CD34 and CD45 were not found on the cell membrane.So,these cells were considered as gastric carcinoma cells escaping from the original focuses and existing in the peripheral blood circulation.Cardnoma cells were found in 25 of 60(41.7%) specimens of peripheral blood from patients with gastric carcinoma, while there were no such cells separated from the blood specimens of chronic gastritis patients.The difference of positive rates of disseminated carcinoma cells between two groups was markedly significant (P<0.005).The expressions of CK20 mRNA in peripheral blood specimens were examinated withRT-PCR. CK20 mRNA was detected from 32 of 60(53.3%) peripheral blood specimens in the group of gastric carcinoma patients,while none of the specimens from patients suffering from chronic gastritis had CK20 mRNA. Significant difference was also found between two groups (P<0.005).Statistic analyses also showed that there was a significant difference between the positive rates of two methods in detecting the disseminated carcinoma cells from the peripheral blood circulation of gastric carcinoma patients (P<0.05).CONCLUSION:The results demonstrated that there were disseminated carcinoma cells in the peripheral blood circulation of some patients with gastric carcinoma.Disseminated carcinoma cells can be detected from the peripheral blood samples with immunomagnetic beads sorting technique and RT-PCR technique.The positive rate of RT-PCR technique is higher than that of immunomagnetic beads sorting technique in detecting micrometastasis. | Xi-MeiChen Guo-YuChen Zhi-RongWang Feng-ShangZhu Xiao-LeiWang XiaZhang | 2004 | World Journal of Gastroenterology2004，10，6： | 32 |
| 2 | Effect of alpha 2b interferon on inducement of mIL-2R and treatment of HCV in PBMC from patients with chronic viral hepatitis C显示文摘AIM: To study the level of membrane interleukin-2 receptor (mIL-2R) on surface of peripheral blood mononuclear cells (PBMC) and the therapeutic efficacy of alpha 2b interferon on the treatment of HCV-RNA in PBMC of patients with chronic hepatitis C and to compare the negative rates of HCV-RNA in PBMC, HCV-RNA and anti-HCV in serum.METHODS: Before and after treatment of alpha 2b interferon, the level of mIL-2R of patients with chronic hepatitis C was detected by biotin-streptavidin (BSA). The therapeutic group (26 cases) was treated with alpha 2b interferon (3 MU/d) and control therapeutic group (22 cases)was treated with routine drugs (VitC, aspartic acid). The total course of treatment with alpha 2b interferon and routine drug was six months and per course of the treatment was three months. The levels of HCV-RNA in PBMC, HCV-RNA and anti-HCV in serum were detected before and after a course of the treatment.RESULTS: Before and after treatment of alpha 2b interferon and routine drugs, the levels of mIL-2R in silence stage were (3.44±0.77)% and (2.95±0.72)%, the levels of mIL-2R in inducement stage were (33.62±3.95)% and (30.04±3.73)%. There was a significant difference between two groups (P<0.01-P<0.05). After treatment of alpha 2b interferon with 