Objective To evaluate the different methods of continent urinary diversion and the ileal orthotopic neobladder after total cystectomy. Methods 4 different kinds of continent urinary diversion were undertaken after total cystectomy for 68 cases.Continence and catheterization, volume and pressure of the reservoirs, image and hydroelectrolyte condition were investigated. Results Of the 3 cases with intussusceptive efferent tract,2 had partial deintussusception resulting in incontinence and had to be reoperated;44 cases with tapered terminal ileal efferent tract were continent and could be catheterized easily with 16～20F catheters, whereas only one case had difficulty in catheterization. 39 cases with detubularized and reshaped intestinal segment reservoirs, including 3 ileal,22 colonic and 14 ileocolonic, all achieved the demand of low intrareservoir pressure, whereas 8 cases operated in the early period had dilated reservoirs with a volume as large as 1 470～1 650 ml;8 cases with detenia cecocolonic reservoir had the volume of 430～600 ml and as intrareservoir pressure of 30～45 cmH 2O with peristalsis waves,2 of them had urine leakage in the early stage after operation.21 cases with ileal orthotopic neobladder had a volume of 350～480 ml and an intrareservoir pressure of 12～20 cm H 2O.1 being incontinent at daytime and 2 at night whereas all the others were continent. Conclusions Continent urinary reservoirs constructed by 30 cm detubularlized cecocolon can achieve the demand of low intrareservoir pressure.Detenial colonic reservoirs had more chance of urine leakage and adhesion, and higher intrareservoir pressure. Tapered terminal ileal efferent tract is better than ileal intussusception efferent tract for its excellent continence, large caliber, easy catheterization and fewer complications. Ileal orthotopic neobladder has the advantages of excellent continence, higher quality of life, but has limited indications.

Objective To determine the efficacy of retrovirus-mediated HSV-TK gene transfer and GCV in a BALB/c mice model with urinary bladder transitional cell carcinoma. Methods A replication defective retroviral vector containing HSV-TK gene was used. In vivo experimental animals were divided into 3 groups.In group A,7 tumors were induced in BALB/c mice by subcutaneous injection of MBT-2 transitional cell carcinoma (TCC);the virus was directly injected into the tumor and the animals received intraperitoneal GCV.In group B, 6 tumors were induced in BALB/c mice by subcutaneous injection of transferred HSV-TK gene TCC and the animals received intraperitoneal GCV.In group C,6 tumors were induced in BALB/c mice by subcutaneous injection of MBT-2 TCC and the animals received intraperitoneal normal saline.Tumor-volume measurement was performed in these 3 groups 10 d and 20 d after treatment, and the volume changes were compared among the 3 groups. Results In vivo experiment indicated that the mean tumor volume of group A [MBT-2 group, (55.37?4.52) mm3] and group B [MBT-2/HSV-TK group,(49.77?4.15)mm3] was reduced significantly compared with that of group C [control group,(146.27?10.46)mm3](P0.05).At 20 d after treatment, the mean tumor volume of group A [(186.75?8.14)mm3] and group B [(72.50?6.70)mm3] was also reduced compared with that of group C [(441.76?41.80)mm3] (P

ObjectiveTo assess the safety and efficacy of retrograde ureteroscopy lithotomy (URSL)assisted antegrade percutaneous nephrolithotomy (PCNL) for complex upper ureteral calculi in semisupine-lithotomy position.MethodsFrom March 2007 to December 2010,a total of 95 patients with complex upper ureteral calculi underwent retrograde URSL assisted antegrade PCNL in semisupine-lithotomy position.Ureteral calculi size was 12 mm × 6 mm to 38 mm × 15 mm,24 cases combined with renal calculus.Firstly retrograde URSL was performed,once the stone fragments moved up to renal pelvis,a 16-22 F PCNL working channel was established under the ultrasound guidance through which lithotripsy was performed using an ureteroscope.Finally a 6-7 F double-J tube was indwelled.ResultsOperations were successfullycompleted in 93 patients.However,in it 2 patients were converted to open surgery because of significantureteral distortion due to previous open surgery.Operative time was(42.7 ± 14.9) min; estimated blood loss was(34.5 ± 26.1 ) ml.The ureteral calculi clearance rate was 100.0％,and renal calculus clearance rate inthose combined with renal calculus was 95.8％ (23/24).There were no major intraoperative and postoperative complications excepted early urinary leakage in 2 cases and fever ≥39℃ in 3 cases.ConclusionsRetrograde URSL assisted antegrade PCNL in semisupine-lithotomy position is safe and feasible for complex upperureteral calculi,especially non-opaque calculi,combined with renal calculus,easily ascending ureteral calculi and large calculi burden which has low calculi clearance rate after URSL.The outcomes are encouraging with fewer complications.It also avoids intraoperative change of patient's position.

