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 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 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.
Objective To systematic review of bladder cancer antigen (BTA) stat and urine cytology (UC) in the diagnosis of bladder cancer. Methods MEDLINE (Jan.1966 to June 2008), EMbase (Jan.1988 to June,2008), Cochrane Library (Issue 1,2008), CMCC (1979 to June, 2008) and CNKI (Jan.1979 to June, 2008) were searched for studies about BTA stat and cytology in the diagnosis of bladder cancer. The search strategy was made according to the Collaborative Review Group search strategy. Quality of included trials wa assessed by quality assessment of diagnostic accuracy studies.Data were extracted by two reviewers using the designed extraction form. The software MetaDiSc1.4 was used to review management and data analysis. Results In total, 71 relevant studies were searched, of which 13 were included and 58 were excluded, with 3 733 patients involved. Heterogeneity (except for threshold effect) was found within these studies. A meta-analysis was performed using random effect model. Pooled accuracy indicators of sensitivity, specificity, positive likelihood ratio (LR) , negative LR and diagnostic odds ratio (dOR) and 95%CI of BTA stat and UC were 0.68 (0.65,0.70), 0.74 (0.72, 0.76), 2.51 (2.04, 3.09), 0.46 (0.38, 0.55), 5.66 (3.87, 8.29) and 0.41 (0.39, 0.44), 0.97 (0.97, 0.98), 12.64 (7.58, 21.08), 0.62 (0.55, 0.71), 22.16 (12.38, 39.66), respectively. The sensitivity of both methods increased as the higher of tumor grade and stage, and the incipient tumor was higher than the recurrence. Area under curve (AUC) of SROC curve of BTA stat and UC were 0.753 5 and 0.711 9, and Q index were 0.696 3 and 0.662 4, respectively. Conclusions The performance of urine BTA stat is moderate in the diagnosis of bladder tumor. It can not replace the traditional urine cytology and diagnose the bladder cancer alone, but which can be an available noninvasive examination and an important adjunct of preoperative detecting and postoperative monitoring of bladder tumor.
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.
Objective To explore an effective method of culturing the canine bladder smooth muscle cells, observe the morphological characteristics of the bladder smooth muscle cells growing on acellular small intestinal submucosa（SIS） and offer an experimental basis for reconstruction of the bladder smooth muscle structure by the tissue engineering techniques. Methods The enzymetreatment method and the explant method were respectively used to isolate and harvest the canine bladder smooth muscle cells, and then a primary culture of these cells was performed. The canine bladder smooth musclecells were seeded on the SIS scaffold, and the composite of the bladder smooth muscle cells and the SIS scaffold were co cultured for a further observation. At 5,7 and 9 days of the co culture, the specimens were taken; the bladder smooth muscle cells growing on the SIS scaffold were observed by the hematoxylin staining, the HE staining, and the scanning electron microscopy. The composite of the bladder smooth muscle cells on the SIS scaffold was used as the experimental group, and the bladder smooth muscle cells with no SIS were used as the control group. In each group, 9 holes were chosen for the seeded bladder smooth muscle cells, and then the cells were collected at 3, 5 and 7 days for the cell counting after the enzyme treatment. Morphological characteristics of the cells were observed under the phase contrast microscope and the transmission electron microscope. Expression of the cell specific marker protein was assessed by the immunohistochemical examinaiton. The proliferation of the cells was assessed by the cell counting after the seeding on the SIS scaffold. Results The primary bladder smooth muscle cells that had been harvested by the enzyme treatment method were rapidly proliferated, and the cells had good morphological characteristics. After the primary culture in vitrofor 5 days, the bladder smooth muscle cells grew in confluence. When the bladder smooth muscle cells were seeded by the explant method, a small amount of the spindleshaped bladder smooth muscle cells emigrated from the explant at 3 days. The cells were characterized by the welldeveloped actin filaments inthe cytoplasm and the dense patches in the cell membrane under the transmissionelectron microscope. The immunohistochemical staining showed the canine bladdersmooth muscle cells with positive reacting α actin antibodies. The bladder smooth muscle cells adhered to the surface of the SIS scaffold, growing and proliferating there. After the culture in vitro for 5 days, the smooth muscle cells covered all the surface of the scaffold, showing a singlelayer cellular structure. The cell counts at 3, 5 and 7 days in the experimental group were（16.85±0.79）×105，（39.74±2.16）×105 and （37.15±2.02）×105, respectively. Thecell counts in the control group were（19.43±0.54）×105，（34.50±1.85）×105 and （33.07±1.31）×105, respectively. There was a significant difference between the two groups at 5 days （P＜0.05）. ConclusionWith the enzyme treatment method, the primarily cultured canine bladder smooth muscle cells can produce a great amount of good and active cells in vitro. The acellular SIS can offer an excellent bio scaffold to support the bladder smooth muscle cells to adhere and grow, which has provided the technical foundation for a further experiment on the tissue engineered bladder reconstruction.
