Objective To observe the effect of isolating and harvesting the chondrocytes from rabbits rib cartilage with the method of three-step enzymatic digestion, and the biological characteristic of the isolated chondrocytes during cultivation in vitro to evaluate their biological activity. Methods The method of three-step enzymatic digestion was designed that the rib cartilage was digested one by one with 1 g/L trypsin and 1 g/L EDTA, 1 g/L hyaluronidase and 2 g/L collagenaseⅠ in the culture medium to isolate chondrocytes. The harvesting and viability rate of the primary chondrocytes were detected. During the passage cultivation in vitro, the changes of the chondrocyte shape and growth were observed, and the changes of the collagen typeⅠ and Ⅱ and aggrecan in the extracellular matrix were detected. Results ① The extracellular matrix of rib cartilage was completely dissolved by the three-step enzymatic digestion, and the chondrocytes were completely isolated from the solid matrix. The number of the harvested chondrocytes from every gram of wet cartilage was (4 295.7)?10~(4) on average,and their viability rate was 97.2% on average. ②The primary and first passage chondrocytes had triangle or multi-angle shape, and became elliptic shape at the growing confluence with the positive immunohistochemical staining of collagen type Ⅱ and the strong heterochromia to toluidine blue. The content of sulfate glycosaminoglycans(GAG) in the extracellular matrix of the primary passage cells was (80.61?11.40) ?g/cm~(2). The chondrocytes after the third passage gradually became spindle shape with the negative staining of collagen typeⅡ and the weak heterochromia to toluidine blue. The content of sulfate GAG of the fourth passage cells was (44.74?10.18) ?g/cm~(2). Conclusion ① The method of three-step enzymatic digestion can make the extracellular matix of rib cartilage to be completely degraded, and has advantages of the high efficiency of harvesting primary chondrocytes with high cellular viability rate and simple manipulation. ②The primary and first passage chondrocytes have fine biological activity.

Objective To study the protein expression of Tspan-1 gene in colorectal carcinoma,and investigate the relationship between the expression and biologic behavior of colorectal carcinoma. Methods Immunohistochemical ELIVISION two-step method was used to detect the protein expression of Tspan-1 gene in 80 cases of colorectal carcinoma,13 cases of colorectal adenoma and 27 cases of normal colorectal tissues. Results The positive expression rate of Tspan-1 in colorectal carcinoma was significantly higher than that in colorectal adenoma and normal colorectal tissues,there was significant difference in the expression among the three groups.The positive expression rate of Tspan-1 was 90% in colorectal carcinoma,23% in colorectal adenoma and 7% in normal colorectal tissues (P ＜ 0.01).The positive expression rate of Tspan-1 in poor and moderately differentiated carcinoma was significantly higher than that in well-differentiated carcinoma (P ＜ 0.01).The expression of Tspan-1 was significantly associated with lymph node metastasis (P＜0.01),and with the 5-year survival rate of colorectal carcinoma patients,with also TNM stage and the depth of tumor invasion (P ＜0.01),and while it was not associated with gender,age,location,tumor size,and type (P ＞ 0.05). Conclusion The expression of Tspan-1 gene could be as a marker predicting the prodnosis of colorectal adenocarcinoma patients,also it may play an important role in the pathogenesis of colorectal adenoma.

Objective To explore risk factors relating to occurrence of spontaneous rupture (SR)of hepatocellular carcinoma (HCC). Methods We retrospectively reviewed medical records of 409 HCC patients treated in our department from Jan. 1st 2005 to Aug. 31st 2009. Clinicopathologic factors were explored for their possible association with occurrence of SR by univariate and multivariate analysis using Logistic regression model. Results Among 409 patients, SR occurred in 40 patients (9. 8% , 40/409).Type of SR was defined according to presentation of acute abdominal emergency, especially unstable hemodynamic status. Nineteen cases (4. 7%, 19/409) were grouped as type of HCC-SR Ⅰ and 21 cases (5. 1% ,21/409) as HCC-SR Ⅱ. Patients in the two groups possessed similar hepatitis infection status,severity of background liver disease and tumoral factors except for tumoral location, while HCC-SR Ⅰ group often needed emergency treatment demonstrating elevated values of aspartate aminotransferase (AST) and WBC, decreased PLT counts and worse Child-Pugh classification. Patients in the HCC-SR Ⅱ group had more opportunities of receiving curative hepatectomy and had longer overall survival. The 1,2 and 3 year survival rates were 26. 3 % (5/19), 5. 3% ( 1/19 ) and 0% (0/19)in the HCC-SR Ⅰ group and 66. 7% ( 14/21 ),42.9% (9/21)and 19. 5% (5/21) in the HCC-SR Ⅱ group, respectively. Differences reached statistical significance( P = 0. 011,0. 009,0. 049). Multivariate analysis identified that severity of concomitant liver cirrhosis , WBC ＞ 10 × 109/L and AST level more than twice normal (2N) as independent risk factors associating with occurrence of HCC-SR Ⅰ , while elevated values of AFP and presence of tumor thrombi in portal vein branch related to HCC-SR Ⅱ. Conclusions HCC-SR includes two different types, since different risk factors predict occurrence of each HCC-SR type.

