Objective To study the effects of advanced glycation end (AGEs) products induced by bovine serum albumin (BSA) on the survival and the morphology of bovine retinal endothelial cells (BREC) and pericytes (BRP). Methods BSA with the final concentration of 50 mg/ml was incubated in PBS, containing 500 mmol/L D-glucose, for 12 weeks under 37℃. AGEs-BSA was purified by Sephacryl S-300 column chromatography and was confirmed by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE). The concentration of AGEs-BSA was determined by the method of commassie protein assay. In order to detect the toxic effects of AGEs-BSA on cultured BREC and BRP, groups of AGEs-BSA and BSA with different concentration and untreated control were set up. Phase contrast microscope was used to observe the effect of AGEs-BSA and BSA (with the concentration of 500mu;g/ml and actuation duration of 48 hours) on morphology of BREC and BRP. Results As the dosage of AGEs-BSA increased, the number of inhibited cells increased. When the concentration of AGEs-BSA was 500mu;g/ml, the inhibited BREC in AGEs-BSA group was (72.8plusmn;15.9)% of which in untreated control group, and the inhibited BRP was (64.8plusmn;9) % of which in untreated control group. AGEs-BSA with low concentration promoted the proliferation of endothelial cells, but there was no significant difference between AGEs-BSA and the control group (P=0.231). Inhibited proliferation and abnormal morphology were seen under the phase contrast microscope while the normal morphology of cells was found in BSA and control group. Conclusion AGEs-BSA with the high concentration may inhibit the growth of both BREC and BRP, which leads the loss of BRP and damage of vascular function. These results suggest that nonenzymatic glycosylation plays a major role in diabetic complications. (Chin J Ocul Fundus Dis, 2006, 22: 11-15)
Objective To probe a selective cultural method for bovine retinal endothelial cells (BREC) and pericytes (BRP) in vitro.Methods With the isolation of active retinal blood vessels, BREC were cultured in a fibronectin coated substrate and Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 10% human serum and 100μg/ml heparin, while homogeneous cultures of retinal pericytes were obtained when isolated microvessels were seeded to uncoated dishes and grown in DMEM supplemented with 20% fetal bovine serum. BREC were identified by acetylated-low density lipoprotein (Dil-Ac-LDL) incorporation and immunohistochemical method of Von Willebrand factor, while BRP were identified by the immunohist ochemical method of α-isoform of smooth-muscle actin. Results The purity of selectively cultured BREC and BRP was more than 98%, being reproducible. BREC got together around the microvessel fragments with the small-cyprinoid-like configuration at first，and could phagocytize Dil-Ac-LDL with the expression of fluorescence in cytoplasm. The expressions of Von Wllebrand factor and α-isoform of smooth-muscle actin were positive and negative in BREC respectively, while were negative and positive in BRP respectively.Conclusion BREC and BRP with high purity can be obtained by using selective culture and coated-dishes respectively which are simple and repeatable methods. (Chin J Ocul Fundus Dis，2004，20：23-26)
ObjectiveTo analyze the correlation between the vaccination status of inpatients with Omicron variant infection and the risk of Omicron critical illness. MethodsA retrospective analysis was performed on the clinical data of patients with Omicron infection admitted to a designated hospital for COVID-19 in Chengdu from December 1, 2022 to January 31, 2023. Patients were divided into critical group and non-critical group according to their condition and the "COVID-19 Diagnosis and Treatment Program (Tenth Edition)". According to the vaccination status, the patients were divided into incomplete vaccination group, full vaccination group and booster vaccination group. Multivariate logistic regression was used to analyze the association between vaccination, symptoms and signs at admission, and the risk of critical illness. ResultsA total of 3 603 inpatients with Omicron infection were included, including 730 cases (20.3%) in the critical group and 2 873 cases (79.7%) in the non-critical group. There were 2 399 people (66.6%) in the incomplete vaccination group, 433 people (12%) in the full vaccination group, and 771 people (21.4%) in the booster vaccination group. Compared with the incomplete vaccination group, the proportion of critical illness in the full vaccination group and booster vaccination group was lower, and the critical illness rate increased with age (P<0.05). After adjusting for age, gender, and underlying diseases, the results of multivariate logistic analysis showed that full vaccination (OR=0.67, 95%CI 0.50 to 0.89) and booster vaccination (OR=0.76, 95% CI 0.61 to 0.94) were significantly associated with a reduced risk of critical illness. ConclusionFull vaccination and booster dose can effectively reduce the risk of critical illness after infection.