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BACKGROUND: Tissue engineering technology was used to construct and form tissue engineering, tubes which can significantly reduce the incidence of various perioperative complications with good anti-urine leakage performance in vitro. OBJECTIVE: To evaluate the feasibility of using adipose stem cells and expanded polytetrafluoroethylene to construct tissue-engineered urine outflow tract in vitro for canine urinary diversion surgery. METHODS: In the experimental group, adipose stem cells were isolated and cultured from four female beagles, and then differentiated into urothelial cells in vitro for 14 days. The differentiated cells were implanted on the inner surface of expanded polytetrafluoroethylene tubes to form a tissue-engineered urinary diversion device. Bladders were removed from dogs ///9only 1.0 cm × 1.0 cm of trigonometric area was retained), and both ends of the tissue engineered urinary diversion device were anastomosed with trigonometric area and abdominal wall respectively to complete the urinary diversion operation. The samples were collected at 1, 2, 4 and 8 weeks postoperatively. The control group (two dogs) underwent urinary diversion with expanded polytetrafluoroethylene engineered tube, and samples were collected 8 weeks after operation. The animal experiment was approved by laboratory animal ethics committee of Renmin Hospital of Wuhan University (approval No. WDRM（Fu）20180810). RESULTS AND CONCLUSION: (1) There were different degrees of adhesion between the two groups of implanted tissue-engineered urinary diversion devices and the abdominal wall or intestinal wall. Two dogs of the experimental group had mild hydronephrosis, and no serious complications such as urinary exosmosis or infection. One dog in the control group developed severe urinary fistula and died 7 weeks after operation. The tissue-engineered urinary diversion device of the other dog was covered with stones. (2) Hematoxylin-eosin staining showed that thin epithelial cells with loose intercellular connections began to grow continuously in the outflow tract lumen of the experimental group 1 week after operation. At 2 weeks postoperatively, the intercellular connections became dense and the number of epithelial cells increased. Four weeks after operation, the cells began to accumulate and thicken, and gradually changed into a multilayer structure. The entire lumen was covered by the epidermis, and neovascularization was observed 8 weeks after surgery. (3) The results showed that it was feasible to construct tissue-engineered urine outflow tract together with expanded polytetrafluoroethylene after adipose stem cells were differentiated into urothelial cells for urinary diversion in beagles. The inner cavity of outflow tract could form a tightly connected urothelial cortex with the function of draining urine and preventing urinary extravasation. [ABSTRACT FROM AUTHOR]

BACKGROUND: Evidence has shown that the treatment of overactive bladder at the level of mucosal structure has a certain advantage, so the study on the possible involvement of mucosa in regulating bladder afferent signal transduction and improving the symptoms associated with overactive bladder has gradually become a hot spot. OBJECTIVE: To review the role of signal pathways and receptors in various layers of mucosa in the pathogenesis of overactive bladder. METHODS: With the key words of “overactive bladder, mucosa, urothelium, lamina propria, mechanisms" in both English and Chinese, CNKI, WanFang and PubMed were searched for relevant literature published from 2000 to 2020, and eventually 49 articles meeting the requirements were reviewed. RESULTS AND CONCLUSION: The pathogenesis of overactive bladder is very complex. At present, there are few reports on whether the myogenic hypothesis and the neurogenic hypothesis interact with the mucosal signaling mechanism. Urothelial signaling pathway is the key of the current research, but the basement membrane layer and the lamina propria related signaling pathways are less reported. The various types of cells in the lamina propria and their functions have not been clear, and the relationship of mucosal signal with spinal cord center and senior center has not been fully research. Therefore, a horizontal study will be done on the relationship between the three types of pathogenesis of overactive bladder, and deeply and vertically explore the upstream or downstream of one of the three signal pathways, so as to provide a clearer target for the treatment of overactive bladder. [ABSTRACT FROM AUTHOR]

BACKGROUND: Adult stem cells are pluripotent stem cells that exist in differentiated tissues. Urine-derived stem cells are newly discovered adult stem cells. They have attracted increasing attentions in the tissue engineering, due to its advantages of convenient sampling, highly proliferative ability and multidirectional differentiation potential. OBJECTIVE: To investigate the separation and extraction method of urine-derived stem cells and to investigate the feasibility of differentiation into urothelial cells and smooth muscle cells in vitro. METHODS: Urine specimens were collected from healthy adults, and urine-derived stem cells were obtained by isolation and culture in vitro. Cell counting kit-8 assay was used to detect cell proliferation and plot cell growth curve. Cell phenotype was detected by flow cytometry. The differentiation into urothelial and smooth muscle cells was induced by special medium respectively in vitro. The cell differentiation was detected by quantitative PCR, immunohistochemical staining, immunofluorescence cell staining and western blot assay. RESULTS AND CONCLUSION: Urine-derived stem cells were successfully isolated from the urine specimens of healthy adults. Urine-derived stem cells possessed high proliferation rate and the cell growth curve exhibited an S-shape. Urine-derived stem cells exhibited high expression of CD29 (98.11%) and CD90 (95.74%), both of which are mesenchymal stem cell surface markers. After 14 days of induction in vitro, urine-derived stem cells were able to express urothelial cell specific genes Cytokeratin 7, Cytokeratin 20, Uroplakin II and smooth muscle cell specific genes α-SMA and SM22. These results suggest that urine-derived stem cells can differentiate into urothelial cells and smooth muscle cells after induction in vitro and can be used as ideal seed cells for urinary tract tissue engineering. [ABSTRACT FROM AUTHOR]

