Objective To investigate the mechanisms of metformin in inducing autophagy of gastric cancer MNK-45 cells.Methods Human gastric cancer MNK-45 cells in logarithmic growth phase were incubated in the culture plates,and were divided into the intervention group [gastric cancer MNK-45 cells were intervened by metformin at different concentrations (2,4,8,16,32,64 mmol/L) for 24,48,72 hours] and the control group (gastric cancer MNK-45 cells were cultured in the DMEM medium).The inhibition rate of gastric cancer MNK-45 cells was detected by MTT method.The IC50 value of metformin on gastric cancer MNK-45 cells was 17 mmol/L.Gastric cancer MNK-45 cells were intervened by metforrnin at 17 mmol/L for 48 hours in the experimental group.Gastric cancer MNK-45 cells in the control group were cultured in DMEM medium at 17 mmol/L for 48 hours.The apoptosis of the gastric cancer MNK-45 cells of the 2 groups were detected by flow cytometry.The mRNA expressions of Bax and Bcl-2 of the 2 groups were detected by RT-PCR.The protein expressions of type Ⅰ LC3b,type Ⅱ LC3b,beclinl,AKT,p-AKT,mTOR,p-mTOR,P70s6k,p-P70s6k of the 2 groups were detected by Western blot.The measurement data were presented as (x) s,and were analyzed using the one-way ANOVA or repeated measures ANOVA.Data of the 2 groups were compared using the t test.Results The inhibition rates of gastric cancer MNK-45 cells were 3.0％ ± 1.1％,8.6％ ± 1.7％,15.9％ ± 1.6％,26.1％ ± 3.4％,37.5％ ± 2.3％,49.7％± 3.6％ after intervention by metformin at concentrations of 2,4,8,16,32,64 mmol/L for 24 hours,5.2％± 1.9％,10.4％±2.1％,26.9％± 1.6％,49.5％± 1.6％,59.1％±2.0％,82.1％±2.2％ after intervention by metformin at concentrations of 2,4,8,16,32,64 mmol/L for 48 hours,and 9.5％ ± 2.2％,17.6％ ± 1.4％,30.6％ ± 2.6％,63.2％ ± 2.6％,78.9％ ± 1.4％,93.3％ ± 2.7％ after intervention by metformin at concentrations of 2,4,8,16,32,64 mmol/L for 72 hours.There were significant differences in the inhibition rates among the 6 groups at the same time points (F =155.174,728.229,743.826,P ＜ 0.05),and significant differences were also observed within the same group at different time points (F =39.420,58.692,166.125,30.383,117.517,311.642,P ＜ 0.05).The apoptosis rates of gastric cancer MNK-45 cells in the experimental group and the control group were 25.4％ ± 1.7％ and 6.9％ ± 0.5％,with significant difference between the 2 groups (t =18.378,P ＜0.05).The relative mRNA expressions of Bax/Bcl-2 mRNA in the experimental group and the control group were 1.88 ± 0.16 and 1.00 ± 0.00,with significant difference between the 2 groups (t =9.743,P ＜ 0.05).The relative protein expressions of LC3 Ⅱ/LC3 Ⅰ and beclin 1 in the gastric cancer MNK-45 cells were 1.65 ± 0.08 and 1.47 ± 0.06 in the experimental group and 0.79 ± 0.03 and 0.56 ± 0.06 in the control group,with significant difference between the 2 groups (t =18.023,18.283,P ＜ 0.05).The relative protein expressions of AKT and P70s6k in the gastric cancer MNK-45 cells were 0.80 ±0.14 and 0.97 t0.21 in the experimental group and 0.96 ±0.17 and 1.37 ±0.23 in the control group,with no significant difference between the 2 groups (t =2.103,1.699,P ＞0.05).The relative protein expressions of mTOR and p-mTOR were 0.58 ± 0.l 1 and 0.57 ±0.15 in the experimental group and 1.88 ±0.23 and 2.36 ±0.25 in the control group,with significant difference between the 2 groups (t =11.293,10.979,P ＜ 0.05).No p-AKT and p-P70s6k expression was detected in the experimental group,and the expressions of p-AKT and p-P70s6k in the control group were 1.00 ± 0.00 and 1.00 ± 0.00,respectively.Conclusions Metformin could induce autophagy,inhibit proliferation and promote apoptosis of gastric cancer MNK-45 cells.The mechanism may be associated with the inhibition of mTOR expression and the expression of mTOR downstream proteins p-P70s6k by mefformin,and then the autophagy of gastric cancer MNK-45 cells happens.

