Objective　To observe the effect of repairing the articular cartilage defects with tissue-engineered cartilaginous grafts in rabbits. 　Methods　 A total of 60 rabbits were divided into the cartilage graft group, the pure carrier control group and the blank control group. In the cartilage graft group, the bone marrow mesenchymal stem cells (MSCs) of the rabbits were obtained by isolating and culturing the bone marrow aspirates in vitro. The culture system that facilitates the chondrogenous differentiation of MSCs in rabbits was established. The tissue engineering cartilaginous graft was composed of chondrogenetic MSCs, bovine type Ⅰ collagen and human fibrin. Then transplantations of the cartilaginous grafts were performed to repair articular cartilage defects in rabbits. 　Results　Hyaloid cartilage was formed within the defects repaired with the grafts at 12 weeks after transplantation by analyzing the content of type Ⅱ collagen and metachromatism. In the control groups, the fibrous cartilage repair was observed first, then the fibrous tissues and bone repairs were found. 　Conclusions　 The cartilaginous graft through tissue engineering is a promising graft for repairing articular cartilage defects.

Objective　To study the feasibility of constructing tissue engineered cartilage by differentiated rabbit bone marrow mesenchymal stem cells(MSC) cultured in vitro and in vivo. Methods　The MSC were isolated from the nucleated cells fraction of autologous bone marrow by density gradient centrifuge, and then induced into chondrogenic differentiation by adding dexamethasone, transforming growth factor-β1(TGF-β1) and ascorbic acid in vitro. After 3 weeks, some cells turned to round shape and secreted metachromatic matrix. The cartilaginoid grafts composed of chondrogenic MSC. Bovine type Ⅰ collagen and human fibrin were cultured within the chondrogenic medium for 2 weeks in vitro or transplanted subcutaneously adjacent to the knee joint for 3 weeks in vivo. Results　The most cells in the grafts were degenerated and disappeared after cultured in vitro. But the residual cells were survival and secreted metachromatic staining proteoglycan with toluidine blue, which was characteristic cartilage matrix. The grafts developed into matured cartilage tissue assessed by histological examination after 3 weeks of transplantation in vivo. Conclusion　MSC can be used as functional cells to constructing tissue engineered cartilage.