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Objective: To analyze the clinical pathologic characteristics of cases with fluorescence in situ hybridization (FISH) positive of exfoliated urothelial cells, so as to evaluate the clinical utility of FISH in the diagnosis of urothelial carcinoma (UC)., Methods: A total of 271 cases of FISH positive in Department of Urology of Peking University First Hospital from Apr. 2012 to Sep. 2015 were recruited in this study. Retrospective analysis was made on their clinical data. For FISH analysis, labeled probes specific for chromosomes 3, 7, 17, and the p16 (9p21) gene were used to assess chromosomal abnormalities indicative of malignancy. The positive predict values (PPV) of all the techniques were analyzed., Results: Of the 271 patients, 207 cases were UC, 7 cases were non-UC, and 57 cases were benign diseases. The PPV of FISH in detecting UC was 76.4%, while the 95% confidence interval (CI) 71.3% to 81.5%. In the cohort of FISH positive, this value was similar to that of urinary cytology (PPV 86.8%, 95% CI: 78.5%-95.0%). The PPV of FISH was lower than that of cystoscopy and ureteroscopy (PPV 96.1%, 95% CI: 91.7%-100.0%). There were significant differences between this study and the PPV of FISH reported abroad (PPV 53.9%, χ 2 =33.048, P<0.001). Of all the UC with FISH positive, bladder cancer showed an earlier pathological stage versus renal pelvic carcinoma and ureteral carcinoma, with significance (χ 2 =5.894, P=0.015, and χ 2 =13.601, P<0.001, respectively). However, no difference was found in the size, pathological stage and pathological grade of tumors between the urinary cytology positive group and the urinary cytology negative group. The rate of high-grade UC in ureteral carcinoma of FISH positive was 92.3%, much higher than that of ureteral carcinoma reported domestically., Conclusion: The PPV of FISH in detecting UC is higher relatively, with a better clinic value for Chinese patients. The ureteral carcinoma with FISH positive obtains a higher pathological grade, which is of great guiding significance for UC.

Objective: To investigate the feasibility of autologous bone marrow mesenchymal stem cells(BMSCs) seeded on bladder acellular matrix graft (BAMG) for bladder reconstruction in a canine model., Methods: This study included 25 mongrel dogs. Five dogs were sacrificed for the preparation of BAMG. Twenty dogs were randomly divided into two groups and received partial cystectomy. In group A, the bladder defect was repaired with unseeded BAMG. In group B, the bladder defect was repaired with autologous BMSCs-seeded BAMG. The bladders were retrieved and studied histologically and immunohistochemically at the time point of 1, 2, 4, 8, 12 weeks after surgery to evaluate tissue regeneration., Results: All dogs survived the procedure. Histopathological examination in group B showed there was urothelium developed at the end of the 2 weeks. By 8 and 12 weeks all bladder wall components were regenerated in the repaired area, which were similar to normal bladder tissue. In group A, urothelium regeneration was observed at the end of the 4 weeks, whereas smooth muscle was still not well-formed by 12 weeks., Conclusion: Autologous BMSCs-seeded BAMG could promote the repair of bladder defect, which is superior to unseeded BAMG in regenerative properties.

Objective: To investigate the construction of urothelial structure by tissue engineering., Methods: Fresh bladder of New Zealand white rabbits were processed to prepare the bladder acellular matrix graft (BAMG) as the scaffold, which was evaluated by Masson's trichrome staining and the scanning electronic microscope. Bladder epithelia were obtained by enzymatic digestion and were proliferated in vitro. Growth of cells was observed under the inverted phase contrast microscope, and cells were identified by immunohistochemical method. The bladder epithelia were seeded on BAMG, and the epithelia/BAMG composites were observed by HE staining and the scanning electronic microscope. The composites fabricated in vitro were implanted into nude rats, and were retrieved in 4 and 8 weeks, which were observed by general observation, histological and immunohistochemical method., Results: White semi-transparent membrane appeared in the prepared BAMG, and a fibre mesh structure of the material without residual cells was observed under the scanning electronic microscope. Bladder epithelia cultured in vitro showed a paving stone structure. Immunohistochemical staining with cytokeratin was performed, and brown cellular plasma staining was observed as positive reaction. After the epithelia were seeded on BAMG in vitro for 7 days, the cells fully covered the surface of the framework, showing a single-layer cellular structure. After being implanted into nude rats for 4 weeks and 8 weeks, the epithelia seeded on the BAMG formed a multi-layer structure., Conclusion: Urothelial structures can be constructed in vitro and in vivo by tissue engineering, which lays a technical foundation for further tissue engineered urinary tract reconstruction experiments.

Objective: To evaluate the feasibility of reconstruction of 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. The experiment 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 markley in experimental group. Immunocytochemical 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,which provides basic understandings for further development of the bladder and ureteral tissue engineered research.