Objective To investigate the clinical application of robot-assisted laparosocopic ureterolithotomy.Methods The clinical data of 28 cases(15 males and 13 females),aged 42(18～72),of laparosocopic ureterolithotomy with AESOP system(computer Motion,USA) was analyzed.Of the 28 cases,18 patients had upper ureter stones and 10 had middle ureter stones.Results All procedures were completely performed by robot assisted laparosocopic ureterolithotomy.The operative time was ranged from 35 to 150 min(averaged 50 min) and the intraoperative blood loss was ranged from 20 to 50 ml(averaged 30ml).The postoperative hospital time was 3 to 5 days and no ureter stricture or recurrent calculus was found during the follow-up period(2～13 months).Conclusion The ZEUS AESOP system has the characteristics of high intelligence and humanization.The robot-assisted laparoscopic ureterolithotomy is effective,safe and precise.

Objective To evaluate the effect of advanced age on sepsis-caused heterogeneity of nicotinic acetylcholine receptors (nAChR) in the cytomembrane of skeleton muscle in rats.Methods Twenty SPF adult rats (aged 4-5 months,weighing 250-280 g) and 20 aged male Sprague-Dawley rats (aged 18-20 months,weighing 550-600 g),were obtained from the Experimental Animal Centre of Chongqing Medical University.The adult rats were randomly divided into control group (CAd group,n =10) and sepsis group (SAd group,n =10).The aged rats were randomly divided into control group (CAg group,n =10) and sepsis group (SAg group,n =10).Sepsis was produced by cecal ligation and puncture.The specimens of anterior tibial muscle were obtained at 24 h after operation for determination of the expression of neuronal nAChR (α7-nAChR) and fetal nAChR (γ-nAChR) using immunohistochemistry and Western blot.Results The expression of γ-nAChR and α7-nAChR in the cytomembrane of anterior tibial muscle was significantly higher in CAg and SAd groups than in CAd group,and in SAg group than in CAg and SAd groups.Conclusion Advanced age can aggravate sepsis-induced heterogeneity of nAChR in the cytomembrane of skeleton muscle in rats.

Objective To investigate the related prognostic factors of stage Ⅳ gastric cancer.Methods The clinical data of 248 patients with stage Ⅳ gastric cancer and intact follow up data in the Tumor Prevention and Treatment Center of Sun Yat-Sen University from 2000 to 2010 were retrospectively summarized.The twelve clinicopathological parameters served as the observation indicators,including age,sex,body mass reduction,H b,CEA,CA19-9,Borrmann type,tumor location,tumor size,pathological pattern,operative mode,metastatic sites and therapeutic model.The survival curve was drawn by using the Kaplan-Meier method.The median survival time was calculated.The univariate analysis was conducted with Log-rank test.The prognosis multivariate analysis was conducted by the Cox's proportional hazards regression analysis.Results MST in the patients of whole group was 254 d.The univariate analysis showed that sex,Borrmann type and therapeutic mode were the related factors afecting gastric cancer prognosis,while the Cox regression model revealed that above 3 indicators were also independent factors affecting the prognosis of the patients with stage Ⅳ gastric cancer in this group(P＜0.05).Conclusion The treatment mode is an important independent factor affecting the survival of stage Ⅳ gastric cancer,the translational medicine model of palliative chemotherapy combined with palliative operation conduces to improve the prognosis in the patients with stage Ⅳ gastric cancer.

Objective To explore the therapeutic efficacy of nanometer high-frequency square pulse light technology targeting kidney cancer.Methods Fifty BALB/c nude mice were vaccinated with human ACHN cell line and randomly divided into 1 control group and 4 therapeutic groups.The 4 therapeutic groups were cured with high-frequency square pulse light and nanometer high-frequency square pulse light.The treatment cycle was 4 weeks.The tumor growth condition and tumor-repres-sion change were observed and compared.Results The tumor volumes of the control group in-creased obviously,whereas the tumor volumes of the therapeutic groups decreased obviously or in-creased gently.The mean tumor volume and the tumor growth curve of the therapeutic groups were significantly lower than those of the control group(P＜0.05),but there was no significant difference between the therapeutic efficacy of the kidney cancer using high-frequency square pulse light and nanometer high-frequency square pulse light(P＞0.05).Synteresis of kidney carcinogenesis experiments results indicated that using high-frequency square pulse light and nanometer high-frequency square Dulse light could prevent the production and development of the kidney cancer(P＜0.05),but the svnteresis efficacy of the 2 methods had no obvious difference(P＞0.05).Conclusion Using highfrequency square pulse light and nanometer high-frequency square pulse light can cure the kidney cancer and,to some extent,prevent the production and development of kidney cancer.

