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Objective:The prokaryotic expression vector of human anti-Siglec-9 Fab fragment antibody was constructed and purified,while was identified. Methods:The variable and conserved regions of heavy chain and light chain were obtained by polymerase chain reaction respectively(PCR),which was combined by overlap extension PCR and was digested with restriction enzyme,and then it was transformed into Escherichia coli BL21 and was purified by His-trap Lambda Fab column and AKTA system. SDS-PAGE,ELISA and Western blot were used for the identification of human anti-Siglec-9 Fab fragment antibody. The effect of human anti-Siglec-9 Fab fragment antibody on regulating the mRNA expression of TNF-α,IL-1,IL-6,IL-8 was detected by real-time PCR. Results:Successfully obtained the chains of heavy and light, while constructed an activation human anti-Siglec-9 Fab fragment antibody which could specifically bind to Siglec-9 protein. The human anti-Siglec-9 Fab fragment antibody could specifically bind to Siglec-9 was confirmed by SDS-PAGE,ELISA and Western blot. The human anti-Siglec-9 Fab fragment antibody inhibited the mRNA expression of TNF-α,IL-1, IL-6,IL-8. Conclusion:Successful prokaryotic expression,purification,character analysis,and suppressed the mRNA expression of in-flammatory cytokines with the human anti-Siglec-9 Fab fragment antibody and lay the biology foundation for the further study.
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Objective: To study the distribution of genotypes 5′-AMP-activated protein kinase alpha-2 subunit gene PRKAA2 rs10889007 (301A>T) and rs2746338 (201A>G) polymorphism in Han population in Chongqing, China, and to explore the relationship between these two single nucleotide polymorphisms (SNPs) and type 2 diabetes mellitus (T2DM).
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<p><b>OBJECTIVE</b>To perform a systematic comparative analysis of two different commercial automated systems using chemiluminescence immunoassay to quantitatively detect hepatitis B virus surface antigen (HBsAg) in patient sera.</p><p><b>METHODS</b>The Elecsys2010 electrical chemiluminescence immunoassay (ECLIA; manufactured by Roche) and the ARCHITECT il000 chemiluminescence magnetic microparticle immunoassay (CMIA; manufactured by Abbott) were used to detect HBsAg in 100 serum samples of individuals who presented at our department with suspected hepatitis infection between January and May 2012. The manufacturer's protocols were strictly followed. The categorical data was analyzed by Chi-squared test, and linear regression analysis was used to compare the results of the two assay systems.</p><p><b>RESULTS</b>The HBsAg detection results from the two different assay systems showed good correlation (r >or= 0.95), and had good correlation at a low (r = 0.966), medium (r = 0.974) and high (r = 0.984) cutoff values. However, the positive detection rate of CMIA was significantly higher than that of ECLIA(94% vs. 88%, P < 0.05). When the HBsAg content was below 0.10 IU/ml, the ECLIA detection rate and sensitivity were slightly higher than those of CMIA.</p><p><b>CONCLUSION</b>The ARCHITECT i1000 and Elecsys 2010 immunoassay systems have good correlation in quantitative detection of HBsAg, but the former may be more sensitive.</p>
Subject(s)
Humans , Hepatitis B Surface Antigens , Blood , Immunoassay , Methods , Luminescent Measurements , Sensitivity and SpecificityABSTRACT
To investigate the inhibitory effect of acetoacetate extract from Celastrus orbiculatus Thumb (COT) on the growth of red fluorescent protein (RFP)-xenografted human hepatocellular carcinoma (HCC) in a nude mouse model. Human HCC HepG2 cells were transduced with RFP and inoculated into the liver of BALB/c nude mice. The tumor-bearing mice were randomly divided into five groups: control group (G1), oxaliplatin positive control group (G2; 25 mg/kg), COT low-dose group (G3; 20 mg/kg), COT high-dose group (G4; 40 mg/kg), and COT early treatment group (G5; 20 mg/kg). The early treatment group received oral COT from day 2 post-tumor implantation. All other mice were treated from day 20 post-tumor implantation. Growth of xenografted tumors was monitored weekly by in vivo real-time fluorescence imaging technology. At the end of the four-week treatment period, all mice were sacrificed and tumor tissues were collected and weighed. The two-sided t-test was used to evaluate intergroup differences in tumor volumes, final tumor weights, and final body weights. Mice treated with COT had significantly smaller xenografted tumors. On day 45 post-implantation, the mean tumor volumes (mm3) in the different groups were: G1, 803.1+/-512.3 ; G2, 83.8+/-23.5; G3, 852.7+/-502.6; G4, 410.0+/-231.6; and G5, 120.5+/-60.1. The mean tumor weights (g) were: G1, 0.95+/-0.49; G2, 0.36+/-0.09; G3, 0.67+/-0.29; G4, 0.48+/-0.15; and G5, 0.38+/-0.11. The differences in tumor weights from G2, G4 and G5 were significantly less than the weight in G1 (P less than 0.05); however, there was no significant differences between the tumor weights in G2, G4 and G5 (P more than 0.05). The tumor weight from the G2 group was significantly less than that of the G3 group (P less than 0.05). COT significantly inhibited the proliferation of human HCC in a nude mouse model. Early treatment with COT produced a more robust inhibitory effect, which was very similar to that achieved with oxaliplatin treatment.
Subject(s)
Animals , Female , Humans , Male , Mice , Acetoacetates , Carcinoma, Hepatocellular , Pathology , Celastrus , Hep G2 Cells , Liver Neoplasms , Pathology , Mice, Inbred BALB C , Mice, Nude , Organoplatinum Compounds , Pharmacology , Plant Extracts , Pharmacology , Xenograft Model Antitumor AssaysABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of lamivudine, interferon alpha and oxymatrine treatment for surviving hepatic failure patients with hepatitis B.</p><p><b>METHODS</b>200 hepatitis B patients, including 100 subacute or acute-on-chronic hepatic failure survivals (group A), and 100 chronic (group B, n=100) hepatic failure survivals, were enrolled in this study. Patients in group A received interferon alpha (n=35), lamivudine (n=33) , or combinational lamivudine and oxymatrine (n=32) therapy for six months; Patients in group B received lamivudine (n=49), or combinational lamivudine and oxymatrine (n=51) therapy for six months, respectively. After the treatment, all patients were followed-up for six months.</p><p><b>RESULTS</b>At the end of follow-up, all patients in group A survived, while in group B three patients (6.1%) receiving lamivudine, and four (7.8%, P>0.05) receiving combinational therapy died; more than 90% of all survivals had their HBV DNA loss. The HBeAg/anti-HBe seroconversion rate in patients of group A treated with interferon alpha (9/17, 52.9%) was higher than that in patients treated with combinational lamivudine and matrine (5/16, 31.3%, P<0.05), which was higher than that in the patients treated with lamivudine alone (1/17, 5.9%, P<0.01), and the Knodell histological activity index score in patients treated with lamivudine (7.2+/-0.8, P<0.05) was lower than that in patients treated with interferon alpha (8.2+/-1.3, P<0.05), and the best efficacy was found in receiving combinational therapy (6.9+/-0.7, P<0.01); Lamivudine or lamivudine in combination with matrine significantly inhibited the intrahepatic inflammatory activities, but had no effect on the existing fibrosis in group B patients.</p><p><b>CONCLUSION</b>Long term nucleotide analogues treatment may delay the progress of fibrosis in hepatitis B-induced hepatic failure survivals, and the administration of matrine in time may further enhance the anti-fibrotic effect of nucleotide analogues.</p>