Objective To assess effects of cerebral ischemic postconditioning(IPost)on neuronal apoptosis and phosphated glycogen synthase kinase-3β(GSK-3β)after cerebral ischemic/reperfusion.Methods The experiment was conducted at the center for animal experiment of Renmin Hospital,Wuhan University.Thirty male Wistar rats were randomly divided into three groups:sham operation(S),ischemic/reperfusion(I/R)and ischemic postconditioning(IPost).Each group contained ten rats.Global brain ischemia was produced with four-VO method.Animals were killed after two days.Apoptosis in neurons in the cortex region were detected by TUNEL assay; infarct areas were detected by TTC ; activity of p-GSK-3β was detected by spectrum assay; Statistical software SPSS13.0 was applicated to perform one-way ANOVA Student-Newman-Keul test; correlation was detected by linear regression.Results Compared with group S,TUNEL-positive cells and infarct areas increased(P ＜0.01),the activity of p-GSK-3β decreased in I/R and IPost groups(P ＜ 0.01).Compared with group I/R,TUNEL-positive cells and infarct areas significantly decreased(P ＜ 0.01),the activity of p-GSK-3β increased in group IPost(P ＜ 0.01).The activity of p-GSK-3β and TUNEL-positive cells had highly correlation,so as infarct areas(P ＜ 0.01).Conclusions IPost can lessen the ischemic/reperfusion injury of Cortex,through increas the activity of p-GSK-3[β and decreasing neuronal apoptosis.

Objective To observe the effects of exogenous sodium phosphocreatine (PCr) on cerebral repeffusion injury of rats after ischemia in order to explore the potential mechanism. Method Thirty-six healthy adult male Wistar rats with body weight 200- 220 g were randomly (random number) divided into sham operation group, ischemic reperfusion (I/R) group and PCr treatment group. The I/R model was established by using electro-cauterizing bilateral vertebral arteries and occluding bilateral common carotid arteries with atraumatic carotid clasps for 10 min, and then the clasps were released for 48 hours reperfusion. In sham operation group, bilateral common carotid arteries were exposed without occlusion. In PCr treatnent group, PCr in dose of 150 mg/kg was administered intravenously 60 min before the occlusion of bilateral common carotid arteries. Normal saline was administered intravenously instead of PCr into rats of I/R group. After reporfusion for 48 hours, the rats were sacrificed and brains removed for detections of neuron apoptosis by using TUNEL, malondialdebyde (MDA) level by using chromtometry and calmodulin (CaM) activity by using ELISA. Results Compared with sham operation group, TUNEL-positive cells, MDA level and CaM activity increased in I/R group and PGr treatment group ( P ＜0.01). Compared with I/R group, TUNEL-positive cells, MDA level and CaM activity were lower significantly in PCr treatment group ( P ＜ 0.01). Conclusions PCr can lessen cerebral ischemic reperfusion injury and neuron apoptosis, the mechanism maybe relates to the attenuation of abnormalities in calcium balance and reduction of oxygen free radicals by PCr.

