Objective To evaluate the role of extracelluar signal-regulated kinase (ERK)-cyclic AMP response element binding protein (CREB) signaling pathway in the spinal cord in naloxone-induced withdrawal response in morphine-dependent rats.Methods Fifty male adult SD rats,aged 2 months,weighing 200-250 g,in which intrathecal catheters were successfully implanted without complications,were randomly divided into 5 groups (n =10 each):group control (group C); group morphine dependence (group MD); group morphine withdrawal (group MW); group U0126 (ERK signaling pathway blocker); group dimethyl sulfoxide (DMSO,solvent for U0126).Morphine dependence was induced by increasing doses of subcutaneous morphine for 6 days.The initial dose of morphine was 10 mg/kg twice a day and was increased by 10 mg/kg twice every other day until 50 mg/kg on 6th day in groups MD,MW,U0126 and DMSO.Morphine withdrawal response was induced by intraperitoneal naloxone 4 mg/kg at 4 h after last morphine administration in groups MW,U0126 and DMSO.U0126 150μg (in DMSO 10 μl) and DMSO 10 μl were administered intrathecally at 30 min before naloxone administration in groups U0126 and DMSO respectively.Morphine withdrawal response (0=no withdrawal response,3 =severe response)and touch evoked agitation (0 =no agitation,2 =severe agitation) were observed and scored during 1 h after naloxone administration.The animals were then sacrificed and the spinal cord was removed for determination of the expression of phosphorylated ERK (p-ERK) and phosphorylated CREB (p-CREB) by immuno-histochemistry and Western blot.Results Morphine withdrawal significantly up-regulated the p-ERK and p-CREB expression in group MW compared with group C ( P ＜ 0.05).Withdrawal response score and touch evoked agitation score were significantly increased in groups MW,U0126 and DMSO as compared with group MD ( P ＜ 0.05).U0126 pretreatment significantly attenuated naloxone-induced increase in withdrawal response score and touch evoked agitation score and down-regulated p-ERK and p-CREB expression in group U0126 as compared with group MW ( P ＜ 0.05).Conclusion ERK-CREB signaling pathway in the spinal cord is involved in morphine withdrawal response in morphine-dependent rats.

Actovegin is a drug,which can improve cellular energy metabolism,especially glycometabolism.It has an insulin-like effect,such as increasing the uptake of glucose and activating glucose metabolic enzyme.In addition,it can inhibit the production of lactic acid.Actovegin was mainly applied to the treatment of Alzheimer disease initially,but now it is indicated more and more.

ObjectiveTo investigate the effects of different concentrations of ketamine on the differentiation of human T helper (Th) cells.MethodsTwenty patients ASA Ⅰ,20-60 years undergoing elective operation under general anesthesia were enrolled in this study.Peripheral venous blood samples were taken before anesthesia.Peripheral blood mononuclear cells were isolated and assigned to one of 3 groups ( n ＝ 20 each).Peripheral blood mononuclear cells were incubated in the presence of 0.9％ NaCl (group C),2 concentrations of ketamine (2.5,25.0 μg/ml) (groups K1,K2) for 24 h,and were then stimulated with phytohaemagglutinin for another 48 h.The perce.ntage of Thl and Th2 cells were detected by four-color flow cytometry.The Thl/Th2 ratio was calculated.The IL-2,IL-4,IL-6,IL-10,IFN-γ and TNF-α concentrations in the supernatant were determined by cytometric bead array.ResultsThere was no significant difference in the IL-2,IL-4,IL-6,IL-10,IFN-γ and TNF-α concentrations in the supernatant,the percentage of Thl and Th2 cells and Thl/Tb-2 ratio among groups C,K1and K2 ( P ＞ 0.05).ConclusionsThe sedative and anesthetic concentrations of ketamine exert no effect on the differentiation of human Th cells in vitro.

