BACKGROUND: Studies on substitute cells of bladder smooth muscle cells are in the early period at present. Mesenchymal stem cell transplantation is an ideal method. Human amniotic mesenchymal cells can differentiate into cardiomyocytes and nerve growth factor and promote local structure repair. OBJECTIVE: To investigate the differentiation of human amniotic mesenchymal cells into smooth muscle cells following transplantation, and the effects on bladder muscle layer. DESIGN, TIME AND SETTING: The cytology in vitro study was performed at the Japan Xinzhou University from October 2006 to April 2007. MATERIALS: Human amnion was obtained from healthy full-term puerperal. A total of 18 clean female Sprague Dawley rats were randomly and equally assigned into normal bladder cell transplantation group, frostbite bladder control group, and frostbite bladder cell transplantation group. METHODS: Amnion was cut and mixed in DMEM containing trypsin. Epithelial cells were removed prior to tissues were incubated in DMEM, supplemented with collagenase and DNA enzyme. Human amniotic mesenchymal cells were harvested for use. Posterior vertex of urinary bladder was frozen using a -70 ℃ iron rod in rats of the frostbite bladder control group and frostbite bladder cell transplantation group. Hematoma appeared at the frostbite region of the bladder three days later. 100 ?L DMEM was injected into the hematoma of the rats of the frostbite bladder control group, while an equal volume of human amniotic mesenchymal cell suspension (105 cells) was injected into the hematoma of the rats of the frostbite bladder cell transplantation group. Human amniotic mesenchymal cell suspension was implanted into the normal rat bladder in the normal bladder cell transplantation group. After three weeks, the bladder tissue including partial urethra was used for bladder sample preparation. MAIN OUTCOME MEASURES: Bladder smooth muscle repair was observed using hematoxylin-eosin staining. The differentiation of human amniotic mesenchymal cells in the wall of urinary bladder was detected using fluorescence immunohistochemistry. RESULTS: Compared with the normal bladder cell transplantation group, normal wall structure of urinary bladder, few fibrosis and good proliferation of smooth muscle cells were detected in the frostbite bladder cell transplantation group, while the wall of urinary bladder was slowly repaired, and disorder muscle structure, fibrosis, scar-like shape, and inflammatory cells were found in the frostbite bladder control group. Three weeks later, human amniotic mesenchymal cells were not seen in the normal bladder tissue. A large number of human amniotic mesenchymal cells was lived and some of them had differentiated into the smooth muscle cells in the frostbite bladder cell transplantation group. CONCLUSION: Human amniotic mesenchymal cells had the potential to differentiate into bladder smooth muscle cells and promoted self-repair of the wall of urinary bladder.

Objective To compare the efficacy of tamsulosin and tolterodine for the adjunctive expulsive therapy in patients with lower ureteral stones.Methods A total of 160 patients with lower ureteral stones(4-10 mm)were included in the study.The patients were divided into 4 groups by block randomization.Group Ⅰ patients received tamsulosin 0.4 mg/d;group Ⅱpatients received tamsulosin 0.4 mg/d plus tolterodine 2 mg(twice a day);group Ⅲ patients received toherodine 2 mg(twice a day);and group Ⅳpatients served as controls.All patients were observed for 2 weeks.Remits The stone expulsion rate of group Ⅰ,Ⅱ,Ⅲ,Ⅳ was 76.9%(30/39),70.0%(28/40),46.2%(18/39)and 42.1%(16/38),respectively.The stone expulsion rate in group ⅠandⅡwas,higher than that in group Ⅲand Ⅳ(P＜0.05).The expulsion time of group Ⅰ,Ⅱ,Ⅲ,Ⅳ was(5.3±2.5),(6.4±2.2),(10.7±1.8),(12.8±3.4)d,respectively,with significant differences between group Ⅰ,Ⅱ andⅢ,Ⅳ,between groupⅢand Ⅳ(P＜0.05).Almost all of the patients tolerated the expulsive therapy and only 4 patients withdrew from treatment.No obvious side effect occurred.Conclusion The use of tamsulosin for the expulsion of lower ureteral stones is effective and safe;however,the use of tolteredine provides no additional advantages.

