Altered expression profiles of microRNAs in a stable hepatitis B virus-expressing cell line / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>MicroRNAs (miRNAs) are highly conserved small non-coding RNAs of 18 - 25 nucleotides (nt) that mediate post-transcriptional gene regulation. Hepatitis B virus (HBV) can cause either acute or chronic hepatitis B, and is a high risk factor for liver cirrhosis and hepatocellular carcinoma. Some mammalian viruses have been shown to modulate the expression of host cellular miRNAs. However, interactions between the HBV and the host cellular miRNAs are largely unknown.</p><p><b>METHODS</b>miRNA microarray and Northern blotting analysis were used to compare the expression profile of cellular miRNAs of a stable HBV-expressing cell line HepG2.2.15 and its parent cell line HepG2. mRNA microarray assay and the miRanda program were used to predict the miRNA targets. A flow cytometric assay was further used to investigate the expression of human leukocyte antigen (HLA)-A.</p><p><b>RESULTS</b>Eighteen miRNAs were differentially expressed between the two cell lines. Among them, eleven were up-regulated and seven were down-regulated in HepG2.2.15 cells. Northern blotting analysis confirmed that the expression of miR-181a, miR-181b, miR-200b and miR-146a were up-regulated and the expression of miR-15a was down-regulated, which was in consistent with the results of the microarray analysis. Furthermore, some putative miRNA targets were predicted and verified to be linked with mRNA expression. The 3'-UTR of HLA-A gene had one partially complementary site for miR-181a and miR-181a might down-regulate the expression of HLA-A.</p><p><b>CONCLUSION</b>HBV replication modulates the expression of host cellular miRNAs, which may play a role in the pathogenesis of HBV-related liver diseases.</p>

Screening and identification of proteins interacting with HCV NS4A via yeast double hybridization in leukocytes and gene cloning of the interacting protein / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To screen proteins interacting with HCV NS4A protein in leukocytes by yeast-double hybridization.</p><p><b>METHODS</b>The bait plasmid pGBKT7-NS4A was transformed into yeast AH109 was transformed, and the expressing of the fusion protein was identified by SDS-page. The transformed yeast was mated with yeast Y187 containing leukocytes cDNA library plasmid in 2xYPDA medium. Diploid yeast was plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) and synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing x-alpha-gal for selecting two times and screening. After extracting and sequencing of plasmid from blue colonies, analysis was conducted by bioinformatics. And, the gene encoding the interesting protein was cloned, and back-cross was performed.</p><p><b>RESULTS</b>Forty-five colonies were sequenced, among them, 29 colonies were human calcium modulating cyclophilin ligand (CAML). The gene encoding CAML was cloned, and the interaction between NS4A and CAML was ensured.</p><p><b>CONCLUSION</b>Seven kinds of proteins interacting with NS4A in leukocytes were successfully screened and the results brought some new clues for studying the pathogenesis of HCV.</p>

Analysis on factors related to X-ray outcomes in 93 SARS patients / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study the related factors of the X-ray outcomes in recovered SARS patients.</p><p><b>METHODS</b>The X-ray results of 93 patients with SARS were studied retrospectively. The possible related factors analyzed were age, sex, body temperature at onset, range of the lesion, glucocorticoid administration time. The data were analyzed by chi square test.</p><p><b>RESULTS</b>Among all the patients with abnormal X-ray result, 19 were male (54.29%), 16 (45.71%) were female, P > 0.01; 7 (58.33%) were above the age of 45; 28 (34.57%) were below the age of 45, P > 0.01; hyperpyrexia (>/= 39), 26 (50.00%), below 39, 9 (21.95%); multiple-lesion, 22 (52.38%), mono-lesion, 13 (25.49%), P < 0.01; glucocorticoid administration time within 5 days, 22 (38.60%) after 5 days, 12 (33.33%), P > 0.01; within 7 days, 21 (30.00%), after 7 days, 14 (60.87%), P < 0.01.</p><p><b>CONCLUSION</b>The X-ray results of SARS were closely related to the severity of the disease (hyperpyrexia and bilateral lung field lesion). There was no significant correlation between X-ray result and the age or sex of the patients. Early use of glucocorticoid (within 5 days after onset), had no remarkable influence on the X-ray result. It was noted, however, the incidence of residual lesion in lung obviously increased if glucocorticoid was administered after 7 days of onset.</p>

IFN or oxymatrine in combination with lamivudine in patients with lamivudine-resistant chronic hepatitis B / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To observe the therapeutic efficacy of IFN or oxymatrine in combination with lamivudine in patients with lamivudine-resistant chronic hepatitis B.</p><p><b>METHODS</b>Forty patients ongoing treatment with lamivudine were randomized to three groups: group A, 14 patients with addition of IFN alpha-2b 3MU to ongoing lamivudine, daily, one month, followed by the same dose given every other day, five months; group B, 15 patients with addition of injectable oxymatrine 60 mg daily, three months, followed by oral oxymatrine every day, three months, and group C, 11 patients ongoing treatment with lamivudine alone. The HBV DNA level in serum, HBeAg seroconversion, and ALT level were detected at the end of the treatment.</p><p><b>RESULTS</b>After 6 months of treatment, HBV DNA became negative in 35.73% patients treated with combination with IFN, and in 13.3% patients treated with combination with oxymatrine. ALT level was normal in 85.71% or 86.66% of patients, respectively. In none of the patients under ongoing treatment with lamivudine alone HBV DNA or HBeAg became negative, and ALT level was normal in 36.36% of patients.</p><p><b>CONCLUSION</b>These data indicated that IFN or oxymatrine in combination with ongoing lamivudine therapy provided effective antiviral therapy in patients with lamivudine-resistant HBV. The addition of IFN or oxymatrine to ongoing lamivudine therapy in lamivudine-resistant patients led to significant inhibition of viral replication and improvement in liver function after 6 months of therapy.</p>

Study on the interaction between hepatitis virus C nonstructural protein 4A and calcium modulating cyclophipin ligand by in vivo coimmunoprecipitation / 中华传染病杂志

Objective To prove the interaction between hepatitis virus C(HCV)nonstruetural protein 4A(HCV NS4A)and calcium modulating cyclophilin tigand(CAML)with yeast-two hybrid- ization and coimmunoprecipitation.Methods The gene encoding CAML was cloned,and subcloned into the yeast expression vector pGADT7 and eucell expression vector pcDNA3.1/His-A.The back- cross test between HCV NS4A and CAML was performed in yeast cells.After that,the pCMV-Myc/ NS4A plasmid and pcDNA3.1/His-A-CAML plasmid were co transfected into 293 cells and,then, coimmunoprecipitation and Western blot were performed.Results The gene encoding CAML was cloned sucessfully,and then the gene was subcloned into yeast expression vectors,pGADT7.After the interaction between NS4A and CAML was ensured in yeast cells,the eukaryotic expression vec- tors of NS4A and CAML were constructed and their interaction was ensured again by Co-immunopre- cipitation.Conclusions The interaction between HCV NS4A and CAML is proved.CAML is one of the proteins involved in Ca~(2+)signaling,which suggests that the interaction of HCV NS4A and CAML may be a new clue of the chronic mechanism of HCV infection.Future studies will be required to de- fine the physiologic significance of this interaction.