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【Objective】 To screen the differentially expressed immune genes between responders （Rs） and non-responders （NRs） in chronic hepatitis B patients receiving interferon alpha （IFN-α） treatment and to explore the molecular basis of IFN-α treatment failure. 【Methods】 The gene expression profile GSE27555 which contained 6 Rs and 7 NRs was obtained from the Gene Expression Omnibus （GEO） database； then differentially expressed genes between liver tissues of Rs and NRs were selected by the R software. The iconic immune gene set consisting of 1793 genes was downloaded from the immunology database and analysis portal （ImmPort）. The immune genes were extracted from the differentially expressed genes to obtain the differentially expressed immune genes. Subsequently， Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analyses of the differentially expressed immune genes were performed by the R software. Protein-protein interaction （PPI） network of the differentially expressed immune genes was constructed using the STRING online tool. The plugin CytoHubba of the Cytoscape software was applied to identify the top 10 genes by using Degree， MCC， MNC， and Closeness algorithms； then the intersection was taken to obtain the hub genes. 【Results】 A total of 88 differentially expressed immune genes， consisting of 13 upregulated and 75 downregulated genes， were identified between Rs and NRs. GO analysis showed that the differentially expressed immune genes were significantly enriched in T cell activation， cell chemotaxis， regulation of cell-cell adhesion， antigen processing and presentation. KEGG pathway analysis suggested that the differentially expressed immune genes were significantly enriched in cytokine-cytokine receptor interactions， Th cell differentiation， antigen processing and presentation， interactions between viral proteins and cytokines and cytokine receptors， chemokine signaling pathways， T cell receptor signaling pathway， IL-17 signaling pathway， natural killer cell-mediated cytotoxicity， Toll-like receptor signaling pathway， and other immune response signaling pathways. The top 7 hub genes， identified by the plugin cytoHubba of the Cytoscape software by using Degree， MCC， MNC and Closeness algorithms， were CD8A， IFNG， CCL2， CCL5， CXCL10， CCL4， and FCGR3A. 【Conclusion】 This study made a comprehensive analysis of the differentially expressed immune genes and signal pathways between Rs and NRs by bioinformatics， and identified 7 Hub genes related to the ineffectiveness of IFN-α treatment in CHB patients. These hub genes may serve as potential biomarkers for predicting the response of IFN-α treatment in CHB patients.
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ObjectiveTo investigate the effect of prohibitin (PHB) on hepatitis C virus (HCV) replication. MethodsHuman hepatocarcinoma Huh-7.5 cells were transfected with the full-length genome HCV RNA in vitro to establish the full-length genome cell model of HCV, and the supernatants were collected 24, 48, 72, and 96 hours later to measure HCV copies. The indirect immunofluorescence assay was used to measure the expression of HCV core proteins, and a transmission electron microscope was used to observe the changes in the ultrastructure of HCV-infected Huh-7.5 cells and identify the full-length genome cell model of HCV. Quantitative real-time PCR and Western Blot were used to measure the expression of PHB in HCV-infected Huh-7.5 cells, and the RNA interference technique was used to measure the replication of HCV RNA. The t-test was used for comparison between groups. ResultsThe results of identification showed that the HCV full-length genome cell model was successfully established. At 24 and 48 hours after Huh-7.5 cells were transfected with HCV RNA, the mRNA expression level of PHB was 13.41±1.35 and 16.45±1.76, respectively, which differed significantly from that in the control group (101±057 and 101±087，t=29540 and 31361，both P＜0.01), and the 24- and 48-hour transfection groups showed a significantly higher expression level of PHB than the control group (both P＜0.01). At 24 and 48 hours after Huh-7.5 cells were transfected with the RNA interference plasmids of PHB, the level of HCV RNA was 64.32±5.49 and 84.45±7.06, respectively, significantly higher than that in the control group (shRNA-control group; 10.52±1.57 and 16.34±2.97, t=29538 and 25908，all P＜0.01). ConclusionIn HCV-infected Huh-7.5 cells, the increased expression of PHB is closely related to HCV RNA infection, and to a certain extent, PHB can inhibit the replication of HCV RNA.
