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.

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.

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.

Objective To study the clinical features and possible pathogenesis of acute ischemic stroke in renal cell cancer patients. Methods The clinical data from in-hospital patients with renal cell cancer who developed acute ischemic stroke were collected, including the patients with renal cell cancer who developed acute ischemic stroke during anti-cancer therapies and those patients with acute ischemic stroke who were firstly diagnosed to have renal cell cancer during anti-stroke therapies between January 2003 and December 2015. Results A total of 2516 patients with renal cell cancer were screened, and there were 36 patients (1.43%) with acute ischemic stroke. Out of the 36 patients, there were 29 men (80.56%) and 7 women (19.44%). Their age ranged from 45 to 68 years, with a average age of (65.11 ± 14.77) years. Eight patients (22.22%) had some conventional cardiovascular risk factors, while the other 28 patients (77.78%) had no such risk factors. Magnetic resonance imaging (MRI) scans at the acute stage of ischemic stroke were carried out for all these patients. Based on the diffusion weighted imaging (DWI) of MRI, 8 patients (22.22%) had single lesion and 28 patients (77.78%) had multiple lesions in different arterial territories in their brains. The pathological types of renal cell cancer were:suprarenal epithelioma (18 patients, 50.00%), papillary cell carcinoma (12 patients, 33.33%) and chromophobe renal cell carcinoma (6 patients, 16.67%). Metastases were found 10 patients (27.78%) out of the 36 patients. Blood biochemical examination showed that 28 patients had elevated plasma D-dimer level, 22 patients had elevated plasma cancer antigen (CA)125 level, and 17 patients had elevated plasma carcinoembryonic antigen (CEA) levels. Conclusions It is suggested that the renal cell cancer associated stroke is characterized by lacking of traditional risk factors and having multiple lesions in brain;and that the elevated plasma D-dimer, CA125 and CEA levels may lead to hypercoagulable state and lead to ischemic stroke eventually .

Objective To compare the efficacy,toxicitis and side-effects of Casodex and Flutamide in the hormonal therapy of advanced prostate cancer patients.Methods One hundred and thirty-six advanced prostate cancer patients were treated with with hormonal therapy.The patients were divided into 3 groups,of which 52 patients (group A) used LHRHa and Casodex as intermittent hormonal therapy;60 patients(group B) used LHRHa and Flutamide as intermittent hormonal therapy;24 patients(group C) were treated with surgical castration only.The difference of clinical symptoms,serum PSA,disease progression risk,survival rate,toxicitis and side-effects of 3 groups were compared.Results The relief rates of group A and B were 80.8% (42/52)and 81.7% (49/60) respectively,higher than 70.8% (17/24) of group C.The mean serum PSA of group A and B decreased from 133.3 ng/ml(17.9-982.8 ng/ml) to 15.8 ng/ml(0.02-28.9 ng/ml),142.6 ng/ml (20.2-1001.0 ng/ml)to 16.1 ng/ml(0.07-53.8 ng/ml),respectively,both better than that of group C,which decreased from 142.3 ng/ml (27.1-988.0 ng/ml) to 27.6 ng/ml(6.0-62.1 ng/ml).The mean chemical recurrence rates of group A and B were 34.7% (18/52) and 36.7% (22/60),respectively,lower than 58.3% (14/24) of group C.The mean chemical recurrence time of group A and B was 22(5-52)months and 22(6-65)months,respectively,longer than 11(5-54)months of group C.The mortality rates of group A and B were 26.9% (14/52) and 31.7% (19/60),respectively,lower than 66.7%(16/24) of group C.88.5% (46/52)of group A were treated continuously,while group B had 66.7% (40/60).The side-effects rate of group A was lower than group B.Conclusions Both Cadosex and Flutamide are effective for prostate cancer,and decrease the disease progression risk.Casodex is more effective and safer as for the treatment of prostate cancer compared to Flutamide.

