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.
目的 探討組織蛋白酶B（CB）在膀胱移行細胞癌（TCC）中的表達以及其與TCC浸潤的關系。 方法 取TCC標本40例，TCC分級Ⅰ級23例，Ⅱ～Ⅲ級17例；表淺型TCC（Tis，Ta，T1期） 25例，浸潤型TCC（T2～4期）15例。另取10例正常膀胱組織作為對照。用鏈霉素抗生物素蛋白-過氧化物酶連接法行CB免疫組織化學染色觀察并計算CB陽性細胞百分率。 結果 正常膀胱組織中基質無明顯著色；在TCC癌組織中，CB可為細胞染色，部分基質亦有染色，部分毛細血管內皮細胞及部分成纖維細胞CB表達陽性，在癌周血管內皮細胞的陽性著色CB表達增強。CB在分級和分期高的癌組織中多為彌散陽性染色。TCC分級Ⅰ級組、TCC分級Ⅱ～Ⅲ級組、TCC分期表淺型組、TCC分期浸潤型組及正常對照組的CB陽性細胞百分比分別為10.53% ± 3.76%、21.52% ± 3.58%、11.32% ±2.69%、20.57% ± 3.25%、0.11% ± 0.18%，TCC各組均高于正常對照組（P＜0.01）；TCC分級Ⅱ～Ⅲ級組高于TCC分級Ⅰ級組，TCC分期浸潤型組高于TCC分期表淺型組，差異均有統計學意義（P＜0.01）。 結論 CB可能成為判斷TCC進展和預后的重要指標。
Objective To explore the effect of hydrostatic pressure on intracellular free calcium concentration ([Ca2+]i) and the gene expression of transient receptor potential vanilloid (TRPV) in cultured human bladder smooth muscle cells (hb-SMCs), and to prel iminarily probe into the possible molecular mechanism of hb-SMCs prol iferation stimulated by hydrostatic pressure. Methods The passage 6-7 hb-SMCs were loaded with Ca2+ indicator Fluo-3/AM. When the hb-SMCs were under 0 cm H2O (1 cm H2O=0.098 kPa) (group A) or 200 cm H2O hydrostatic pressure for 30 minutes (group B) and then removing the 200 cm H2O hydrostatic pressure (group C), the [Ca2+]i was measured respectively by inverted laser anningconfocal microscope. When the hb-SMCs were given the 200 cm H2O hydrostatic pressure for 0 hour, 2 hours, 6 hours, 2 hours, and 24 hours, the mRNA expressions of TRPV1, TRPV2, and TRPV4 were detected by RT-PCR technique. Results The [Ca2+]i of group A, group B, and group C were (100.808 ± 1.724), (122.008 ± 1.575), and (99.918 ± 0.887) U, respectively; group B was significantly higher than groups A and C (P lt; 0.001). The [Ca2+]i of group C decreased to the base l ine level of group A after removing the pressure (t=0.919, P=0.394). The TRPV1, TRPV2, and TRPV4 genes expressed in hb-SMCs under 200 cm H2O hydrostatic pressure at 0 hour, 2 hours, 6 hours, 12 hours, and 24 hours, but the expressions had no obvious changes with time. There was no significant difference in the expressions of TRPV1, TRPV2, and TRPV4 among 3 groups (P gt; 0.05). Conclusion The [Ca2+]i of hb-SMCs increases significantly under high hydrostatic pressure. As possible genes in stretch-activated cation channel, the TRPV1, TRPV2, and TRPV4 express in hb-SMCs under 200 cm H2O hydrostatic pressure. It is possible that the mechanical pressure regulates the [Ca2+]i of hb-SMCs by opening the stretch-activated cation channel rather than up-regulating its expression.
