Objective:To investigate the effects of hepatitis B virus(HBV)X protein(HBx)on the expression of tumor necrosis factor-α(TNF-α)in glomerular mesangial cells(GMCs)and the underlying intracellular signal pathways.Meth...Objective:To investigate the effects of hepatitis B virus(HBV)X protein(HBx)on the expression of tumor necrosis factor-α(TNF-α)in glomerular mesangial cells(GMCs)and the underlying intracellular signal pathways.Methods:The plasmid pCI-neo-X that carries the X gene of hepatitis B virus was transfected into cultured GMCs.HBx expression in the transfected GMCs was assessed by Western-blot.TNF-αprotein and mRNA were assessed by ELISA and semi-quantitative RT-PCR,respectively.Three kinase inhibitors-U0126,an inhibitor of extracellular signal-regulated kinases(ERKs);lactacvstin,an inhibitor of nuclear factor-κB(NF-κB);and SB203580,a selective inhibitor of p38 MAP kinase(p38 MAPK)were used to determine which intracellular signal pathways may underlie the action of HBx on TNF-αexpression in transfected GMCs.Results:A significant increase in HBx expression in pCI-neo-X transfected GMCs was detected at 36 h and 48 h,which was not affected by any of those kinase inhibitors mentioned above.A similar increase in the expression of both TNF-αprotein and mRNA was also observed at 36 h and 48 h,which was significantly decreased in the presence of U0126 or lactacytin,but not SB203580.Conclusions:HBx upregulates TNF-αexpression in cultured GMCs,possibly through ERKs and NF-κB pathway,but not p38 MAPK pathway.展开更多
The X gene of HBV encodes a 17-kD protein, termed HBx, which has been shown to function as a transcriptional trans-activator of a variety of viral and cellular promoter/enhancer elements. The aim of this study was to ...The X gene of HBV encodes a 17-kD protein, termed HBx, which has been shown to function as a transcriptional trans-activator of a variety of viral and cellular promoter/enhancer elements. The aim of this study was to investigate the effect of HBx on gene expression of interleukin （IL）-1β and IL-6, and proliferation of rat mesangial cells in vitro. The X gene of HBV was amplified by PCR assay, and inserted into the eukaryotic expression vector pCI-neo. The structure of recombinant pCI-neo-X plasmid was proved by restrict endonuclease digestion and sequencing analysis. pCI-neo-X was transfected into cultured rat mesangial cell line in vitro via liposome. HBx expression in transfected mesangial cells was detected by Western blot. The IL-1β and IL-6 mRNA expression in those cells was assayed by semiquantitative RT-PCR. Mesangial cell proliferation was tested by MTT. The results showed that HBx was obviously expressed in cultured mesangial cell line at 36th and 48th h after transfection. The expression of IL-1β and IL-6 mRNA was simultaneously increased. The cell proliferation was also obvious at the same time. It was concluded that HBx gene transfection could induce IL-1β and IL-6 gene expression and mesangial cell proliferation. HBx may play a critical role in mesangial cell proliferation through upregulation of the IL-1β and IL-6 gene expression.展开更多
With the introduction of the new highly effective antiviral therapies, there has been a dramatic increase in the use of the hepatitis C virus(HCV)-positive livers in HCV-positive recipients. In the majority of studies...With the introduction of the new highly effective antiviral therapies, there has been a dramatic increase in the use of the hepatitis C virus(HCV)-positive livers in HCV-positive recipients. In the majority of studies, HCV positivity was defined as a donor testing HCV Ab positive. In 2015, all Organ Procurement Organizations were mandated to perform and report HCV Nucleic Acid Amplification Testing(NAT) results on all deceased and living donors. Studies are not yet available on how organs are being utilized based on NAT status and whether NAT status affects recipient outcomes. Further studies are needed to maximize the use of these organs.展开更多
