Objective The study consisted of two parts: in part Ⅰ the effects of propofol or/and procaine on CD18, CD62L expression and superoxide anion (SOA) release of phorbol 12-myristal 13-acetate(PMA) stimulated human neutrophils (PMNs) were studied in vitro; in part Ⅱ the effects of propofol or/and procaine on IL-6 and TNF-?production in endotoxin-stimulated human whole blood were studied. Methods PMNs were separated from the whole blood obtained from healthy 20-40yr old subjects. Part Ⅰ consisted of 9 groups: in group 1 (control) phosphate buffer solution was added to PMNs; in group 2 PMNs were stimulated by PMA 100mg?ml-1 ; in group 3-5 different concentrations of propofol (0.4, 4.0,40?g?ml-1) were added to PMA stimulated PMNs; in group 6-8 different concentrations of procaine (1.5,15,150?g?ml-1 ) were added to PMA stimulated PMNs; in group 9 propofol 2?g?ml-1 and procaine 8?g?ml-1 were added to PMA stimulated PMNs. Part II also consisted of 9 groups: in groupl whole blood was mixed with normal saline; in group2 lipopolysaccharide (LPS) l?g?ml-1 was added to whole blood; in group 3-5 different concentrations of propofol (0.4,4.0,40?g?ml-1) were added to LPS stimulated whole blood; in group 6-8 different concentrations of procaine (1.5, 15, 150?g?ml-1 ) were added to LPS stimulated whole blood; in group 9 propofol 2?g?ml-1 and procaine 8?g?ml-1 were added to LPS stimulated whole blood. CD18, CD62L expressions were measured by flow cytometry. SOA release was determined by cytochrome C reduction in the presence or absence of superoxide dismutase (SOD) . IL-6 and TNF-? production were measured by using radioimmunoassay. Results Propofol or/and procaine depressed CD18 up-regulation, CD62L shedding and SOA release of PMA-stimulated PMNs and propofol was more effective than procaine. Propofol enhanced but procaine inhibited the increased production of TNF-? and IL-6 in the LPS stimulated whole blood but when propofol and procaine were used in combination. The effectof procaine predominated. Conclusions Propofol or/and procaine can attenuate tissue damage induced by neutrophils by inhibiting the function of neutrophils.

ObjectiveTo evaluate the effect of sufentanil preconditioning on acute lung injury induced by intestinal ischemia-reperfusion (I/R) in rats and the role of opioid receptors.MethodsForty-eight male Wistar rats weighing 200-250 g were randomly divided into 6 groups ( n =8 each):sham operation group (group S) ; I/R group; sufentanil preconditioning group (group SPC) ; COTP (μ receptor antagonist) + SPC group; NTD (δ receptor antagonist) + SPC group and nor-BNI (κ receptor antagonist) + SPC group.Intestinal I/R was produced by occlusion of the superior mesenteric artery for 45 min followed by 2 h reperfusion in groups I/R,SPC,COTP +SPC,NTD + SPC and nor-BNI + SPC.Sufentanil 10 μg/kg was injected intravenously at 10 min before ischemia in groups SPC,COTP + SPC,NTD + SPC and nor-BNI + SPC.COTP 1 mg/kg and NTD 5 mg/kg were injected intravenously at 10 min before sufentanil injection in groups COTP + SPC and NTD + SPC respectively,while nor-BNI 5 mg/kg was injected intravenously at 15 min before sufentanil administration in group nor-BNI + SPC.The animals were sacrificed at 2 h of reperfusion,the intestinal tissue was removed for microscopic examination and intestinal damage was assessed and scored according to Chiu.Left lung tissue was also removed for microscopic examination (1 =normal,4 =severely injured).The apoptosis in lung cells was detected using TUNEL and apoptosis index (AI) was calculated.The expression of Bcl-2 and Bax proteins in lung tissues was detected using immuno-histochemistry and Bcl-2/Bax ratio was calculated.ResultsCompared with group S,Chiu's score,lung injury score and AI were significantly increased,expression of Bcl-2 and Bax proteins was up-regulated,Bcl-2/Bax ratio was significantly decreased in the other groups ( P ＜ 0.01 ).Compared with group I/R,Chiu' s score,lung injury score and AI were significantly decreased,Bcl-2 protein expression was up-regulated,Bax protein expression was downregulated,and Bcl-2/Bax ratio was significantly increased in group SPC (P ＜ 0.01 ).Compared with group SPC,Chiu's score,lung injury score and AI were significantly increased,Bcl-2 protein expression was down-regulated,Bax protein expression was up-regulated,and Bcl-2/Bax ratio was significantly decreased in groups COTP + SPC,NTD + SPC and nor-BNI + SPC ( P ＜ 0.01 ).ConclusionSufentanil preconditioning can attenuate lung injury induced by intestinal I/R by activation of opioid receptors in rats.