Objective To explore the effect of low-dose ketamine combined with propofol anesthesia on expres?sion of glutamine receptor subunit 1 (GluR1) and 2 (GluR2) in the hippocampus of depressed rats after electroconvulsive therapy. Methods Healthy adult male Sprague-Dawley rats, weighing 200~250 g, were used in this study. Mental depres?sion was induced by chronic unpredictable mild stress. Thirty-two depressed rats were randomly divided into 4 groups (n=8): metal depression group (group A), ECT group (group B), ECT+propofol group (group C) and ECT+propofol+ket?amine group (group D). Eight normal rats served as control group. Control group received no treatment. Group A received intraperitoneal injection of normal saline 8 mL/kg plus sham ECT. Group B, C and D received ECT once a day for 7 con?secutive days following intraperitoneal injection of normal saline 8 mL/kg, propofol 80 mg/kg and propofol 80 mg/kg +ketamine 10mg/kg, respectively. Sucrose preference test and Morris water maze were performed to assess depressed be?havior and learning and memory function, respectively. RT-PCR and Western-blot assay were used to detect the expres?sion of GluR1 , GluR2 and their mRNA expression. Results After ECT, compared with control group and group A, changes of SPP in group B, C and D were obvious. The change of SPP in group D was much higher than all other groups (P<0.05). Rats in group B showed prolonged escape latency and shortened space exploration time, which were significantly different from all other groups (P<0.05). Rats in group D showed the most shortened escape latency and prolonged space exploration time (P<0.05). The expression of GluR1 was significantly increased in group B, C and D compared with group A (P<0.05). The expression of GluR2 and mRNA was significantly decreased in group B and C (P<0.05). The difference in GluR2 and mRNA expression was not significant among group A, D and control group (P>0.05). Conclusion Low-dose ketamine combined with propofol anesthesia exert effective antidepressive action and improve learning and memory function of depressed rats after electroconvulsive therapy. The beneficial effects of the ketamine combined with propofol anesthesia may be related to up-regulation expression of GluR1 and GluR2 in hippocampus.

Objective To investigate the role of autophagy and synaptophysin (SYP) in cognitive impairment in de?pressed rats receiving electroconvulsive shock (ECS). Methods Clean and healthy adult male Sprague-Dawley rats were acclimatized to a standard laboratory environment for 7 days. The chronic unpredictable mild stress (CUMS) was used to establish the rat model of depression. Behavior tests were conducted before and after CUMS to evaluate the depression and cognition level of rats. After establishment of the model, 24 rats were randomly divided into ESC group (group E) and depression group (group D) with 12 rats in each group. The rats in group E were administered 80 mg/kg of propofol (10 mg/mL) by intraperitoneal injection, followed by ECS treatment. The rats in group D were administered propofol by intra?peritoneal injection, followed by sham-ECS treatments. The above interventions were conducted daily for 7 consecutive days. After the interventions, rats underwent behavior tests as before. Subsequently, rats were killed and specimens were collected for measurements. Immunohistochemistry was performed to examine autophagy markers such as Beclin 1 and LC3Ⅱand ELISA was used to detect SYP in the hippocampus. Results Group E after ECS significantly increased the percentage of sucrose preference (68.2%±8.7%), rearing times (7.0±1.9), total horizontal distance [(569.5±70.0) cm], es? cape latency [(21.9±5.3)s] and space exploration time [(20.5±3.9)s] compared with group D or group E before ECS. There was no significant difference in these index between groups before ECS or in group E between before and after ECS(P>0.05). Compared with group D, group E had upregulated protein expression levels of Beclin 1 and LC3Ⅱin CA1, CA3, DG as well as the area near the hippocampus and increased SYP contents (P<0.05). Conclusions Cognitive impairment in depression rats following ECS correlates with activated autophagy and increased SYP by ECS.