ObjectiveTo prepare a composite scaffold using bladder acellular matrix (BACM) and polyurethane (PU) for bladder repair and regeneration, and to evaluate its mechanical properties and biocompatibility. MethodsFresh bladder tissues were obtained from New Zealand rabbits and then treated with 1%SDS and 1%Triton X-100 to obtain BACM. The BACM was combined with PU to fabricate PU-BACM composite scaffold. The tensile strength and elongation at break of BACM and PU-BACM scaffolds were tested. Scaffolds and extracts of scaffolds were prepared to evaluate the biocompatibility. For cell-proliferation analysis, cell counting kit 8 method was used at 1, 3, 5, and 7 days after co-culture of human bladder smooth muscle cell (HBSMC) and scaffolds. The cell cycle was tested by flow cytometry after HBSMC co-cultured with extracts of scaffolds and DMEM culture medium (control group) for 24 hours. Finally, 12 New Zealand rabbits were used to establish the model of bladder repair and regeneration. Incision of 5 mm was made on the bladder, and PU-BACM scaffold was sutured with the incision. The rabbits were sacrificed at 10, 20, 40, and 60 days after surgery to observe the inflammatory cell infiltration, new tissues formation, and regeneration of epithelium by HE staining. ResultsThe tensile strength of BACM and PU-BACM composite scaffold was (5.78 ± 0.85) N and (11.88 ± 3.21) N, and elongation at break was 14.46%±3.21% and 23.14%±1.32% respectively, all showing significant diffeence (t=3.182, P=0.034；t=4.332, P=0.012). The cell-proliferation rates of controls, PU, BACM, and PU-BACM were 36.78%±1.21%, 30.49%±0.89%, 18.92%±0.84%, and 22.42%±1.55%, it was significantly higher in PU-BACM than BACM (P<0.05). In the bladder repair and regeneration experiment, inflammatory cell infiltration was observed at 10 days after operation, and reduced at 20 days after implantation. In the meanwhile, the degradation of scaffolds was observed in vivo. The regeneration of epithelium could be observed after 40 days of implantation. At 60 days after implantation, in situ bladder tissue formed. ConclusionPU-BACM composite scaffold has higher mechanical properties and better biocompatibility than BACM scaffold. PU-BACM composite scaffold will not lead to strong immune response, and new bladder tissue can form in the in vivo rabbit bladder repair experiment. These results can provide research basis and theoretical data for 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.
ObjectiveTo analyze the disease burden of prostate, bladder and kidney cancers attributable to smoking in China from 1990 to 2019. MethodsBased on the global burden of disease study 2019, the current situation of the disease burden of prostate, bladder and kidney cancers attributable to smoking was analyzed by using the population attributable fraction (PAF), deaths and disability-adjusted life years (DALYs). Furthermore, the annual percent change (APC) and the average annual percent change (AAPC) were calculated by joinpoint regression analysis to describe the long-term trends of the smoking-attributable burden of these three cancers from 1990 to 2019. ResultsThere were an estimated 18 800 cases of deaths and 393 106 person-years of DALYs for bladder cancer caused by smoking in 2019. The age-standardized mortality and DALY rate decreased by 0.41% and 0.39% per year from 1990 to 2019, respectively. For prostate cancer, smoking was estimated to have caused 5 016 cases of deaths and 98 276 person-years of DALYs in 2019. The age-standardized mortality and DALY rate decreased by 0.28% and 0.25% per year from 1990 to 2019, respectively. For kidney cancer, the deaths and DALYs attributable to smoking were 4 935 cases and 120 620 person-years, respectively. The standardized mortality and DALY rates increased by 3.03% and 2.98% per year from 1990 to 2019. Additionally, males suffered from a higher disease burden of these three cancers attributable to smoking than females. The elderly population had a higher smoking-attributable disease burden than the younger population. ConclusionThe situation of the disease burden of bladder, prostate and kidney cancers attributable to smoking is still serious in China, which has substantial disparities in different groups. Specifically, males and the elderly are the high-risk groups for the smoking-attributable burden. Among the three cancers, bladder cancer has the highest burden and kidney cancer has the largest burden increase during 1990-2019.
ObjectivesTo analyze the trend of incidence and mortality of bladder cancer from 1990 to 2017 and the effects of age, time period and birth cohort on bladder cancer incidence and mortality.MethodsData on age-standardized incidence rate (ASIR) and age-standardized death rate (ASDR) of bladder cancer from 1990 to 2017 were extracted from the Global Burden of Disease 2017 (GBD 2017) database. Joinpoint regression model was used to analyze the average annual percentage change of ASIR and ASDR of bladder cancer. The age-period-cohort model was established to analyze the age, period and birth cohort effects on ASIR and ASDR of bladder cancer.ResultsFrom 1990 to 2017, both ASIR and ASDR of bladder cancer decreased slightly. ASIR decreased from 6.42 per 100 000 in 1990 to 6.04 per 100 000 in 2017, with an average annual percentage change of −0.9% (−1.0% to −0.8%), and ASDR decreased from 3.15 per 100 000 in 1990 to 2017 2.57/100 000, with an average annual percentage change of −0.4% (−0.4% to −0.3%). The age-period-cohort model results showed that as age increased, the risk of bladder cancer incidence and mortality increased; as the birth cohort progressed, the risk of bladder cancer morbidity and mortality decreased. The time period had little effect on the incidence and mortality of bladder cancer.ConclusionsThe incidence and mortality of bladder cancer are declining globally. On the other hand, the increase of the aging global population could reverse the incidence and mortality trend, active measures should be taken to address the adverse effects of aging.