Objective To construct a lentiviral vector for RNA interference(RNAi)of MACC1 gene and to detect the best trans-fection condition by transfected into MB-231 cells .Methods The siRNA was designed and converted into cDNA of shRNA (small hair pin RNA) of siRNA for MACC1 gene .The cDNA was synthesized and inserted into pMAGic lentiviral plasmid vector which was linearized by enzyme Age Ⅰ and EcoRⅠ .The recombinant plasmid was transformed into competent E .coli DH5α cells .The positive recombinant colony was selected by ampicillin medium agar and identified by DNA sequencing .The recombinant lentivirus was packaged into mature lentivirus by 293FT cells and used to infect MB-231 cells .To detected the transfection condition of high efficiency of infection and low multiplicity of infection .Results PCR and sequencing verified that the recombinant lentivirus plasmid MACC1-shRNA was successfully constructed .The best transfection condition was MOI=40 by transfected into MB-231 .Conclu-sion The lentiviral RNAi expression vector targeting MACC 1 gene is successfully constructed and it can infect MB-231 cells effi-ciently ,which lays the experimental foundation for the research on the changes of malignant biological activity of tumor cell lines and gene therapy .

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 compare the clinical and pathological features of Chinese young breast cancer(age ≤ 35)with elder patients(> 35)using Meta analysis .Methods Published studies concerning clinical and pathological features of young breast cancer in China were searched systemically and assessed .Stata12 .0 software was used for data analyzing and calculating OR and its 95%CI .Results Totally 31 studies were selected for Meta analysis ,and most of them were classified as 6 - 7 scores ,which showed the quality of articles was high .The risk factors of breast cancer and its pooled odds ratio values with statistical significance were as fol‐lows 6 .42(95% CI :4 .22 - 9 .79) ,0 .61(95% CI :0 .50 - 0 .74)when clinical staging of 0 - Ⅱ phase or Ⅰ - Ⅱ phase ,2 .25(95% CI :1 .69 - 2 .99)when histological type of Invasive carcinoma ,1 .73(95% CI :1 .23 - 2 .43)when histological grade of III grade ,1 .80 (95% CI :1 .23 - 2 .43)when positive of lymph node metastasis .Conclusion Compared with elder breast cancer ,the clinical and pathological characteristics of young breast cancer were mainly for the high misdiagnosis rate ,the late clinical stage ,the high pro‐portion invasive carcinoma ,the poor histological differentiation and the lymph node metastasising easily ,the hint of young breast cancer screening and treatment may be different principles and measures should be adopted .

Objective To understand the human resource status of the community rehabilitation management in Shanghai and provide evidence for the construction of this human resource. Methods Census institutions of community rehabilitation management in Shanghai were investigated with self-questionnaire. Results Shanghai has initially established personnel of community rehabilitation management,which contains rehabilitation coordinators and street community administrator. However, the structure and number of the personnel has yet to be improved. Conclusion Personnel building must focused on optimizing the personnel structure of rehabilitation coordinator, moderately expanding the street community administrators, filling the community rehabilitation service gap between city and urban in next stage.

This study primarily explored and discussed‘generality and individuality’mode in clinical teaching of congenital heart disease(Congenital heart disease,CHD)from the aspects of teaching methods,comparison with traditional method,requirements for teachers and problems existed under present conditions.At last the paper summarized advantages and disadvantages of this model in clinical teaching of CHD and offered a new approach for clinical teaching of cardiology.

Objective　To evaluate the technique and clinical outcomes of modified transperitoneal laparoscopic radical prostatectomy.　Methods　A total of 285 patients received the operation with mean age of 67 years (50-76 years) from January 2008 to April 2012.Mean level of PSA was 15.7 μg/L (1.8 -50.0 μg/L),and mean prostatic volume was 44 ml (26 -74 ml). No lymph node or seminal vesicle involvement was found by CT or MR and radionuclide bone scan revealed no metastasis.271 cases were confirmed diagnosis by prostatic biopsy and 14 were detected through pathological studies of TURP specimens.Gleason score ranged from 6 to 8.14 cases were in clinical stage T1b,29 cases in T1c,214 cases in T2 and 28 cases in T3a.Transperitoneal approach and modified technique involving bladder neck dissection,nervesparing technique and vesicoureteral anastomosis were applied on patients.　Results　Mean operative time was 105 min (55 -150 min).Mean intraoperative estimated blood loss was 240 ml (50-800 ml).Rectal injures occurred in 2 cases and were repaired under laparoscopy.Drainage tube and urinary catheter were removed 48 -72 h and 5 -8 d postoperatively.Postoperative hospital stay was 7 d (5 - 11 d).Positive surgical margin was present in 58 patients.Mean follow-up time was 29 months (3 -50 months).Complete continence were found in 208 patients immediately after catheter removal.68 patient recovered continence within 3 months and 9 patients remained incontinence 3 months after surgery. Normal erection presented in 42 of the 57 cases with nerve-sparing.　Conclusions　Transperitoneal laparoscopic radical prostatectomy is safe and efficient.Higher efficiency and lower complication rate have been achieved through modified laparoscopic technique involving bladder neck dissection,nerve-sparing technique and vesicoureteral anastomosis.