Case-based teaching (CBT) on the basis of treatment guidelines was carried out to develop standard medical behavior, improve learning efficiency and students' ability of solving clini-cal problems. In clinical internship teaching, guidelines for common and frequently-occurring respira-tory diseases and CBT were organically combined. Cases were carefully selected and questions were meticulously set. Guidelines were thoroughly analyzed and teaching was reasonably evaluated. There-fore, students' learning interests were initiated and learning efficiency was improved. It not only trained evidence-based medicine thinking of intern students, but also improved the teaching ability of clinical teachers.

Objective To analyze C4d deposition in the patients with late acute renal allograft rejection,and explore the role of C4d in grafts survival and grafts loss. Methods Thirty-six patients clinical and pathologically diagnosed as having acute rejection more than one year post-transplant were selected. C4d was detected by immunohistochemistry in renal allograft biopsies. The effect of C4d deposition on long-term graft survival was studied. Results Among 36 recipients with late acute renal allograft rejection, 16 cases were positive for C4d (44.4 %) and 20 negative for C4d (55.6 %). Five cases experienced graft loss in C4d positive group (31.3 %), while 6 cases in C4d negative group (30.0%). There was no significant difference in the graft loss rate between C4d-positive group and C4d-negative group. Log-Rank test demonstrated there was no significant difference in graft survival between C4d-positive group and C4d-negative group. The count of the interstitial infiltrated eosinophils in renal allograft was (9.4 + 4.5) and (2.6 + 1.8) respectively in the C4d-positive group and C4dnegative group (P＜0.05). Conclusion C4d deposition in peritubular capillary of the recipients with late acute renal allograft rejection might not be a prognostic marker for graft outcome.

Objective To compare MR,CT,and radiography in the detection of arthropathies in patients with hemophilia.Methods Forty-one symptomic joint images in the 14 patients with hemophilia, aged from 11 to 24 years,were used in this study.Each joint had the examinations of radiography,CT and MR within one day.The severity of each joint was staged using conventional radiographic classification. Severe HA patients with stage 5 were excluded from the study.Imaging findings of soft tissue swelling, osteoporosis,epiphyseal overgrowth,joint erosion,cyst,joint space narrowing,bone marrow,joint effusion, hemorrhage,synovial hypertrophy,widened intercondylar notch as well as anterior and posterior crueiate ligaments(only for knee joint)were used for the all imaging comparison.Results The 41 symptomatic joints in 14 patients with hemophilia were classified by radiographic criteria into stage 0(n=5),stage 1(n=7),stage 2(n=6),stage 3(n=8)and stage 4(n=15).Soft tissue swelling or joint effusion was observed in 33 joints by radiographs,in 34 joints by both CT and MR.Joint erosions were demonstrated in 34 joints by MR,in 33 joints by CT and 20 joints by radiographs.Joint cysts were shown in 21 joints by MR,in 18 joints by CT and 9 joints by radiographs.Significant differences in detection of erosion and cyst were found between radiography with either CT(P0.05).MR showed improvement for detecting nlore loci of both erosion and cyst than CT and radiography,and also CT showed the improvement than radiography.Bone marrow edema 14 joints, hemon'hage in 34 joints and synovial hypertrophy in 27 joints were revealed on MR images.Conclusion MRI is superior to CT and conventional radiography in detecting the abnormal changes and should be considered as the first choice among the imaging modafities in evaluating hemophilic arthropathies.

BACKGROUND: Pancreas-kidney transplantation is an effective treatment for diabetes combined with final stage renal disease. However, as the patients suffer diabetes for a long period of time, and cardiovascular system disease is complex, pre- and post-transplantation treatment is very important for successful pancreas-kidney transplantation.OBJECTIVE: To discuss immunosuppressant, coagulant, perioperative and postoperative treatment during pancreas-kidney transplantation to provide some clinical experience for pancreas-kidney transplantation.METHODS: Clinical data of 5 cases undergoing simultaneous pancreas-kidney transplantation in Department of Urinary Surgery, the 309 Hospital of Chinese PLA General Hospital between 2003 and 2008 were retrospectively analyzed to summarize the application of immunosuppressants and anticoagulant drugs and perioperative clinical monitoring focus. RESULTS AND CONCLUSION: There were 5 male patients with an average age of 43 years, and suffered type I diabetes mellitus complicated with final stage renal disease. The preoperative insulin dosage was 1.5-2.4 U/(kg·d). One case had diabetic retinopathy and fundus oculi hemorrhage for many times; two cases showed apparent coronary atherosclerotic heart disease with preoperative cardiac ejection fraction of 52% and 50%. Exocrine of transplanted pancreas had been considered by the intestinal fluid drainage. A total of 3 cases were complete rehabilitation. Of them, 1 case developed acute rejection in the first seven days after operation, but renal function restored with the hormones impact; 1 case had postoperative acute rejection of transplanted duodenum as well as intestinal fistula, eventually, transplanted pancreas was ectomized, but transplanted kidney was preserved; two cases succeeded in restoring and no complications occurred; 1 had postoperative gastrointestinal bleeding and died from multiple organ failure. Simultaneous pancreas-kidney transplantation is the most effective way to treat the diabetes mellitus with terminal nephropathy. Because of complications in the transplanted exocrine pancreas with bladder drainage, it has been replaced by the enteric drainage. Recovery of the transplanted kidney function is important for successful transplantation. After operation, oral FK should be taken when the serum creatinine returned to 300 umol/L. The application of clotting drug is one of the important factors for recovery of transplanted pancreatic function. Jejunostomy is an important therapeutic measure to prevent the reflux of intestinal juice to the transplanted pancreas in perioperative period. In the follow-up period cathartic drugs are recommended to prevent constipation and reduce the occurrence of acute pancreatitis caused by intestinal fluid reflux.

