ObjectiveTo assess the feasibility of seeding adipose-derived stem cells (ADSCs) onto bladder acellular matrix grafts (BAMGs) for bladder reconstruction in a rabbit model.MethodsAutologous ADSCs were isolated,expanded and identified by flow cytometry.In the experimental group,ADSCs were seeded onto BAMGS for reconstructing bladder defects in 12 male rabbits.Unseeded BAMGs were used for bladder reconstruction in the control group of 12 rabbits.Cystography was performed at 24 weeks after grafts implantation.Following cystography,the animals were scarified and grafts were harvested； H&E and immunohistochemical staining were performed with cytokeratin AE1/AE3,smooth muscle α-actin and S-100 markers.ResultsFlow cytometry demonstrated that the ADSCs expressed CD90,CD44,CD105,CD166 and CD34,but not CD45 or CD106.The cells demonstrated good biocompatibility with BAMGs.At 24 weeks,in the experimental group,the reconstructed bladders reached a mean volume of (94.68 ± 3.31 )％ of the precystectomy bladder capacity.Complete regeneration of smooth muscle and nerve tissue was evident.Regenerated SMCs,urothelium and nerve cells stained positively for α-smooth muscle actin,AE1/AE3 and S100.In the control group,the mean bladder volume was (69.33 ± 5.05 )％ of the pre-cystectomy volume.Histologically,the control group was characterized by multi-layered urothelium without evidence for organized muscle or nerve tissue.Conclusion The tissue engineering bladder constructed by ADSCs and BAMG can be used as an ideal biomaterial to replace and repair the bladder.

AIM: The present study was to investigate the time course of ?-adrenoceptor desensitization and changes in the calcium transient in rat ventricular myocytes following chronic hypoxia. METHODS: With the spectrofluorometric method, the intracellular calcium( i) transient and its response to ?-adrenoceptor stimulation were determined in the single right ventricular myocytes, loaded with Fura-2.RESULTS: After 2-3 weeks of chronic hypoxia, the amplitudes of electrically induced i transient and caffeine-induced i transient started to decrease and the duration of i transient prolonged. The enhanced electrically induced i transient evoked by isoprotrenol was also decreased. After 3 or 4 weeks of chronic hypoxia, these changes aggravated gradually. After 8 weeks of chronic hypoxia, the changes of all these parameters were convalescent, meanwhile there was no significant difference compared with that of 4 weeks group. CONCLUSIONS: After 2-4 weeks of chronic hypoxia, the ?-adrenoceptor desensitization occurs, the underlying mechanism is related to the decreased function of L-type of calcium channel, ryanodine receptor-operated calcium channel and calcium ATPase, which is responsible for the decreased cardiac functionl. At 8 weeks of hypoxia, the heart is in adaptation and compensatory process.

Objective To evaluate the feasibility and efficiency of endoluminal catheter-based ultrasonography on diagnosis of upper urinary tract diseases. Methods From January 1997 to December 2006,the clinical data of upper urinary tract of 68 cases who examined by endoluminal uhrasonography with a 10 MHz miniature ultrasound transducer and a 8 F outer catheter were reviewed.There were 30 males and 38 females aged 17-81 years with an average of 58 years.The renal pelvis and its surrounding structures were examined in 48 cases and the ureter and its surrounding structures were examined in 20 cases.The total of 73 sides had experienced by this technique.The accuracy of diagnosis with endoluminal catheter-based ultrasonography was evaluated by the pathology and clinical follow-up. Results The total success rate of the ultrasound probe introduced into ureter was 91%in 68 cases.In the pelvis group it was 96%(46/48)and the ureter group 80%(16/20).In renal pelvis abnomalities,the detectable rate of abnormal by ultrasonography was 89%(41/46).The total detectable rate in ultrasound probe group was significantly higher than that in other equipment groups(P＜0.01).The detectable rate of ultrasonography on ureter disease was 75%(12/16). Conclusion Endoluminal catheter-based ultrasonography is an effective,safe technique for the diagnosis of diseases in the upper urinary tract.