Objective To investigate the role of sufentanil on intestinal ischemia-reperfusion model of liver cell apoptosis and bel-2/Bax expression,and the opioid receptor-mediated effects in the process.Methods Seventy two healthy adult Wistar rats,weight 200-250 g,either male or female,were randomly divided into 9 groups(n=8),sham operation group(S),ischemia-reperfusion group(IR),sufentanil treated group (SPC),three kinds of opioid receptor antagonist,NTD(δ receptor antagonist,),nor-BNI(κ receptor antagonist),CTOP(μ receptor antagonist)in the intervention groups(NTD + SPC,nor-BNI + SPC,CTOP + SPC),three kinds of opioid receptor antagonist control groups(NTD,nor-BNI,CTOP).by clamping the superior mesenteric artery production model of intestinal ischemia-reperfusion injury,after the end of the experiment,immediately take the left lobe of liver specimens.Immunohistochemical detection of bcl-2/Bax protein expression,terminal transferase labeling(TUNEL)assay of liver cell apoptosis index(AI).Results Compared with IR group,SPC Group bcl-2 expression increased(0.277±0.014 vs 0.230±0.013)(P＜0.05),bcl-2/Bax ratio increased(1.375±0.113 vs 0.819±0.102)(P＜0.05),Bax down regulation(0.202±0.013 vs 0.283±0.025)(P＜0.05),liver cell AI reduced(6.491±1.191vs10.094±1.051)(P＜0.05).Compared with the SPC group,NTD + SPC group,nor-BNI +SPC group,CTOP+SPC group bcl-2 expression decreased(F=59.698,P＜0.05),bcl-2/Bas ratio decreased(F=40.349,P＜0.05),Bax expression was elevated(F=53.765,P＜0.05),liver cells AI elevated(F=76.728,P＜0.05).Conclusion Sufentanil pretreatment inhibit rat intestinal ischemia-reperfusion liver cell apoptosis.δ,κ,μthree kinds of opioid receptor mediate the protective effect of sufentanil,and the effects are equal.

Objective To evaluate the effects of delayed remote limb ischemic preconditioning (DRLIP) on myocardial injury in patients undergoing cardiac valve replacement under cardiopulmonary bypass (CPB).Methods A total of 41 patients of both sexes,aged 55-70 yr,weighing 50-75 kg,scheduled for elective cardiac valve replacement under CPB,of ASA physical status Ⅱ or Ⅲ (NYHA Ⅱ or Ⅲ),were randomly divided into 2 groups using a random number table:control group (group C,n =20) and group DRLIP (n =21).In group DRLIP,the patients underwent three 5-min cycles of unilateral lower limb ischemia,induced by a manual cuff-inflator placed on the left thigh and inflated to 200 mmHg starting from 24 h prior to surgery.Blood samples were taken from the central vein before aortic clamping,at 6 h of reperfusion and at 24 h after surgery for determination of plasma cardiac troponin Ⅰ concentrations.Before aortic clamping and at the end of CPB,myocardial tissues were obtained from the right auricle for determination of the expression of caspase-3 and cell apoptosis.Apoptotic index was calculated.The recovery of spontaneous heart beats was recorded.Results Compared with group C,plasma troponin Ⅰ concentrations were significantly decreased at 6 h of reperfusion,and the expression of caspase-3 was downregulated,apoptotic index was decreased at the end of CPB,and no significant change was found in the incidence of recovery of spontaneous heart beats in group DRLIP.Conclusion DRLIP can protect myocardium against injury in the patients undergoing cardiac valve replacement under CPB,and inhibition of cell apoptosis is involved in the mechanism.

Objective To evaluate the cardioprotection induced by combination of dexmedetomidine and limb ischemic preconditioning in the patients undergoing coronary artery bypass grafting (CABG) with cardiopulmonary bypass (CPB).Methods Eighty American Society of Anesthesiologists physical starus Ⅱ or Ⅲ patients of both sexes,aged 52-64 yr,weighing 51-78 kg,with New York Heart Association Ⅱ or Ⅲ,scheduled for elective CABG with CPB,were divided into 4 groups (n =20 each) using a random number table method:control group (group C),limb ischemic preconditioning group (group L),dexmedetomidine group (group D) and dexmedetomidine plus limb ischemic preconditioning group (group DL).Limb ischemic preconditioning was induced by 3 cycles of 5-min unilateral lower limb ischemia followed by 5-min reperfusion starting from 30 min before aortic clamping in L and DL groups.Dexmedetomidine was injected via the central vein in a loading dose of 1 μg/kg after induction of anesthesia,followed by an infusion of 0.4 μg · kg-1 · h-1 until the end of operation in D and DL groups.Venous blood samples were obtained immediately before aortic clamping,at the end of CPB and at the end of operation for determination of plasma concentrations of cardiac troponin Ⅰ (cTnI) by enzyme-linked immunosorbent assay.Myocardial tissues were obtained from the right auricle immediately before aortic clamping and at the end of CPB for determination of the expression of Bcl-2 and Bax (by immunohistochemistry) and apoptosis index (AI) (using TUNEL).The restoration of spontaneous heart beat was recorded.Bcl-2/Bax ratio was calculated.Results Compared with group C,the plasma cTnI concentrations were significantly decreased,the Bcl-2 expression was up-regulated,the Bcl-2/Bax ratio was increased,Bax expression was down-regulated,and AI was decreased in the other three groups (P＜0.05).Compared with L and D groups,the plasma cT-nI concentrations were significantly decreased,the Bcl-2 expression was up-regulated,the Bcl-2/Bax ratio was increased,Bax expression was down-regulated,and AI was decreased in group DL (P＜0.05).The rate of restoration of spontaneous heart beat was significantly increased in group DL as compared with the other three groups (P＜0.05).Conclusion Combination of dexmedetomidine and limb ischemic preconditioning can mitigate myocardial injury,it provides better efficacy than either alone,and the mechanism is related to inhibiting cell apoptosis in the patients undergoing CABG with CPB.

