ObjectiveTo explore the expression of programmed death-1 ligand (PD-L1) on peripheral blood monocytes in septic mice, and analyze the difference between the mild and severe septic mice. MethodsFirstly, thirty C57 mice were randomly divided into a sham group, a mild sepsis group and a severe sepsis group. Sepsis model was induced by cecal ligation and puncture (CLP). The severity of sepsis was distinguished by length of cecum ligated. Survival rate was recorded after CLP and compared between three groups. Then sixty C57 mice were randomly divided into a sham group, a mild sepsis group and a severe sepsis group. Peripheral blood was obtained at 6 h and 24 h to detect PD-L1 expression on monocytes in peripheral blood by flow cytometry. Tumor necrosis factor-α(TNF-α) and interleukin-10 (IL-10) in serum were detected by enzyme-linked immunosorbent assay. ResultsThe survival rate in the mild sepsis group was higher than that in the severe sepsis group. TNF-αand IL-10 levels in the mild and severe sepsis groups were higher than those in the sham group at 6 h and 24 h (P < 0.01). PD-L1 expression on monocytes in the mild and severe sepsis groups was higher than that in the sham group at 6 h and 24 h (P < 0.05). The expression of PD-L1 in the severe sepsis group was higher than that in the mild sepsis group, however, there was no statistical difference between two groups (P > 0.05). ConclusionsPD-L1 expression on monocytes is increased in septic mice. PD-L1 expression tended to increase in severe sepsis compared with the mild sepsis.
ObjectiveTo investigate whether the miR-33s negatively regulates LPS-induced production of inflammatory cytokines by targeting p38 MAPK. MethodsHuman monocytes THP-1 cells were cultured in vitro and transfected with miR-33s mimic (25 nmol/L) or miR-33s inhibitor (25 nmol/L)by TransIT-X2® Dynamic Delivery System for 24 h. Then the transfected THP-1 cells were stimulated by LPS of 10.0 ng/mL for 24 h. The expression of miR-33s and p38 MAPK protein were measured by semi-quantitative RT-PCR. The concentrations of TNF-α,IL-6 and IL-1β in the cultured supernatant were assessed by ELISA. ResultsThe transfection of miR-33s mimic significantly increased the release of TNF-α,IL-6 and IL-1β(P<0.05). The expression of p38 MAPK protein was also significantly reduced(P<0.05). However,the pre-treatment of miR-33s inhibitor reversed the LPS-induced release of TNF-α,IL-6,and IL-1β,and the expression of p38 MAPK protein of THP-1 cells. ConclusionmiR-33s may play an important role in the regulation in inflammatory factors released from THP-1 cells by targeting p38 MAPK.
Objective To explore risk factors of stroke-associated pneumonia (SAP) for elderly stroke patients in ICU, and analyze the predictive value of human leukocyte antigen-DR (HLA-DR) on monocytes for SAP. Methods During January 2015 to August 2016, 155 elderly patients with stroke were recruited. The level of monocyte HLA-DR expression was measured after admission and the incidence of SAP was recorded. The risk factors for SAP were analyzed by univariate and multivariate analysis. ROC curve was drawn to analyze prognostic value of HLA-DR. Results SAP occurred in 75 cases with occurrence rate of 48.4%, including 42 early-onset cases and 33 later-onset cases. Age (OR=11.532), Glasgow Coma Scale (OR=7.124), dysphagia (OR=8.846), mechanical ventilation (OR=15.184), atrial fibrillation (OR=7.869), smoking history (OR=11.784), diabetes (OR=7.185) were independent risk factors (all P<0.05). The expression rate of monocyte HLA-DR in the SAP patients was significantly lower than those in the patients without SAP (allP<0.05). Through the ROC curve analysis, the expression rate of HLA-DR that below 78.65% was the optimum cut-off value for prediction of SAP with the area under ROC curve of 0.922, the sensitivity of 80.0% and the specificity of 85.0%. The sensitivity to predict early-onset SAP was 90.5% (38/42), and to predict later-onset SAP was 66.7% (22/33). Conclusions Age, severe coma, dysphagia, mechanical ventilation, atrial fibrillation, smoking history and diabetes are risk factors for SAP in elderly stroke patients in ICU. The detection of monocyte HLA-DR has reference value for early prediction of SAP especially for early-onset SAP with higher sensitivity.