BACKGROUND: Recently, lithium was reported shown neuroprotective effect against apoptosis induced by a variety of insults in vitro and in vitro,but the precise mechanisms underlying its neuroprotective effect remain unknown.OBJECTIVE: To observe the effect of lithium chloride on neuronal apoptosis and the expression of P53 or nuclear factor kappa B (NF-κB) protein in the CA1 region of the hippocampus after global ischemia in gerbils.DESIGN: A randomized controlled experimental research.SETTING: Department of Anatomy of Nanjing Medical University.MATERIALS: Fifty-four healthy male gerbils weighing 50-70 g, clearing grade, were purchased from Experimental Animal Center of Zhejiang Province.METHODS: Totally 54 gerbils were randomly divided into three groups namely: sham-operation group (SH group), ischemia-reperfusion group (IR group) and lithium chloride group (LI group), with 18 in each group. SH group, IR group and LI group were further divided into 3 subgroups respectively (SH1d, SH3d, SH7d; IR1d, IR3d, IR7d; LI1d, LI3d, LI7d), according to the time of reperfusion, with 6 gerbils in each. Gerbils in LI group were injected intraperitoneally with lithium chloride 3 mEq /kg, once a day for 7consecutive days before operation. Normal saline was used instead of lithium in SH group and IR group as vehicle control. Forebrain ischemia was induced at 24 hours after the last injection of lithium chloride. After gerbils being anesthetized, the bilateral common carotid arteries were blocked with micro aneurysm clips for 5 minutes, and the micro aneurysm clips were removed and the cerebral blood flow restored. Sham-operation animals were underwent the same operation except occlusion of bilateral common carotid arteries. Gerbils in each group were killed at every time points.4 μm coronal sections at 1.7-4.0 mm visual cross were cut at the level of the dorsal hippocampus. The apoptosis cells were assayed with in situ Cell Death Detection Kit, and assay of positive cell in cell apoptosis, P53 and positive NF-κB was performed with immunohistochemistry staining. The total number of TUNEL positive cells, P53 or NF-κB positive cells per image (area of 1 mm2) was counted.MAIN OUTCOME MEASURES: Neuronal apoptosis and expression of P53 or NF-κB protein in the CA1 region after cerebral ischemic reperfusion.apoptosis cell in cerebral hippocampus CA1 region: No TUNEL positive cells were detected in SH group, a large majority of TUNEL positive cells were detected in the CA1 region in IR group on the 3rd day after reperfusion [(552.0±145.5, 142.4±103.5) pcs/mm2, t= 5.623, P ＜ 0.01], and TUNEL positive cells declined on the 7th day after reperfusion. The numbers of TUNEL positive cells in the CA1 region of LI3d, LI7d group were significantly lower than those of IR3d, IR7d group [(408.0±119.8, 156.0±108.2) pcs/mm2,CA1 region: In IR group, the expression of P53 protein was increased on the 1st, 3rd and 7th day after reperfusion compared with that in SH group and cerebral hippocampus CA1 region: No NF-κB protein was expressed in SH group. In IR group, the expression of NF-κB protein was increased on the 1st day after reperfusion (78.5±25.2)/mm2, significantly increased on the 3rd day after reperfusion (176.5±35.5)/mm2 and on the 7th day after reperfusion, the expression of NF-κB protein disappeared. There were no significant statistical difference between LI group and IR group on the 1st day after reperfusion. The expression of NF-κB protein in LI group was significantly lower than that in IR group on the 3rd day after reperfusion [(64.5±30.8)/mm2,t=5.824, P ＜ 0.01].CONCLUSION: Lithium chloride can significantly suppress neuronal apoptosis after global ischemia in gerbils. The down-regulation of expression of P53 or NF-κB protein is one of the mechanisms of the neuroprotective effect by lithium chloride.

Objective To investigate the effects of thoracic epidural anesthesia and analgesia on cellular immune function and erythrocytes glycometabolism in the patients undergoing thoracic surgery.Methods Forty esophageal carcinoma patients,classified as ASA Ⅰ or Ⅱ,scheduled for elective thoracic surgery were randomly divided into two groups with 20 cases each：group A underwent general anesthesia plus thoracic epidural anesthesia （TEA） during thoracic surgery and received patient-controlled epidural analgesia （PCEA） with fentanyl and ropivacaine postoperatively;group B received general anesthesia during thoracic surgery and patient-controlled intravenous analgesia （PCIA） postoperatively. Venous blood samples were collected for the measurement of Th1,Th2 and the activities of PFK,G-6PD and AR before the induction（T0）,2 h after the initiation of the incision（T1）,and 4 h（T2）,24 h（T3）and 48 h（T4）after surgery. Results The Th1/Th2 ratio in both groups were decreased significantly after completion of surgery compared with baseline levels （P0.05）. At T2,T3 and T4 the Th1/Th2 ratio in group A were higher than group B. Compared with these before operation,the activity of PFK was decreased significantly and the activities of G-6PD and AR in erythrocytes were increased markedly at T3 in group B（P0.05）.But erythrocytes PFK,G-6PD and AR activity slightly changed in group A.Conclusion These findings show that thoracic epidural anesthesia and PCEA may inhibit Th0 cells to differentiate into Th2 cells,protect cellular immune function and moderate erythrocyte glucose metabolism changes.