Objective To investigate the effects of different duration of sevoflurane anesthesia in the neonatal period on the long?term cognitive function and hippocampal synaptic plasticity in rats. Methods Twenty?four pathogen?free healthy Sprague?Dawley rats of both sexes, aged 7 days, weighing 12-16 g, were divided into 3 groups ( n=8 each) using a random number table: control group ( group C) , sevoflu?rane anesthesia for 2 h group ( group S1 ) , and sevoflurane anesthesia for 6 h group ( group S2 ) . Group S1 and group S2 inhaled 2% sevoflurane for 2 and 6 h, respectively. Morris water maze test was performed at 30 days after the end of anesthesia ( postnatal day 37) to assess the cognitive function. After the end of the test, the rats were sacrificed, and hippocampi were isolated for determination of the expression of brain?de?rived neurotrophic factor ( BDNF) , postsynaptic density?95 ( PSD?95) and synapsin 1 in hippocampal tis?sues by Western blot. Results Compared with group C, the escape latency on 4th and 5th days of the test in group S1 and on 2nd-5th days of the test in group S2 was significantly prolonged, and the frequency of crossing the original platform was significantly decreased, and the time of staying at the platform quadrant was significantly shortened in S1 and S2 groups, the expression of BDNF, PSD?95 and synapsin 1 in hipp?ocampal tissues was significantly down?regulated in group S2 (P<0?05), and no significant change was found in the expression of BDNF, PSD?95 and synapsin 1 in hippocampal tissues in group S1 ( P>0?05) . Compared with group S1 , no significant change was found in the escape latency and frequency of crossing
the original platform (P>0?05), the time of staying at the platform quadrant was significantly shortened, and the expression of BDNF, PSD?95 and synapsin 1 in hippocampal tissues was significantly down?regula?ted in group S2 ( P<0?05) . Conclusion Short?time and long?time sevoflurane anesthesia both can induce long?term cognitive dysfunction in the neonatal period, and the severity is aggravated with prolonged anes?thesia; the partial mechanism is related to inhibition of the synaptic plasticity of hippocampal neurons of rats.

Objective To evaluate the changes in the expression of DJ-1 protein during myocardial ischemia-reperfusion (I/R) in diabetic rats.Methods Fifty male Sprague-Dawley rats,weighing 220-280 g,were used in this study.Type 1 diabetes mellitus was induced by intraperitoneal streptozotocin 65 mg/kg and confirmed by fasting blood glucose ＞ 16.7 mmol/L.Forty animals with type 1 diabetes mellitus were randomly divided into 3 groups:diabetes group (group D,n =10),diabetic sham operation group (group DS,n =15) and diabetic I/R group (group DIR,n =15).Another 10 non-diabetic rats in which citrate buffer 6 ml/kg was injected intraperitoneally were served as control group (group C).Myocardial I/R was produced by occlusion of the anterior descending branch of left coronary artery for 30 min followed by 120 min reperfusion in group I/R.At 120 min of reperfusion,5 rats were sacrificed and myocardial specimens were c(on)tained for determination of infarct size in groups DS and DIR,and 10 rats were sacrificed and myocardial specimens were obtained for microscopic examination and for determination of cell apoptosis,malondialdehyde (MDA) content,superoxide dismutase (SOD) activity and expression of DJ-1 and phosphatase and tensin homologue (PTEN) protein.Apoptotic index (AI) was calculated.Linear correlation between the expression of DJ-1 protein and MDA content,SOD activity,AI and expression of PTEN protein was analyzed.Results Compared with group DS,the myocardial infract size was significantly increased in group DIR (P ＜ 0.05).Compared with group C,MDA content and AI were significantly increased,SOD activity was decreased,the expression of DJ-1 was down-regulated,and the expression of PTEN protein was up-regulated in groups D,DS and DIR (P ＜ 0.05).Compared with groups D and DS,MDA content and AI were significantly increased,SOD activity was decreased,the expression of DJ-1 was down-regulated,and the expression of PTEN protein was up-regulated in group DIR (P ＜ 0.05).There was no significant difference in the parameters mentioned above between groups D and DS (P ＞ 0.05).There was linear correlation between the expression of DJ-1 protein and MDA content,SOD activity,AI and expression of PTEN protein and the correlation coefficients (r) were-0.734,0.593,-0.818,and-0.812 in turn.Conclusion Down-regulation of DJ-1 protein expression is involved in myocardial I/R injury in diabetic rats via decreasing anti-oxidative stress responses and upregulating PTEN protein expression.

