The study included 200 thalassemia patients, 120 （60%） males and 80 （40%） females, at age 2-30 years the mean age 16 years and 100 persons from blood donors at age 20-50 years, the mean age 35 years, as control gr...The study included 200 thalassemia patients, 120 （60%） males and 80 （40%） females, at age 2-30 years the mean age 16 years and 100 persons from blood donors at age 20-50 years, the mean age 35 years, as control group. The authors used ELISA HCV 3.0 to detect anti-HCV in serum of thalassemia patients and blood donors. The results show 17% positive, 76% negative and 7% equivocal in serum ofthalassemia patients, but show 1% positive, 98% negative, 1% equivocal in serum of blood donors. The results show significant difference in P ≤ 0.05 between prevalence of anti-HCV antibodies among thalassemia patients and blood donors, the confirmatory tests by recombinant immunoblot assay version 3.0 （RIBA） show 17.5% positive, 79% negative and 3.5% equivocal in serum of thalassemia patients but show 2% positive, 98% negative in serum of blood donors. The thalassemia patients in the present study region had high seroprevalence of anti-HCV.展开更多
Objective:To evaluate the antiviral activity and phytochemicals of selected plant extracts and their effect on the mitogen-activated protein kinase(MAPK)signaling pathway modulated by hepatitis C virus(HCV)nonstructur...Objective:To evaluate the antiviral activity and phytochemicals of selected plant extracts and their effect on the mitogen-activated protein kinase(MAPK)signaling pathway modulated by hepatitis C virus(HCV)nonstructural protein 5 A(NS5A).Methods:A total of ten plant extracts were initially screened for their toxicities against Hep G2 cells.The non-toxic plants were tested for their inhibitory effect on the expression of HCV NS5A at both m RNA and protein levels using real-time PCR and Western blotting assays,respectively.The differential expression of the genes associated with MAPK pathway in the presence of NS5A gene and plant extract was measured through real-time PCR.Subsequently,the identification of secondary metabolites was carried out by phytochemical and HPLC analysis.Results:The phytochemical profiling of Berberis lyceum revealed the presence of alkaloids,phenols,saponins,tannins,flavonoids,carbohydrates,terpenoids,steroids,and glycosides.Similarly,quercetin,myricetin,gallic acid,caffeic acid,and ferulic acid were identified through HPLC analysis.The methanolic extract of Berberis lyceum strongly inhibited HCV RNA replication with an IC50 of 11.44μg/m L.RT-PCR and Western blotting assays showed that the extract reduced the expression of HCV NS5A in a dosedependent manner.Berberis lyceum extract also attenuated NS5A-induced dysregulation of the MAPK signaling pathway.Conclusions:Our findings suggest that Berberis lyceum extract strongly inhibits HCV propagation by reducing HCV NS5A-induced perturbation of MAPK signaling.展开更多
Twenty patients with chronic hepatitis c were investigated during the treatment with interferon to explore the changes of antibodies to HCV (anti-RCV). Anti-HCV was tested with recombinant immunoblot assay (RIBA) by u...Twenty patients with chronic hepatitis c were investigated during the treatment with interferon to explore the changes of antibodies to HCV (anti-RCV). Anti-HCV was tested with recombinant immunoblot assay (RIBA) by using three antigens (C22, C33c, and C100-3) encoded by different regions of HCV genome. The changes of individual anti-HCV and ALT were compared with the change of HCV RNA. The results showed that persistent disappearance of serum HCV RNA was closely related to the changes of anti-C33c (P<0. 01) and anti-C100-3 (P<0. 005), but there was no relation between persistent ALT normality and HCV viremia clearance (P<0. 05). In conclusion, monitoring anti-C33c and anti-C100-3 could indicate the changes or HCV viremia. The normalization of ALT after interferon treatment did not indicate disappearance of HCV viremia.展开更多