ObjectiveTo investigate the effect of propofol on IL-1β and TNF-α release from BV-2 microglia cells induced by lipopolysaccharide (LPS) and the role of Toll-like receptor 4 (TLR4).MethodsBV-2 microglia cells were seeded in 96-well plates and randomly divided into 4 groups ( n =12 each):control group,LPS group,propofol group (group P) and LPS + propofol group.In group LPS,the cells were incubated with LPS 1 μg/ml for 24 h.In group P,the cells were incubated with propofol 30 μmol/L for 24 h.In group LPS + propofol,the cells were incubated with LPS 1 μg/ml and propofol 30 μmol/L for 24 h.The concentrations of TNF-α ( at 6 h of incubation) and IL-1β (at 24 h of incubation) in the supernatant were detected by ELISA.TLR4 mRNA expression was detectedat at 6 h of incubation by RT-PCR.TLR4 protein expression was detected at 24 h of incubation by Western blot.ResultsCompared with control group,IL-1β and TNF-α release was significantly increased,and the expression of TLR4 mRNA and protein up-regulated in groups LPS and LPS + propofol ( P ＜ 0.05).Compared with group LPS,IL-1β and TNF-α release was significantly decreased,and the expression of TLR4 mRNA and protein down-regulated in group LPS + propofol (P ＜ 0.05 ).Conclusion Propofol can inhibit IL-1β and TNF-α release from BV-2 microglia cells induced by LPS and inhibition of TLR4 expression may be involved in the mechanism.

Objective The purpose of this study was to examine the effect of preoperative glucose infusion on glucose transport-4 mRNA ( Glut-4 mRNA) expression in skeletal muscle before and after thoracic operation. Methods Twelve male ASA Ⅰ - Ⅱ patients undergoing elective radical surgery for esophagus cancer were randomly divided into 2 groups : (Ⅰ ) control group; (Ⅱ) glucose infusion group. Patients with hypertension, hyperlipidemia or diabetes mellitus were excluded. The patients in both groups were premedicated with phenobarbitol and atropine. Anesthesia was induced with midazolam 0.1 mg ? kg-1 , propofol 1.5 mg ?kg-1 , fentanyl 4?g?kg-1 and vecuronium 0.15 mg?kg-1 . The patients were mechanically ventilated after trachea! intubatioa Anesthesia was maintained with continuous iv infusion of propofol (80?g ? kg-1? min-1 ) and vecuronium (1.0 ?g?kg-1?min-1) supplemented with droperidol 0.1 mg?kg-1 and fentanyl 6 ?g ?kg-1 given in several boluses. In group Ⅱ 5 % glucose (0.25 g?kg-1) was infused 2 h before operation. Blood samples and 1 g of skeletal muscle were obtained while chest was being opened and closed for determination of plasma glucose and insulin levels and Glut-4 mRNA expression in muscle. The total RNA of the muscle cells was extracted by trizol one-step template. The Glut-4 mRNA amplification products were determined using RT-PCR with ?-action mRNA as an internal control. The Glut-4 mRNA expression was expressed by desired gene/ ?-action? 100% .Results Glut-4 mRNA expression was significantly decreased ( P

Objective To study the effects of allogeneic blood transfusion on expression of CD40L on peripheral blood monocytes (PBMCs) in patients undergoing radical gastrectomy for gastric carcinoma. Methods Thirty patients classified as ASA physical status Ⅰ- Ⅱ undergoing radical gastrectomy for gastric carcinoma were randomly divided into three groups with 10 patients in each group : group A received no allogeneic blood; group B received leukodepleted blood; group C received allogeneic whole blood during operation or within 12 h after operation. The patients were premedicated with intramuscular phenobarbital sodium 0.1 g and atropine 0.5 mg. Anesthesia was induced with midazolam 0.06 mg ? kg-1 , fentanyl 2 ?g ?kg-1 , propofol 1.5 mg ?kg-1 and vecuronium 8mg and maintained with continuous infusion of propofol (100?g?kg-1?min-1) and vecuronium (5?g ?kg-1?min-1) and intermittent inhalation of 1l%-2% isoflurane. The patients were mechanically ventilated after tracheal intubation. Blood samples were taken before operation and 2, 5 and 10 days after operation. The PBMCs and plasma were separated from peripheral blood by Ficoll-Hypaque centrifugation. The PBMCs were washed and incubated with the patients own plasma (final-concentration 10% ) and PHA (final concentration 20 ?g?ml-1 ) at 37℃ in a 5% CO2 atmosphere for 48h. CD40Lexpression on PBMCs was quantified by flow-cytometry.Results The demographic data including sex, age, bodyweight and duration of operation and intraoperative blood loss were comparable among the three groups. There was no significant difference in the CD40L expression before operation among the 3 groups. In group A there was no change in CD40L expression after operation. In group B CD40L expression on PBMCs increased significantly on the 2nd postoperative day, but returned to preoperative level on the 5th postoperative day. In group C the CD40L expression on PBMCs kept increasing on the 2nd and 5th postoperative day and did not return to preoperative level on the 10th day. The increase in CD40L expression was significantly larger in group C than that in group B ( P