The study was aimed at establishing an indirect ELISA for epidemiological investigation of hepatitis E viral(HEV) infection in pigs from the major pig-producing regions of Zhejiang Province. The gene fragment covering...The study was aimed at establishing an indirect ELISA for epidemiological investigation of hepatitis E viral(HEV) infection in pigs from the major pig-producing regions of Zhejiang Province. The gene fragment covering the immunogenic epitopes of HEV ORF2 was synthesized and expressed in Escherichia coli.Western blot analysis revealed that the purified protein reacted to HEV positive sera,but not to positive sera from some other common viruses that infect pigs.An indirect ELISA system was then developed using truncated HEV ORF2 protein as the coating antigen and had diagnostic accuracy of 91.5% as compared with a diagnostic kit for HEV antibodies.A total of 1 330 serum samples were collected from 46 pig farms in Zhejiang Province from 2005-2008 and tested for sero-prevalence of HEV using the indirect ELISA.The average HEV-positive rate was 55.7%(741/1 330).The positive rate of 2005 and 2006 was 62.7%(175/279) and 61.4%(301/490) respectively,much higher than those of 2007(43.0%,59/137) and 2008(48.6%,206/424).Pigs aged 66-100 days had a statistically higher positive rate(58.2%,110/189) when compared to that of pigs aged 30-65 days(39.7%,73/184) or 101-160 days(44.1%,83/188).Only three herds in the study were HEV antibody-negative,indicating high sero-prevalence of HEV in the pig populations in Zhejiang Province.展开更多
On the global scale, hepatitis B virus(HBV) infection is the main cause of hepatocellular carcinoma(HCC) especially in regions of Asia where HBV infection is endemic. Epidemiological studies show that the incidence of...On the global scale, hepatitis B virus(HBV) infection is the main cause of hepatocellular carcinoma(HCC) especially in regions of Asia where HBV infection is endemic. Epidemiological studies show that the incidence of inflammation-driven HCC in males is three times as high as in females. Recent studies suggest that sex hormones have a crucial role in the pathogenesis and development of HBV-induced HCC. We found that the estrogen/androgen signaling pathway is associated with decreased/increased transcription and replication of HBV genes and can promote the development of HBV infections by up/downregulating HBV RNA transcription and inflammatory cytokines levels, which in turn slow down the progression of HBV-induced HCC. Additionally, sex hormones can also affect HBV-related HCC by inducing epigenetic changes. The evidence that both morphology and function of the human liver are affected by sex hormones was found over 60 years ago. However,the underlying molecular mechanism largely remains to be elucidated. This review focuses mainly on the molecular mechanisms behind the sex difference in HCC associated with HBV and other factors. In addition, several potential treatment and therapeutic strategies for inflammation-driven HCC will be introduced in this review.展开更多
基金Supported by National Nature Science Foundation of China(GrantNo.30772360)Nature Science Foundation of Health Department of Hubei Province,China(No.JX4B48)Fund of Yangtze University for Doctor(No.2009001)
文摘Objective:To investigate the effects of hepatitis B virus(HBV)X protein(HBx)on the expression of tumor necrosis factor-α(TNF-α)in glomerular mesangial cells(GMCs)and the underlying intracellular signal pathways.Methods:The plasmid pCI-neo-X that carries the X gene of hepatitis B virus was transfected into cultured GMCs.HBx expression in the transfected GMCs was assessed by Western-blot.TNF-αprotein and mRNA were assessed by ELISA and semi-quantitative RT-PCR,respectively.Three kinase inhibitors-U0126,an inhibitor of extracellular signal-regulated kinases(ERKs);lactacvstin,an inhibitor of nuclear factor-κB(NF-κB);and SB203580,a selective inhibitor of p38 MAP kinase(p38 MAPK)were used to determine which intracellular signal pathways may underlie the action of HBx on TNF-αexpression in transfected GMCs.Results:A significant increase in HBx expression in pCI-neo-X transfected GMCs was detected at 36 h and 48 h,which was not affected by any of those kinase inhibitors mentioned above.A similar increase in the expression of both TNF-αprotein and mRNA was also observed at 36 h and 48 h,which was significantly decreased in the presence of U0126 or lactacytin,but not SB203580.Conclusions:HBx upregulates TNF-αexpression in cultured GMCs,possibly through ERKs and NF-κB pathway,but not p38 MAPK pathway.