Objective: To evaluate the effects of ketamine anesthesia on proteome in hippocampus in aged rats. Methods: Thirty healthy male Wistar rats, aged 20 months, weighing 560-610 g, were divided into 2 groups (n=15 each) using a random number table method: control group (group C) and ketamine group (group K). In group K, ketamine 80 mg/kg was intraperitoneally injected, additional 1/2 initial dose was given when the righting reflex was recovered, and anesthesia was maintained for 3 h. Morris water maze test was performed starting from 1st day after the end of anesthesia.Five rats were selected at days 1 and 7 after the end of anesthesia and sacrificed, and hippocampal tissues were obtained to extract proteins.Proteins extracted from rat hippocampi were identified by 2-dimensional electrophoresis (2-DE). The differentially expressed proteins were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and biological information system. Results: Compared with group C, the escape latency and total swimming distance to find the submerged platform in Morris water maze at the 1st day after anesthesia were significantly prolonged in group K (P<0.05 or 0.01). The MALDI-TOF-MS analysis showed that there were 21 differentially expressed proteins at 1st day after ketamine anesthesia, of which 6 proteins (involving maintenance of intracellular protein homeostasis, energy metabolism, etc.) presented with up-regulated expression and 15 proteins (involving synaptic vesicle transport efficiency, synaptic structural and functional plasticity, maintenance of intracellular protein homeostasis, NMDA-mediated Ca²⁺ signal transport, energy metabolism, etc.) presented with down-regulated expression.There were 8 differentially expressed proteins at 7th day, including 3 proteins with up-regulated expression and 5 proteins with down-regulated expression (P<0.05). Conclusion Ketamine anesthesia can induce 21 differentially expressed proteins in hippocampi of aged rats, involving synaptic vesicle transport efficiency, synaptic structural and functional plasticity, intracellular protein homeostasis, NMDA-mediated Ca²⁺ signal transport, energy metabolism, and etc.which may be involved in the mechanism of ketamine-induced temporary cognitive dysfunction.

Objective To evaluate the effects of ketamine anesthesia on proteome in hippocampus in aged rats.Methods Thirty healthy male Wistar rats,aged 20 months,weighing 560-610 g,were di-vided into 2 groups(n＝15 each)using a random number table method: control group(group C)and ket-amine group(group K).In group K,ketamine 80 mg/kg was intraperitoneally injected,additional 1/2 ini-tial dose was given when the righting reflex was recovered,and anesthesia was maintained for 3 h.Morris water maze test was performed starting from 1st day after the end of anesthesia.Five rats were selected at days 1 and 7 after the end of anesthesia and sacrificed,and hippocampal tissues were obtained to extract proteins.Proteins extracted from rat hippocampi were identified by 2-dimensional electrophoresis(2-DE).The differentially expressed proteins were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF-MS)and biological information system.Results Compared with group C,the escape latency and total swimming distance to find the submerged platform in Morris water maze at the 1st day after anesthesia were significantly prolonged in group K(P＜0.05 or 0.01).The MAL-DI-TOF-MS analysis showed that there were 21 differentially expressed proteins at 1st day after ketamine an-esthesia,of which 6 proteins(involving maintenance of intracellular protein homeostasis,energy metabo-lism,etc.)presented with up-regulated expression and 15 proteins(involving synaptic vesicle transport ef-ficiency,synaptic structural and functional plasticity,maintenance of intracellular protein homeostasis,NMDA-mediated Ca2+signal transport,energy metabolism,etc.)presented with down-regulated expres-sion.There were 8 differentially expressed proteins at 7th day,including 3 proteins with up-regulated ex-pression and 5 proteins with down-regulated expression(P＜0.05).Conclusion Ketamine anesthesia can induce 21 differentially expressed proteins in hippocampi of aged rats,involving synaptic vesicle transport efficiency,synaptic structural and functional plasticity,intracellular protein homeostasis,NMDA-mediated Ca2+signal transport,energy metabolism,and etc.which may be involved in the mechanism of ketamine-induced temporary cognitive dysfunction.