BACKGROUND:Bladder acellular matrix graft (BAMG) is frequently used for domains of tissue engineering scaffold due to its great biocompatibility and cell adhesion. OBJECTIVE:To verify the biological characteristics of BAMG after frozen and lyophilized processing. DESIGN,TIME AND SETTING:A biocompatibility experiment was performed at Shanghai Tissue Engineering Research and Development Center and Experimental Animal Department of the Sixth People's Hospital of Shanghai between May and November 2008. MATERIALS:Two New Zealand rabbits were used in this study for BAMG preparation. METHODS:After midsection of rabbit bladder,mucous membrane of urinary bladder was isolated and dipped in three-distilled water for 24 hours. Thereafter,the samples were incubated with acellular solution containing 0.1% Triton X-100 and 0.15% aqueous ammonia for 14 days. The culture medium was changed regularly. The samples in the control group were stored in 75% ethanol,while samples in the experimental group were frozen for 24 hours at -80 ℃,vacuum-dried for 24 hours,and stored in 75% ethanol. MAIN OUTCOME MEASURES:Biological characteristics of BAMG were detected using hematoxylin and eosin staining,Masson staining,and scanning electron microscopy; biological characteristics were compared between the two groups using cell adhesion test,MTT assay,and subcutaneously embedding test. RESULTS:Hematoxylin and eosin staining and Masson staining revealed that no residual cells were detected in the BAMG,and collagen was intact. Scanning electron microscopy demonstrated that cells exhibited a slit-shaped structure mainly containing collagen which was beneficial for cell adhesion. Mechanical test revealed that the BAMG after frozen and lyophilized processing not only reserved the mechanical properties of the raw BAMG,but also had a great elongation. MTT assay confirmed that cytotoxicity was grade 0,and BAMG had a good compatibility to smooth muscle cells. After subcutaneously embedding for one month,BAMGs had good adhesions to subcutaneous tissues,and muscular adhesion and vascular proliferation were observed. CONCLUSION:BAMG after frozen and lyophilized processing reserves original biocompatibility and has great elongation; therefore,it will become a useful and ideal biomaterial for tissue engineering scaffold.

Objective To investigate the feasibility of constructing tissue engineering urethral substituted graft using human lingual keratinocytes and natural derived scaffold.Methods From Oct.2009to Jan.2010,ten patients with anterior urethral stricture were enrolled in this study.A 0.5 × 0.8 cm lingual mucosa was harvested during the operation.Lingual keratinocytes were then isolated and cultured from the mucosa.AE1/AE3 antibody was used to identify the lingual keratinocytes.Keratinocytes were collected and seeded onto three types of scaffold including dehydrated BAMG,liquid stored BAMG and 4-layer SIS product,with the density of 1 × 107/ml at passage three.After being cultured for seven days in vitro,H&E staining and Electronic Scan Microscopy were used to evaluate the compound matrixes.Results No complication occurred in the patients after operation.The primary passage of confluence lingual keratinocytes appeared in a typical cobblestone shape after being cultured for 14 days in vitro.The proliferation rate of these cells increased rapidly during the three passages.However,it decreased significantly in the 4th passage.H&E and Electronic Scan Microscopy examinations showed that few cells grew on the surface of the liquid stored BAMG.Nevertheless,multiple keratinocytes layers could be seen in dehydrated BAMG and 4-layer SIS.Meanwhile,cellular infiltration could be observed in SIS sections.Conclusions Human lingual keratinocytes could be the alternative of seeding cells for constructing tissues for engineering the urethra.These cells exhibited good compatibility and are adhesive to the SIS or BAMG.The compound matrix,which used human lingual keratinocytes and natural derived scaffold,may meet the clinical need of urethral disease in the future.