Objective To study the changes in the polyol pathway of glucose metabolism and oxidative stress responses induced by general thoracic surgery by measuring erythrocyte aldose reductase (AR), phosphofructokinase (PFK) and glucose-6 phospate dehydrogenase (G-6PD) activities and plasma MDA and GSH levels before and after surgery performed under different anesthetic techniques. Methods Thirty-two ASA Ⅰ-Ⅱ patients aged 45-71 yrs weighing 55-70 kg were randomly allocated to one of two groups with 16 patients in each group : group Ⅰ received isoflurane inhalation for maintenance of anesthesia and group Ⅱ isoflurane inhalation combined with epidural block. Patients with endocrine and glucose metabolism-related diseases were excluded. The patients were premedicated with intramuscular phenobarbital 0.1 g and atropine 0.5 mg. Anesthesia was induced with intravenous midazolam 0.1 mg?kg-1 , propofol 1.0-1.5 mg?kg-1 , fentanyl 0.1-0.15 mg, droperidol 1-2 mg and vecuronium 0.1 mg?kg-1 in both groups. The patients were mechanically ventilated (VT 8-10 ml?kg-1, RR 10-12 bpm) after tracheal intubation. Anesthesia was maintained with isoflurane inhalation at 1.2-1.6 MAC in group Ⅰ or isoflurane inhalation (0.6-0.8 MAC) combined with epidural block (T9-10) with 1 % lidocaine in group Ⅱ supplemented with fentanyl 5-10 ?g?kg-1 and droperidol 0.1-0.2 mg?kg-1 in divided doses and vecuronium infusion at 1-2 ?g? kg?min-1 . Venous blood samples were obtained before anesthesia (baseline, T0), 90 min after skin incision (T1), 60 min after surgery (T2 ) and on the 1 st and 2nd postoperative days (T3 , T4 ) for determination of erythrocyte AR, PFK and G-6PD activities and plasma glucose, MDA and GSH levels. Results In group Ⅰ plasma MDA level and AR and G-6PD activities in erythrocyte were significantly increased, while plasma GSH level and PFK activity in erythrocyte were significantly decreased on the 1st postoperative day compared to the baseline values before anesthesia (T0) (P 0.05). Conclusion Isoflurane inhalation combined with epidural block can effectively attenuate the effects of surgical trauma on glucose metabolism in theerythrocyte and the oxidative stress responses of the body.

Objective To investigate and compare the effects of pretreatment with different concentrations of trarnadol and morphine on the differentiation of hunan helper T cells (Th) in vitro. Methods Fifteen ASA Ⅰ patients of both sexes (8 males, 7 females) aged 23-59 yrs scheduled for minor day-care surgery were enrolled in this study. The patients had never taken morphine and tramadol before. Blood samples were taken from peripheral vein. Whole blood and mononuclear cells isolated from peripheral blood (PBMCs) were incubated with morphine 10, 100 or 1000 ng?ml-1(group M1 , M2 , M3) or tramadol 50, 500, 5000 ng?ml-1(group Q1, Q2, Q3) for 24 h. Then irrntants was added and incubated for another 5 or 24 h. In the whole blood the level of IL-2 and IFN-?(as markers of Th1 cells) and IL-4 and IL-10 (as markers of Th2 cells) were determined using three-color flow cytometry. In PBMCs the expression of CD4+ and CCR5+ (as markers of Thl cells) and CD4+ and CCR3+ (as markers of Th2 cells) were determined. The Th1/Th2 ratio was calculated. Results Pretreatment with morphine or tramadol significantly increased the number of Th2 cells while the number of Thl cells significantly decreased both in a dose-dependent manner leading to a significant decrease in Th1/Th2 ratio. Conclusion Tramadol and morphine can inhabit in vitro the cellular immune function in dose-dependent manner. The immunosuppressive effect of tramadol is less than that of morphine.