Objective To explore an effective method to culture and purify canine bladder transitional epithelial cells.Methods Bladder tissue was obtained from healthy puppy under sterile conditions. Bladder mucosa was removed from the remaining tissue with fine scissor and minced into small pieces, and then were dissociated into single cell suspensions with 0.125% trypsin. The bladder epithelial cells were cultured in defined keratinocyte serum free medium. The cells were passaged and purified by 0.05% trypsin and 0.02% EDTA. Morphological characterization were studied under inverted phase contrast microscope and transmission electron microscope. Expression of cell specific marker protein was assessed by immunohistochemistry. Results Canine bladder transitional epithelial cells could be efficiently cultivated and expanded in serum-free medium without fibroblast contamination. The cells could be passaged 4-6 times without a distinguished decrease in cell proliferation. The cells were characterized by well-developed micro filament and desmosome junction under transmission electron microscope. Immunohistochemical staining with broadly reacting anticytokeratin antibodies (AE1/AE3) confirmed the epithelial phenotype of the cells.Different generations of cells showed diploid cells. Conclusion A large number of bladder transitional epithelial cells can be obtained from small bladder tissue with our digestion method. The cultured bladder epithelial cells can be proliferated to sufficient quantities for further reconstructive purposes.
Objective To evaluate the feasibility of reconstructionof urothelium tissue in vivo using tissue-engineering technique. Methods The urothelium cells were obtained from young rabbit, bladder by mechanical and enzyme digested methods. After expanded in vitro, the 4th to 5th generation urothelium cells were seeded onto the surface of 8 Polylatical／glycolic acid copolymer polymer，the polymer matrix without seeding cells served as control group. A total of 8 cell-polymer scaffolds and 4 simply scaffolds were separately implanted into subcutaneous pockets of athymic mice. Theexperiment groups included cell-polymer scaffolds 4 weeks and cell-polymer scaffolds 8 weeks. The control group included simply scaffold 4 weeks and simply scaffold 8 weeks.After 4 and 8 weeks, the specimens were obtained and examined by gross inspection, histologically and immunohistochemically. Results The results of HE and Masson staining showed that the polymer were covered by urothelium cells layers and cells layers increased markly in experimental group. Immuocytochemical studies revealed that the cells were stained positively for anti-cytokeratins (AE1/AE3) in experimental group. Fiber tissue deposition were found on the surface of polymers in control group by HE and Masson staining. Immunocytochemical staining of implants showed the negative result for cytokeratins in control group. Conclusion It is feasibility that reconstruction of urothelium tissue using tissue-engineering -technique,whichprovides basic understandings for further development of the bladder and ureteral tissue engineered research.
Objective To review the study on adi pose derived stem cells (ADSCs) in the therapy of urological diseases. Methods The recent l iterature concerning ADSCs in bladder repair, urethral reconstruction, incontinence treatment, and erectile dysfunction treatment was reviewed. Results The appl ication of tissue engineering using ADSCs has made significant achievements in the treatment of urological diseases and in animal studies, and has been initially used in cl inicaland has achieved a good therapeutic effect. Conclusion Tissue engineering using ADSCs has good prospects in the study on urological diseases, and is expected to widely used in the treatment of urological diseases.
Objective To evaluate the urine cytology silver staining combined with ultrasonography（USG）in the detection of bladder transitional cell carcinoma （TCC） recurrence after transurethral resection of bladder tumor（TURBT）in terms of sensitivity and specificity. Methods Cystoscopy was used as “gold standard”. Urine cytology combined with USG or cystoscopy was measured separately and blindly. AgNORs protein stained by silver were used in cytology with Kappa of inter-observers 0.81. For the USG, the patients were scanned with trans-rectal probe with Kappa of inter-observers 0.76. The results of urine cytology combined with USG （Positive when urine cytology and/or USG positive. Negative when both urine cytology and USG negative） were compared with “gold standard”. Results The 148 consecutive superficial TCC patients with TURBT one year previously were included in this study. Fifty seven recurrenced cases were detected. Recurrence rate was 38.51%. The sensitivity and specificity of urine cytology silver stain were 89.47% （95% CI 0.82 to 0.98） and 87.91% （95% CI 0.81 to 0.95）. Area under ROC curve was 82.22%. The sensitivity and specificity of USG were 57.90% （95% CI 0.45 to 0.71 ） and 90. 11% （ 95% CI 0.84 to 0.96）. Area under ROC curve was 73.13% . The sensitivity was improved to 94. 74% （95% CI 0.89 to 1.00） when cytology combined with USG. But specificity decreased to 84. 62% （95% CI 0.77 to 0.92 ）. Area under ROC curve was improved to 98.28%. Conclusions Urine cytology silver stain combined with USG improves the high sensitivity for follow-up TCC patients after TURBT. The non-invasive protocol is suggested.
