AIM To study persistence and replication ofheltitis C virus (HCV) in patients' peripheralblood mononuclear cells (PBMC) cultured invitro.METHODS Epstein-Barr virus (EBV) was usedto transform the hepatitis C virus ...AIM To study persistence and replication ofheltitis C virus (HCV) in patients' peripheralblood mononuclear cells (PBMC) cultured invitro.METHODS Epstein-Barr virus (EBV) was usedto transform the hepatitis C virus from a HCVpositive patient to permanent lymphoblastoidcell lines (LCL). Positive and negative HCV RNAstrands of the cultured cells and growth mediawere detected by reverse transcriptase-polymerase chain reaction ( RT-PCR ) eachmonth. Core and NS5 proteins of HCV werefurther tested using immunohistochemical SPmethod and in situ RT-PCR.RESULTS HCV RNA positive strands wereconsistently detected the cultured cells for oneyear. The negative-strand RNA in LCL cells andthe positive-strand RNA in supernatants wereobserved intermittently. Immunohistochemicalresults medicated expression of HCV NS3 and Cproteins in LCL cytoplasm mostly. The positivesignal of PCR product was dark blue and mainlylocalized to the LCL cytoplasm. The RT-PCRsignal was eliminated by overnight RNasedigestion but not DNase digestion.CONCLUSION HCV may exist and remainfunctional in a cultured cell line for a longperiod.展开更多
AIM: To determine the prevalences of TTV and HGV infections among blood donors and patients with chronic liver disease in Korea, to investigate the association of TTV and HGV infections with blood transfusion, and to ...AIM: To determine the prevalences of TTV and HGV infections among blood donors and patients with chronic liver disease in Korea, to investigate the association of TTV and HGV infections with blood transfusion, and to assess the correlation between TTV and HGV viremia and hepatic damage.METHODS: A total of 391 serum samples were examined in this study. Samples were obtained from healthy blood donors (n= 110), hepatitis B surface antigen (HBsAg)-positive donors (n=112), anti-hepatitis C virus (anti-HCV)-positive donors (n=69), patients with type B chronic liver disease (n=81), and patients with type C chronic liver disease (n= 19).TTV DNA was detected using the hemi-nested PCR. HGV RNA was tested using RT-PCR. A history of blood transfusion and serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were also determined.RESULTS: TTV DNA was detected in 8.2 % of healthy blood donors, 16.1% of HBsAg-positive donors, 20.3 % of antiHCV-positive donors, 21.0 % of patients with type B chronic liver disease, and 21.1% of patients with type C chronic liver disease. HGV RNA was detected in 1.8 % of healthy blood donors, 1.8 % of HBsAg-positive donors, 17.4 % of anti-HCV-positive donors, 13.6% of patients with type B chronic liver disease, and 10.5% of patients with type C chronic liver disease. The prevalence of TTV and HGV infections in HBV- or HCV-positive donors and patients was significantly higher than in healthy blood donors (P<0.05),except for the detection rate of HGV in HBsAg-positive donors which was the same as for healthy donors. There was a history of transfusion in 66.7% of TTV DNA-positive patients and 76.9% of HGV RNA-positive patients (P<0.05). No significant increase in serum ALT and AST was detected in the TTV- or HGV-positive donors and patients.CONCLUSION: TTV and HGV infections are more frequently found in donors and patients infected with HBV or HCV than in healthy blood donors. However, there is no significant association between TTV or HGV infections and liver injury.展开更多
Forty-two patients with hepatocellular carcinoma (HCC) were examined for hepatitis C virus (HCV) RNA in liver tissues by reverse transcription-polymerase chain reaction (RT-PCR) with primers deduced from 5'-non-co...Forty-two patients with hepatocellular carcinoma (HCC) were examined for hepatitis C virus (HCV) RNA in liver tissues by reverse transcription-polymerase chain reaction (RT-PCR) with primers deduced from 5'-non-coding region (5'-NCR). HCV liver samples of展开更多