Objective To explore the relationship between dyslipidemia and urinary stone formation.Methods The clinical data of 427 patients diagnosed with urolithiasis in our hospital during January 2015 to May 2016 were collected.Among them,272 men accounting for 63.7%,155 women account for 36.3%.The average age were 53 (43-63).218 cases were kidney stones,accounting for 35.6%;158 cases were ureteral calculi, accounting for 25.8%; 23 cases were kidney stones with ureteral calculi, accounting for 3.8%;28 cases were bladder calculi, accounting for 4.6%.At the same time,950 age and gender matched healthy controls were collected.Among them,570 men accounting for 60%, 380 women account for 40%.The average age were 53 ( 48-60 ).All of them had undergone renal ultrasound to excluded urolithiasis.The difference between lipid level and incidence of dyslipidemia in patients with urolithiasis were observed. The relationship between lipid level and serum UA, urine pH and stone composition was evaluated and analyzed with logistic regression.Results The average serum TC,TG,HDL-C levels of patients with urolithiasis were 4.34mmol/L, 1.38mmol/L, 1.25mmol/L, which levels were 4.32mmol/L,1.09mmol/L,1.40mmol/L in healthy controls.Significant difference were seen between the two groups ( P <0.05 ).The average serum LDL-C was 2.63mmol/L in patients with urolithiasis and 2.65mmol/L in healthy controls.No difference were seen between the two groups ( P=0.241).31.6% of patients with urolithiasis had different degree of dyslipidemia.The average serum UA levels,urine pH value of patients with dyslipidemia were 392μmol/L and 5,which were 339μmol/L and 6 in patients with normal lipid level.Significant difference were seen between the two groups ( P<0.05 ).Among 193 patients who had stone composition analysis, 130 cases had normal lipid level, accounting for 67.4%; 63 cases had dyslipidemia, accounting for 32.6%.In 63 patients with calculi who had dyslipidemia,31 cases had uric acid calculi,accounting for 49.2%.In 130 patients with calculi who had normal lipid level,40 cases had uric acid calculi, accounting for 30.8%.Significant difference were seen between the two groups ( P =0.013 ).Multivariate logistic regression showed TG was the independent risk factor of urinary stone formation ( P=0.001).Conclusion Dyslipidemia is closely related to urinary stone formation,especially concerning the for hypertriglyceridemia.

Objective To construct survivin-targeting siRNA-expressing plasmid.Methods DNA sequence correspond to siRNA targeting survivin was designed and synthesized,and cloned into plasmid pRNAT-U6.1/Neo to produce surviving-targeting plasmid.Two oligos in the template with cohesive BamHⅠ and HindⅢ sites were prepared and annealled to form the insert fragment for siRNA vector.The vector was cut with BamHⅠ and HindⅢ and ligated with the insert fragment using T4 ligase.The recombinant vector was confirmed by restriction digestion and DNA sequencing,and then was transfected into T24 cells with Lipofectamine TM2000 and the expression of survivin was detected by real-time quantitive PCR.Results DNA sequencing for the PCR product showed that the recombinant vector pRNAT-U6.1/Neo-survivin was successfully constructed without any base pair mutation.The plasmid pRNAT-U6.1/Neo-survivin could efficiently reduce the expression of survivin and confer G-418 resistance in T24 cells.Conclusion The siRNA-expressing plasmid which were successfully constructed and transfected into T24 cells in this study may facilitate the application of RNA interference technique,and lay foundation for further studies on the function of survivin.

Objective To improve the diagnosis and treatment of upper urinary tract hematuria. Methods A total of 121 patients with hematuria who had undergone B-utrasonography,KUB plus IVU,CT and cystoscopy were suspected of hematuria from upper urinary tract.For these cases ureterorenoscopy was performed to establish the diagnosis and to conduct specific therapies. Results The diagnostic accordance rate was 92%(111/121).Among these cases,ureteral small stones in middle and lower segments were found in 45 cases;renal pelvis and ureteral tumors in 32 cases;renal hemorrhagic diseases in 19 cases and ureteral polyps in 15 cases.19 cases who had renal hemorrhagic diseases and 10 who had no definite lesions received specific therapies were followed up for 6 to 8 months.The long-term successful rate was 79%(23/29). Conclusions The application of ureterorenoscopy for the management of upper urinary tract hematuria is quite effective and worthy of widespread application.