Objective To investigate the safety, efficacy and morbidity of onestage urethroplasty by using bladder mucosa for treatment of hypospadias. Methods From August 1991 to August 2003, 38 cases of congenital hypospadias were given bladder mucosa flap procedure and one stage urethroplasty. Results Thirty-eight cases of hypospadias treated with one stageurethroplasty by using bladder mucosa were followed up 6 months-9 years afterthe procedure. The success rate of the operation was 95%. Three cases of urethral fistula after the procedure were surgically repaired again, 2 cases of urethral stricture recovered after distension. The complication markedly lessened, micturation became normal with the reconstructed meatussituated at the proper site on the glands. Conclusion one stage urethroplastyby using bladder mucosa for treatment of hypospadias is a simple, effective andsafe surgery.
To introduce a micturition alert device dedicated to neurogenic bladders. Methods The design and mechanism of the micturition alert device were explained, the effectiveness was tested in a cranine experiment. Results The micturition alert device consisted of a permanent magnet sutured on the anterior bladder wall and a warning unit sutured on theinferior abdominal wall. The warning unit was assembled with a compass-l ike switch, a power supply, a buzzer and a power switch. Bladder volume determined the position of the magnet which determined the magnetic field at the point of the warning unit. The change of magnetic field was read by the warning unit. With increasing bladder volume from initial state to 200 mL in 8 dogs, the magnet moved cranially 32.8 mm averagely (from 31.3 mm to 34.1 mm) and the hand of warning unit turned 52° (from 47° to 57°). The value of the warning unit was correlated positively to the bladder volume (r =1.0, P lt; 0.01）. If the desired bladder volume was determined as 150 mL to activate the warning unit to alarm in advance, the fullness of bladder was 147.6 mL averagely from135 mL to 160 mL, with an error less than 15 mL (10%). Conclusion The micturition alert device including a warning unit and permanent magnet could monitor bladder volume continuously and alarm in time for the patients with loss of micturition desire. It is simple, easily-made, cheap and conveniently used. It is worth of further study.
Objective To evaluate the cytocompatibility of collagenmembraneswith transitional cells of rabbit in vitro and to discuss the possibility of the collagen membranes as urologic tissue engineering scaffolds. Methods Primary cultured transitional cells isolated from New Zealand rabbits were implantedon collagen membranes at 1×105 cells/cm2. The changes of cell adhering were observed by inverted microscope and scanning electron microscope 2, 12 and 24hours later. The experiment was divided into 4 groups: non-cell group (black control) culture medium group(negative control), extract medium from Polyvinyl chloride group(positive control) and extract medium from collagen membranes group(experimental group). The cells of generations 2 to 4 were implanted in 96-hole-plank at 1×104 cells every hole. And every group had 5 holes. Then absorption coefficient were detected at the wave length of 490 nm by MTT assay. Then the cytotoxicity and cytocompatibility were evaluated by comparison of the numbers of absorptioncoefficient.Results The bladder transitional cells began to adhere to the collagen membrane 2 hours after implanting, and the number of the adhered cells increased with time.The actual absorption coefficient of experimental groups was 0.590±0.024,1.065±0.040 and 1.129±0.074 after 24, 72 and 120 hours. The actual absorption coefficient of negative control group was 0.639±0.068,1.022±0.044 and 1.087±0.111. The actual absorption coefficient of positive control group was 0.302±0.029,0.653±0.083 and 0.694±0.031. There was significantdifference between the experimental group and positive control (Plt;0.01), and no significant difference between the experimental group and negative control(Pgt;0.05).Conclusion Collagen membrane has good cytocompatibility withtransitional cells and no cytotoxity. It can be used as scaffolds of urologic tissue engineering.
OBJECTIVE To establish an artificial bladder reflex arc in canines to reinnervate the neuropathic bladder and restore bladder function after spinal cord injury. It involves a somatic reflex arc with a modified efferent branch which passes the somatic motor impulses to the bladder and initiates autonomic bladder detrusor contraction. METHODS Intradural microanastomosis of the right L5 ventral root to S2 ventral root was performed to maintain the right L5 dorsal root intact. After axonal regeneration, the new patellar ligament-spinal cord center-bladder artificial bladder reflex pathway was established, and micturition was induced by knocking the patellar ligament. The early and final function of the reflex arc was observed by electrophysiological examinations, bladder pressure tests and detrusor electromyograms(EMG) at 6 months and 18 months postoperatively. RESULTS Single stimuli (115 mV, 1.0 ms) of the right L5 dorsal root resulted in evoked potentials recorded from the right S2 ventral root distal to the anastomosis site before and after the spinal cord was transected horizontally at the T10 segment level in all 6 canines. Bladder contraction was very quickly initiated by trains of stimuli(1,000 mV, 10 Hz, 2 s) of the right L5 dorsal root and bladder pressures increased rapidly to 65% of normal, and bladder contraction induced by knocking the right patellar ligament was increased to 51% of normal through the new reflex arc in 4 canines after 6 months of operation. Bladder pressures were increased by the same stimuli to average 84% of normal and to 62% of normal by knocking the patellar ligament in 2 canines after 18 months of operation. Stimuli(3.8 mA, 1.0 Hz) of the right L5 dorsal root and femoral nerve resulted in EMG similar to normal EMG could be recorded from the detrusor in 2 canines after 18 months postoperatively. CONCLUSION The somatic motor axons can be regenerated into the parasympathetic endoneurial tubes of autonomic nerve. Using the survived somatic reflex under the horizon of spinal cord injury to reconstruct the bladder autonomic reflex arc by intradural microanastomosis of ventral root is practical in the canine model and may have a potential of clinical application.