ObjectiveTo evaluate the efficacy,safety and clinical value of anterior urethral fusion to treat diphallia.MethodsA 17-year-old male patient with complete penile diphallia was treated in March 2011.The physical examination showed two completely separated mature penis; urethrography,and urethroscope showed the two urethras were completely separated and entered the bladder respectively.Two anterior urethras were incised respectively at ventral sides ( from the meatus to bulbar urethra) and then two incised anterior urethras were splintered by a continuous suture with 4-0 polyglactin sutures in side and side.Two penises were splintered into one.The nocturnal penile tumescence test and the penis vibration thresholds were assessed before,and after surgery..ResultsThe patient was followed up for 12 months.There was no hematoma,urinary fistula,urinary incontinence and other complications postoperatively.The penis was recovered with normal appearance; urination and morning erection of penile were normal.6 months after surgery,there was no significant difference in the nocturnal penile tumescence and penis vibration thresholds detection ( P ＞ 0.05).ConclusionThe operation of anterior urethral fusion is a safe and efficient surgical treatment to treat partial complete penile diphallia with less complication and no worse effect on urination and erection of penile.

Objective To investigate and assess the best seeding method for constructing three dimensional urethral tissue in vitro. Methods High speed agitation decellular method was used for preparing the porcine acellular corporous spongiosum matrix (ACSM). Before seeding, the matrix was sterilized via soaking compound iodine solution. Rabbit tongue epithelial cells and cavernosal smooth muscle cells were isolated and cultured. Three different groups of seeding method was used in this study. Group A (sandwich seeding group): The smooth muscle cells and epithelial cells were seeded onto the different side of ACSM by static method. Group B (injection seeding group): The smooth muscle cells were injected into the scaffold. Then, epithelial cells were seeded onto the urethral surface of ACSM by static method. Group C (agitation seeding group): The smooth muscle cells were seeded into the scaffold by agitation method. Then, epithelial cells were seeded onto the urethral surface of ACSM by static method. After being seeded, all matrixes were cultured in vitro for 14 d. HE and immunoassay staining were used to examine the results of seeding. Results Looser matrix was obtained after using high speed agitation decellular method. Compound iodine solution could not only sterilize efficiently but also reserve the original structure of biomaterial. An intact epithelial cellular layer onto the surface of scaffold could be observed in HE staining section after 14 d culturing in vitro.Few smooth muscle cells could be found in big space of biomaterial in group A. In group B, smooth muscle cells were restrained in some regions of the matrix. Smooth muscle cells were well distributed into the scaffold in group C. Conclusions After using high speed agitation decellular method, an ideal matrix with three dimensional structure can be obtained. Combined with agitated seeding method, three dimensional urethral tissue can be constructed.

ObjectiveTo investigate the etiology and management of male urethral stricture at 8 medical centers in China during the period from 2004 to 2009 years,and to investigate whether there were any changes in etiology and management of urethral stricture with time change.MethodsThe database on 3455 male patients with urethral stricture who underwent treatment at 8 medical centers in China between January 2004 and December 2009 were prospectively collected.The databases were analyzed for possible cause of stricture and treatment techniques for urethral stricture,and for the changes in etiology and management with time change.ResultsThere were 3455 operations for urethral stricture during the study period.The main causes of urethral strictures were traumas in 1833 patients (53.05％),among which pelvic fractures were in 1327 (38.41％) and perineal trauma in 506 (14.65％).The second cause was iatrogenic causes in 1181 patients (34.18％ ),among which transurethral operations or examinations were in 602 (17.42％),hypospadias surgery in 291 (8.42％) and urethral catheterization in 164 (4.75％ ).Less common causes were urethritis in 201 patients (5.82％),lichen sclerosus in 149 (4.31％),undefined in 91 (2.63％).The treatments of urethral strictures were endourological surgery including internal urethrotomy and dilation and open urethroplasty including end-to-end urethroplasty and the substitude urethroplasty etc.The ratios of using various techniques in total number of patients were obviously different by time.The most application technique for treatment of urethral stricture was endourological surgery ( 709 ) during 2004 -2006 and occupied 52.67％ in total number of patients.It was gradually decreased during 2007 -2009 (726) and only occupied 34.42％ (P ＜0.01 ).Open urethroplasty gradually increased during 2007 -2009 ( 1243,58.94％ ) compared with the first three years (563,41.83％ ) (P ＜ 0.01 ). Conclusions During the recent years there was an increase in the incidence of urethral stricture being trauma and iatrogenic causes.The main treatments of urethral strictures were endourological surgery and open urethroplasty.Endourological surgery was significantly decreased in total number of patients,while open urethroplasty were significantly increased during the late three years.

