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.

Objective To assess the quality of randomized controlled trials(RCT)abstracts published in Journal of Clinical Anesthesiology by CONSORT for Abstracts.Methods Articles invol-ving human RCTs published from 2012 to 2013 were reviewed and searched through WanFang Med Online.Trials involving animal experiments,in vitro,RCTs without abstract,and meta-analyses were excluded.According to the CONSORT checklists for abstracts,the quality of abstracts for RCT were assessed.Results In total,392 RCT abstracts were analysed.The median word counts of ab-stracts was 364 (IQR 306-444),sample size was 60 (IQR 40-80).Almost all abstracts provided an appropriate description of conclusions (100%), numbers randomized (99.0%) and objective (99.0%).The majority of abstracts described interventions (94.6%)and participants (82.4%).Re-quirements present in less than 50% of the abstracts were details regarding trial design (46.2%)and harms (48.7%).The descriptions of randomization (13.3%),blinding (1.8%),methods-outcome (3.6%)and results-outcome (9.7%)were very low.Moreover,title,recruitment and numbers ana-lysed were not reported.Conclusion The quality of RCT abstracts and adherence to the CONSORT checklist for abstracts remains poor,and the CONSORT for Abstracts should be endorsed to improve the quality of RCT abstracts as early as possible.

Objective Surgical stress induces a series of endocrine and metabolic changes including glucose metabolism and insulin-resistance. The purpose of the present study was to investigate the changes in glucose transporter-4 (Glut-4) mRNA expression in skeletal muscle after cholecystectomy under epidural block. Methods One gram of the rectus abdominis muscle was taken while abdomen was being opened and closed in patients undergoing cholecystectomy under epidural block. Total RNA of the muscle cells was extracted by trizol one-step template. RT-PCR was used to determine the Glut-4 mRNA amplification products with ?-actin mRNA as an internal control. The Glut-4 mRNA expression was expressed by (desired gene/ ?-actin?100% . The plasma glucose and insulin levels were determined at the same time. Results Glut-4 mRNA expression was significantly reduced (P 0.05 ) . Conclusions The results indicated that the synthesis of Glut-4 is suppressed by surgical stress of cholecystectomy under epidural block leading to insulin resistance.

Objective To investigate the effect of intraperitoneal local anesthetic on patients undergone laparoscopic cholecysteetomy (LC).Methods Thirty-six patients (ASAⅠ-Ⅱ),who were undergoing selective LC were randomly divided into there groups.GroupⅠreceived preoperational anesthetic spary with 30 ml of 0.5% ropivacaine.GroupⅡwas given the anesthetic at a same dosage after the operation.While groupⅢwas set as a control by using 0.9% NaCl instead of ropivacaine.The LC was completed under general anesthesia.After the operation,visual analog scale (VAS) was recorded at 6,24,and 48 hours to evaluate the degree of postoperative pain.Meanwhile,the number of the patients who received anesthetics after the surgery,as well as the incidence rates shoulder or back pain and nausea or vomiting were recorded.Results Postoperative VAS of the groupⅠwas significantly lower than that of the other two groups,while the VAS of groupⅡwas significantly lower than that in groupⅢ(both P0.05). Conclusions Intraperitoneal local anesthetic can significantly reduce postoperative pain after LC.It is more effective to give local anesthetic at the end of the procedure than using it before operation.

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.

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.

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: To investigate the effects of preconditioning with lithium on the expressions of HSP70 in gerbil after global ischemia. Methods: The transient ischemia model of gerbil global was established by clamping the bilateral common carotid arteries for 5 min.36 gerbils were randomly divided into group lithium(n=18) and group control(n=18).Gerbils in group lithium were injected with lithium intraperitoneally,5 mmol/kg,once a day for 5 consecutive days.Normal saline was used instead of lithium in group control as vehicle control.Both group lithium and control underwent ischemia after preconditioning,and were then further divided into three subgroups(n=6,in each subgroup).The garbles in lithium 1 or control 1 subgroup were sacrificed 1 day after ischemi;lithium 3 or control 3 sacrificed in 3 days;and lithium 7 or control 7 were sacrificed in 7 days.The brains were made into paraffin sections and were used for immunohistochemistry against HSP70. Results: Compared with the vehicle control,the HSP70 expression in subgroup lithium1 was inhibited in the hippocampus CA1(P