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.

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 obtain shRNA sequences that can stably block the expression of Nuclear Factor kappa- B (p65) in the prostate cancer cell line LNCaP and construct the lentivirus vector.And validate the gene function of p65 in the cell line. Methods According to p65 genetic information, we design siRNA1, siRNA2, siRNA3 those three siRNA sequences targeting the ods area of p65 gene and then form the corresponding four pairs of complementary single strand DNA of shRNA, including the sense strand and the antisense strand. The synthetic shRNA sequence was inserted into the empty pSIH1-H1-copGFP shRNA Vector, and after transfecting the prostate cancer cells , the inhibitory effect of p65 mRNA by different sequences was detected through real-time PCR, and the inhibitory effect of p65 protein expression was detected by Western-blotting. Thus we can obtain highly effective shRNA sequences in the inhibition of p65 in prostate cancer cells. MTT, flow cytometry, transwell were chosen to test the cell growth, migration and invasive power in vitro to compare the difference of the experimental group, control group and negative group. Results The third shRNA sequence had the best inhibitory effect and the inhibitory effect of p65 mRNA in prostate cancer cell line was 59 % and the protein was 81%. It's position locates in p65 (NM_021975 ) 1096-1113 and it's stemloop sequence is 5'-GATCCGCCCTATCCCTTTACGTCATTCAAGAGATGACGTAAAGGGATAGGGCTTTTTG-3'. After transfecting, the prostate cancer cell line had the low expression of p65 stably. Through MTT, we got the growth curve, which showed that the growth ability of experimental group was significantly decreased compared with the control group and the Logarithmic growth didn't appear in the first 96 hours. Flow cytometry test displayed that the percentage of G0-G1-phase cells in experimental group was 61.49%, and the control group was 44.89%, idle group was 41.52%, which was increasing oberviously. The S-phase cells in the experimental group was 28.58%, compared with the 47.36% and 46. 10% diminished. The results of transwell showed that the experimental group had 16. 5000±6. 62076 cells and the other two groups had 45. 6333 13. 54159 and 36. 8333±5. 68412 cells, which showed the invasive power of experimental group was significantly declined(P＜0.05).Conclusions It's successful to obtain shRNA sequences that can stably block the expression of p65 in the prostate cancer cell line LNCaP and construct the lentivirus vector. p65 can positively regulates the biological behavior of prostate cancer LNCaP cell line in the cell growth, migration and invasive power.

Objective To present our initial experience in laparoscopic radical prostatectomy performed through an umbilical incision using a home-made multichannel port. Methods From August 2009 to March 2010, we performed single-port laparoscopic radical prostatectomy in 11 patients with localized prostate cancer. A home-made multichannel port was inserted extraperitoneally through a 3-cm umbilical incision. The single port extraperitoneal procedures included obturator fossa lymphadenectomy, radical prostatectomy and urethro-vesical anastomosis, while the urethro-vesical anastomosis was performed by a slip-knot running suture technique. Data were collected and analyzed prospectively. Results All cases were completed successfully, without conversion to a standard laparoscopic approach or open surgery except adding an additional port in one case. The average operative time was 256 minutes (range195-315), and the mean blood loss was 90 ml (range 20- 180), without any blood transfusion. The postoperative hospital stay was 15.4 days (range13- 24), and the Foley catheter was removed 12 days after surgery. No intraoperative complications occurred. One patient developed a vesico-rethralanastomosis leakage, 2 had lymphatic leakage and 1 had urinary tract infection,all of the cases were managed successfully with conservative treatment. Histopathological results showed negative surgical margine and negative lymph node dissection. All patients had no biochemical relapse after an average follow-up of 7 months. Conclusions Single-port laparoscopic radical prosta tectomy is feasible, cosmetic and minimally invasive with a low complication rate and good short-term outcome. Additional investigation is needed to evaluate the long-term safety and oncologic adequacy of this new approach.