Objective To observe the effect of selective sacral rhizotomy in treating spastic bladder after spinal cord injury and to explore the mechanism and the best surgical method of different sacral rhizotomies. Methods The spastic bladder models were established in 12 male dogsand were divided into 4 groups according to the different rhizotomies of the sacral nerve as the following: rhizotomy of the anterior root of S2(group A), rhizotomy of the anterior root of S2 and half of the anterior root of S3(group B), rhizotomy of the anterior roots of S2 and S3(group C), and total rhizotomy of the nerve roots of S2-4 (group D). By urodynamic examination and electrophysiological -observation, the changes of all functional data were recorded and comparedbetween pre-rhizotomy and post-rhizotomy to testify the best surgical method. In clinical trial, according to the results of the above experiments, rhizotomy of the anterior root of S2 or one of the halfanterior root of S3 were conducted on 32 patients with spastic bladder after spinal cord injury. The mean bladder capacity, the mean urine evacuation and the mean urethra pressure were (120±30), (100±30)ml and (120±20) cm H2 O, respectively before rhizotomy. Results After rhizotomy, the bladder capacity in 4 groups amounted to (150±50), (180±50), (230±50), and (400±50) ml, respectively; and the urine evacuation volume were (130±30), (180±50), (100±50) and (50±30)ml, respectively. In the treated 32 patients, the mean bladder capacity were raised to 410 ml, and the mean urine evacuation volume were also increased to 350 ml. Incontinence of urine disappeared in all patients. After 22-month follow-up on 13 patients, no recurrence was observed. Conclusion The effectof selective sacral rhizotomy in treating spastic cord injury is significant and worthy of further studies.
Bladder cancer is one of the most common cancers of the urinary system. Baesd on the involvement of the blandder muscle or not, bladder cancer can be generally classified into muscule-invasive bladder cancer (MIBC) and non-MIBC. Cisplatin-based neoadjuvant chemotherapy followed by radical cystectomy is the standard treament recommended by current guidelines for MIBC. Based on the good efficacy of immunocheckpoint inhibitors in advanced bladder cancer. More and more studies have explored the safety and efficacy of immunotherapy in MIBC neoadjuvant therapy, and analyzed biomarkers to explore the benefit groups. This article reviews the latest progress of various neoadjuvant immunomonotherapy in MIBC, and prospect the future direction of development.
Objective To evaluate tissue regeneration, body reaction, and biological safety of xenogeneous bladder acellular matrix (BAM) that can be used to repair rabbit bladder. Methods Porcine BAM was prepared through physical, chemical, and enzymatic methods, and the effects of acellularization and the structure were observed with HE staining and scanning electron microscope (SEM). Eighteen New Zealand white rabbits (weighing, 2.5-3.0 kg) undergoing partial cystectomy were randomly divided into 2 groups. After partial (about 30%) cystectomy, the porcine BAM was used to replace partial rabbit bladder in the experimental group (n=12), and the incision was directly sutured as control group (n=6). The survival condition of animals was observed after operation. At 15 days, 1, 2, 3, and 6 months after operation, the blood routine, renal function, and electrolyte were tested by collecting the blood samples. At 1, 2, 3, and 6 months after operation, maximum bladder capacity, bladder leak point pressure, and bladder compliance were measured through urodynamic studies. Then gross observation was performed for regeneration of bladder, and the specimens of the bladder were harvested for HE staining and immunohistochemical staining. The surrounding organs and local lymphoid tissues were harvested for gross observation and HE staining. Results Cell components were completely removed in the porcine BAM, showing three-dimensional porous structure under SEM. All the animals survived during the experiment. At 15 days after operation, white blood cell count increased, and then returned to normal level in 2 groups, showing no significant difference between 2 groups (P gt; 0.05). The tests of renal function and electrolyte suggested no significant difference between 2 groups (P gt; 0.05). The level of serum creatinine showed a tendency of increase, but it remained within normal range at 6 months after operation. The maximum bladder capacity and compliance in experimental group were significantly higher than those in control group at 3 and 6 months after operation (P lt; 0.05), but no significant difference in bladder leak point pressure at each time point between 2 groups (P gt; 0.05). The urothelial regeneration, smooth muscle regeneration, and blood vessel regeneration were seen by histological observation in 2 groups. In the 2 groups, chronic inflammatory cells infiltration could be observed at 1 month postoperatively, and then chronic inflammatory cells decreased significantly (P lt; 0.05), until complete disappearance. There was no significant difference in score of chronic inflammatory cell infiltration between 2 groups at 3 and 6 months after operation (P gt; 0.05). The α-smooth muscle actin expression was significantly increased with time passing in 2 groups (P lt; 0.05), and it was significantly higher in control group than in experimental group at each time point (P lt; 0.05). In addition, gross and HE staining observations showed no abnormalities in surrounding organs and local lymphoid tissues. Conclusion No immune rejection response occurs when porcine BAM is used for xenotransplantation. It is indicated that porcine BAM is relative safety for xenotransplantation.