3 MU/d for two courses of the treatment,the total negative rates of HCV-RNA in the PBMC and HCVRNA, anti-HCV in serum were 42.31% (11/26), 57.69%(15/26), 65.38%(17/26) respectively. After the treatment of routine drug, the negative rates of HCV-RNA in PBMC and HCV-RNA, anti-HCV in serum were 13.64% (3/22),22.73% (5/22), 27.27% (6/22) respectively. There was high significant difference in the group treated with alpha 2b interferon and the group treated with routine drugs (P<0.01-P<0.05).CONCLUSION: The mIL-2R can be induced by alpha 2b interferon during the treatment. The alpha 2b interferon has a definite effect on the treatment of HCV-RNA in PBMC.The curative effect of alpha 2b interferon is better than that of the routine drugs. | JianWang Gui-JuXiang Bing-XiangLiu | 2003 | World Journal of Gastroenterology2003，9，4： | 8 |
| 3 | Involvement of TLR2-My D88 in Abnormal Expression of mi R-146a in Peripheral Blood Monocytes of Patients with Chronic Hepatitis C显示文摘miR-146 a is an immunoregulatory micro RNA closely associated with viral infection. This study investigated the expression changes of mi R-146 a in peripheral blood monocytes of HCV-infected patients and the mechanism by which the THP-1 cells were stimulated with HCV core protein in vitro. It was found that in the peripheral blood monocytes of HCV-infected patients, mi R-146 a expression was upregulated. After treated by interferon/ribavirin, mi R-146 a expression was decreased when HCV RNA became undetectable. HCV core could directly stimulate THP-1 cells to produce mi R-146 a. Silencing TLR2 and My D88 could significantly inhibit the expression of mi R-146 a. It was concluded that the expression of mi R-146 a in peripheral blood monocytes of HCV-infected patients was abnormally increased. The TLR2-My D88 signaling pathway may take part in the overexpression of mi R-146 a in monocytes stimulated with HCV core protein. | 张文静 王华 童巧霞 揭盛华 杨东亮 彭程 | 2015 | Journal of Huazhong University of Science and Technology(Medical Sciences)2015，35，2： | 5 |
| 4 | HCVNS3蛋白对正常人源肝细胞生长及MAPK磷酸化的影响显示文摘目的:研究丙型肝炎病毒非结构区3(HCVNS3)蛋白对正常人源肝细胞的转化及MAPK磷酸化调节的作用.方法:利用脂质体介导转染技术和G418筛选得到稳定表达NS3蛋白的正常人源性肝细胞QSG7701,PCR和免疫组化S-P法检测细胞中NS3的表达;细胞记数和软琼脂实验鉴定其生物学性质;抗磷酸化MAPK抗体和抗MAPK抗体Westernblot检测转染细胞MAPK活性及表达.结果:HCVNS3转染的QSG7701肝细胞,其NS3蛋白过度表达于细胞质;真核表达质粒pRcHCNS3-5’转染细胞倍增时间为12h,较pRcHCNS3-3’、pRcCMV转染细胞和未转染的QSG7701明显缩短(分别24h,26h和28h).pRcHCNS3-5’、pRcHCNS3-3’和pRcCMV转染细胞及正常肝细胞在软琼脂中的克隆形成率分别为33%、1.33%、1.46%、1.11%,pRcHCNS3-5’形成的克隆明显高于其他三种细胞(P<0.01).pRcHCNS3-5’转染细胞MAPK磷酸化程度明显高于其他三种细胞(分别为8858±877,5612±656,2212±245和989±188).(P<0.01),而MAPK的表达量没有差异(P>0.05).结论:人源性正常肝细胞QSG7701是研究HCVNS3蛋白致病机制的较好的细胞系;HCVNS3蛋白N端多肽具有促进细胞增生和改变细胞表型的作用;HCVNS3蛋白N端多肽能上调MAPK活性,不影响MAPK的表达量. | 孙意 程瑞雪 冯德云 欧阳小明 郑晖 | 2003 | 世界华人消化杂志2003，11，2： | 4 |
| 5 | IRES特异性IRNA对HCV IRES启动蛋白翻译细胞内抑制作用显示文摘目的:研究核糖体内部进入位点(internalribosomeentrysite,IRES)特异性抑制性RNA(inhibitorRNA,IRNA)细胞内对丙型肝炎病毒(HCV)IRES介导蛋白翻译的抑制作用.方法:体外应用脂质体细胞转染法,将IRNA及其突变体mIRNA真核表达载体pcRz-IRNA/pcRz-mIRNA转染人肝癌细胞株(HHCC),经G418筛选4wk后建立IRNA及mIRNA表达株;以相同的方法构建pcHCVcluc转染株;以脂质体介导细胞转染法将pCMVNCRLuc转染IRNA及mIRNA细胞株,于转染后48h检测荧光素酶表达量;将IRNA及mIRNA真核表达体转染pcHCVcLuc表达株,于转染后不同时间检测荧光素酶表达量.结果:HCVIRES介导蛋白翻译在IRNA表达株明显受到抑制,同样IRNA对HCV翻译复制子的蛋白翻译作用有明显抑制性;突变体mIRNA表达株和空载体对照株中未见相似的抑制性.结论:pcHCVcluc在HHCC细胞中获得有效表达;IRES特异性IRNA能有效的抑制HCVIRES介导细胞内蛋白翻译作用. | 梁雪松 连建奇 周永兴 聂青和 郝春秋 | 2003 | 世界华人消化杂志2003，11，2： | 4 |
