Objective To investigate the relationship between some autoantibodies including anti-? 2-glycoprotein Ⅰ (anti-? 2GPⅠ) and chronic liver disease. Methods A cohort of patients with chronic hepatitis B (CHB) and post-hepatitis B cirrhosis (PHBC) was studied for the serum anti-? 2GPⅠ levels by ELISA with purified ? 2GPⅠ as antigen. The serum dsDNA, smooth muscle antibody (SMA) and ribonucleoprotein antibody (RNPA) were also detected. Results High positive rate was observed in patients with CHB or PHBC (20.9%, 9/43; 49.3%,35/75) comparing with that in control group (3.1%, 1/32) ( P

Objective:Human sera ACA was detected by ELISA method and some factors including the dilution rate of sera, blocking materials, temperature,ionic strength and interval time between blocking and detection,which could probablly influence the detective level of sera ACA were investigated.Methods:Human sera ACA was detected by ELISA.Results:10% NBS/PBS was more feasible than 1% bovine albumin/PBS for blocking non specific binding;the ACA titer of the same sample detected at 37℃ was lower than that of 22℃ and 25℃,the dilution rate 1∶50 for sera was suitable for the detection; time internal between blocking and detection could influence ACA level when it exceeding 2 weeks;the blinding ability of Ag with Ab was signicantly inhibited when the NaCl inonic strength was higher than 1.5 mol/L.Conclusion:The detected results of human sera ACA influenced by the dilution rate of sera,blocking materials,temperature ionic strength and internal time between blocking and detection.

Objective:To investigate oxidative modification of β2-glycoprotein Ⅰ in vit ro.Methods:Rat β2-glycoprotein Ⅰ was purified and characterized,then oxidize d by hypoxanthine plus xanthine oxidase as a supreroxide free radical generating system;carbonl groups of β2-glycoprotein Ⅰ were detected by the reaction w ith 2,4-dinitrophenylhudrazine.Results:There was a significant increase of carbonyl groups formation in β2- glycoprotein Ⅰ oxidized in comparison with native β2-glycoprotein Ⅰ (P <0.05). Conclusion:Carbonyl groups have been formed in vitro on rat β2-glycoprotein Ⅰ after oxidative modification using hypoxanthine plus xanthine oxidase system.

OBJECTIVETo clarify the binding character between Beta-2-glycoprotein I (Beta-2-GPI) and HBsAg.

METHODSBeta-2-GPI was purified from human plasma and labelled with biotin. Solid phase enzyme linked absorbance assay was used to investigate its binding with HBsAg.

RESULTSBiotinylated Beta-2-GPI was found to bind HBsAg and the reaction could be inhibited by excess unlabelled Beta-2-GPI.

CONCLUSIONSBeta-2-GPI may play a role in hepatitis B virus infection.

Binding Sites ; Biotinylation ; Enzyme-Linked Immunosorbent Assay ; methods ; Glycoproteins ; isolation & purification ; metabolism ; Hepatitis B Surface Antigens ; metabolism ; Hepatitis B virus ; chemistry ; metabolism ; Humans ; beta 2-Glycoprotein I

OBJECTIVESTo study the interaction of beta-2-glycoprotein I (beta 2GPI) with the membrane of hepatocytes and determine whether beta 2GPI participates in HBV infection.

METHODSLigand blotting, fluorescence microscopy, and fluorescence activated cell sorter (FACS) analysis were used to detect the specific interaction of beta 2GPI with the hepatoma cell line smmc7721, the gastric carcinoma cell line SGC7901, and the lymphoma cell line HL-60.

RESULTSA specific 40 kDa beta 2GPI band was observed by ligand blotting in the case of smmc7721 cells. No such band was observed in SGC7901 or HL-60 cells. Fluorescence microscopy also revealed specific binding of FITC-beta 2GPI to smmc7721 cells, but neither to SGC7901 nor HL-60 cells. FACS analysis demonstrated that the binding rate of FITC-beta 2GPI to smmc7721 cells was significantly higher than these in SGC7901 and HL-60 cells (P < 0.01). The binding rate to smmc7721 cells did not increase with increasing amounts of FITC-beta 2GPI.