AIM： To evaluate the efficacy of a new hepatitis C virus （HCV） core antigen assay developed in China. METHODS： After the determination of HCV infection, 49 serial samples were selected from II regular plasma donor...AIM： To evaluate the efficacy of a new hepatitis C virus （HCV） core antigen assay developed in China. METHODS： After the determination of HCV infection, 49 serial samples were selected from II regular plasma donors in 5 different plasma stations. To compare the performance of HCV core antigen detection and HCV PCR, these samples were genotyped, and each specimen was analyzed by ELISA for the detection of HCV core antigen and by qualitative HCV PCR. RESULTS： Among all of the sequential samples, the original 23 specimens were HCV RNA-negative, and 36 samples were HCV RNA-positive. Twenty-seven samples （75%） were HCV core antigen-positive from these HCV RNA-positive specimens. Conversely, 27 samples （93.2%） were found HCV RNA-positive in HCV core antigen- positive samples. Intervals between HCV RNA and HCV core antigen-positive, as well as between HCV core antigen-positive and HCV antibody-positive were 36.0 and 32.8 d, respectively. CONCLUSION： This HCV core antigen assay, developed in China, is able to detect much of anti-HCV-negative, HCV RNA-positive preseroconversion window period （PWP） plasma donations.展开更多
Two hundred and ninety-six samples of patient serum and 28 samples of donor serum were tested for anti-HCV with second generation domestic made testing kit,and for HCV RNA with PCR assay.The purpose of the study was
Hepatitis C virus is associated with Hepatitis C which can be acute or chronic. Acute hepatitis C is usually short term illness that can lead to chronic infection, while chronic hepatitis C is a chronic liver disease ...Hepatitis C virus is associated with Hepatitis C which can be acute or chronic. Acute hepatitis C is usually short term illness that can lead to chronic infection, while chronic hepatitis C is a chronic liver disease leading to cirrhosis, hepatocellular carcinoma and even death. The mode of transmission is mainly parenteral and vertical. Hepatitis C is the leading cause of morbidity and mortality and a serious public health problem, worldwide as well as in Pakistan. There is no vaccine available for hepatitis C so the only way to prevent hepatitis C is by changing behaviors which can spread the disease. Pakistan is also facing a huge burden of this disease. In Pakistan, the frequency of hepatitis C virus infection ranges from 8% - 15% in the general population with variations in different parts of the country. The general method of detecting infection with HCV is to observe the presence of antibodies to the virus. The aim of this study was to determine frequency of hepatitis C among general population and to see the increasing frequency in our part. Selection of Patients: All patients visiting outpatient clinics, Hepatitis clinic and indoor patients at Civil Hospital Karachi, Pakistan from January 1, 2010 to December 31, 2014 were screened for anti HCV. Methods: Blood samples were collected from the patients in Red Topped vaccutainers and allowed to clot, then centrifuged and serum was screened for anti HCV by Immunochromatography (ICT) method. Results: A total of 156,990 patients were screened for anti-HCV, 23,511 (14.98%) patients were reported positive. During the year 2010, total 11,058 patients were screened for anti HCV, 1791 (16%) were positive, while in the year 2011 total 26,921 were screened and 3908 (14.5%) were positive, during the year 2012, total 23,321 were screened, 3036 (14%) were positive, in the year 2013 we screened total 40,483 patients 6070 (15%) were positive, while in 2014, total 55,484 patients were screened and 8584 were positive (15.5%). The most affected age group is between 21 - 40 years and males are affected more than females. Conclusion: Hepatitis C in increasing in our population affecting the young generation between the age of 21 - 40 years and male population is affected more than females so a mass screening is needed to know the exact position of the disease as well as awareness programs should be started using media.展开更多