| 6 | A small yeast RNA inhibits HCV IRES mediated translation and inhibits replication of poliovirus in vivo显示文摘AIM: To investigate the anti-virus infection activity of internal ribosome entry site (IRES) specific inhibitor RNA (IRNA).METHODS: IRNA eukaryotic vector pcRz-IRNA or mIRNA eukaryotic vector pcRz-mIRNA was tansfected into human hepatocarcinoma cells (HHCC), then selected with neomycin G418 for 4 to 8 weeks, and then infected with polio virus vaccinas line. The cytopethogenesis effect was investigated and the cell extract was collected. At last the polio virus titer of different cells was determined by plaque assay.RESULTS: Constitutive expression of IRNA was not detrimental to cell growth. HCV IRES-mediated capindependent translation was markedly inhibited in cells constitutively expressing IRNA compared to control hepatoma cells. However, cap-dependent translation was not significantly affected in these cell line. Additionally, HHCC cells constitutively expressing IRNA became refractory to infection of polio virus.CONCLUSION: IRES specific IRNA can inhibit HCV IRES mediated translation and poliovirus replication. | Xue-Song Liang Jian-Qi Lian Yong-Xing Zhou Qing-He Nie Chun-Qiu Hao The center of diagnosis and treatment for infectious diseases of Tangdu Hospital of Military Medical University of PLA,Xi’an 710038,Shaanxi Province,China | 2003 | World Journal of Gastroenterology2003，9，5： | 2 |
| 7 | 热休克蛋白70与IL-2对小鼠肝癌移植模型的治疗比较显示文摘目的:比较分析肿瘤热休克蛋白70(HSP70)与IL-2对小鼠肝癌移植模型的治疗作用,评价HSP70的疗效,为应用HSP70治疗人类恶性肿瘤提供有重要参考价值的实验依据。方法:细胞培养、蛋白质纯化技术、电泳技术、Westernblot法、动物实验等。结果:IL-2及HSP70对小鼠肝癌移植模型均有治疗作用,其中IL-210万U、HSP7010μg效果最佳,IL-25万U及HSP705μg有部分治疗作用,10万U的IL-2与5μgHSP70治疗效果相当,HSP7010μg治疗后有40%小鼠移植肿瘤全部消退,长期生存790d,与对照组及其他各组相比,差异具有显著性(P<0.01,P<0.05)。结论:HSP70具有良好的抗肿瘤免疫活性,其疗效明显优于IL-2,本研究对于应用HSP70治疗人类的恶性肿瘤有十分重要的借鉴意义。 | 傅庆国 沈晓东 孟凡东 郭仁宣 | 2003 | 世界华人消化杂志2003，11，4： | 2 |
| 8 | Inhibitory effect of miR-125b on hepatitis C virus core protein-induced TLR2/My D88 signaling in THP-1 cells显示文摘AIM: To investigate the role of mi R-125 b in regulating monocyte immune responses induced by hepatitis C virus(HCV) core protein.METHODS: Monocytic THP-1 cells were treated with various concentrations of recombinant HCV core protein, and cytokines and mi R-125 b expression in these cells were analyzed. The requirement of Tolllike receptor 2(TLR2) or My D88 gene for HCV core protein-induced immune responses was determined by the transfection of THP-1 cells with gene knockdown vectors expressing either TLR2 si RNA or My D88 si RNA. The effect of mi R-125 b overexpression on TLR2/My D88 signaling was examined by transfecting THP-1 cells with mi R-125 b mimic RNA oligos.RESULTS: In response to HCV core protein stimulation, cytokine production was up-regulated and mi R-125 b expression was down-regulated in THP-1 