基金a grant from National Natural Science Foundation of China (No. 30772360)
文摘The X gene of HBV encodes a 17-kD protein, termed HBx, which has been shown to function as a transcriptional trans-activator of a variety of viral and cellular promoter/enhancer elements. The aim of this study was to investigate the effect of HBx on gene expression of interleukin （IL）-1β and IL-6, and proliferation of rat mesangial cells in vitro. The X gene of HBV was amplified by PCR assay, and inserted into the eukaryotic expression vector pCI-neo. The structure of recombinant pCI-neo-X plasmid was proved by restrict endonuclease digestion and sequencing analysis. pCI-neo-X was transfected into cultured rat mesangial cell line in vitro via liposome. HBx expression in transfected mesangial cells was detected by Western blot. The IL-1β and IL-6 mRNA expression in those cells was assayed by semiquantitative RT-PCR. Mesangial cell proliferation was tested by MTT. The results showed that HBx was obviously expressed in cultured mesangial cell line at 36th and 48th h after transfection. The expression of IL-1β and IL-6 mRNA was simultaneously increased. The cell proliferation was also obvious at the same time. It was concluded that HBx gene transfection could induce IL-1β and IL-6 gene expression and mesangial cell proliferation. HBx may play a critical role in mesangial cell proliferation through upregulation of the IL-1β and IL-6 gene expression.
文摘With the introduction of the new highly effective antiviral therapies, there has been a dramatic increase in the use of the hepatitis C virus(HCV)-positive livers in HCV-positive recipients. In the majority of studies, HCV positivity was defined as a donor testing HCV Ab positive. In 2015, all Organ Procurement Organizations were mandated to perform and report HCV Nucleic Acid Amplification Testing(NAT) results on all deceased and living donors. Studies are not yet available on how organs are being utilized based on NAT status and whether NAT status affects recipient outcomes. Further studies are needed to maximize the use of these organs.
基金supported by Technological Project of Zhejiang Emtry-Exit Inspection and Quarantine Bureau(ZK200611)Technology Innovation Service Platform of Inspection and Quarantine for Safe Frontier (2006C1704)
文摘The study was aimed at establishing an indirect ELISA for epidemiological investigation of hepatitis E viral(HEV) infection in pigs from the major pig-producing regions of Zhejiang Province. The gene fragment covering the immunogenic epitopes of HEV ORF2 was synthesized and expressed in Escherichia coli.Western blot analysis revealed that the purified protein reacted to HEV positive sera,but not to positive sera from some other common viruses that infect pigs.An indirect ELISA system was then developed using truncated HEV ORF2 protein as the coating antigen and had diagnostic accuracy of 91.5% as compared with a diagnostic kit for HEV antibodies.A total of 1 330 serum samples were collected from 46 pig farms in Zhejiang Province from 2005-2008 and tested for sero-prevalence of HEV using the indirect ELISA.The average HEV-positive rate was 55.7%(741/1 330).The positive rate of 2005 and 2006 was 62.7%(175/279) and 61.4%(301/490) respectively,much higher than those of 2007(43.0%,59/137) and 2008(48.6%,206/424).Pigs aged 66-100 days had a statistically higher positive rate(58.2%,110/189) when compared to that of pigs aged 30-65 days(39.7%,73/184) or 101-160 days(44.1%,83/188).Only three herds in the study were HEV antibody-negative,indicating high sero-prevalence of HEV in the pig populations in Zhejiang Province.
基金supported by the State Key Project for Liver Cancer(2012ZX10002009)National Natural Science Foundation of China(81301811,81521091,81422032,81272212,81672860,81372674,and 91529303)+1 种基金Science Foundation of Shanghai(134119a3700)Strategic Priority Research Program of the Chinese Academy of Sciences(XDA12010201)
文摘On the global scale, hepatitis B virus(HBV) infection is the main cause of hepatocellular carcinoma(HCC) especially in regions of Asia where HBV infection is endemic. Epidemiological studies show that the incidence of inflammation-driven HCC in males is three times as high as in females. Recent studies suggest that sex hormones have a crucial role in the pathogenesis and development of HBV-induced HCC. We found that the estrogen/androgen signaling pathway is associated with decreased/increased transcription and replication of HBV genes and can promote the development of HBV infections by up/downregulating HBV RNA transcription and inflammatory cytokines levels, which in turn slow down the progression of HBV-induced HCC. Additionally, sex hormones can also affect HBV-related HCC by inducing epigenetic changes. The evidence that both morphology and function of the human liver are affected by sex hormones was found over 60 years ago. However,the underlying molecular mechanism largely remains to be elucidated. This review focuses mainly on the molecular mechanisms behind the sex difference in HCC associated with HBV and other factors. In addition, several potential treatment and therapeutic strategies for inflammation-driven HCC will be introduced in this review.