ObjectiveTo systematically evaluate the efficacy and safety of simultaneous transurethral resection of bladder cancer and prostate (TURBT+TURP) in the treatment of bladder cancer with benign prostatic hyperplasia (BPH). MethodsWe searched PubMed, EMbase, The Cochrane Library, Web of Science, CBM, CNKI, WanFang Data and VIP from inception to January 2015, to collect randomized controlled trials (RCTs) and cohort studies investigating the efficacy and safety of TURBT with TURP in the treatment of bladder cancer with BPH. Two reviewers independently screened literature, extracted data, and assessed the risk bias of included studies, and then meta-analysis was performed using RevMan 5.3 software. Results3 A total of 3 RCTs (n=137) and 10 retrospective cohort studies (n=998) were included. The results of meta-analysis showed that there were no significant differences between the simultaneous resection group and the control group in the overall recurrence rate (RCT:OR=0.55, 95% CI:0.24 to 1.24, P=0.15; retrospective cohort study:OR=0.78, 95% CI:0.60 to 1.01, P=0.06), postoperative recurrence rate in the prostatic fossa/urethra (RCT:OR=1.40, 95% CI:0.28 to 7.60, P=0.68; retrospective cohort study:OR=1.36, 95% CI:0.49 to 3.74, P=0.55), progression rate (OR=0.93, 95% CI:0.53 to 1.61, P=0.79) and overall perioperative complication rate (RCT:OR=0.35, 95% CI:0.08 to 1.55, P=0.17; retrospective cohort study:OR=0.1.75, 95% CI:0.44 to 6.98, P=0.43). ConclusionCompared with only TURBT or sequential TURBT and TURP, simultaneous TURBT and TURP do not increase the overall recurrence rate, postoperative recurrence rate in the prostatic fossa/urethra, progression rate and overall postoperative complication rate. However, due to the limited quality and quantity of included studies, larger sample size and higher quality RCTs are needed to verify the above conclusion.
Objective It is a thorny problem to reconstruct long ureteral defect in urinary surgery. To investigate the feasibil ity of intestinal sero-muscular segment with autograft of bladder mucosa as a replacement material for reconstructionof long ureteral defect. Methods Twelve adult Beagle dogs (weighing 6.5-9.3 kg and being male or female) were randomlydivided into 3 groups, each group including 4 dogs. In group A, lower segment of ureter was reconstructed by autograft of bladder mucosa to the intestinal sero-muscular segment; furthermore, the proximal and distal reconstructed ureter were anastomosed to the bladder and the upper ureter, respectively. In group B, upper segment of ureter was reconstructed by the same method as that of group A, the proximal and distal reconstructed ureter anastomosised with pelvic and lower ureter, respectively. In group C, whole ureter was reconstructed by the same method as that of group A, the proximal and distal reconstructed ureter were anastomosised with pelvic and bladder, respectively. Blood urea nitrogen, Cr2+, K+, Na+, Cl-, Ca2+ and carbon dioxide combining power were detected before operation, the general state, drainage volume, heal ing of wound, and compl ications were observed after operation. At 6 weeks, the blood biochemical indexes and intravenous urography (IVU) were detected, and the gross and histological observations of ureter were done. Results In group B, urine leakeage and infection occurred in 1 dog 2 days after operation because ureter stent prolapsed; other dogs had no complications. There was no significant difference in the biochemical indexes between before operation and 6 weeks after operation. IVU showed: in group A, hydronepherosis and ureterectasia occurred on the operation side of 1 dog; in group B, anastomotic stricture between the reconstructed ureter and lower ureter and hydronepherosis occurred in 1 dog; and in other dogs of all groups, renal function was good and the reconstructed ureter had peristalsis function. The histopathological observation showed that the reconstructed ureter had similar structure to normal ureterat 6 weeks in 3 groups; the inflammatory cells infiltrating of the reconstructed ureter was observed in 1 dog of groups A and C, respectively. Conclusion Reconstruction of ureter by intestinal sero-muscular segment with autograft of bladder mucosa has similar structure and function to the normal ureter. The results might provide an experimental basis for cl inical use.