Objective To detect the differentially expressed microRNAs (miRNAs) in bladder cancer tissue and normal bladder tissue. Methods Total RNA was extracted from bladder cancer tissue and normal bladder tissue by Trizol, and then miRNAs were isolated and enriched from the total RNA. Mammalian miRNA microarrays were used to analyze the differentially expressed miRNAs between the bladder cancer tissue and normal tissue. Data analysis was performed by software of LuxS-can3. 0 and SAM version 2. 1. Choose let-7 gene which was interesting to us, and validation of mi-croarray results was carried out by northern blotting. Results Compared with normal bladder tissues, there were 71 differentially expressed miRNAs in bladder cancer tissue, of which there were 38 down-regulated ones and 33 up-regulated ones. Among these miRNAs, 26 miRNAs were the most significant with 12 up-regulated and 14 down-regulated. The expression of let-7 gene in bladder cancer was down-regulated to normal bladder tissue by northern blotting, which was in agreement with the results of the miRNA microarrays. Conclusion There are differentially expressed miRNAs between bladder cancer tissue and normal bladder tissue, and let-7 gene is probably as a tumor suppressor in bladder cancer.

AIM: To identify the non-steroid transcription factors upregulating the expression of L-plastin in hormone-independent prostate cancer, and partly elucidate the mechanism of hormone-refractory prostate cancer. METHODS: TF SEARCH software was used to analysis the possible binding sites of transcription factors in the 3’ end of L-plastin promoter that had been identified as important part of regulation response elements. Gel shift assay and supershift assay were used to confirm the transcription factors binding the speculated response elements. PCR site-mutagenesis technique was performed to delete the binding site of transcription factor and luciferase activity assay was carried out after deletion of the binding site. RESULTS: SP-1 respond element GGTGGGGCGGGGA located at -54- -41 of L-plastin promoter was identified with the TF SEARCH software. Gel shift assay and supershift assay confirmed that SP-1 was the transcription factor binding to GGTGGGGCGGGGA. Mutant deleted the SP-1 binding-site had low-luciferase activity than that of the naive. CONCLUSION: SP-1 plays an important role in the up-regulation of L-plastin expression in hormone-independent prostate cancer.

AIM: To construct eukaryotic expression vector of small interfering RNA(siRNA) specific to bcl-2 and investigate the effect of recombinant plasmid on suppressing bladder cancer cell growth.METHODS: siRNA of bcl-2 gene was designed according to the principle of RNAi-based medicine, and was converted into cDNA coding expression of small hairpin RNAs(shRNA) of siRNA. The cDNA was synthesized and inserted into plasmid pGenesil-1. The recombinant eukaryotic expression vectors of pGenesil-1545 and pGenesil-1555 were controlled by the U6 promoter of RNA polymerase Ⅲ, identified by the restriction map and the sequence analysis, and transfected into T24 cells. After T24 cells were transfected for 72 h, expression of bcl-2 mRNA was assayed by RT-PCR; and MTT was used to observe the proliferation of T24 cells.RESULTS: The recombinant plasmids of pGenesil-1545 and pGenesil-1555 were identified by the restriction map and the sequence analysis. The sequences completely coincided with the designs. The expression of the bcl-2 mRNA in T24 cells transfected with recombinant plasmid decreased nearly 80%, and the growth of T24 cells was suppressed significantly.CONCLUSION: The siRNA eukaryotic expression vector against bcl-2 gene is successfully constructed. It effectively downregulates the expression of bcl-2 in T24 cells and suppresses the cell growth.

BACKGROUND:Smooth muscle cells and transitional epithelial cells were traditionally used to construct tissue-engineered bladder and to perform double-sided implantation of scaffold.However,double-sided implantation is difficult to perform,because smooth muscle cells are difficult to isolate or culture in vitro and passage is limited.OBJECTIVE:To verify the feasibility of tissue-engineered bladder reconstruction with bone marrow mesenchymal stem cells(BMSCs)and bladder acellular matrix(BAM).DESIGN:A basic empirical study.SETTING:Linbaixin Medical Research Center,Second Affiliated Hospital,Sun Yat-sen University.MATERIALS:Experiments were performed at the Linbaixin Medical Research Center,Second Affiliated Hospital,Sun Yat-sen University from March 2006 to Mav 2007.The laboratory was the Opening Laboratory of Hospital Affiliated to Health Department of China.One-month old SD rats of either sex,weighting 80-100 g were provided by Animal Experimental Center of Sun Yat-sen University.Fresh porcine bladders were offered by Animal Experimental Center of Southern Medical University.METHODS:Whole bone marrow culture and successive adherence method was used to culture rat BMSCs in vitro.Flow cytometry was employed to detect surface antigen.Eradicator washing method was applied to prepare porcine BAM and measure its purity and characteristies.Third passage of BMSCs were inoculated in BAM and cultured in a medium containing vascular endothelial growth factor(VEGF165)(25 ng/L)in vive and in vitro to test compatibility.Cells cultured alone were considered to be controls for the in vivo trial,and materials non-implanted with cells were considered to be controls for in vitro trial.Suitable microenvironment was simulated to induce the differentiation of BMSCs.Four weeks and eight weeks later,compound materials were respectively removed to perform tissue section test.Simultaneously,immunohistochemistry keratin staining was conducted to examine regeneration of epithelial cells.MAIN OUTCOME MEASURE:Biocompatibility of BMSCs and BAM.RESULTS:①BMSCs were cultured by whole bone marrow method.Flow cytometry demonstrated that third passage of cells were positive for CD29(99.43%).②BAM had good biological characteristics.Homogen matrix and byssoid collagen appeared under a microscope.Compatibility trials showed good compatibility of BMSCs and BAM and well-growth cells.③Four weeks later,histological section test confirmed inflammatory cell infiltration,closely-arranged collagen and elastic fiber.Immunohistochemistry keratin staining showed lamellar and discontinuous simple epithelium.Eight weeks later,no inflammatory cell infiltration was found,and closely-arranged collagen and elastic fiber were detected.Immunohistochemistry keratin staining showed lamellar and continuous multiple epitheliums.CONCLUSIoN:With good compatibility,BMSCs and BAM appear to be an ideal material for bladder tissue engineering.