cells. The modulatory effect of HCV core protein on cellular events was dose-dependent and required functional TLR2 or My D88 gene. Forced mi R-125 b expression abolished the HCV core protein-induced enhancement of tumor necrosis factor-α, interleukin(IL)-6, and IL-10 expression by 66%, 54%, and 66%, respectively(P < 0.001), by inhibiting My D88-mediated signaling, including phosphorylation of NF-k Bp65, ERK, and P38.CONCLUSION: The inverse correlation between mi R-125 b and cytokine expression after HCV core challenge suggests that mi R-125 b may negatively regulate HCVinduced immune responses by targeting TLR2/My D88 signaling in monocytes. | Cheng Peng Hua Wang Wen-Jing Zhang Sheng-Hua Jie Qiao-Xia Tong Meng-Ji Lu Dong-Liang Yang | 2016 | World Journal of Gastroenterology2016，22，17： | 2 |
| 9 | Monocyte chemotactic protein-1 and soluble adhesion molecules as possible prognostic markers of the efficacy of antiviral treatment in chronic hepatitis C显示文摘AIM: To explain the role of Monocyte chemotactic protein-1 (MCP-1) and soluble adhesion molecules in chronic hepatitis C during the treatment of interferon alpha (IFNα) 2 b and ribavirin (RBV).METHODS: Concentrations of MCP-1, soluble adhesion molecules intercellular adhesion molecule-1 (sICAM-1), sPselectin, interleukin (IL) 6, and ILl0 in serum were estimated in the group of 40 patients with chronic hepatitis C treated with IFNalpha2 b and RBV in 0, 16, 32, 48 wk of the therapy.RESULTS: In chronic hepatitis C, before and during the treatment, the serum levels of MCP-1 and sP-selectin in responders were similar to those of healthy subjects. In nonresponders (NR), MCP-1 increased in the course of IFN(α+RBV treatment, differences were statistically significant as compared to responders. MCP-1 correlated statistically with the activity of periportal inflammation (r= 0.35, P<0.05) but not with staging of liver fibrosis, sICAM-1 positively correlated with inflammatory activity and fibrosis in NR. sP-selectin did not correlate with histological findings in the liver. The MCP-1 correlated with the soluble form of sP-selectin concentrations (r= 6, P<0.001) and with IL-10 level in NR (r = 0.4, P<0.05). There was no correlation observed between the concentration of MCP-1 and sICAM-1, IL-6 during the treatment.CONCLUSION: MCP-1 concentration may be a prognostic marker of the efficacy of IFN+RBV therapy in patients with chronic hepatitis C. | AnatolPanasiuk DanutaProkopowicz BozenaPanasiuk | 2004 | World Journal of Gastroenterology2004，10，24： | 1 |
| 10 | HCV核蛋白羧基端信号肽基因缺失突变体的构建、鉴定和真核表达显示文摘目的:构建羧基末端缺失核蛋白的真核表达载体并在小鼠肥大细胞瘤细胞株P815细胞中表达.方法:用RT-PCR方法从西安地区丙型肝炎患者血清中扩增HCVC区及部分E1区基因并进行克隆、测序,以此为模板扩增羧基末端缺失突变核蛋白基因片段(C507),将其定向克隆入真核表达载体pCI-neo中并转染P815细胞,经间接免疫荧光染色、激光共聚焦显微镜检测细胞中突变核蛋白(P169)的表达.结果:从患者血清中成功克隆出HCVCcDNA,构建了编码羧基末端缺失核蛋白的重组质粒pCI-507,并经酶切鉴定和测序证实;间接免疫荧光染色表明P169主要在P815细胞胞质中表达,少数可见于细胞核内.结论:重组体pCI-507构建正确,其表达产物P169在P815中得到有效表达,为在此基础上的DNA免疫研究提供了实验依据. | 洪沙 黄长形 杨为松 陈红梅 李光玉 王平忠 陈伟红 张岩 李羽 | 2005 | 世界华人消化杂志2005，13，7： | 1 |