ObjectiveTo evaluate a second endoscopy for prevention of delayed bleeding after ESD.MethodsData of 67 patients with gastric epithelial neoplasms undergoing ESD from May to November 2011 were reviewed.The median age was 63 ( 31 ～ 84) years.All patients were followed up by endoscopy on the first and the third day after ESD.ResultsOf 67 lesions,5 were located at cardia,6 at gastric body,3 at fundus,35 at antrum,16 at gastric angle,and 2 at residual stomach.The mean maximum diameter of the lesions was 3.73±4 1.24 (2.0 ～ 7.0) cm.There were no intraoperative complications.Post-ESD delayed bleeding was detected by endoscopy in 6 (9.0％ ) patients,with 5 on the third day and 1 on the fourth day.Forrest grading showed 2 cases of Ⅰ b,and 4 of Ⅱ b.All 6 cases were cured by endoscopy.The incidence of postoperative bleeding was far more than that evaluated based on the patients' clinical manifestations only.But therapeutic effect and saffety were the same according to the follow-up results.ConclusionIncidence of post-ESD bleeding is high,but there are no symptoms or severe consequences,so a second endoscopy after gastric ESD may contribute little to the prevention of delayed bleeding.

Objective To explore the clinical value of endoscopic treatment by summarizing the experience of endoscopic treatment of duodenal submucosal tumor (SMT).Methods The data of SMT patients with endoscopic treatment from May 2006 to May 2011 at Endoscopy Center of Zhongshan Hospital Fudan University were studied retrospectively,including the lesions characters,the procedure of performance,complications and recurrence after the treatment.Results A total of 67patients received 69 times of endoscopic treatment,including 36 males and 31 females.The median agewas 55 years,and the average maximum diameter of the lesions was (1.34±0.50) cm.Of these 69lesions,38 lesions located at the bulb,12 at the ball and descending junction,and 19 at the descending part.All lesions were treated by endoscopic treatment successfully.Eleven leisions were treated by polypectomy,12 by endoscopic mucosal resection (EMR),45 by endoscopic submucosal dissection (ESD) and 1 by nylon rope.Complication rate was 14.5％ (10/69),including 1 case of active bleeding,2 cases of perforation,3 cases of delayed bleeding,3 cases of transient increase in amylase level and 1 case of delayed perforation.Total 67 cases were with pathological diagnosis,and brunner′s glands adenoma (36 cases) was the most common.Sixty patients in all were followed up,and the median follow-up time was 13 months.After ESD,one carciniod case was indicated tumor cells in vascular by the pathologic diagnosis and then received extended resection.One case of brunner′s glands adenoma recurred 1 year after EMR and improved after ESD.Conclusion Endoscopic treatment is safe,minimally invasive and effective,which may be used of duodenal SMT.

Objective To explore the therapeutic effect,advantages and shortcomings of pleural effusion controlled by interrupted drainage via central venous catheter.Methods The research group had 52 cases with pleural effusion controHed by interrupted drainage via central venous catheter,and the control group had 50 cases with pleural effusion controlled by conventional pleuracentesis.Pleural effusion disappearing time,average hospitalization time,pleuracentesis frequency,related therapy expenses and complications of two groups were observed and compared.Results The pleuracentesis frequency,pleural effusion disappearing time,average hospitalization time,related therapy expenses and per capita incidence of complications of the research group were(1.06±1.30)times,(4.31±2.20)days,(9.87±2.30)days,(264.77±37.20)yuan and 9.62%and in the control group were(4.20±2.60)times,(9.92±3.70)days,(15.08±5.80)days,(487.62±55.56)yuan and 38.00%.The differences between two groups were particularly remarkable.Conclusion The method of pleural effusion controlled by interruptod drainage via central venous catheter is safe,eonvenient,economical and microtraumatic,and it is worth using more widely.