Infection with hepatitis C virus （HCV）, a major viral cause of chronic liver disease, frequently progresses to steatosis and cirrhosis, which can lead to hepatocellular carcinoma. HCV infection strongly induces host...Infection with hepatitis C virus （HCV）, a major viral cause of chronic liver disease, frequently progresses to steatosis and cirrhosis, which can lead to hepatocellular carcinoma. HCV infection strongly induces host responses, such as the activation of the unfolded protein response, autophagy and the innate immune response. Upon HCV infection, the host induces the interferon （IFN）-mediated frontline defense to limit virus replication. Conversely, HCV employs diverse strategies to escape host innate immune surveillance. Type I IFN elicits its antiviral actions by inducing a wide array of IFN-stimulated genes （ISGs）. Nevertheless, the mechanisms by which these ISGs participate in IFN-mediated anti-HCV actions remain largely unknown. In this review, we first outline the signaling pathways known to be involved in the production of type I IFN and ISGs and the tactics that HCV uses to subvert innate immunity. Then, we summarize the effector mechanisms of scaffold ISGs known to modulate IFN function in HCV replication. We also highlight the potential functions of emerging ISGs, which were identified from genome-wide siRNA screens, in HCV replication. Finally, we discuss the functions of several cellular determinants critical for regulating host immunity in HCV replication. This review will provide a basis for understanding the complexity and functionality of the pleiotropic IFN system in HCV infection. Elucidation of the specificity and the mode of action of these emerging ISGs will also help to identify novel cellular targets against which effective HCV therapeutics can be developed.展开更多
Background and Aims:Ras-related nuclear(RAN)protein is a small GTP-binding protein that is indispensable for the translocation of RNA and proteins through the nuclear pore complex.Recent studies have indicated that RA...Background and Aims:Ras-related nuclear(RAN)protein is a small GTP-binding protein that is indispensable for the translocation of RNA and proteins through the nuclear pore complex.Recent studies have indicated that RAN plays an important role in virus infection.However,the role of RAN in hepatitis C virus(HCV)infection is unclear.The objective of this study was to investigate the role and underlying mechanisms of RAN in HCV infection.Methods:Huh7.5.1 cells were infected with the JC1-Luc virus for 24 h and then were incubated with complete medium for an additional 48 h.HCV infection and RAN expression were determined using luciferase assay,quantitative reverse transcription-PCR and western blotting.Small interfering RNA was used to silence RAN.Western blotting and immunofluorescence were used to evaluate the cytoplasmic translocation of polypyrimidine tract-binding(PTB),and coimmunoprecipitation was used to examine the interaction between RAN and PTB.Results:HCV infection significantly induced RAN expression and cytoplasmic redistribution of PTB.Knockdown of RAN dramatically inhibited HCV infection and the cytoplasmic accumulation of PTB.Colocalization of RAN and PTB was determined by immunofluorescence,and a direct interaction of RAN with PTB was demonstrated by coimmunoprecipitation.Conclusions:PTB in the host cytoplasm is directly associated with HCV replication.These findings demonstrate that the involvement of RAN in HCV infection is mediated by influencing the cytoplasmic translocation of PTB.展开更多
文摘The study included 200 thalassemia patients, 120 （60%） males and 80 （40%） females, at age 2-30 years the mean age 16 years and 100 persons from blood donors at age 20-50 years, the mean age 35 years, as control group. The authors used ELISA HCV 3.0 to detect anti-HCV in serum of thalassemia patients and blood donors. The results show 17% positive, 76% negative and 7% equivocal in serum ofthalassemia patients, but show 1% positive, 98% negative, 1% equivocal in serum of blood donors. The results show significant difference in P ≤ 0.05 between prevalence of anti-HCV antibodies among thalassemia patients and blood donors, the confirmatory tests by recombinant immunoblot assay version 3.0 （RIBA） show 17.5% positive, 79% negative and 3.5% equivocal in serum of thalassemia patients but show 2% positive, 98% negative in serum of blood donors. The thalassemia patients in the present study region had high seroprevalence of anti-HCV.
