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Objective: To analysis the effects of laser and lucentis treatment of diabetic retinopathy. Methods:Fifty-three patients with DR were treated with laser and lucentis in our hospital from October 2012 to October 2013 as observer group. Forty-nine patients with DR were treated with laser in our hospital from September 2011 to September 2012 as control group. Results:The results indicated that visual acuity improved rate was 67.9%and decreased vision rate was 13.2% in observer group. Vision conditions in the observation group was better than that in the control group with significant difference (x2=4.60, x2=3.87; P<0.05). Simultaneously, improve time of hemorrhage, exudates, edema and macular thickness in the observation group was better than that in the control group with significant difference (t=4.17, t=3.92, t=4.06; P<0.05). Conclusion: The laser and lucentis treatment of diabetic retinopathy can significantly improving symptoms of retinal and promoting visual recovery.
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Objective To prepare polyclonal antibody of transactivated protein 5 of hepatitis C virus nonstructural 5A (NA5ATP5) and to explore its expression in the liver tissues. Methods In Escherichia coil BL21, the prokaryotic expression vector pET32a(+)-NS5ATP5 was induced by isopropyl-β-D-thiogalactoside (IPTG), and it was analyzed with sodium dodecyl sulfate-polyaerylamide gel electrophoresis (SDS-PAGE) and Western blotting. And the purified protein was used to immunize the rabbit to prepare polyelonai antibody, with which we studied the function of NSSATP5 by determining the different liver tissues with the streptavidin-perosidase (SP) immunohistochemistry method. Results Recombinant NS5ATP5 (molecular weight: 65 kD) and polyclonal antibody were successfully prepared. NS5ATP5 expression in the liver of patients with chronic HCV infection was much higher than that of a normal person, and it was detected mainly in the cytoplasm. Conclusion The findings of the expression difference between HCV patients and normal people led to a novel diagnostic marker to detect HCV infection.
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Objective To evaluate the methylation status at CpG site -55 in the interferon-gamma (IFN-7) gene promoter and its effect on IFN-7 expression in chronic hepatitis B. Method The authors recruited 30 patients with UBeAg-positive chronic hepatitis B (CHB), 30 HBeAg-negative CHB patients, and 30 healthy blood donors. Pyrosequeneing was used to determine the methylation status at CpG site -55 in the IFN-γ gene promoter following bisulfite treatment of DNA in peripheral blood mononuclear cells (PBMCs). The expression of IFN-γ was analyzed by real-time RT-PCR and ELISA. HBV DNA in PBMCs was detected by nested PCR. Results The methylation level at CpG site -55 in the IFN-γ gene promoter was significantly increased, resulting in subsequent down-regulation of the expression of this cytoldne in CHB. The methylation level at CpG site -55 was significantly higher in HBeAg-positive patients than in HBeAg-negative ones (P<0.01) and was also significantly higher in PBMCs from HBV DNA-positive patients than from HBV DNA-negative ones (P<0.01) ; the methylation level at CpG site -55 was positively correlated with the amount of HBV DNA in serum (P<0.01). Oonclusion IFN-γ gene expression appears to be regulated by methylation of the IFN-γ gene promoter in CHB; the methylation level at CpG site -55 is associated with HBV infection.
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Objective To explore the roles of cytokines in the pathogenesis of hemorrhagic fever with renal syndrome(HFRS). Methods Double-antibody sandwich ELISA was used to determine serum interleukin (IL)-6, urine tumor necrosis factor (TNF), IL-6 and IL-8 levels in 56 patients with HFRS. Results Serum IL-6, urine TNF, IL-6 and IL-8 concentrations in HFRS patients were significantly higher than those in control group, respectively (P＜0.001). The concentrations increased at fever stage, then continued to increase during hypotension stage and peaked at oliguria stage. The concentrations of serum IL-6, urine TNF, IL-6 and IL-8 increased in accord with the severity of the disease and differed greatly among different types of the disease. Serum IL-6 had remarkable relationships with serum specific antibodies. It was positively related to serum β2-microglobulin (β2-MG), blood ureanitrogen (BUN) and creatinine (Cr). Significant positive relationships were also found both between urine IL-6 and TNF, and between IL-6 and IL-8 (r=0.5768, P＜0.05; r=0.3760, P＜0.01). Conclusion TNF, IL-6 and IL-8 activated during the course of the disease. IL-6 is associated with the immunopathological lesions caused by the hyperfunction of humoral immune response. IL-6, IL-8 and TNF are involved in the renal immune impairment. Determining them might, in certain extent, be used in predicting the prognosis and outcome of patients with HFRS.