| 11 | 联合用药治疗慢性乙、丙型肝炎显示文摘目前治疗慢性乙、丙型肝炎不尽人意,要提高其应答率,减少肝硬化和肝细胞癌的发生,联合用药 是比较可行的方案。本文综述了近年来治疗慢性乙、丙型肝炎病毒感染遇到的困惑和联合应用胸腺素α1治疗的 进展,分析了治疗失败的原因,提出了增强疗效的建议。 | 邓永岳 陈紫榕 | 2004 | 国外医学(流行病学.传染病学分册)2004，31，3： | 1 |
| 12 | 丙型肝炎病毒非结构蛋白NS3上调硫氧还蛋白还原酶1基因的表达显示文摘目的:探讨丙型肝炎病毒(HCV)非结构蛋白NS3对硫氧还蛋白还原酶1(TXNRD1)启动子转录的激活作用. 方法:以HCV非结构蛋白NS3反式调节基因的基因表达谱芯片结果为基础,利用生物信息学技术确定TXNRD1的启动子区域(TXNRD1p),聚合酶链反应(PCR)扩增TXNRD1p, 克隆至真核报告载体pCAT3-Basic中,构建pCAT3- TXNRD1p报告载体;以该质粒转染肝癌细胞系HepG2细胞系,用酶联免疫黏附法(ELISA)检测氯霉素乙酰转移酶(CAT)的表达活性;并与pcDNA3.1(-)-NS3共转染HepG2 细胞系,用ELISA法检测CAT的表达活性. 结果:成功获得TXNRD1启动子的正确克隆CATB-TXNRD1p 和pcDNA3.1(-)-NS3瞬时转染的HepG2细胞的CAT表达活性是pCAT3-Basic空载体的9倍,pCAT3-TXNRD1p的2.0倍,进一步验证了利用基因表达谱技术研究HCV NS3 蛋白反式激活作用的结果. 结论:TXNRD1启动子有顺式激活下游基因的活性;HCV的NS3蛋白具有对TXNRD1的反式激活作用. | 纪冬 成军 郭江 董菁 王建军 刘妍 | 2004 | 世界华人消化杂志2004，12，7： | 1 |
| 13 | Inhibitor RNA blocks the protein translation mediated by hepatitis C virus internal ribosome entry site in vivo显示文摘AIM:To investigate the inhibitory effect of hepatitis C virus internal ribosome entry site (HCV IRES) specific inhibitor RNA(IRNA) on gene expression mediated by HCV IRES in vivo.METHODS:By using G418 screening system, hepatoma cells constitutively expressing IRNA or mutant IRNA (mIRNA) were established and characterized, and HCV replicons containing the 5′ untranslated region (5′UTR) were constructed by using the same method. Cotransfection of pCMVNCRluc containing HCV 5′UTR-Iuc fusion genes and eukaryotic vector of IRNA into human hepatic carcinoma cells (HepG2) was performed and the eukaryotic expression plasmid of IRNA was transfected transiently into HCV replicons, pCMVNCRluc or pCDNA-luc was cotransfected with pSV40-β Gal into IRNA expressing hepatoma cells by using lipofectamine 2000 in vitro. Then the reporting gene expression level was examined at 48h after transfection by using a luminometer and the expressing level of HCV C antigen was analysed with a confocal microscope.RESULTS: Transient expression of IRES specific IRNA could significantly inhibit the expression of reporter gene and viral antigen mediated by HCV IRES by 50% to 90% in vivo, but mIRNA lost its inhibitory activity completely. The luciferase gene expression mediated by HCV IRES was blocked in the HHCC constitutively expressing IRNA. At 48h after transfection, the expression level of reportor gene descreased by 20%, but cap-dependent luciferase gene expression was not affected. IRNA could inhibit the HCV replicon expression 24h after transfection and the highest inhibitory activity was 80% by 72h, and the inhibitory activity was not increased until 7d after transfection.CONCLUSION:IRNA can inhibit HCV IRES mediated gene expression in vivo. | Xue-SongLiang Jian-QiLian Yong-XingZhou Mo-BinWan | 2004 | World Journal of Gastroenterology2004，10，5： | 0 |
| 14 | 白细胞介素-12与肝病显示文摘 | 何登明 王宇明 | 2003 | 肝脏2003，8，2： | 0 |
| 15 | 丙型肝炎病毒非结构蛋白 NS3反式激活研究进展显示文摘丙型肝炎病毒非结构区3(HCV NS3)基因位于HCV基因组非结构区3420-5312核苷酸区段.HCV NS3具有多种潜在生物学功能,如蛋白酶、解旋酶活性,介导细胞免疫反应、反式激活端粒酶、调节p53活性及参与蛋白激酶A和STAT信号转导等[1,2],从而影响宿主细胞功能,导致细胞增生、凋亡,甚至癌变.对于NS3蛋白反式激活作用研究,可为进一步研究HCV致病(癌)的分子生物学机制和探索新的丙型肝炎治疗技术提供帮助. | 荣义辉 纪冬 赵军 | 2005 | 传染病信息2005，18，4： | 0 |
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