Objective To explore the relationship of post-transplant major histocompatibility complex class I chain-related gene A(MICA)antibody status and renal allograft function in clinical stable phase.Methods Fifty-seven patients accepted renal allografts followed up for at least 6 months were detected with the levels and specialties of MICA antibodies by Flow PRATM beads.Simultaneously,their serum ereatinine levels were tested as well.The impact of MICA antibody status on renal allograft function was assessed.Results Among the 57 patients,38 cases showed no HLA and MICA antibody.11 cases had HLA antibodies but not MICA antibody,8 cases had MICA antibodies and 3 cases had both MICA and HLA antibodies.There were 5 patients with MICA019 antibodies.3 patients with MICA027 antibodies,2 patients with MICA018 antibodies,while 1 patient with MICA004 and MICA017 antibodies,respectively.There were 9 patients with antibody positive score higher than 6,accounting 75%(9/12).Except age,there was no significant difference between patients with positive and negative MICA antibodies in the aspects of blood transfusion history,CDC,and cold ischemia time(P＞0.05).The average ages were(32.5±7.9)years for MICA antibodypositive patients and were(43.0±1 0.4)years for MICA antibody-negative patients(P=0.008).MICA antibody-positive patients without HLA antibody had higher serum creatinine level[(117.20±12.30)μmol/L]than MICA and HLA antibody-negative patients[(89.40±28.95)μmol/L,P＜0.05].Conclusions The measurement of MICA antibodies has prognostic value in the assessment of patients without HLA antibodies after renal transplantation.MICA antibody positive has clear association with chronic renal allograft function decline.

Objective To detect the gene expression of PCA3 and PSA in peripheral blood and urine simultaneously to investigate whether PCA3 combining PSA gene could become new markers for diagnosis of Pca. Methods From June 2009 to December 2009,the initial urine after prostatic massage and the peripheral blood specimens were collected from 37 patients with PCa and 68 patients with BPH that were pathologically confirmed,g patients with urinary stone were used as normal control,the expression of PCA3 and PSA mRNA of mononuclear cells in urine sediments and peripheral blood were detected by fluorescence real-time quantitative PCR,with β-actin mRNA as internal control. Results The sensitivity and specificity of the expression of PCA3 mRNA in peripheral blood for diagnosis of prostate cancer were 48.6％ and 100％ respectively.ROC curve analysis was performed for the PCA3 score and the area under the ROC curve was 0.908.Using 64.6 as the cutoff,the sensitivity was 81.1％ and the specificity was 86.8％.In group with serum tPSA value ＜4 pg/L,the positive rate and negative rate of urinary PCA3 score for diagnosing prostate cancer were 80％ (4/5) and 89.4％ (20/22) respectively.In group with serum tPSA value 4 - 10 μg/L,the positive rate and negative rate of urinary PCA3 score were 66.7％ ( 2/3 ) and 84.2％(16/19) respectively.In group with serum tPSA value ＞ 10 μg/L,the positive rate and negative rate of urinary PCA3 score were 82.8％ (24/27) and 81.5％ (22/27) respectively.The sensitivity of simultaneous detection of PCA3 mRNA in peripheral blood and urinary PCA3 score was 86.5％. Conclusions The expression of PCA3 mRNA in peripheral blood was a specific marker for the diagnosis of PCa.The simultaneous detection of PCA3 mRNA in peripheral blood and urinary PCA3 score could increase the sensitivity for the diagnosis of PCa.

BACKGROUND: Studies have demonstrated that incidence rate of acute rejection in renal transplant recipients with pre-production of major histocompatibility complex class I chain-related gene A (MICA), including parts of autoantibody, before transplantation in body, is obviously greater than that of recipients with negative antibody. OBJECTIVE: To investigate effects of exogenous antigen on MICA expression in endothelial cells. METHODS: The endothelial cells were cultured with exogenous recombinant MICA protein (group M5, M10 and M25) and heat shock protein-70 (group H5, H10 and H25) with dosages of 5, 10 and 25 μg/L, respectively, for 48 hours. Same volume of phosphate buffer saline was added into the control groups. RESULTS AND CONCLUSION: At 48 hours after induction, the expressions of MICA mRNA and protein were increased significantly in each experimental group (M5, M10 and M25) than that of the control group with significant (P < 0.05). The expression of MICA mRNA and MICA protein of group M5 and group M10 were remarkably higher than group M25 (P < 0.05); however, there was no significant difference between group M5 and M10 (P > 0.05). The expression of MICA membrane protein in the group M10 was obviously greater than that of the group M5 and M25 (P < 0.05). The level of soluble MICA (sMICA) in experimental groups (M5, M10 and M25) was decreased obviously comparing with that of the control group. These differences had statistical significances (P < 0.05). But there was no significant difference among the experimental groups (P > 0.05). However, the expression of MICA gene and sMICA level did not change after heat shock protein-70 stimulation. The exogenous MICA antigen up-regulates the expression of MICA mRNA and protein, especially increases the expression of membrane protein on the cell surface significantly, but sMICA in supernatant was dramatically decreased.