Objective To verify the efficiency of a new laparoscopic urethrovesical anastomosis training model by comparing it with the chicken skin model. Methods Chicken posterior trunks and porcine colons were used to construct the training model. The posterior trunk of a chicken was used to simulate a human pelvis, and a 3-mm cloacal stump was used to simulate a human urethral stump. A 15-cm segment of porcine colon with a 1-cm orifice was used to simulate a human bladder or neobladder. An imitation urethrovesical anastomosis was performed with laparoscopic instruments in a laparoscopic training box. Forty trainees with no laparoscopic experience were randomized into 2 groups.The trainees in group A (n=20) practiced using this new model for 8 h, while those in group B (n=20) practiced using the chicken skin model for 8 h. The trainees' skills were assessed using the porcine model before and after training. Results Compared with the chicken skin model, this new training model more accurately resembled the structure and characteristic of human pelvis, urethral stump, and bladder (neobladder). After the training sessions, both groups improved in anastomosis time [GroupA: (64±11)min vs. (123±20)min, P＜0.05; Group B: (77±12)min vs. (121±17)min, P＜0.05] and quality (Group A: 8.8± 1.0 vs. 3.8 ± 1.2, P＜0. 05 ; Group B: 7.7 ± 0.9 vs.3. 7± 1.1, P＜0. 05). Compared with trainees in group B, trainees in group A required less time and achieved a higher quality score (P＜0.05). Conclusions This new training model can help urologic surgeons to reduce learning curve of this technique and improve their suturing skills. It is an effective,convenient training model for laparoscopic urethrovesical anastomosis.

Objective To approach the clinical presentation,treatment and diagnosis of primary seminal vesicle carcinoma.Methods The records of 4 patients who diagnosed seminal vesicle carcinoma were retrospectively reviewed,including the symptoms signs and examination results as well as operation program,postoperative therapy.Considered to the literature reports.Bilateral seminal vesicles,bladder, prostate and rectum were totally removed in one case.Seminal vosiculectomy and partial cystoprostotectomy were performed in 2 cases,and the another one,bilateral lower ureterectomy and ileum substitute bladder was be done.Results Followed up for 3 months to 5 years,and no recurrence had been observed so far, one died of colon cancer after 2 years.Conclusions Early symptoms of primary seminal vesicle carcinoma are unobvious,so that early diagnosis of seminal vesicle carcinoma is difficult and the misdiagnosis is so usually.Thus,accurate recognition is important for early diagnosis.Radical surgery appears to offer the best chance and different approaches can be selected according to tumor stage and invasive condition of the circumambient organ.Comprehensive treatment like hormonal therapy,along with the 5-fluorouracil,paclitaxel,and oxaliplatin chemotherapy regimen appears to be effective against adenocarcinoma.

Objective To study the feasibility and safety of performing nephrectomy together with the removal of complicated inferior vena cava tumor thrombus under profound hypothermia and arrested circulation. Methods After made the median thoraco-abdominal incision, the exploration of the abdominal organs was done. The right kidney, inferior vena cava and renal pedicle were well exposed then. After the whole body heparinization, cannulas were put into ascending aorta, superior vena cava, aortic root and right superior pulmonary vein. The body temperature was reduced to 20℃ with cardiopulmonary bypass unit and the extracorporeal circulation was stopped then. Cut open the inferior vena cava at vena renalis dextra ingress and the F16 urinary catheter was inserted into atrum dextra through inferior vena cava and inflated. The tumor thrombus was pulled out and the right kidney was removed. The inferior vena cava incision was sutured to close and the extracorporeal circulation was resumed and patient was re-warmed.Results The operation time was 330 min and the extracorporeal circulation time was 90 min, while the profound hypothermia with circulatory arrest time was 20 min. The estimated blood loss during operation was 400 ml and 6 unit red cells and 600 ml blood plasm were transfused. The patient was awaked 2.5 h after the operation, food intake resumed 4 days after operation and the patient was discharged on day 10 post-operatively. After 6 months'follow-up, there were no local recurrence and metastasis occurred. Conclusion The technique of profound hypothermia and circulation arrest could improve the safety and efficacy in the treatment of renal cell carcinoma with suprahepatic (level Ⅲ) caval tumor thrombus.