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Objective To compare the difference of the HBsAg detected results between the Roche cobas e601 electrochemilumi-nescence immunoassay instrument and the Abbott Architect i2000chemiluminescent microparticle immunoassay instrument.Methods The HBsAg positive specimens with the quantitation results of lower than 250IU/mL detected by the Abbott Architect i2000 were selected and simultaneously detected by the Roche cobas e601.The differences of detected results were compared and per-formed the linear correlation and analysis regression.Results 46 clinical specimens were detected.The detected results had best correlation between the two instruments by getting rid of 1 specimen with unconformable reactivity of detected results.15 speci-mens had the HBsAg detected result of 0.05-1.00 IU/mL by the Abbott Architect i2000,the linear regression equation was Y =17.49X+0.843(r=0.979);15 specimens had the HBsAg detected result of 1.1 -10.00 IU/mL IU/mL by the Abbott Architect i2000,the linear regression equation was Y =15.72X +21.06(r=0.952);15 specimens had the HBsAg detected result of 11 -250 IU/mL by the Abbott Architect i2000,the linear regression equation was Y =29.17X -129(r=1.000).Conclusion The detected results have better correlation between the two instruments and can be mutually converted by the formulas.
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@#Objective To explore the effect of ischemic postconditioning on serum superoxide dismutase (SOD) and malondialdehyde (MDA) in acute myocardial infarction (AMI) and clinical significance. Methods 101 AMI patients accepted emergency percutaneous coronary intervention (PCI) were divided into postconditioning group (n=46) and control group (n=55) according to the treatment they accepted.The concentration of serum SOD and MDA were observed 4, 8, 12, 16, 20, 24, 36, and 48 h after PCI, as well as the grade of Thrombolysis in Myocardial Infarction (TIMI) and TIMI Myocardial Perfusion Grades (TMPG), serum creatine kinase-MB (CK-MB) peak value, scoring of nuclide distribution 10 d after PCI, and frequence of cardiac events within 30 d after PCI. Results Compared with the control group, serum SOD increased (P<0.01) and MDA decreased (P<0.001) respectively 2, 4, 8, 12, 16, 20, 24, 36 and 48 h after the PCI, especially the valley of SOD and peak of MDA value in the postconditioning group; while the patient with TIMI flow of grade 3 and TMPG of grade 3 increased (P<0.05), the peak of serum CK-MB decreased (P<0.01), and the score of nuclide distribution decreased (P<0.05). After the operation for 30 days, the frequence of cardiac events was less in the postconditioning group than in the control group (P<0.05). Conclusion Ischemic postconditioning can reduce the peroxidation after PCI, to increase myocardial perfusion, reduce infarct area, and improve prognosis in acute ST-segment elevated myocardial infarction
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Objective To obtain I50 anti-idiotype antibody and identify its activity in vitro.Methods I50 anti-idiotype (Id) antibody gene was amplified from the template of fuse 5-I50 by PCR to construct a prokaryotic expression vector pET25b-I50. The expression of pET25b-I50 in E. coli BL21(DE3) was induced by isopropylthio-β-D-galactopyranoside (IPTG) and was confirmed by SDS-PAGE and Western blot with Ab1(FC2) monoclonal antibody and an anti-hexahistidine tag antibody. The method of dialysis refolding was used to restore the activity of I50 anti-Id antibody, which was measured by Dot-ELISA and lymphocyte proliferation assay. Results The recombinant vector was successfully constructed and the recombinant protein was successfully expressed and purified with 90% purity. The relative molecular weight of the expressed protein was 15 kD, which was in accordance with expectation. The activity of I50 anti-Id antibody could be restored and could promote the proliferation of lymphocyte in a dose-dependent manner. Conclusion These results suggested that I50 anti-Id protein vaccine is likely an option in the therapy against nasopharyngeal carcinoma in vivo.
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Objective To express and purify the human scFv antibody,SA3,against the hepatoma fused to enhanced green fluorecsent protein,and to observe the targeted capacity of fusion protein EGFP-SA3 in vivo.Methods SA3 and EGFP genes were cloned into plasmid pET-25b( + )to construct the recombinant plasmid EGFP-SA3/pET-25b ( + ),followed by DNA sequencing.Then it was transformed into E.coli BL21 ( DE3 ) and induced for fusion expression of EGFP-SA3with IPTG.The expressed fusion protein EGFP-SA3 was purified and detected with SDS-PAGE.HepG2 cells were incubated with the fusion protein EGFP-SA3 in vitro,and the binding bioactivity was observed under the fluorecsent microscope.Further more,we injected the EGFP-SA3 by caudal vein into nude mice planted by hepatoma and observed the whole body fluorescence image of EGFP.Results SA3 and EGFP genes were successfully cloned into pET-25b( + ),which was confirmed by restriction enzyme Nco I-Xho I or Nco I-Eco RI.A band migrated at the position 750 bp,same to EGFP gene,emerged when recombinant plasmid was digested by restriction enzyme Nco I-Eco RI.Similarly,a band,about 1 500 bp,emerged when digested by Nco I-Xho I.The open-reading frame was confirmed by DNA sequencing.Fusion protein EGFP-SA3 was expressed as inclusion body.After purification and refolding,the result of immunofluorecsence detection verified that EGFP-SA3could specifically bind to HepG2 cells and maximum tumor penetration was at 24 h after the injection.Conclusion The purified fusion protein EGFP-SA3 has strong binding capacity to HepG2cells,indicating the scFv SA3 has a potential value as a targeting molecule for diagnosis and targeted therapy for liver cancer.
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Objectives Construct a subtractive library of Caski cell line induced by exposing to the space environment by suppression subtractive hybridization and pave the way to explain the molecular mechanisms of the changes at the gene level. Methods Super SMART cDNA synthesis and suppression subtractive hybridization (SSH) were performed to isolate differentially expressed cDNA fragments from strains subclonal 48A9 cell line. cDNA from the 48A9 cell line were used as " tester" , and the other from the control Caski cell line as "driver". Subtractive products were directly inserted into T/A cloning vector, and then transformed into host bacteria to set up a subtractive cDNA library of specially or highly expressed genes in strains subclonal 48A9 cell line. Results mRNA were directly extracted and purified with good quality. Double strand cDNA were reverse transcripted integratedly, and then cut by Rsa I into even length short segments. Liga-tion was identified as high effective. After two hybridizations, a subtractive library of differentially expressed genes in strains subclonal 48A9 cell line was successfully constructed by SSH. Conclusion SSH is an effective approach to isolate differentially expressed genes.