Objective To investigate the mechanism of myocardial ischemia and reperfusion-induced acute lung injury (ALl) and protective effect of ischemic postconditioning.Methods Forty SD rats were allocated to sham group,myocardial ischemia/reperfusion group (reperfusion group),ischemic postconditioning group (postconditioning group),and ischemic postconditioning + phosphatase and tensin homolog deleted on chromosome ten (PTEN) inhibiting group (inhibitor group) according to the random number table,with 10 rats per group.Myocardial ischemia/reperfusion was induced by left anterior descending coronary artery occlusion.Postconditioning was performed within 1 minute before reperfusion consisting of 3 10 s cycles of reperfusion followed by 10 s occlusion.Lung was immediately removed 120 minutes after reperfusion for HE stain,immunohistochemical detection of inflammatory factors and apoptosis factors,TUNEL assay of cell apoptosis,and Western blot of protein kinase B (Akt),phospho-Akt (p-Akt),glycogen synthase kinase-3β (GSK-3β),and phospho-GSK-3β (p-GSK-3β).Results Down-regulated B-cell lymphoma-2 (Bcl-2) and IL-10 and up-regulated Bcl-2 associated X protein (Bax),cysteinyl aspartate specific proteinase-3 (Caspase-3),IL-6 as well as IL-8 were observed in other 3 groups compared with sham group (P ＜0.01).Moreover,down-regulated Bax,Caspase-3,IL-6,IL-8 as well as TUNEL and up-regulated Bcl-2 as well as IL-10 were observed in reperfusion group compared to postconditioning group and tensor group (P ＜ 0.01).No statistical differences were found among the four groups in levels of Akt,p-Akt,and GSK-3β,but level of p-GSK-3β was significantly down-regulated in reperfusion group compared to other 3 groups(P ＜ 0.01).Conclusion Development of ALI may relate to down-regulation of p-GSK-3β evoked directly by the release of inflammation factors in early period of myocardial ischemia/reperfusion and ischemic postconditioning may attenuate the condition.

Objective To investigate the analgesic efficacy and spinal neurotoxicity of intrathecal (IT) different doses of dexmedetomidine in rats. Methods Sixty male SD rats weighing 180-220 g were randomly divided into 5 groups ( n = 12 each): groupnormal control (group C); group IT normal saline (group N); different doses of dexmedetomidine groups received IT dexmedetomidine 0.75, 1.50 and 3.00 μg/kg respectively (groups D1.3). Paw withdrawal threshold to mechanical stimulation (PWMT)with yon Frey filaments and tail flick latency (TFL) to a thermal nociceptive stimulus were measured before (To, baseline) and at 30 or60 rin after IT dexmedetomidine or normal saline administration (T1, T2 ) and the percentage of the maximum possible effect ( MPE ) was calculated. Lumbar segment of the spinal cord ( L4-6 ) was removed for microscopic examination and determination of c-Fos expression (by immuno-histochemistry) at 7, 24 and 48 h after IT dexmedetomidine or normal saline administration. Results PWMT, TFL and the percentage of MPE were significantly increased after IT dexmedetomidine as compared with the baseline values at T0 in groups D1-3 ( P ＜ 0.05). PWMT was significantly higher at T1 and TFL and the percentage of MPE were higher at T2 in groups D1-3 than in groups C and N,and in group D3 than in groups D1,2 ( P ＜ 0.05). At 7,24 h after IT dexmedetomidine c-Fos protein expression was significantly higher in group D3 than in groups C and N( P ＜ 0.05). There was no significant difference in c-Fos expression at 48 h after IT dexmedetomidine between group D3 and groups C and N ( P ＞ 0.05 ). At 24 h after IT dexmedetomidine c-Fos protein expression was significantly higher in group D3 than in other 4 groups( P ＜ 0.05). Slight spinal cord injury was observed at 24 h after IT dexmedetomidine in group D3. Conclusion IT dexmedetomidine has antinociceptive effect. High dose dexmedetomidine IT can produce transient reversible toxicity to the spinal cord.

Objective To observe inhibiting effect of endostatin on subcutaneous transplant tumor of breast carcinoma, and to illuminate the therapeutic effect of endostatin in the cancer. Methods The effect of en-dostatin on MCF-7 cell proliferation was studied by MTr. The model of MCF-7 cell transplant tumor on nude mice was constructed. Endostatin was injected intradermally around the transplant tumor. Inhibition effect on the tumor was observed. Results Endostatin with the concentration of 10 μ/mL and 15 μg/mL can inhibitMCF-7 cell proliferation effectively (P < 0. 05 ). After endostatin injection, tumor weight, volume and mi-crovessel density decreased significantly(P < 0.05 ). Conclusion Endostatin can inhibit breast carcinoma proliferation through inhibiting angiogenesis and the tumor cell itself.

