1.Effects of a novel inhibitor of nitric oxide synthase on chronic arterial allograft rejection in rats
Jun OUYANG ; Duangai WEN ; Cunying YAN
Chinese Journal of Organ Transplantation 2005;0(10):-
Objective To evaluate the effects of a novel inducible nitric oxide synthase (iNOS) inhibitor (FR260330) in prevention of chronic rejection in a model of rat aortic allograft and to investigate the mechanism of the arterial wall lesion of chronically rejecting solid organ grafts. Methods Male Lewis (LEW, RT1~ l) rats received male ACI (RT1~ a) aorta allografts or LEW aorta isografts. Seven groups (n≥12) were involved in this study. FR260330 and/or tacrolimus were administered orally for 14 or 90 days according to protocol. The degree of intimal proliferation of graft aorta was determined by a computerized image system.Results Both low and high doses of FR260330 or tacrolimus treated grafts showed significantly decreased intima/(intima+media) ratios at day 90 compared with placebo controls. Combined therapy of low-dose of FR260330 with low-dose of tacrolimus produced a significant decrease of intima/(intima+media) ratios with intact endothelium as compared with placebo controls. Anti-?-actin immunohistochemical staining demonstrated that one of the mechanisms of intimal proliferation was related to migration of vascular smooth muscle cells. The intima in iNOS inhibition groups was more smooth than in placebo control group and low-dose FK506 treated groups.Conclusions A selective inhibitor of nitric oxide synthase, FR260330 plays a protective role in chronic aortic allograft rejection in the rat. Combined therapy of low-dose of FR260330 with tacrolimus produces significant protection of immune injury and may serve to improve long-term graft survival and function.
2.Application of highly selective alpha 1-blockers after extracorporeal shock wave lithotripsy for ureteral stone
Kelin DU ; Jun OUYANG ; Caibin FAN ; Duangai WEN
Chinese Journal of Postgraduates of Medicine 2011;34(8):6-9
Objective To investigate the application of highly selective alpha 1-blockers in treatment of ureteral stone after extracorporeal shock wave lithotripsy (ESWL). Methods One hundred and twenty patients with ureteral stone who accepted ESWL were divided into three groups by random digits table,each 40 cases. Tamsulosin group received tamsulosin (0.4 mg,once daily) after ESWL,doxazosin group received doxazosin (4 mg,once daily) ,control group were given no ureteral smooth musclar relaxant served. All patients were observed for 2 weeks. Results During the 2 weeks, only 4 patients withdrew due to adverse drug reactions. In tamsulosin group and doxazosin group, the stones expulsion rate [89.7%(35/39), 83.8%(31/37) respectively] were significantly higher than control group [65.0%(26/40)] (P<0.05), the expulsion time [(3.1-1.2), (3.7 ± 1.4) d] were significantly lower than control group [(6.5 ±1.1) d] (P <0.05),the incidence of renal colic [12.8%(5/39), 21.6%(8/37)] and the stone street formation rate [7.7% (3/39), 13.5% (5/37)] were significantly lower than control group [45.0% (18/40) and 40.0% (16/40)] (P < 0.05). But there was no significant difference between tamsulosin group and doxazosin group (P > 0.05). Orthostatic hypotension occurred in 1 patient in tamsulosin group, but 7 patients experienced orthostatic hypotension in doxazosin group,the difference was significant (P < 0.05).Conclusions Highly selective alpha 1-blockers can improve the stone-free rate of ureteral stone after ESWL,reduce expulsion time,decrease renal colic rate,and it is safe and tolerated. It can be regarded as an auxiliary clearance method after ESWL for ureteral stone.
3.Comparative evaluation of efficacy of tamsulosin versus tolterodine for the management of lower ureteral stones
Yunyan WANG ; Jianquan HOU ; Duangai WEN ; Jun OUYANG
Chinese Journal of Postgraduates of Medicine 2010;33(12):21-23
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.
4.Expression of Bmi-1 and p16 gene in transitional cell carcinoma of bladder
Qingsong ZHAO ; Ye LIU ; Zhanpeng LU ; Duangai WEN ; Jun HE
Chinese Journal of Postgraduates of Medicine 2014;37(11):4-6
Objective To detect the expression of Bmi-1 and p16 gene in transitional cell carcinoma of bladder(TCC) tissue and explore its clinical significance.Methods The expression of Bmi-1 and p16 gene were detected by real-time quantitative polymerase chain reaction in 61 cases of TCC tissue and 12 cases of normal bladder tissue.Results The expression of Bmi-1 gene in TCC tissue was significantly higher than that in normal bladder tissue (0.242 ± 0.129 vs.0.031 ± 0.011),and the expression of p16 gene was significantly lower than that in normal bladder tissue (0.059 ± 0.021 vs.0.165 ± 0.029),there was significant difference (P < 0.05).The expression of Bmi-1 and p16 gene were highly correlated with pathological grades,clinical stages and tumor recurrence (P < 0.05 or < 0.01).But there were not correlated with age and gender (P > 0.05).There was a negative correlation between the expression of Bmi-1 gene and p16 gene in TCC tissue(rs =-0.714,P< 0.05).Conclusions Bmi-1 gene high expression and p16 gene low expression may be involved in the occurrence and development process of TCC.Bmi-1 may decrease the expression of p 16 gene in some ways,and then lead to the occurrence and development of TCC.
5.Construction and identification of survivin-targeting siRNA-expressing plasmid
Xiaolin WANG ; Jianquan HOU ; Jun HE ; Shenmin YANG ; Duangai WEN
Chinese Journal of Clinical Laboratory Science 2006;0(01):-
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.