目的:通过研究原发性肝癌组织中野生型雌激素受体(wild type Estrogen receptor,wER)以及第5外显子缺失而变异的雌激素受体(variant estrogen receptor,vER)的表达情况,期望对原发性肝癌进行内分泌治疗的可能性进行分析。方法:收...目的:通过研究原发性肝癌组织中野生型雌激素受体(wild type Estrogen receptor,wER)以及第5外显子缺失而变异的雌激素受体(variant estrogen receptor,vER)的表达情况,期望对原发性肝癌进行内分泌治疗的可能性进行分析。方法:收集我院原发性肝癌手术切除的标本28例,应用 RT-PCR 的方法分析 wER 与 vER 在 mRNA 水平的表达,同时通过免疫组织化学分析对 ER 在蛋白水平的表达进行检测。结果:免疫组织化学显示原发性肝癌组织中 ER 的表达率为39.3%(11/28);RT-PCR 显示所有标本中均检测到雌激素受体的表达,89.3%(25/28)的原发性肝癌病人表达 wER,96.4%(27/28)的肝癌表达 vER,其中 wER 和 vER 联合表达的病人为85.7%(24/28),仅表达wER 的病人为3.5%(1/28),而仅表达 vER 的为10.7%(3/28)。结论:在原发性肝癌的形成过程中,96.4%(27/28)的肝癌出现 ER 的变异,提示 ER 的变异与原发性肝癌的形成有密切的关系。展开更多
基金The paper was support by a grant from the Ministry Youth Research of China,No.98-1-269
文摘AIM To study persistence and replication ofheltitis C virus (HCV) in patients' peripheralblood mononuclear cells (PBMC) cultured invitro.METHODS Epstein-Barr virus (EBV) was usedto transform the hepatitis C virus from a HCVpositive patient to permanent lymphoblastoidcell lines (LCL). Positive and negative HCV RNAstrands of the cultured cells and growth mediawere detected by reverse transcriptase-polymerase chain reaction ( RT-PCR ) eachmonth. Core and NS5 proteins of HCV werefurther tested using immunohistochemical SPmethod and in situ RT-PCR.RESULTS HCV RNA positive strands wereconsistently detected the cultured cells for oneyear. The negative-strand RNA in LCL cells andthe positive-strand RNA in supernatants wereobserved intermittently. Immunohistochemicalresults medicated expression of HCV NS3 and Cproteins in LCL cytoplasm mostly. The positivesignal of PCR product was dark blue and mainlylocalized to the LCL cytoplasm. The RT-PCRsignal was eliminated by overnight RNasedigestion but not DNase digestion.CONCLUSION HCV may exist and remainfunctional in a cultured cell line for a longperiod.
文摘AIM: To determine the prevalences of TTV and HGV infections among blood donors and patients with chronic liver disease in Korea, to investigate the association of TTV and HGV infections with blood transfusion, and to assess the correlation between TTV and HGV viremia and hepatic damage.METHODS: A total of 391 serum samples were examined in this study. Samples were obtained from healthy blood donors (n= 110), hepatitis B surface antigen (HBsAg)-positive donors (n=112), anti-hepatitis C virus (anti-HCV)-positive donors (n=69), patients with type B chronic liver disease (n=81), and patients with type C chronic liver disease (n= 19).TTV DNA was detected using the hemi-nested PCR. HGV RNA was tested using RT-PCR. A history of blood transfusion and serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were also determined.RESULTS: TTV DNA was detected in 8.2 % of healthy blood donors, 16.1% of HBsAg-positive donors, 20.3 % of antiHCV-positive donors, 21.0 % of patients with type B chronic liver disease, and 21.1% of patients with type C chronic liver disease. HGV RNA was detected in 1.8 % of healthy blood donors, 1.8 % of HBsAg-positive donors, 17.4 % of anti-HCV-positive donors, 13.6% of patients with type B chronic liver disease, and 10.5% of patients with type C chronic liver disease. The prevalence of TTV and HGV infections in HBV- or HCV-positive donors and patients was significantly higher than in healthy blood donors (P<0.05),except for the detection rate of HGV in HBsAg-positive donors which was the same as for healthy donors. There was a history of transfusion in 66.7% of TTV DNA-positive patients and 76.9% of HGV RNA-positive patients (P<0.05). No significant increase in serum ALT and AST was detected in the TTV- or HGV-positive donors and patients.CONCLUSION: TTV and HGV infections are more frequently found in donors and patients infected with HBV or HCV than in healthy blood donors. However, there is no significant association between TTV or HGV infections and liver injury.
文摘Forty-two patients with hepatocellular carcinoma (HCC) were examined for hepatitis C virus (HCV) RNA in liver tissues by reverse transcription-polymerase chain reaction (RT-PCR) with primers deduced from 5'-non-coding region (5'-NCR). HCV liver samples of
文摘目的:通过研究原发性肝癌组织中野生型雌激素受体(wild type Estrogen receptor,wER)以及第5外显子缺失而变异的雌激素受体(variant estrogen receptor,vER)的表达情况,期望对原发性肝癌进行内分泌治疗的可能性进行分析。方法:收集我院原发性肝癌手术切除的标本28例,应用 RT-PCR 的方法分析 wER 与 vER 在 mRNA 水平的表达,同时通过免疫组织化学分析对 ER 在蛋白水平的表达进行检测。结果:免疫组织化学显示原发性肝癌组织中 ER 的表达率为39.3%(11/28);RT-PCR 显示所有标本中均检测到雌激素受体的表达,89.3%(25/28)的原发性肝癌病人表达 wER,96.4%(27/28)的肝癌表达 vER,其中 wER 和 vER 联合表达的病人为85.7%(24/28),仅表达wER 的病人为3.5%(1/28),而仅表达 vER 的为10.7%(3/28)。结论:在原发性肝癌的形成过程中,96.4%(27/28)的肝癌出现 ER 的变异,提示 ER 的变异与原发性肝癌的形成有密切的关系。