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Objective To analyze the changes of thyroid function of healthy primipara before 20 weeks of gestation to establish normal gestational age-specific reference interval of thyroid hormones,and to investigate the prevalence of maternal thyroid disorders during the first half of pregnancy.Methods A total of 1605 healthy primipara without risk factors of thyroid diseases before 20 gestational weeks and 200 non-pregnant healthy women who accepted pre-conception care in Beijing Friendship Hospital from September 2010 to June 2011 were tested for serum thyroid stimulating hormone (TSH),free thyroxine (FT4) and thyroid peroxidase antihody (TPOAb) by chemiluminometric immunoanalysis.One thousand two hundred and fourty-three pregnant women among them with negative thyroid antibooly and without previons thyroid diseases were selected as the standard population for normal interval.Gestational age-specific percentile categories for TSH and FT4 were calculated.The prevalence of maternal thyroid disorders was examined by gestational agespecific intervals.Results (1) Compared with non-pregnant women,the median value of serum TSH in pregnant women decreased by 29.56％ to the value of 0.91 mU/L; while that of FT4 rose by 7.79％ to the value of 11.33 pmol/L before 12 weeks; and TSH increased while FT4 decreased during 13 to 20 weeks.(2) The median values and reference intervals (2.5th percentile,97.5th percentile) for TSH were 1.59 mU/L (0.15 mU/L,5.19 mU/L) in no-pregnant women,1.12 mU/L (0.03 mU/L,3.67 mU/L) at 8-12+6 gestational weeks,1.21 mU/L (0.05 mU/L,3.74 mU/L) at 13-16+6 gestational weeks,1.50 mU/L (0.31 mU/L,4.33 mU/L) at 17-19+6 gestational weeks; and the median values and reference intervals (2.5th percentile,97.5th percentile) for FT4 were 9.91 pmol/L (6.69 pmol/L,14.03 pmol/L),10.68 pmol/L (7.98 pmol/L,18.66 pmol/L),10.04 pmol/L (6.18 pmol/L,16.22 pmol/L),9.40 pmol/L (6.44 pmol/L,13.51 pmol/L) respectively.(3) According to gestational age-specific reference intervals,the general prevalence of maternal thyroid disorders,including hyperthyroidism,hypothyroidism,subclinical hypothyroidism and hypothyroinemia,was 3.55％ (57/1606).At 8-12+6 gestational weeks,13-16+6 gestational weeks and 17 19+6 gestational weeks,the occurrence of hyperthyroidism was 0.00％,0.13％ and 0.00％;that of hypothyroidism was 0.00％,0.13％ and 0.00％; the incidence of subclinical hypothyroidism was 3.60％,2.76％ and 3.00％; the occurrence of hypothyroxinemia was 0.16％,0.26％ and 0.86％,respectively.The positive rate of TPOAb at 8-12+6,13-16+6 and 17-19+6 gestational weeks were 22.91％ (140/611),16.56％ (126/761) and 15.45％(36/233),and the total positive rate of TPOAb was 18.82％ (302/1605).The median level of TPOAb was 38.90,41.87 and 39.10 mU/L,respectively.Conclusions Before 20 gestational weeks,specific changes occur in maternal thyroid function.TSH level decreases during 8 to 12 gestational weeks,and then increases gradually; while FT4 level increases during 8 to 12 weeks,and then decreases gradually.Thyroid dysfunction during pregnancy is common and subclinical hypothyroidisum is the leading problem in thyroid disorders.Screening for thyroid function during early pregnancy is suggested.
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Objective:To investigate the FSP1mRNA, TGF-?mRNA and HGFmrNA expressinn of lung fibroblasts being affected by alveolar typeⅡcells in pingyangmycin-induced pulmonary fibrosis in rats and its possible mechanism. Methods: Twenty-eight SD rats were randomly divided into two groups as the Pingyangmycin group and saline group. Pulmonary fibrosis was induced by intralracheal instillation with pingyangmycin in the Pingyangmycin group, and the saline group was treated with normal saline, Then ten rats in Pingyangmvcin group and four rats in saline group were sacrificed respectively at the 7th,28th day after intratracheal instillation. Three sacrificed rats in the Pingyangmyein group and three sacrificed rats in the saline group were abstracted alveolar typeⅡcells. The six sacrificed rats in the Pingyangmycin group were abstracted lung fihroblasts. When the two kinds of cells were at the good stale, the experiment group:put the supernate fluid of alveolar typeⅡcells of Pingyangmycin group into the lung fihroblasts of Pingyangmycin group;the control group: put the supernate fluid of alveolar typeⅡcells of saline group into the lung fibroblasts of Pingyangmycin group, then coculture 48 hours, the levels of FSP1mRNA,TGF-?mRNA, HGFmRNA of lung fibroblasts were detected by demi-quantitate RT-PCR. The rest rats of Pingyangmycin grnup and the saline group were used HE stained. Results:(1)The expression of FSP1mRNA, TGF-?mRNA, HGFmRNA in experiment group were higher than the control group at the 7 th,28th day (P0.05),except the level of HGFmRNA in experiment group at the 28th was higher than at the 7th (P