6.Significant prolongation of renal allograft survival by delayed combination therapy of FK778 with tacrolimus in non human primates
Jun OUYANG ; Chunyin YAN ; Duangai WEN ; Al ET ;
Chinese Journal of Organ Transplantation 2003;0(05):-
Objective To investigate the effectiveness of malononitrilamide 715 (FK778) in combination with tacrolimus in prevention of acute renal allograft rejection in Vervet monkeys. Methods Eleven groups ( n ≥4/group) were involved in this study. FK778 and tacrolimus were administered orally for 60 days according to protocol. Proliferation assay was used to evaluate the effect of FK778 plus tacrolimus on monkey lymphocytes, after activation with T or B cell specific mitogens. Results Naive controls rejected renal graft with a median survival time (MST) of 8.0 days in group 1. When recipient monkeys were treated with tacrolimus 1.0 mg?kg -1 ?d -1 in group 2 or FK778 2.5 mg?kg -1 ?d -1 in group 3, the MST was 16.0 days ( P = 0.001 ) and 11.0 days ( P = 0.266 ), respectively. Combination therapy of these two agents at the same doses immediately after transplantation resulted in a MST of 25.0 days ( P = 0.016 ) in group 4. When tacrolimus was initiated immediately after transplantation and FK778 treatment was delayed until day 7 after surgery in group 5, recipient survivals were significantly prolonged to 38.0 days ( P = 0.02 ). These results were repeatable when FK778 5.0 mg?kg -1 ?d -1 ( 9.0 days, P = 0.544 in group 6) was combined with tacrolimus 1.0 mg?kg -1 ?d -1 immediately after transplantation ( 8.0 days, P = 0.339 ) in group 7, or when FK778 was delayed 7 days ( 60.0 days, P = 0.002 ) in group 8. Furthermore, it was also repeatable when FK778 10 mg?kg -1 ?d -1 was combined with tacrolimus 1.0 mg?kg -1 ?d -1 with a 7 day delay. Proliferation assay in the combination groups revealed that 88.8 % (8/9) produced additive to synergistic effects in B cells, while 66.6 % (6/9) produced moderate antagonistic effects in T cells. Conclusion A significant prolongation of renal allograft survival was produced when FK778 administration was delayed by 7 days combined with tacrolimus in Vervet monkeys. And the combination of FK778 with tacrolimus in vitro produces synergistic inhibition on B cells proliferation, but not on T cells.
7.Gene expression of PCA3 in peripheral blood and urine and the significance of urine PCA3 score indiagnosis of prostate cancer
Longya LIU ; Duangai WEN ; Jun HE ; Jianquan HOU ; Jiannong CEN ; Jinxian PU
Chinese Journal of Urology 2012;33(4):278-281
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.
8.Effect of exogenous antigen on expression of major histocompatibility complex class I chain-related gene A in endothelial cells***☆
Yunyan WANG ; Jianquan HOU ; Jun HE ; Xiaoni YUAN ; Jianglei ZHANG ; Duangai WEN
Chinese Journal of Tissue Engineering Research 2011;15(53):9999-10002
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.
9.Value of renal parenchymal volume and thickness by non-contrast spiral CT in evaluating differential glomerular filtration rate for chronic obstructed kidneys
Gang LI ; Quanqi LIU ; Jinxian PU ; Chunyin YAN ; Jin ZHANG ; Weiguo CHEN ; Jianquan HOU ; Duangai WEN
Chinese Journal of Urology 2011;32(7):442-445
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.
10.Effect of transurethral feedback microwave thermotherapy in high risk patients with benign prostate hyperplasia
Yuhua HUANG ; Chunyin YAN ; Duangai WEN ; Jianquan HOU ; Jinxian PU ; Yangjun OU ; Gang LI ; Xiang DING
Chinese Journal of Urology 2010;31(2):113-115
Objective To evaluate the effect of transurethral feedback microwave thermotherapy with the ProstaLund CoreTherm Device(PLFT) in high risk patients with benign prostate hyperplasia (BPH). Methods Sixty-six high risk patients diagnosed with BPH, including aged ≥80 in 32 pa-tients, hypertension in 31 patients, diabetes in 5 patients, heart failure in 8 patients, chronic obstruc-tive pulmonary disease in 8 patients, cerebral infarction in 11 patients, fracture, amputation or joint stiffness unsuitable for lithotomy position in 3 patients, abnormal blood coagulation in 4 patients, pan-creatitis in 2 patients, cardiac arrhythmia in 6 patients and malignant tumor in 3 patients, were treated with PLFT using individual power at urethral local anesthesia, resulting in coagulation necrosis in 15%-30% of prostate tissue around urethra. Meanwhile, real-time monitoring the temperature of prostate and the tissue around it was used. All patients were evaluated by comparing volume of pros-tate, maximal urinary flow (Q_max), international prostate symptom score (IPSS) and quality of life questionnaire (QOL) in pre-treatment and three months after respectively. Results All of patients well tolerated PLFT. There was bleeding lightly, infection lightly and temporary incontinence. There was no severe surgical complication. After three months, the volume of prostate reduced from 62. 2 ml to 44.5 ml; IPSS decreased from 23. 4 to 11.7; QOL decreased from 4.5 to 2.4; Q_max rised from 4, 2 ml/s to 11.2 ml/s. All differences reached significance. Conclusion PLFT is one of effective and safe treatments for patients with BPH especial BPH complicating with severe conditions.