Objective To explore the safety and feasibility of vagus nerve-preserving Da Vinci robotassisted radical gastrectomy for gastric cancer.Methods The retrospective cross-sectional study was conducted.The clinicopathological data of 12 gastric cancer patients who underwent vagus nerve-preserving Da Vinci robotassisted radical gastrectomy at the Southwest Hospital of the Third Military Medical University from January 2015 to November 2016 were collected.All patients underwent vagus nerve-preserving Da Vinci robot-assisted radical gastrectomy for gastric cancer.During operation,lymph node dissection of the pyloric region,the right side of the cardia and the superior margin of the pancreas were noticed,and other surgical procedures were the same as the traditional Da Vinci robot-assisted radical gastrectomy.Observation indicators:(1) intra-and post-operative situations:surgical methods,digestive tract reconstruction,operation time,volume of intraoperative blood loss,number of lymph node dissected,results of postoperative pathological examination,recovery time of gastrointestinal function,time for liquid diet intake,duration of postoperative hospital stay,short-term surgery-related complications (postoperative bleeding,anastomotic fistula,obstruction and intra-abdominal infection);(2)follow-up situations:postoperative long-term complications (gastric retention,alkaline reflux gastritis,dumping syndrome,gallbladder disease and cholelithiasis),postoperative quality of life (diet,upper abdominal discomfort,nausea,vomiting and diarrhea),postoperative nutritional status [body weight,hemoglobin (Hb),total protein (TP),albumin (Alb)] and tumor recurrence.Follow-up using telephone interview and outpatient examination was performed up to December 2016.Telephone interview included detecting diet of patients,digestive tract symptoms and body weight.Routine blood test,liver and kidney functions,tumor markers,chest X-ray,abdominal computed tomography (CT) or color Doppler ultrasound and gastroscopy of outpatient examinations were performed to detect tumor recurrence and metastasis.Measurement data with normal distribution were represented as x±s and measurement data with skewed distribution were described as M (range).Results (1) Intra-and post-operative situations:all the 12 patients underwent successful vagus nerve-preserving Da Vinci robot-assisted radical gastrectomy for gastric cancer,without conversion to laparoscopic surgery or open surgery,including 2 patients with D1 lymphadenectomy,2 patients with extended D1 lymphadenectomy and 8 patients with D2 lymphadenectomy.Five and 7 patients underwent respectively Billroth Ⅰ anastomosis and Billroth Ⅱ anastomosis of digestive tract reconstruction.Operation time,volume of intraoperative blood loss and number of lymph node dissected of 12 patients were (247± 34) minutes,(94 ± 23) mL and 27 ± 7,respectively.Results of postoperative pathological examination showed that distal and proximal surgical margins of 12 patients were negative and achieved R0 resection;326 lymph nodes were dissected,6 patients didn't have lymph node metastasis and 18 positive lymph nodes were detected in 6 patients.Recovery time of gastrointestinal function,time for liquid diet intake and duration of postoperative hospital stay in 12 patients were (57±14)hours,(64± 14)hours and (7.3±0.9)days,respectively.There was no occurrence of short-term surgery-related complications.(2) Follow-up situations:12 patients were followed up by telephone interview (10 receiving outpatient exaninations) for 9 months (range,1-20 months).Of 12 patients with long-term complications,2 had loss of appetite,1 had diarrhea,without occurrence of cholelithiasis,cholecystitis,gastric retention and dumping syndrome.Of 10 patients receiving outpatient examinations,body weight,Hb,TP and Alb were (56± 12) kg,(126± 10) g/L,(69.9±5.1) g/L,(43.2±3.3)g/L at 1 month postoperatively and (52±13)kg,(126±10)g/L,(72.1±2.4)g/L,(45.2±1.6)g/L at 3 months postoperatively,respectively,with negative carcinoembryonic antigen.There was no tumor recurrence and metastasis in 12 patients.Conclusion Vagus nerve-preserving Da Vinci robot-assisted radical gastrectomy is safe and feasible for gastric cancer,which has not affected the lymph node dissection and incidence of surgeryrelated complications,and it also can improve the postoperative quality of life and maintain good nutritional status.

Objective: To study the effect of enalapril on inducible atrial fibrillation(AF) in old rats. Methods: Old male Wistar rats were randomly divided into control group(n = 12) and experimental group(n = 13). Rats in control group were fed routinely. Rats were fed with enalapril besides normal diet in experimental group for three months. Rats were then anesthetized, thoracotomy was performed and pericardium was opened to expose heart. Right atrium effective refractory period(ERP) was measured. Sinus conduction time (SCT) and sinus recovery time (SRT) were measured for evaluating sinus function. Interatrial conduction time(IACT) and atrium response to burst pacing were evaluated in vivo. Plasma angiotensinⅡ level and atrial tissue angiotensinⅡ level were determined by radioimmunoassay. Sections were cut from the tissue of atrium and stained with Masson trichrome. The ratio of the area occupied by interstitial to the total area was measured. Results: Contrast to control group,IACT and SRT were shorter in experimental group(P ＜ 0.01 and P ＜ 0.05 respectively). AF were induced in 9 rats in control group and 4 rats in experimental group(P ＜ 0.05). AngiotensinⅡconcentration was significantly decreased in right and left atrium tissues of experimental group compared with that in control group(P ＜ 0.01). A significant decrease in interstitial atrial fibrosis was presented in experimental group compared with that of control group(P ＜ 0.01). Conclusion: Inducible atrial fibrillation rate was decreased in old rats after treatment with enalapril. This effect maybe resulted from the inhibited local atrium renin-angiotensin system and improved sinus node function by enalapril.

