ABSTRACT
Objective:To investigate the effect of stress-induced protein Sestrin2 (SESN2) on necroptosis of mouse dendritic cell (DC) induced by lipopolysaccharide (LPS) combined with zVAD, a panaspartate-specific cysteine protease (caspase) inhibitor.Methods:The DC2.4 cell line derived from the bone marrow of mouse in the 3rd to 10th generations was cultured. The cells were stimulated with LPS for 0 hour, 6 hours, 12 hours, and 24 hours, and grouped according to the stimulation time points. Western blotting was performed to determine the protein expression of SESN2 in each group. Overexpression empty lentivirus (NC), SESN2 gene overexpression RNA sequence lentivirus (SESN2 LV-RNA), small interfering empty lentivirus (NS), and SESN2 gene small interfering RNA sequence lentivirus (SESN2 siRNA) were transfected into DC2.4 cells. After 72 hours of transfection, cell fluorescence expression was observed under the inverted fluorescence microscope. Cells in each transfection group were stimulated with LPS for 24 hours. The blank control groups were set up and cultured with phosphate buffered saline (PBS) for 24 hours. Western blotting was performed to measure SESN2 protein expression. In the same groups as above, cells were stimulated with LPS+zVAD for 24 hours. The blank control groups were set up and cultured with PBS for 24 hours. Western blotting was used to determine the expression of mixed lineage kinase domain-like protein (MLKL) and phosphorylated-MLKL (p-MLKL). The p-MLKL levels and the number of positive cells were observed using laser scanning confocal microscopy. The necroptotic cell ratios were assessed by both flow cytometry and Hoechst staining.Results:Compared to the LPS 0 hour group, the expression of SESN2 in the LPS 24 hours group showed a significant increase. Therefore, 24 hours was chosen as the subsequent stimulation time point. After successful lentivirus transduction and 24 hours of cultivation, the MLKL phosphorylation level in the SESN2 siRNA+LPS+zVAD group was significantly higher than that in the NS+LPS+zVAD group. The MLKL phosphorylation in the SESN2 LV-RNA+LPS+zVAD group was significantly lower than that in the NC+LPS+zVAD group. The MLKL phosphorylation levels in both the NS+LPS+zVAD group and the NC+LPS+zVAD group were obviously higher than those in the NS+PBS group and the NC+PBS group, respectively. Laser scanning confocal microscopy showed that the trends in quantity and fluorescence intensity of p-MLKL protein expressions were consistent with the above results. The results from flow cytometry analysis and Hoechst staining showed that the rates of cell necrotic apoptosis in SESN2 siRNA+LPS+zVAD group were significantly higher than those in NS+LPS+zVAD group [flow cytometry analysis: (30.800±1.153)% vs. (20.800±1.114)%, Hoechst staining: (75.267±0.451)% vs. (46.267±3.371)%, both P < 0.05], indicating that knocking down SESN2 further exacerbated the occurrence of necroptosis. The necrotic apoptosis rates in SESN2 LV-RNA+LPS+zVAD group were significantly lower than those in NC+LPS+zVAD group [flow cytometry analysis: (7.160±0.669)% vs. (19.240±2.322)%, Hoechst staining: (32.433±3.113)% vs. (48.567±4.128)%, both P < 0.05], indicating that overexpressing SESN2 reversed such response and markedly reduced the proportion of necroptotic cells compared to the corresponding empty vector group. Conclusion:SESN2 exhibits an inhibitory effect on necroptosis of DC in sepsis. Targeted SESN2 expression may regulate the process of DC-mediated immune response in sepsis.
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Objective To study effects of bivalirudin and heparin plus tirofiban on plasma P-selectin in patients with ST-segment elevation acute myocardial infarction (STEMI) undergoing emergency percutaneous coronary intervention (PCI) and compare the clinical effects on bleeding and thrombosis.Methods 260 hospitalized patients with acute STEMI who accepted emergency PCI were enrolled in the study.They were randomly divided into the bivalirudin group (129 cases) and the heparin plus tirofiban group (131 cases).Blood samples were taken before operation,during operation (about 20 minutes after medicacion injection) and after operation (three hours after the end of operation).Serum P-selectin levels were measured.All patients were followed-up for 30 days.The main events included hemorrhage within 30 days,acquired thrombocytopenia and stent thrombosis.Results The level of P-selectin increased significantly in the heparin group during operation compared to the preoperative level and remained statistically higher at three hours after the end of operation than preoperative level (P ＜ 0.05).Compared with preoperative level,the P-selectin level of the bivalirudin group was significantly lower than that of heparin group (P ＜ 0.05).After 30 days of follow-up,bivalirudin had lower rates bleeding events compared to the heparin group(5.4％ vs.15.3％,P =0.009).The need of medical intervention for bleeding events (BRAC2-5 hemorrhage) was less in the bivalirudin group(0.8％ vs.5.3％,P =0.029).There was no significant difference in the major bleeding events between the 2 groups (BRAC3-5 hemorrhage) (0 vs.0.8％,P =0.32).No significant difference found between the two groups in acquired thrombocytopenia (P ＞ 0.05).Conclusion Bivalirudin may reduce P-selectin levels in STEMI patients undergone emergency PCI during perioperative period.Heparin increases perioperative P-selectin.Bivalirudin may reduce the bleeding events.
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ObjectiveTo investigate the prevalence and distributing feature of atrial fibrillation (AF) in Xinjiang Kazaks adult population.MethodsFour-stage selected random samples aged 30-89 years from Tacheng,Yili and Aletai were analyzed.An epidemical study of AF was performed including inquiring medical history,recording electrocardiogram and auscultation. ResultsA total of 22 514 adults were surveyed.The prevalence of AF in Xinjiang Kazaks adult population was 0.37％,which was increasing with aging.The prevalence was higher in men than in women (0.5％ vs 0.2％,P ＜0.01 ).In AF patients,23 was valvular AF.Ischemic stroke was the most frequent type and the stroke rate in the patients with AF was significantly higher than that without AF (6.0％ vs 1.2％,P ＜ 0.01 ).ConclusionsThe prevalence of AF in Xinjiang Kazaks adult population is lower than the reported national prevalence but patients with AF in this population would not like to take the necessary medicine. Therefore,the control of AF need to be reinforced.