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Objective To investigate the effects of microRNA-294 (miR-294) on tumor necrosis factor-α (TNF-α),interleukin-6 (IL-6) and high mobility group box 1 (HMGB 1) secretion in sepsis by targeting triggering receptor expressed on myeloid cell-1 (TREM-1).Methods miRNA-294 was predicted to regulate TREM-1 specially through bioinformatics analysis.Mice macrophage cell lines RAW264.7 were cultured in vitro,the cells were divided into non-inflammatory stage and inflammatory stage,and the cells in the two stages were subdivided into five groups as follows:normal control (NC),NC mimic transfection (NCm),NC inhibitor transfection (NCi),miR-294 mimic transfection (miR-294m) and miR-294 inhibitor transfection (miR-294i) groups.The ability of miR-294 was confirmed with dual-luciferase activity assay.At non-inflammatory stage,the cells were transfected with mimic or inhibitor of miR-294 or NC using TurboFectTM siRNA Transfection Reagent for 48 hours,mRNA expression of TREM-1 was detected by real-time reverse transcription-polymerase chain reaction (RT-PCR).At inflammatory stage,6 hours after stimulation by lipopolysaccharide (LPS,1 mg/L),the concentrations of TNF-α,IL-6 and HMGB1 were determined by enzyme linked immunosorbent assay (ELISA),the protein expression of TREM-1 was determined by Western Blot.Results ① Dual-luciferase activity assay demonstrated that TREM-1 was the target of miR-294.② In non-inflammatory stage,the expression of TREM-1 mRNA (2-ΔΔ~) in miR-294m group was significantly lower than that of the NC and NCm groups (0.673 ± 0.049 vs.1.000 ± 0.003,0.915 ± 0.039,t1=2.184,t2=5.421,both P＜0.001),the expression of TREM-1 protein (gray scale) was (50.00 ± 1.19)％ of NCm group (t=41.586,P＜0.001).③ In inflammatory stage,the concentrations of TNF-α (ng/L) in miR-294m group was significantly lower than that of the NC group (1 547.18 ±47.18 vs.2 702.11 ± 327.20,t=4.212,P=0.010),the concentrations of IL-6 (ng/L) was significantly lower than that of the NC and NCm groups (505.28 ± 33.33 vs.837.66 ± 69.43,918.72 ± 119.39,t1 =4.382,P1=0.015; t2=5.451,P2=0.021),the level of TREM-1 protein (gray scale) was (51.33 ±0.88)％ of NCm group (t=63.368,P＜0.001).Conclusion miR-294 reduce TNF-α and IL-6 secretion in LPS-induced RAW264.7 through inhibiting the expression of TREM-1 specifically.
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Objective This trial was to observe the effect of Ulinastatin on coagulation functions in patients during operation period,and to investigate the protective mechanisms of Ulinastatin.Methods Forty patients were randomly divided into Ulinastatin group (Group U,n =20) and control group (Group C,n =20).Group U was infused intravenously ulinastatin 4000 U/kg (diluted with saline to 30 ml,20min losers) after anesthesia and before cutting skin,while Group C received the same volume of normal saline.All patients were phlebotomized 1 ml peripheral blood before administered (T0) and after 1 hour (T1),respectively.Coagulation activation time (SonACT),clot rate (CR) and platelet function (PF)were detected by sonoclot coagulation analyzer and platelet function analyzer.Results Compared with group C (controlled group),SonACT of Group U was prolonged significantly at T1 (P ＜ 0.05),and PF were increased at T1 (P ＜ 0.05) ; Compared with T0,SonACT and PF were increased at T1,respectively (P ＜ 0.01).Conclusions Ulinastatin can improve perioperative coagulation function and platelet function.It may reduce intraoperative micro-thrombosis syndrome and postoperative deep vein thrombosis.
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Objective To evaluate influence of general anesthesia on postoperative cognition function in the elderly patients by using different methods and choose a better anesthesia method for elderly patients. Methods Forty elderly patients undergoing selective abdominal surgery were selected and divided into two groups: anesthesia was maintained with general anesthesia combined with epidural anesthesia in observe group and with general anesthesia singlely in control group. Vital signs were observed and recorded during operation and blood pressure varied within the extent of 20% of basic level. Cognition function of each patient 24 hours before and 24,48 hours after operation was e-valuated by MMSE method. Results All the patients maintained steady vital signs and there was no significant differ-ence in MMSE scores between two groups at all the time points. There were 11 cases(55% ) who had acute cognitive dysfunction in observe group and 13 cases(65%) in control group 24h after operation(P>0.05) ,5 cases(25%) in observe group and 9 cases(45%) in control group 48h after surgery(P>0.05). Conclusion Compared with single general anesthesia, general anesthesia combined with epidural anesthesia uses less general anesthetics and has less negative effect on postoperative cognition funetion in the elderly patients,it maybe better in elderly patients undergoing non-cardiac operation.
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Objective To observe the effect of intravenous anesthesia (IVA) with propofol (P),ketamine (K) and fentanyl (F) in different ways in children. Methods 90 pediatric patients were divided into three groups ( n =30) randomly according to the drug used for IVA. Ⅰgroup used compounds of P, F and K; Ⅱgroup used compounds of P and F; Ⅲ group used compounds of P and K. All the drugs were injected intravenously with micro-pump. Total dosage of anaesthetics, awaken time after operation and variation of circulation and breath were observed and sedative and analgesic effect during operation were evaluated. Results Compared with Ⅱ or Ⅲ group, Ⅰgroup showed less dosage, shorter awaken time and smaller degree of change in circulation and breath with complete sedation and analgesia ( P
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Objective To investigate the effects of various concentrations of lidocaine on the N-methyl-D-aspartate ( NMDA)-mediated calcium currents in cultured rat hippocampal neurons. Methods Hippocampal neurons were obtained from newborn Wistar rats (0-24 h after birth) . The hippocampal neurons cultured for 12-14 days were divided into control group and 5 lidocaine groups in which lidocaine was added to the extracellular solution achieving the final concentrations of 10-3 , 10-4 , 10-3 , 10-2 and 10-1 . The neuronal cells were voltage clamped at - 80 mv. The currents evoked by NMDA 100 ?mol?L-1 were recorded. The NMDA-mediated Ca currents were isolated by blocking Na-currents with TTX, K-currents with CsCL and TEACL and non-NMDA receptor (to a large degree AMPA receptor) with CNQX. Current density was calculated (pA / pF) .Results Lidocaine significantly reduced the density of NMDA-mediated calcium currents at concentrations of 10-3- 10-1 ?mol?L-1 compared with that in the control group (P