Objective To assese the healing of stoma after magnetic anastomosis for the reconstruction of biliary-enteric continuity under severe inflammation.
Methods Acute bile duct injury was constructed as a bile peritonitis model in mongrel dogs (n=32). Magnetic anastomosis (group A, n=16) and traditional suture anastomosis (group B, n=16) were performed to reconstruct the biliary-enteric continuity in one stage. Half of the dogs in each group were euthanized on the 30th postoperative day, and the other half on the 90th postoperative day to harvest the stoma region. The healing conditions of the stoma after the 2 anastomotic approaches were observed with naked eyes, under light microscope and scanning electron microscope.
Results The stoma leakage rate (50%versus 0%on the 30th postoperative day, 37.5%versus 12.5%on the 90th postoperative day, both P<0.05) and stenosis degree (13.9%±0.3%versus 7.1%±0.3%on the 30th postoperative day, 17.2%±0.4%versus 9.4%±0.4%on the 90th postoperative day, both P<0.01) were significantly higher in group B than in group A. Compared with traditional manual anastomoses, the histological analysis under light and electron microscope showed a more continuous stoma with more regular epithelium proliferation and collagen arrangement, less inflammation in group A.
Conclusions Magnetic anastomosis stent ensures better healing of the stoma even under the circumstance of severe inflammation.

Objective:To evaluate the application of biological mesh in laparoscopic anti-reflux procedure for gastroesophageal reflux disease ( GERD ).Methods: The clinical data of 20 consecutive GERD patients underwent anti-reflux surgery in Sir Run Run Shaw Hospital from December 2012 to April 2014 were retrospectively analyzed . The laparoscopic hiatal repair with 360 fundoplicaiton was performed and the biological mesh ( Biodesign TM , Surgsis ? ) was implanted for reinforcement of hiatal repair . Results: All laparoscopic procedures were successful , no conversion and no intra-operative complications occurred .The pre-operative complains were relieved in all patients, and no recurrence was observed during 3-18 month of follow-up.Six patients got dysphagia after operation; 5 of them were controlled through medication and psychological induction;1 received esophageal dilatation by bougie .Conclusion: The application of biological mesh in laparoscopic anti-reflux procedure for gastroesophageal reflux disease is satisfactory .

OBJECTIVETo evaluate the relation between different therapy and survival rate of liver metastasis of colorectal cancer (LMCC).

METHODSClinical data of 669 LMCC patients,collected from Fudan University Zhongshan Hospital from January 2000 to July 2008, were analyzed retrospectively.

RESULTSOf the 669 cases, 379 cases were synchronous liver metastases(SLM) and 290 cases were metachronous liver metastases(MLM). There were no significant differences in age, gender and position of primary tumor between SLM and MLM groups(P>0.05), but as to liver metastasis characteristics(liver lobe involved, focus number and maximal focus diameter) and CEA, CA19-9 before therapy,there were significant differences(P<0.05). Two hundred and fifty-three cases underwent curative hepatic resection, including 123 cases in SLM and 130 cases in MLM. Until October 31, 2008, all the cases were followed up. The median survival time of SLM was(11+/-1) months and of MLM(23+/-2) months(P<0.01). Five-year survival rate of SLM was 6.4% and of MLM 11.4%(P<0.01). As to different treatments, median survival time and 5-year survival rate of curative hepatic resection group were 37 months and 35.6%, and of non-operation groups(i.e. intervention, chemotherapy, radiofrequency therapy and percutaneous ethanol injection) were 5 to 26 months and 0 to 3.6% respectively(P<0.05).

CONCLUSIONSCurative hepatic resection is the first choice of liver metastasis of colorectal cancer, which can improve the survival rate. Resection rate and survival of MLM are better than those of SLM.

Adult ; Aged ; Colorectal Neoplasms ; pathology ; therapy ; Female ; Follow-Up Studies ; Hepatectomy ; Humans ; Liver Neoplasms ; secondary ; therapy ; Male ; Middle Aged ; Neoplasm Metastasis ; Prognosis ; Retrospective Studies ; Survival Rate

Abnormal proliferation and migration of vascular smooth muscle cells (VSMCs) are the major cause of in-stent restenosis (ISR). Intervention proliferation and migration of VSMCs is an important strategy for antirestenotic therapy. Roscovitine, a second-generation cyclin-dependent kinase inhibitor, can inhibit cell cycle of multiple cell types. We studied the effects of roscovitine on cell cycle distribution, proliferation and migration of VSMCs in vitro by flow cytometry, BrdU incorporation and wound healing assay, respectively. Our results showed that roscovitine increased the proportion of G0/G1 phase cells after 12 h (69.57±3.65 vs. 92.50±1.68, P=0.000), 24 h (80.87±2.24 vs. 90.25±0.79, P=0.000) and 48 h (88.08±3.86 vs. 88.87±2.43, P=0.427) as compared with control group. Roscovitine inhibited proliferation and migration of VSMCs in a concentration-dependent way. With the increase of concentration, roscovitine showed increased capacity for growth and migration inhibition. Roscovitine (30 μmol/L) led to an almost complete VSMCs growth and migration arrest. Combined with its low toxicity and selective inhibition to ISR-VSMCs, roscovitine may be a potential drug in the treatment of vascular stenosis diseases and particularly useful in the prevention and treatment of ISR.
Animals ; Cell Cycle ; drug effects ; Cell Line ; Cell Movement ; drug effects ; Graft Occlusion, Vascular ; drug therapy ; metabolism ; pathology ; Muscle, Smooth, Vascular ; metabolism ; pathology ; Myocytes, Smooth Muscle ; metabolism ; pathology ; Protein Kinase Inhibitors ; pharmacology ; Purines ; pharmacology ; Rats