Objective: To explore the value of magnetic resonance imaging (MRI)-based whole tumor texture analysis in differentiating borderline epithelial ovarian tumors (BEOTs) from FIGO stage I/II malignant epithelial ovarian tumors (MEOTs). Materials and Methods: A total of 88 patients with histopathologically confirmed ovarian epithelial tumors after surgical resection, including 30 BEOT and 58 MEOT patients, were divided into a training group (n = 62) and a test group (n = 26).The clinical and conventional MRI features were retrospectively reviewed. The texture features of tumors, based on T2-weighted imaging, diffusion-weighted imaging, and contrast-enhanced T1-weighted imaging, were extracted using MaZda software and the three top weighted texture features were selected by using the Random Forest algorithm. A non-texture logistic regression model in the training group was built to include those clinical and conventional MRI variables with p value < 0.10. Subsequently, a combined model integrating non-texture information and texture features was built for the training group. The model, evaluated using patients in the training group, was then applied to patients in the test group. Finally, receiver operating characteristic (ROC) curves were used to assess the diagnostic performance of the models. Results: The combined model showed superior performance in categorizing BEOTs and MEOTs (sensitivity, 92.5%; specificity, 86.4%; accuracy, 90.3%; area under the ROC curve [AUC], 0.962) than the non-texture model (sensitivity, 78.3%; specificity, 84.6%; accuracy, 82.3%; AUC, 0.818). The AUCs were statistically different (p value = 0.038). In the test group, the AUCs, sensitivity, specificity, and accuracy were 0.840, 73.3%, 90.1%, and 80.8% when the non-texture model was used and 0.896, 75.0%, 94.0%, and 88.5% when the combined model was used. Conclusion MRI-based texture features combined with clinical and conventional MRI features may assist in differentitating between BEOT and FIGO stage I/II MEOT patients.

OBJECTIVE To provide scientific basis for establishing the evaluation index system of professional and specialized drug inspectors in China. METHODS Through the method of literature research， group discussions and behavioral event interview， combined with the practice situation of drug inspection at home and abroad， in accordance with the requirements of Chinese laws and regulations and department rules， the evaluation indexes for drug inspectors were extracted and screened. In addition， the Delphi method was used to conduct correspondence with experts in relevant fields so as to determine the evaluation index system of drug inspectors. RESULTS & CONCLUSIONS Finally， 62 experts completed two rounds of anonymous online questionnaires， including 29 experts in the first round and 33 experts in the second round. The questionnaire recovery rates were 96.67% and 94.29% respectively， and the expert authority coefficients were 0.81 and 0.79， indicating a high level of authority among experts. After expert consultation， evaluation index system for professional and specialized drug inspectors had been constructed， including five first-level indexes （personal characteristics， professional knowledge， professional skills， professional behavior and professional ethics） and their corresponding 27 second-level indexes. The average score of the importance of second-level indexes was 4.224- 4.879， the full score ratio was 81.8%-100%， the coefficient of variation was 0.067-0.177， and Kendall’s W was 0.643 （P＜0.001）. It indicates that this evaluation index system is relatively reliable and is expected to become a management tool for promoting the construction of professional and specialized drug inspector team and ensuring the quality of drug inspection.