基金supported by the CEMB-TWAS Postgraduate Fellowship(FR number 3240286682,2015)granted to Mr.Koloko Brice Landry
文摘Objective:To evaluate the antiviral activity and phytochemicals of selected plant extracts and their effect on the mitogen-activated protein kinase(MAPK)signaling pathway modulated by hepatitis C virus(HCV)nonstructural protein 5 A(NS5A).Methods:A total of ten plant extracts were initially screened for their toxicities against Hep G2 cells.The non-toxic plants were tested for their inhibitory effect on the expression of HCV NS5A at both m RNA and protein levels using real-time PCR and Western blotting assays,respectively.The differential expression of the genes associated with MAPK pathway in the presence of NS5A gene and plant extract was measured through real-time PCR.Subsequently,the identification of secondary metabolites was carried out by phytochemical and HPLC analysis.Results:The phytochemical profiling of Berberis lyceum revealed the presence of alkaloids,phenols,saponins,tannins,flavonoids,carbohydrates,terpenoids,steroids,and glycosides.Similarly,quercetin,myricetin,gallic acid,caffeic acid,and ferulic acid were identified through HPLC analysis.The methanolic extract of Berberis lyceum strongly inhibited HCV RNA replication with an IC50 of 11.44μg/m L.RT-PCR and Western blotting assays showed that the extract reduced the expression of HCV NS5A in a dosedependent manner.Berberis lyceum extract also attenuated NS5A-induced dysregulation of the MAPK signaling pathway.Conclusions:Our findings suggest that Berberis lyceum extract strongly inhibits HCV propagation by reducing HCV NS5A-induced perturbation of MAPK signaling.
文摘Twenty patients with chronic hepatitis c were investigated during the treatment with interferon to explore the changes of antibodies to HCV (anti-RCV). Anti-HCV was tested with recombinant immunoblot assay (RIBA) by using three antigens (C22, C33c, and C100-3) encoded by different regions of HCV genome. The changes of individual anti-HCV and ALT were compared with the change of HCV RNA. The results showed that persistent disappearance of serum HCV RNA was closely related to the changes of anti-C33c (P<0. 01) and anti-C100-3 (P<0. 005), but there was no relation between persistent ALT normality and HCV viremia clearance (P<0. 05). In conclusion, monitoring anti-C33c and anti-C100-3 could indicate the changes or HCV viremia. The normalization of ALT after interferon treatment did not indicate disappearance of HCV viremia.
基金Supported by the National Key Technologies R&D Program of China during the 10th Five-Year Plan, No. 2001BA705B06 National High Technology Research and Development Program of China (863 Program), No. 2006AA020907
文摘AIM： To evaluate the efficacy of a new hepatitis C virus （HCV） core antigen assay developed in China. METHODS： After the determination of HCV infection, 49 serial samples were selected from II regular plasma donors in 5 different plasma stations. To compare the performance of HCV core antigen detection and HCV PCR, these samples were genotyped, and each specimen was analyzed by ELISA for the detection of HCV core antigen and by qualitative HCV PCR. RESULTS： Among all of the sequential samples, the original 23 specimens were HCV RNA-negative, and 36 samples were HCV RNA-positive. Twenty-seven samples （75%） were HCV core antigen-positive from these HCV RNA-positive specimens. Conversely, 27 samples （93.2%） were found HCV RNA-positive in HCV core antigen- positive samples. Intervals between HCV RNA and HCV core antigen-positive, as well as between HCV core antigen-positive and HCV antibody-positive were 36.0 and 32.8 d, respectively. CONCLUSION： This HCV core antigen assay, developed in China, is able to detect much of anti-HCV-negative, HCV RNA-positive preseroconversion window period （PWP） plasma donations.
文摘Two hundred and ninety-six samples of patient serum and 28 samples of donor serum were tested for anti-HCV with second generation domestic made testing kit,and for HCV RNA with PCR assay.The purpose of the study was
文摘Hepatitis C virus is associated with Hepatitis C which can be acute or chronic. Acute hepatitis C is usually short term illness that can lead to chronic infection, while chronic hepatitis C is a chronic liver disease leading to cirrhosis, hepatocellular carcinoma and even death. The mode of transmission is mainly parenteral and vertical. Hepatitis C is the leading cause of morbidity and mortality and a serious public health problem, worldwide as well as in Pakistan. There is no vaccine available for hepatitis C so the only way to prevent hepatitis C is by changing behaviors which can spread the disease. Pakistan is also facing a huge burden of this disease. In Pakistan, the frequency of hepatitis C virus infection ranges from 8% - 15% in the general population with variations in different parts of the country. The general method of detecting infection with HCV is to observe the presence of antibodies to the virus. The aim of this study was to determine frequency of hepatitis C among general population and to see the increasing frequency in our part. Selection of Patients: All patients visiting outpatient clinics, Hepatitis clinic and indoor patients at Civil Hospital Karachi, Pakistan from January 1, 2010 to December 31, 2014 were screened for anti HCV. Methods: Blood samples were collected from the patients in Red Topped vaccutainers and allowed to clot, then centrifuged and serum was screened for anti HCV by Immunochromatography (ICT) method. Results: A total of 156,990 patients were screened for anti-HCV, 23,511 (14.98%) patients were reported positive. During the year 2010, total 11,058 patients were screened for anti HCV, 1791 (16%) were positive, while in the year 2011 total 26,921 were screened and 3908 (14.5%) were positive, during the year 2012, total 23,321 were screened, 3036 (14%) were positive, in the year 2013 we screened total 40,483 patients 6070 (15%) were positive, while in 2014, total 55,484 patients were screened and 8584 were positive (15.5%). The most affected age group is between 21 - 40 years and males are affected more than females. Conclusion: Hepatitis C in increasing in our population affecting the young generation between the age of 21 - 40 years and male population is affected more than females so a mass screening is needed to know the exact position of the disease as well as awareness programs should be started using media.