CONCLUSIONSThere is a specific beta 2GPI-binding protein on the membrane of hepatoma cells in cell line smmc7721 which as the beta 2GPI receptor may participate in HBV infection of hepatocytes.

Animals ; Carcinoma, Hepatocellular ; Cell Membrane ; metabolism ; Flow Cytometry ; Glycoproteins ; metabolism ; Hepatocytes ; metabolism ; Liver Neoplasms, Experimental ; metabolism ; Platelet Glycoprotein GPIb-IX Complex ; analysis ; Platelet Membrane Glycoproteins ; Tumor Cells, Cultured ; beta 2-Glycoprotein I

Objective:To detect precancerous lesions of gastric cancer and biopsy tissue vascular endothelial growth factor (VEGF)and mutant p53 gene(mtp53)expression,to explore the development of clinical significance of VEGF and mutant p53 gene in gastric cancer.Methods:19 cases by endoscopic biopsies of normal gastric tissues,22 cases of intestinal metaplasia,47 cases of gastro-intestinal mucosal dysplasia, 54 cases of gastric cancer samples by immunohistochemical staining to detect the expression levels of VEGF and mtp53′s.Results: The expression levels of VEGF, mtp53 in normal gastric mucosa, intestinal metaplasia, dysplasia, and gradually increased gastric cancer was the law.mtp53 of VEGF expression in gastric carcinoma and compared with normal gastric tissue,intestinal metaplasia was significantly higher(P<0.05),but with atypical hyperplasia was no significant difference(P>0.05). Conclusion: The abnormal expression of VEGF and mutant p53 may be related to the degree of deterioration of the stomach tissue lesions related.

OBJECTIVETo study the role of outer membrane protein T (OmpT) in the pathogenesis of uropathogenic Escherichia.coli.

METHODSIn cultured human bladder epithelial cell line 5637, we examined the adhesion ability of wild-type (CFT073), ompT gene knockout (COTD), and revertant (pST) strains of E.coli to the cells and the extracellular matrix (ECM). The expressions of the adhesion gene iha and virulence gene iroN were detected by real-time PCR. Murine models of urinary tract infection with the 3 strains were established to evaluate the bacterial burden of the bladder and kidney tissue and bacterial counts in blood. We also detected the expressions of interleukin-6 (IL-6) and IL-8 in the bladder and kidney tissues of the mice.

RESULTThe COTD strain showed a significantly lower cell adhesion rate than CFT073 strain [(4.62∓0.39)% vs (8.81∓1.13)%, P<0.05] with also a lower ECM-adhesion rate [(4.95∓0.59)% vs (8.85∓0.79)%, P<0.05]. The mRNA expressions of iha and iroN in CFT073 strain were 2.1 and 3.8 times that of COTD strain. In the mouse model, the mean bacterial load of CFT073 strain in the bladder tissue was 6.36∓0.06, significantly greater than that of COTD (6.01∓0.07) and revertant (6.29∓0.06) strains (P<0.05); the bacterial load of CFT073 strain in the kidney tissue was also significantly higher than that of COTD strain (6.25∓0.05 vs 5.87∓0.06, P<0.05). In mice infected with the wild-type, knockout, and revertant strains, the detection rates of IL-6, which were identical to those of IL-8, in the inflammatory bladder and kidney tissues were 60%, 12.5%, and 50%, respectively.

CONCLUSIONSOmpT may regulate the expression of the adhesion gene iha and the transferrin gene iroN to affect the adhesion of uropathogenic E.coli to host cells.

Animals ; Bacterial Adhesion ; Bacterial Load ; Bacterial Outer Membrane Proteins ; metabolism ; Cell Line, Tumor ; Escherichia coli Infections ; pathology ; Escherichia coli Proteins ; metabolism ; Gene Knockout Techniques ; Humans ; Inflammation ; Interleukin-6 ; metabolism ; Interleukin-8 ; metabolism ; Kidney ; microbiology ; Mice ; Peptide Hydrolases ; metabolism ; Receptors, Cell Surface ; metabolism ; Urinary Bladder ; microbiology ; Urinary Tract Infections ; microbiology ; pathology ; Uropathogenic Escherichia coli ; pathogenicity