Objective To evaluate the effect of small-dose ketamine on the onset time and course of modified electroconvulsive therapy (MECT) in mentally depressed rats.Methods Sixty SPF adult male SpragueDawley rats,aged 2-3 months,weighing 220-250 g,were randomly divided into 6 groups (n =10 each) using a random number table:normal control group (group C),depression group (group D),ECT group,propofol + ECT group (group PE),ketamine + ECT group (group KE) and ketamine + propofol + ECT group (group KPE).The depression model was established by chronic unpredictable mild stress (CUMS).Mter CUMS,C,D and ECT groups received intraperitoneal normal saline 8 ml/kg,group PE received intraperitoneal propofol 100 ml/kg,group KE received intraperitoneal ketamine 10 ml/kg,and group KPE received intraperitoneal ketamine 10 ml/kg + propofol 80 ml/kg.All the groups received ECT once a day for 7 consecutive days starting from the time point when righting reflex was lost except C and D groups.Open-field test was performed before CUMS,at 1 day after CUMS and at the end of each ECT (T0 8).The total distance and the number of standing on the back legs were recorded.Morris water maze test was performed at 2 days after CUMS and 1 day after the end of therapy,and the escape latency and time of staying at the original platform quadrant were recorded.Results Compared with group C,the total distance was shortened and the number of standing on the back legs was reduced,the escape latency was prolonged,and the time of staying at the original platform quadrant was shortened at T1-8 in D,ECT,PE and KE groups and at T1 5 in KPE group,and no significant was found in KPE group in the total distance,number of standing on the back legs,escape latency,and time of staying at the original platform quadrant at T6-8.Compared with group D,the total distance was prolonged and the number of standing on the back legs was increased at T6-8 in ECT and PE groups and at T4-8 in KE and KPE groups,the escape latency was prolonged,and the time of staying at the original platform quadrant was shortened in ECT group,and the escape latency was shortened,and the time of staying at the original platform quadrant was prolonged in KPE group.Compared with ECT and PE groups,the total distance was prolonged and the number of standing on the back legs was increased at T4-7 in group KE and at T4-8 in group KPE,and the escape latency was shortened,and the time of staying at the original platform quadrant was prolonged in KPE group.Compared with group KE,the total distance was prolonged and the number of standing on the back legs was increased at T6.7,the escape latency was shortened,and the time of staying at the original platform quadrant was prolonged in KPE group.Conclusion Small-dose ketamine can shorten the onset time and course of MECT in mentally depressed rats.

OBJECTIVE:To explore the preventive effect and safety of levetiracetam combined with sodium valproate of diaze-pam on recurrent febrile seizures(FS). METHODS:A total of 90 children with recurrent FS were randomly divided into observa-tion group and control group with 45 cases in each group. Control group was treated with sodium valproate or diazepam orally. On the basis of control group,observation group additionally received levetiracetam orally,with initial dose of 15 mg/kg,bid,for 7 d,and then decreasing gradually;decreasing to 10 mg/kg on 8th-12th day,bid;decreasing to 5 mg/kg on 13th-15th day,bid;drug withdrawal on 16th day. The children of 2 groups were followed up for 1 years,and received routine test every 2 months. The times of fever,the rate of recurrent convulsion,the conversion of epilepsia and the incidence of ADR were recorded in 2 groups during follow-up period. The serum levels of NSE and S-100β protein were determined in 2 groups before treatment and 6 months after treatment. The intelligence and behavior ability of 2 groups were scored by Chinese Modified Wechsler Intelligence Scale for Children and Children’s Adaptive Behavior Rating Scale. RESULTS:3 children of observation group and 2 of control group were failure in follow-up. During the follow-up period,fever times and the rate of recurrent convulsion in observation group were signifi-cantly lower than in control group,with statistical significance(P<0.05). There was no statistical significance in the rate of epilep-sia conversion and the incidence of ADR between 2 groups (P>0.05). After treatment,serum levels of NSE and S-100β in 2 groups were decreased significantly,and the observation group was significantly lower than the control group,with statistical signif-icance(P<0.05). To the end of follow-up,verbal IQ,performance IQ and total IQ score of observation group were significantly higher than those of control group,and the cognition factor,social factor and behavior ability scores of observation group were sig-nificantly higher than those of control group,with statistical significance (P<0.05). CONCLUSIONS:Levetiracetam combined with sodium valproate or diazepam can prevent the occurrence of recurrent FS,relieve cerebral injury and improve the intelligence and behavior ability of the children,so as to improve the life quality of Children.

10.3969/j.issn.1007-3969.2013.05.006