Objective To investigate the morphological change and SHP -2(src -homology domain contai-ning protein tygosine phosphatase type 2) gene expression in auditory cortical neuron after auditory deprivation in rat .Methods A total of 48 SD rats were randomly divided into 2-week group ,4-week group ,6-week group ,8-week group and 4 corresponding control groups with 6 rats in each group .Bilateral cochlear ablation was done to traumatized groups to ensure their postoperative ABR threshold was above 90 dB SPL .Then paraffin sectioning and HE and Nissl staining were used to detect morphological change of auditory cortical neuron ,simultaneously the RT-PCR was used to measure the expression of SHP -2 gene in auditory cortical neuron of each group ,and relative quantitative analysis was used .Results The HE and Nissl staining revealed that apoptotic shape of auditory cortical neuron became serious by time ,with the diversified cellular morphology .The relative quantity of SHP -2 gene showed statistical differences between any two groups by the method of one -way ANOVA ,showing a rising trend by time (P<0 .05) .Conclusion Along with time ,auditory cortex ,auditory deprivation showed increasing neuronic apoptosis and neuronic proliferation .Apoptosis was the final result from the mutual antagonism by the 2 factors .

Objective To investigate the production of carbapenemase and 16S rRNA methylase in five isolates of pan-drugs resistant E.cloacae recovered in Ruijin hospital.Methods MICs of the five isolates to 10 antibiotics were determined by E test.Six kinds of 16S rRNA methylase genes and a series of β- lactamase genes were amplified by PCR.Shotgun cloning was performed to detect carbapenem resistance determinant.The conjugal transfer of carbapenemase gene and 16S rRNA methylase gene was performed in broth culture with E.coli J53 as the recipient.Pulsed-field gel electrophoresis (PFGE) was carried out to analyse the genotyping.IEF was performed to detect β-1actamases.Southern blot was performed to determine the location of carbapenem resistance determinant.Results The MICs of 10 antibiotics were ＞32 mg/L.Four β-1actamases with pIs of 5.4 ( TEM-1 ),6.7 ( KPC-2 ),8.2 ( SHV-12 ),8.4 (CTX-M-14) were determined.The insertion sequence in the recombinant plasmid was blaKPC-2 flanked by a transposon.blaKPC-2 was located on a large non-conjugative plasmid whereas armA was located on an other conjugative plasmid.PFGE patterns of 5 isolates were identical.Conclusion KPC-2 was responsible for carbapenem resistance in pandrugs resistant Enterobacter cloacae.There was no relationship between blaKPC-2 and armA.Although pandrug resistant Enterobacteriaceae remain rare,the emergence of this group of organism merits monitoring.

Objective · To design and synthesize five new tropane compounds, and test their antagonistic activity against M3 receptor and inhibition activity to neutrophil elastase (NE), of which the structure-activity relationship were preliminarily investigated. Methods · The five compounds, A1-A3,B1 and C1, were prepared with 3α-hydroxy-tropane (A0) as the starting material by modifying the structure in C-3α position and N atom on the tropane skeleton. The antagonistic activity of the compounds to muscarinic M3 receptors on tracheal rings of guinea pigs was evaluated by functional assays in vitro. The hydrolysis of PGlu-Pro-Val-PNA as substrate was catalyzed by NE to get colorful nitroaniline (PNA). The NE inhibition activity of the tropane compounds was obtained by determining the absorbance [(D(405 nm)] of PNA. Results · The five new tropane compounds generated strong antagonistic activity against M3 receptors. Among them, A2 had the greatest activity [antagonistic parameter pA2(M3)=9.004], and elicited obvious inhibitory effect to NE (inhibition ratio YA2=20.29%). Conclusion · Introducing strong electron-attraction group, such as sulfuryl and hydrophobic group with large volume into C-3α position on the tropane skeleton can improve the M3 receptor antagonistic activity as well as the NE inhibition activity.