文摘Infection with hepatitis C virus （HCV）, a major viral cause of chronic liver disease, frequently progresses to steatosis and cirrhosis, which can lead to hepatocellular carcinoma. HCV infection strongly induces host responses, such as the activation of the unfolded protein response, autophagy and the innate immune response. Upon HCV infection, the host induces the interferon （IFN）-mediated frontline defense to limit virus replication. Conversely, HCV employs diverse strategies to escape host innate immune surveillance. Type I IFN elicits its antiviral actions by inducing a wide array of IFN-stimulated genes （ISGs）. Nevertheless, the mechanisms by which these ISGs participate in IFN-mediated anti-HCV actions remain largely unknown. In this review, we first outline the signaling pathways known to be involved in the production of type I IFN and ISGs and the tactics that HCV uses to subvert innate immunity. Then, we summarize the effector mechanisms of scaffold ISGs known to modulate IFN function in HCV replication. We also highlight the potential functions of emerging ISGs, which were identified from genome-wide siRNA screens, in HCV replication. Finally, we discuss the functions of several cellular determinants critical for regulating host immunity in HCV replication. This review will provide a basis for understanding the complexity and functionality of the pleiotropic IFN system in HCV infection. Elucidation of the specificity and the mode of action of these emerging ISGs will also help to identify novel cellular targets against which effective HCV therapeutics can be developed.
基金The work was supported by the Anhui Provincial Natural Science Foundation(1608085QH172)States S&T Projects of 13th Five Year(2018ZX10302206),Chinese Founda-tion for Hepatitis Prevention and Control(TQGB20200151,TQGB20180367)Anhui Key Program of Medical Scien-tific Research of China(#2010A010).
文摘Background and Aims:Ras-related nuclear(RAN)protein is a small GTP-binding protein that is indispensable for the translocation of RNA and proteins through the nuclear pore complex.Recent studies have indicated that RAN plays an important role in virus infection.However,the role of RAN in hepatitis C virus(HCV)infection is unclear.The objective of this study was to investigate the role and underlying mechanisms of RAN in HCV infection.Methods:Huh7.5.1 cells were infected with the JC1-Luc virus for 24 h and then were incubated with complete medium for an additional 48 h.HCV infection and RAN expression were determined using luciferase assay,quantitative reverse transcription-PCR and western blotting.Small interfering RNA was used to silence RAN.Western blotting and immunofluorescence were used to evaluate the cytoplasmic translocation of polypyrimidine tract-binding(PTB),and coimmunoprecipitation was used to examine the interaction between RAN and PTB.Results:HCV infection significantly induced RAN expression and cytoplasmic redistribution of PTB.Knockdown of RAN dramatically inhibited HCV infection and the cytoplasmic accumulation of PTB.Colocalization of RAN and PTB was determined by immunofluorescence,and a direct interaction of RAN with PTB was demonstrated by coimmunoprecipitation.Conclusions:PTB in the host cytoplasm is directly associated with HCV replication.These findings demonstrate that the involvement of RAN in HCV infection is mediated by influencing the cytoplasmic translocation of PTB.
