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 Pulmonary embolism is a common cardiovascular emergency with the characteristics of high incidence,high mortality.This study compared pulmonary artery catheter thrombolysis with peripheral intravenous thrombolysis for acute pulmonary embolism in the mid-term clinical efficacy.Methods From June 2011 to September 2015,68 patients were given pulmonary artery interventional therapy,54 cases received peripheral intravenous thrombolysis;discharged patients were followed up for 3-12 months to evaluate the curative rate,effective rate,recurrence rate and the incidence of complications.Results Follow up rate was 79.5％.The cure rate of the treatment group was higher than that of the control group (47.3 ％ vs 23.8 ％,P ＜ 0.05);The significant effective rate of the treatment group was higher than that of the control group (81.8％ vs 52.4％,P ＜ 0.05);The incidence of Chronic pulmonary hypertension (1.8％) was lower than that of the control group (16.7％);There was no significant difference in improvement rate,recurrence rate and mortality.Conclusion The medium-term efficacy of pulmonary artery catheter directed thrombolysis is superior to that of peripheral intravenous thrombolysis;it can effectively reduce the incidence of chronic thromboembolic pulmonary hypertension.

Objective To evaluate the effect of therapeutic hypercapnia preconditioning on lung ischemia-reperfusion (I/R) injury in rats.Methods Forty healthy adult Sprague-Dawley rats of either sex,aged 2 months,weighing 250-300 g,were divided into 5 groups (n=8 each) using a random number table:sham operation group (group S),grouP I/R and preconditioning with therapeutic hypercapnia of different level groups (group THP1-3).Lung I/R injury was induced by clamping the left hilum of lung for 45 min followed by 120 min of reperfusion.In THP1-3 groups,the respiratory parameters were adjusted at 5 min of stability after isolating the left hilum of lung to make PETCO2 reach 55-65,65-75 and 75-85 mmHg respetively and maintained at this level for 5 min,normal ventilation was then used to make PETCO2 restore the normal level,continuously repeating for 3 circles,and then the left hilum of lung was blocked for 45 min followed by 120 min of reperfusion.The bronchoalveolar lavage fluid (BALF) was collected at the end of reperfusion for determination of the total protein (TP) concentration using Coomassie brilliant blue staining.Lung tissues were obtained at the end of reperfusion for examination of pathological changes after haematoxylin and eosin staining (under a light microscope) and for determination of wet/dry weight ratio (W/D ratio),malondialdehyde (MDA) content,superoxide dismutase (SOD) activity,interleukin-8 (IL-8) and IL-10 contents (by enzyme-linked immunosorbent assay),tumor necrosis factor-alpha (TNF-α) expression (by immunohistochemistry) and expression of TNF-α mRNA (by real-time polymerase chain reaction).Results Compared with group S,the TP concentration in BALF,W/D ratio and contents of MDA,IL-8 and IL-10 in lung tissues were significantly increased,the SOD activity was decreased,the expression of TNF-α mRNA was up-regulated (P＜0.05),strong positive expression of TNF-α was found,and the pathological changes of lung tissues were aggravated in group I/R.Compared with group I/R,the TP concentration in BALF,W/D ratio and contents of MDA and IL-8 in lung tissues were significantly decreased,the SOD activity was increased,the expression of TNF-α mRNA was down-regulated (P＜0.05),no significant change was found in IL-10 content (P＞0.05),the staining range and intensity of TNF-α were decreased,and the pathological changes of lung tissues were significantly attenuated in THP1-3 groups.Conclusion Therapeutic hypercapnia preconditioning can reduce lung I/R injury in rats,and the mechanism is related to inhibiting inflammatory responses and oxidative stress responses.

Objective: To investigate the incidence of adverse events following immunization (AEFI) of human papillomavirus (HPV) vaccines in Hangzhou City from 2017 to 2021, so as to provide insights into safety monitoring and evaluation for HPV vaccines. Methods: The AEFI caused by immunization of bivalent (HPV2), quadrivalent (HPV4) and nonavalent HPV vaccines (HPV9) reported in Hangzhou City from 2017 to 2021 were captured from the AEFI Surveillance Module of Chinese Disease Control and Prevention Information System, and HPV vaccination data were captured from the Zhejiang Municipal Immunization Information Management System. The incidence, temporal distributions and clinical symptoms of AEFI were analyzed. Results: Totally 922 310 doses of HPV vaccines were immunized in Hangzhou City from 2017 to 2021, and 232 cases with AEFI were reported, with an overall incidence rate of 25.15/105 doses. The reported incidence rates of AEFI caused by HPV2, HPV4 and HPV9 vaccination were 31.13/105 doses, 25.93/105 doses and 22.01/105 doses, respectively. General reactions and abnormal reactions were predominant AEFI, and the reported incidence rates of general reactions and abnormal reactions were 21.58/105 doses and 2.60/105 dose, respectively. AEFI predominantly occurred 0 to 1 day post-immunization (165 cases, 71.12%), and the main clinical symptoms included local swelling of injection sites, hard tubercle and fever, with reported incidence rates of 10.30/105 doses, 5.96/105 doses and 6.18/105 doses, respectively. Conclusions Low incidence of AEFI was reported following HPV vaccination in Hangzhou City from 2017 to 2021, and all AEFI were mild. The safety of HPV2, HPV4 and HPV9 remains high.

Diffusion tensor imaging (DTI), which responds with the diffusion of water molecules in spinal cord white matter, may be used to detect the integrity of the spinal cord fiber bundles and the pathological changes after injury. It is sensitive in acute and chronic spinal cord injury, such as cervical spondylotic myelopathy, multiple sclerosis, brain damage secondary spinal cord injury, spinal nerve root damage, and so on. In basic studies, DTI can reveal the microstructure and pathological changes of the injured spinal, and be correlated with behavioral assessment, which involved mice, monkeys, calves, cats, swines, dogs, and so on.

Objective To explore the feasibility and effectiveness of lateral gastrocnemius muscle branch nerve transferring for deep pe-roneal nerve injury. Methods Thirty-two adult female Sprague-Dawley rats were divided into control group (n=8), sham group (n=8), nerve direct repairing group (n=8) and nerve transferring group (n=8). Twelve weeks after the anastomosis, the nerve anastomosis was observed vi-sually, the length of lateral of gastrocnemius muscle branch (L1), the diameter at the point of entering muscle (D1), the maximum detachable length of nervus peroneus communis (L2), the diameter of deep peroneal nerve (D2) and the distance between branch point and neck of fibu-la (S) were measured. The peroneal nerve functional index (PFI), the amplitude of compound muscle action potential (CMAP), nerve con-duction velocity (NCV), the weight of the tibialis anterior and the creatine kinase (CK) activity of theanterior tibial were compared among groups. Results L10.05). Conclusion It is feasible that lateral head muscular branches of gastrocnemius nerve transferring can repair deep peroneal nerve injury, which is needed to separate superficial peroneal nerve and deep peroneal nerve in the epineurium without damaging nerve for tension free neuroanastomosis. Lateral head muscular branches of gastrocnemius nerve transferring can repair the func-tion after deep peroneal nerve injury.