OBJECTIVE To investigate antimicrobial resistance and molecular epidemiology profiles of meticillin-resistant Staphylococcus aureus(MRSA)sampled from lower respiratory tract.METHODS Totally 107 MRSA strains were isolated from lower respiratory tract specimens at Shanghai Pulmonary Hospital between Dec 2005 and Dec 2006.PVL genes were detected by PCR.The genotypes of SCCmec were identified by multiplex PCR.The antimicrobial resistance of MRSA were tested by Kirby-Bauer agar dilution.We also performed the homology of 32 MRSA strains using pulsed-field gel electrophoresis(PFGE).RESULTS All of the 107 MRSA strains were negative in the PVL locus detection and the most frequent SCCmec types were type Ⅲ(81.3%),the others including type Ⅱ(15.9%),type Ⅳ(2.8%),type Ⅰ and type Ⅴ were not found in this group.Those 3 different types of SCCmec were all resistant to ?-lactam antibiotics,less resistant to rifampin,and susceptible to vancomycin,teicoplanin and daptomycin.The resistant rate of those 3 types were different to the non-?-lactam antimicrobial drugs such as trimethoprim/sulfamethoxazole,clindamycin,erythromycin,gentamicin,levofloxacin,and tetracycline,the resistant rate in the types Ⅱ and Ⅲ was significantly higher than the type Ⅳ.PFGE analyses assorted the 32 MRSA strains into 4 PFGE patterns:pulsotype A(25 strains),including subtypes A1(17strains),A2(1 strain)and A3(7 strains);pulsotype B(5 strains),pulsotype C(1 strain),and pulsotype D(1 strain).CONCLUSIONS This study does not found positive PVL locus in the MRSA strains in our hospital,the most frequent SCCmec types are type Ⅲ and some are type Ⅱ.PFGE presented that there are outbreaks of MRSA in ICU ward and TB ward No 5 at that time and the pandemic strains are subtypes A1 and A3,most of these MRSA strains are multiple resistant,which deserves attention from both the clinical staff and infection-control department of the hospital.

Objective:To examine the roles of follicular helper T(Tfh)cells,serum IL-21 and B cells in the pathogenesis of alcohol abuse non-traumatic osteonecrosis of the femoral head ( NONFH ) .Flow cytometry was used to measure the frequencies of peripheral blood inducible Tfh cells and B cells in alcohol abuse NONFH patients and healthy controls.The disease progression and the extent of femoral head collapse,the serum IL-21 were quantified.Methods: A significantly higher percentages of CD19+B cells(t=3.765,P=0.0005),CD86+CD19+B cells(t=5.506,P<0.0001),and CD95+CD19+B cells(t=4.152,P=0.0002) in patients than those in controls was found.The percentages of CD86+CD19+B cells were positively associated with the index of femoral head collapse in alcohol abuse NONFH(P<0.0001,r=0.536).Results: The frequencies of Tfh cells (t=7.611,P<0.0001),and IL-21+Tfh cells (t=5.281,P<0.0001) were higher than those in controls;The frequencies of Tfh cells were positively associated with the percentages of CD19+B cells(P=0.0002,r=0.455),IL-21+Tfh cells were positively associated with the percentages of CD86+CD19+B cells(P=0.0002,r=0.447).Conclusion: Tfh cells and B cells may participate in the pathogenesis of alcohol abuse NONFH,and increased CD86+CD19+B cells may be associated with the extent of femoral head collapse,the interaction of Tfh cells and B cells may have an im-portant role in pathogenesis of alcohol abuse NONFH.

Objective: To investigate the effects of trentment with hydroxythy starch(130/0.4) on the serum albumin and immunologic function in postoperative patients with obstructive jaundice.Methods: 24 patients with obstructive jaundice were randomly divided into the control group(n=12) and hydroxythy starch(130/0.4) (HS) group(n=12). The serum ALB was detected 1d, 3d, 5d, 7d after the operation,and the immunologic index were detected 1d and 7d after the operation.Results: The ALB content of control group and HS group were not significantly different in the postoperative 1d and 7 d(P>0.05) and had significant differences in the postoperative 3 d, 5 d (P<0.05).All immunologic index had no significant differences in the postoperative 1d, 7d (P>0.05).Conclusion: The ALB content of patients with obstructive jaundice may decrease postoperatively. Treatment with hydroxylthy starch(130/0.4) can alleviate the drop of the ALB content. But it has no effects on immunologic function.