Objective To evaluate the value of renal parenchymal volume and thickness by non-contrast spiral CT in evaluating the differential glomerular filtration rate (GFR) for chronic obstructed kidneys, and to compare the correlations between the two morphologic indices of renal parenchyma and the GFR for chronic obstructed kidneys. Methods Seventy-one patients who had a diagnosis of unilateral chronic upper urinary tract obstruction were included in this analysis. (1) The renal parenchymal volume was mea-sured by non-contrast spiral CT. Both kidneys were scanned by non-contrast spiral CT. The renal parenchymal area of each section was marked manually. Renal parenchymal volume was calculated as the sum of renal parenchymal area multiplied by the width of each section. The volume percentage of obstructed kidney (%CTvol) was also calculated. (2) Renal parenchymal thickness was measured on the first and last non-contrast CT image levels from the anterior, posterior and lateral locations of the kidney that clearly contained the collecting system. The mean of these measurements was defined as the renal parenchymal thickness. The differential renal parenchymal thickness of the obstructed kidney (%CTt) was defined as the percentage of the obstructed renal parenchymal thickness to the total renal parenchymal thickness for both kidneys. GFR was determined with 99Tcm-DTPA dynamic imaging system by Gates method. The differential GFR for obstructed kidney (%GFR) was the GFR percentage of obstructed kidney to the total GFR for both kidneys. The Pearson relation test was carried out between the %CTvol, %CTt and the %GFR respectively. Results %CTvol and %CTt correlated well with %GFR in chronic obstructed kidneys among the 71 test group patients. Pearson correlation coefficient r was 0.80 (t=11.20, P＜0.05) and 0.66 (t=7.24, P＜0.05), respectively. The linear correlation equation respectively was %GFR=0.05+0.80×%CTvol (F=125.48, P＜0.05) and %GFR=0.12+0.66×%CTt (F=52.36, P＜0.05). Conclusions Renal parenchymal volume and thickness by non-contrast spiral CT might be used as clinical practical parameters to evaluate the differential GFR for chronic obstructed kidneys. Renal parenchymal volume is more accurate than renal parenchymal thickness.

Objective To investigate the value of B7-H3 in expressed prostatic secretions (EPS) in differential diagnosis of patients with inflammatory elevation of PSA in t-PSA gray zone (4-10 ng/ml). Methods One hundred and sixteen patients from the ages of 19 to 80 years (mean, 40 years) were stu-died. In the group there were 91 chronic prostatitis (CP) patients (mean age 31 years, 19-49 years), including 11 chronic bacterial prostatitis (type II) patients, 26 inflammatory nonbacterial prostatitis (IIIA) patients and 54 noninflammatory nonbacterial prostatitis (IIIB) patients. Transrectal ultrsound guided prostate biopsy was performed on 25 patients (mean age 71 years, 62-80 years) with t-PSA in gray zone (7.21±2.60 ng/ml). Five had positive results, Gleason score was 6 in two cases, 7 in two cases and 8 in one case. Twenty patients had negative results, of whom 11 patients had inflammatory cell infiltration. EPS was collected by transrectal massage, and Enzyme-linked immunosorbent assays (ELISA) were performed for B7-H3 detection. In addition, 11 normal male controls with a mean age of 30 years (24-46 years) were recruited into the study. Volunteers were excluded if they had a history of genitourinary symptoms or surgery.Results The EPS B7-H3 levels of controls, II, IIIA, IIIB groups were 49.81±11.54, 19.33±13.90, 17.67±15.76, 25.14±13.44 ng/ml, respectively. The levels of EPS B7-H3 in positive biopsy, noninflammatory negative biopsy and inflammatory negative biopsy groups were 26.30±16.32, 30.23±18.42, 10.11±5.42 ng/ml, respectively. The highest levels were found in the control group (P<0.01). Compared to the IIIB, B7-H3 levels in II and IIIA groups were significantly lower (P<0.05). There was no significantly difference between II and IIIA groups (P>0.05). The EPS B7-H3 levels in the inflammatory negative biopsy group were statistically lower than in positive biopsy and noninflammatory biopsy groups (P<0.05). But no significant differences were found among inflammatory negative biopsy, II and IIIA groups (P>0.05). Receiver operating curve (AUC=0.883, P=0.001) utilizing EPS B7-H3 levels≤16.24 ng/ml identified patients with inflammatory elevation of PSA with a sensitivity of 78.6% and a specificity of 81.8% from patients with t-PSA in gray zone. Conclusion The EPS B7-H3 detection provides a new way for differential diagnosis of patients with inflammatory elevation of PSA in t-PSA gray zone resulting in a reduction of unnecessary prostate biopsy.