Objective:To evaluate the effect of post-conditioning in brain injury induced by myocardial I/R on inflammatory factor and GFAP.Methods:Male Sprague-Dawley rats were randomly allocated into 3 groups (n=8):group Sham,group IR,group IPost.Myocardial IR was induced by occlusion of the anterior descending branch of the left coronary artery for 30 min.group IPost received 3 cycles of 10 s reperfusion followed by 10 s ischemia at the end of myocardial ischemia.The rats were sacrificed at 120 rain of reperfusion and the brains were removed for microscopic examination,inflammatory factors and GFAP.Results:Compared with group Sham,IL-6,IL-8 were significantly increased,IL-10 was down-regulated in group IR(P＜0.01).Post-conditioning can decrease IL-6,IL-8 and up-regulated IL-10(P＜0.01).When compared with group Sham,the expression of GFAP was higher in group IR(P＜0.05),however,the GFAP in group IPost is the most among these three groups(P＜0.01).Conclusion:Post-conditioning could protect brain by decreasing inflammatory factors,increasing GFAP,which both from brain injury induced by myocardial ischemia reperfusion.

Objectives To observe the clinical significance and application value of autologous blood transfusion in neurosurgery of primary hospital. Methods Four hundred and fourteen patients who underwent the neurosurgery operation and were subjected to intraoperative blood transfusion were selected, among whom 97 patients were subjected to autologous blood transfusion (observation group), and 317 patients were subjected to heterogenous blood transfusion (control group). The condition of intraoperative blood transfusion, changes of hemoglobin and hematocrit, blood transfusion related cost were compared between 2 groups. Results There were no statistical differences in operation time, infusion volume, rate of transfusion related complications and postoperation hemoglobin, hematocrit between observation group and control group (P>0.05). The patients in control group were infused with 189 000 ml, and the transfusion liquid volume proportion of total blood transfusion was 79.22%(189 000/238 580);13 patients in observation group were used the heterogenous blood transfusion with 5 400 ml, and the transfusion liquid volume proportion of total blood transfusion was 10.30%(5 400/52 430). Eighty-six patients (88.66%, 86/97) in observation group performed autologous blood collection and transfusion, the volume of autologous collection was 80 650 ml, and the volume of transfusion was 47 020 ml. Eleven patients in observation group did not perform autologous blood transfusion, among whom 6 patients was because of operational and mechanical reasons, and 5 patients performed collection but did not transfuse. The cost of heterogenous concentrated suspension red blood cell over 6 U was significantly higher than the cost of disposable material and injection of autologous blood:(2 287.06 ± 243.52) yuan vs. (1 595.08 ± 133.95) yuan, and there was statistical difference (P<0.05). The rate of heterogenous concentrated suspension red blood cell 6 U in control group was 14.83%(47/317), and the rate of over 6 U was 6.62%(21/317). Conclusions The autologous blood transfusion is safe and effective, and it is worth popularizing in neurosurgery of primary hospital. But in the process of its application, it is necessary to strengthen the user′s operating skills and ensure the quality of autologous blood transfusion.

Objective To test and verify the performance of analyzing IgG using nephelometry assay,and discuss reasonable model of performance verification of this system.Methods According to related documents and standards,this study verified the precision,accuracy,assay measurement range(AMR)and reference interval.Results The within-run precision in low level was 2.24%,while it was 2.73% in high level.The overall precision in low level was 2.25%,while it was 2.68%.The relative bias between the results of analyzing the calibrator with a different lot from that used for calibrating and its concen-tration printed was 5.18%.The AMR of the original dilution was 2.44~33.5 g/L.The results of reference interval verifica-tion identified with what the manufactur declares.Conclusion The major performances of analyzing IgG by this system are identifies with the manufactur declares.The reference interval offered by the manufactur is acceptable.The verification and calculation methods are simple and convenient,with strong operability.