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<p><b>OBJECTIVE</b>To study the effect and mechanism of the peripheral blood mononuclear cell (PBMC) invasion by HBV on artificial immunization in newborns.</p><p><b>METHODS</b>Fifty-two newborns of HBsAg positive mothers were immunized with HBIG (hepatitis B immunoglobulin) and HBVac (hepatitis B vaccine) and were followed up for 7 months. The newborns' HBV-DNA in serum and in the PBMCs was detected with nested-PCR; anti-HBs was tested with solid phase radioimmunoassay (SP-RIA). PBMCs isolated from newborn peripheral blood were incubated in the presence of PHA or purified HBsAg. Interleukin-2 (IL-2) level in culture supernatants of activated cells was detected by ELISA.</p><p><b>RESULTS</b>The failure rate of immunization was higher in infants with positive HBV-DNA in PBMCs than those with negative HBV-DNA (P < 0.05); IL-2 level in PBMC culture supernatants was lower in former than in the latter and in normal controls (P < 0.05). The level of IL-2 in the immunization failure newborns was lower than that in the successfully immunized newborns and in normal controls (P < 0.05).</p><p><b>CONCLUSIONS</b>Intrauterine invasion of PBMCs by HBV is one of the important reasons for immunization failure in newborns. IL-2 production is closely related to the invasion of PBMCs by HBV, which may contribute to the failure of artificial immunization in newborns.</p>
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Humans , Infant, Newborn , DNA, Viral , Blood , Hepatitis B Surface Antigens , Allergy and Immunology , Hepatitis B Vaccines , Hepatitis B virus , Physiology , Immunization , Immunoglobulins , Allergy and Immunology , Interleukin-2 , Blood , Leukocytes, Mononuclear , VirologyABSTRACT
Objective To explore the clinical features of hepatitis G virus (HGV) infection and HGV replication sites in vivo. Methods A sensitive and specific reverse transcription-nested polymerase chain reaction (RT-nested PCR) was set up to detect the plus-stranded and minus-stranded HGV RNA in sera of 129 high risk population as well as in peripheral blood mononuclear cells (PBMCs) from 59 cases of them. The clinical features of HGV infection were analyzed in the meantime. Results Among 9 patients with single HGV infected, 3 cases were diagnosed as acute hepatitis, 3 cases as chronic hepatitis and the other 3 cases as healthy carriers. The detection rate of HGV RNA in PBMCs is much lower than that of HCV RNA (35.7 % vs. 77.3 % , P＜0.05). In all 59 PBMCs and 129 serum samples no minus-stranded HGVRNA was found. Conclusion HGV infection can cause acute or chronic hepatitis with mild clinical manifestation. HGV can invade PBMCs but can not replicate in PBMCs, and the ability of HGV to invade PBMCs is lower than that of HCV.
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Objectives To study the effect and the mechanism of peripheral blood nuclear cells (PBMC) invaded by hepatitis B virus (HBV) on the artificial immunization in newborns Methods Fifty two newborns, whose mothers were hepatitis B surface antigen (HBsAg) positive, were immunized with hepatitis B immunoglobulin and hepatitis B vaccine (HBVac), and then followed for 7 months The newborns′ serum and PBMC HBV DNA was detected by nested PCR, hepatitis B surface antibody (HBsAb) was tested with solid phase radioimmunoassay PBMC from newborn were incubated with PHA and HBsAg The supernatant interleukin 2 (IL 2) level was mesured by enzyme linked immununosorbent assay (ELISA) Results The rate of vaccination failure was higher in the infants with PBMC HBV DNA positive than those with negative ( P
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Objective To study the interaction between HCV core protein and HCBP6 in mammlian cells using CheckMateTM Mammalian Two-Hybrid System.Methods cDNA fragments encoding HCV core protein and HCBP6 were amplified by PCR and subsequently cloned into pGEM-T vector.After verified by sequencing,the target fragments were subcloned into mammalian two-hybrid plasmids,pBIND and PACT,respectively.The recombinant plasmids,pBIND-core and pACT-HCBP6 were co-transfected into HepG2 cells with reporter plasmid pG5luc.pBIND+pACT were induced as background controls,pBIND-Myod+pACT-Id as positive controls,and pBIND-core+pACT,pBIND+pACT-HCBP6 as blank controls.The expression of G5luc,which indicated the interaction between HCV core protein and HCBP6 in mammlian HepG2 cells,was assayed through Dual-Luciferase Report Assay System and Turner Biosystems Veritas Microplate Lunimometer.Results The recombinant vectors pBIND-core and pACT-HCBP6 were successfully constructed.When co-transfected into HepG2 cells with reporter plasmid pG5luc,there were significant differences in the luciferase activity in the pBIND-core and pACT-HCBP6 groups compared with every control group.Conclusions HCBP6 can interact with HCV core protein in HepG2 cells,which provides clues for further study on the function of HCBP6 and core proteins,and on the mechanism of HCV core in cell apoptosis and cancer transformation.
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0.05),the difference of ALT,AST level and AST/ALT ratio between high viral load (serum HCV RNA≥8.5?10~5 IU/ml) group and low viral load (serum HCV RNA