Objective To summarize the recent research situation and progress of decellularized matrix in tissue engineered trachea transplantation and to forecast the possible perspects. Methods Recent original articles about study and application for decellularized matrix in tissue engineered trachea were reviewed. The application and study of different decellularized matrices involved in animals or patients with tracheal lesions were elaborated. Results Decellularized matrices researched and applied in tissue engineered trachea include jejunum, urinary bladder, aorta, and trachea. Conclusion Decellularized urinary bladder matrix and jejunal matrix appears to be efficacious method for the patch repair of partial circumferential tracheal defects. The application of decellularized aortic matrix may need more study, and decellularized tracheal matrix has a bright future in long tracheal defects.
目的:探討基層醫院前列腺增生并膀胱結石的微創治療方法。方法:聯合經尿道等離子雙極電切與恥骨上小切口治41例前列腺增生癥并膀胱結石。結果:手術時間40~110min, 平均55min,術后3d拔造瘺管, 第5~6天拔除尿管,排尿通暢, 無電切綜合征(TURS)、大出血等并發癥,住院時間7±1.5天。數字疼痛評分0~6,平均3.5。結論：等離子體雙極電切結合恥骨上小切口是治療前列腺增生并膀胱結石的一種快速、安全有效、微創的手術方法,值得在基層醫院推廣。
ObjectivesTo evaluate the effects of Q-syte separating film needleless closed transfusion connector in flushing chamber of three-cavity urethral catheter.MethodsTo retrospectively analyze the patients who underwent transurethral resection of bladder tumor for non muscle-invasive bladder cancer from January 2015 to July 2016 in Zhongnan Hospital of Wuhan University. After terminating the continuous bladder irrigation, the observed group used Q-syte separating film needleless closed transfusion connector to seal the flushing chamber of three-cavity urethral catheter, and control group used conditional approach to connect drainage bag. The degree of comfort and satisfaction of patients, urinary tract infection, time of stopping bladder irrigation and bladder perfusion time between two groups were assessed.ResultsA total of 88 patients were included involving 63 (72%) males and 25 (28%) females with a mean age of 60.2±4.7 years. There were no significant differences between two groups in age, gender, BMI, and complications (P>0.05). Compared to control group, case group had higher level of comfort degree (mild discomfort: 86.4% vs. 25.0%, P<0.001; moderate discomfort: 13.6% vs. 52.3%, P<0.001; severe discomfort: 0.0% vs. 22.7%, P=0.001), satisfaction degree (97.9±2.1 vs. 84.5±3.9, P<0.001), and lower rates of urinary tract infection (11.4% vs. 29.5%, P=0.034). In addition, the case group spent shorter time in terminating bladder irrigation (50.48±1.78 vs. 207.74±5.41, P<0.001) and bladder perfusion (141.47±3.25 vs. 205.35±5.17, P<0.001). All differences were statistical significance.ConclusionsApplication of Q-syte separating film needleless closed transfusion connector for sealing flushing chamber of three-cavity urethral catheter after continuous bladder irrigation could promote the degree of comfort and satisfaction of patients, and decrease the rate of urinary tract infection, as well as the working efficiency of health care professionals.