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 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 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.

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 ATP content of CD4+ T lymphocytes in the diagnosis of infection and its correlation with drug concentrations in renal transplant recipients.Methods 45 renal transplant recipients were reviewed from May 2010 to October 2011.There were 33males and 12 females,aged from 21 to 58 years old.The recipients were divided into non-infection group (n =34) and infection group (n =11) according to their clinical manifestation.11 cases of infection were diagnosed by the chest X-ray,CT imaging manifestations and etiological examination,among them 5 cases were pulmonary infection,4 cases were upper respiratory infection,1 case was urinary tract infection and 1 case was perineal abscess.23 healthy volunteers were enrolled as the control group.They were detected ATP content of CD4+T lymphocytes by Immuknow method.Thetrough concentrations of the FK506 and CsA were detected by microparticle enzyme immunoassay and fluorescence polarization immunoassay,respectively.The hs-CRP concentration was detected by immunoturbidimetry.Results The ATP content of CD4+ T lymphocytes of the control group,non infection group and the infection group were (295±74) μg/L,(35± 189) μg/L and (212± 155) μg/L respectively.The levels of ATP of infection group were obviously lower than the control group and non-infection group.There were statistically differences (P ＜0.05).24 recipients were followed up dynamicly.There were 4 cases whose ATP value was lower than the postoperative average levels in 5 infection recipients.The hs-CRP concentration of infection group were (12.4±4.8) mg/L,obviously higher than the non infection group's (3.3 ± 4.7) mg/L and the control group' s (0.5 ± 0.5) mg/L.There were statistically differences (P＜0.05).The ATP content of CD4+ T lymphocytes were no significant associated with drug trough concentrations (P＞0.05).Conclusions Low ATP level after renal transplantation is a risk factor for infection recipients.Immuknow cell function assay can make up for the inadequacy of the drug concentration monitoring,reduce the risk of infection,and guide clinical immunosuppressive adjustment.

[Abstract] Objective To define the age-specific normal reference values of prostate specific antigen (PSA) and related parameters in Chinese middle-aged and elderly men.Methods From April 2007 to November 2011,serum PSAs of over 22 055 men aged more than 40 years old in our medical examination center were statistically analyzed.The men was divided into five groups by a 10-year-old interval.Total PSA (tPSA),free PSA (fPSA) and prostate ultrasound results were recorded.The free-total PSA ratio (f/t),PSA density (PSAD) and PSA velocity (PSAV) were calculated.By convention,the 95th percentile (P95)was used as the upper limit value,and the 5th percentile (P5) as the lower limit value.Results The tPSAs were positively correlated with age (r=0.349,P＜0.001).f/t was negatively correlated with age (r=-0.154,P＜0.01).Although f/t was significantly different (P＜0.001) among each age group,P5 of all groups were 0.18.PSAD was significantly different (P＜0.001) between men over and under 70 years,with P95 as 0.09 and 0.15,respectively.PSAD had a positive correlation with age (r =0.263,P＜0.01).The significant difference of PSAV raised between men over and under 60 years,with P95 as 0.21 and 0.58,respectively.PSAV was positively correlated with age (r=0.130,P＜0.01).Conclusions PSA,PSAD and PSAV are positively correlated with age,while f/t is negatively correlated with age.The normal range of f/tis 0.18-1.00 for Chinese men over 40 years old.PSAD's normal ranges are ＜0.09 and ＜0.15 in Chinese men over and under 70 years,respectively.The normal range of PSAV are ＜0.21 and ＜0.58 for Chinese men over and under 60 years,respectively.

Objective To assess the clinical significance of serum high sensitive C-reactive protein (hs-CRP) in patients undergone prostate biopsy.Methods A total of 273 consecutive patients were enrolled,aged 44-95 years (mean,69 years).All the patients underwent prostate biopsy.The pathological findings showed 96 cases with prostate cancer (PCa) and 177 cases with benign prostate hyperplasia (BPH).The difference of hs-CRP level between patients with PCa and those with BPH was analyzed.The positive prostate biopsy rate was compared between the patients with high hs-CRP level and those with normal hs-CRP level.Logistic regression was used to evaluate the effect of factors such as hs-CRP,tPSA,PSA density,prostate volume and age on prostate biopsy.Results The medians (interquartile range) of hs-CRP were 3.22 mg/L (1.22-9.84 mg/L) in patients with PCa and 1.24 mg/L (0.55-2.76 mg/L) in those with BPH,respectively,with significant difference(P＜0.05).The positive prostate biopsy rate in patients with high hs-CRP (＞ 3 mg/L)was 55％ (51/92),higher than that in those with normal hs-CRP (≤ 3 mg/L).The odds ratio of hs-CRP was larger than that of all other factors analyzed including tPSA,prostate volume and age according to the Logistic regression analysis.Conclusions Elevated serum hs-CRP level is associated with increased positive prostate biopsy.Serum hs-CRP acts as an independent factor increasing the positive prostate biopsy rate in patients undergone prostate biopsy.

Objective To research the consistency of testing results with three different antimajor histocompatibility complex class Ⅰ-related chain A(MICA) specific antibody reagents in order to evaluate their clinical application's value.Method An collaborative study of 18 laboratories was undertaken at the 16th International HLA and Irnmunogenetics Workshop.Total of 16 sera(4 batchs)were tested for anti-MICA antibodies by Luminex method with three different reagents (Kit-A,-B and -C).Result Anti-MICA antibodies were found in 15 sera,except one sera(no.S04) ; No.S10 sera showed positive results in all the laboratories.The anti-MICA antibodies were divided into MICA-G1 group (MICA01,02,07,12,17 and 18) and MICA-G2 group (MICA 04,06,08/27,09 and 19).MICA-G1 group specific antibodies were detected in 5 sera with Kit-A and-B reagent; but there were false-positive results of anti-MICA08/27 and MICA19 antibodies in this 5 sera with Kit-C.MICA-G2group specific antibodies can be detected in other 5 sera with Kit-A and-B,But the MICA specific antibodies testing gave different results with Kit-A,-B and-C in all the last 5 sera samples.Testing of MICA08/27 showed highest consistency results (86.67％,13/15) with Kit-A,-B and-C; and testing of MICA19 showed lowest consistency results (40％,6/15) with this 3 reagents.There were 80％ consistency results of anti-MICA specific antibodies in 13 sera with Kit-B.Conclusion There are the same effect to judgment positive or negative result for anti-MICA antibodies with 3 different reagents,but the results of anti-MICA specific antibodies are not the same.Therefore,it's better to use two or more reagents to test anti-MICA specific antibodies,or choose reagent with wide detection range.