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1.
Article in Chinese | WPRIM | ID: wpr-447768

ABSTRACT

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.

2.
Chinese Journal of Urology ; (12): 120-122, 2012.
Article in Chinese | WPRIM | ID: wpr-420785

ABSTRACT

Objective To evaluate the effect of transurethral feedback microwave thermotherapy with the ProstaLund CoreTherm Device (PLFT) in benign prostate hyperplasia ( BPH ) patients with high risk factors 24 months after treatment.MethodsSixty-two BPH cases with high risk factors of aged ≥ 80or complicating severe conditions of no less than one organ or system,were treated with PLFT under urethral local anesthesia.The average pre-treatment prostate volume,international prostate symptom score (IPSS),quality of life score (QOL) and maximal urinary flow (Qmax) were 62.03 ml,23.19,4.58 and 4.33 ml/s,respectively.The changes of prostate volume,IPSS,QOL and Qmax at 3 months,12 months and 24 months after treatment were analyzed.ResultsAll patients tolerated well of PLFT performed in common therapy room except lightly bleeding,minor infection and temporary incontinence.There was no severe surgical adverse event.After 3 months,the prostate volume reduced to 43.85 ml,IPSS decreased to 11.63,QOL decreased to 2.44,Qmax rose up to 11.44 ml/s; The average values were 45.10 ml,12.23,2.61 and 10.91ml/s at 12 months after treatment.The corresponding values were 45.80 ml,12.37,2.66 and 10.82 ml/s,respectively at 24 months after treatment.Compared with pre-treatment,all the parameters showed significant improvement ( P < 0.01 ).ConclusionsPLFT is one of the effective and safe treatment options for BPH patients with high risk factors.It can be safely used on day-surgery patients.The best effect appears at 3 months after treatment.

3.
Chinese Journal of Urology ; (12): 278-281, 2012.
Article in Chinese | WPRIM | ID: wpr-418690

ABSTRACT

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.

4.
Cancer Research and Clinic ; (6): 316-317,327, 2012.
Article in Chinese | WPRIM | ID: wpr-598118

ABSTRACT

Objective To evaluate the feasibility and effectiveness of bladder-preserving local resection combined with intra-artery chemotherapy for patients with T1G3 bladder cancer.Methods Thirty five cases with T1G3 bladder cancer were analyzed retrospectively. Patients were all treated by bladder-preserving local resection combined with intra-artery chemotherapy. Results Thirty five cases were followed up. The time of follow-up ranged from 7 to 116 months,and mean time was(66.0±18.3)months.The 5 year recurrence and bladder-preserving rate were 48.6 %(17/35)and 68.6 %(24/35),respectively.The overall and tumor specific survival at 5 years was 77.1% (27/35)and 82.9 % (29/35).The effect of bladder-preserving local resection plus intra-artery chemotherapy was satisfactory. Moreover, intra-artery chemotherapy had no obvious side effects.Conclusion Bladder-preserving local resection combined with intra-artery chemotherapy not only can retain normal bladder function, decrease in the recurrence, but also does not reduce the survival rate. It is an effective treatment for some patients with T1G3 bladder cancer instead of cystectomy over-treatment.

5.
Article in Chinese | WPRIM | ID: wpr-423921

ABSTRACT

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 membrane protein in the group M10 was obviously greater than that of the group M5 and M25 (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.

6.
Chinese Journal of Urology ; (12): 750-753, 2011.
Article in Chinese | WPRIM | ID: wpr-422774

ABSTRACT

ObjectiveTo evaluate the value of intraoperative frozen section examination (IFS) in the diagnosis and surgical procedures selection for renal occupying lesions. MethodsFrom January 2006 to December 2010,IFS was used in 114 men and 81 women with renal occupying lesions.The mean age was 52 years (range 17 -78).In 104,89,and 2 patients,lesions were in the right,left and bilateral kidneys,respectively.All patients underwent physical examination,129 were asymptomatic at presentation while clinical symptoms were observed in 66.The largest dimension of the tumors were 4 cm or less in 128 patients,4- 7 cm in 49,and larger than 7 cm in 18,respectively.The outcomes between IFS and postoperative routine paraffin section examination were compared.In cases with renal tumors nephrectomy or partial nephrectomy was performed.The results of IFS were compared between the 2 groups. ResultsThe sensitivity and specificity of IFS for renal malignant lesions was 96.6% and 100%,respectively.The total accuracy rate of IFS for renal occupying lesions was 97.4%.By subgroup analysis,the accuracy rate of clear cell carcinoma,papillary cell carcinoma,chromophobe cell carcinoma,sarcomatoid cancer,nephroblastoma,benign tumor and urothelial cancer was 94.3%,25.0%,16.7%,0,0,97.6% and 100.0%,respectively.Partial nephrectomy and nephrectomy were performed in 57 and 123 patients with renal tumors,respectively.The surgical procedures selection was significantly associated with the lesion size (4 cm or less for 80.7% vs 62.6%,P =0.015) and the malignant lesion diagnosed by IFS (31.6% vs 93.5%,P<0.001). Conclusion The accuracy of frozen section analysis for renal malignant lesions during surgery is reliable and significantly high,and the results can exert an important impact on surgical procedures selection.

7.
Chinese Journal of Urology ; (12): 442-445, 2011.
Article in Chinese | WPRIM | ID: wpr-416798

ABSTRACT

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.

8.
Article in Chinese | WPRIM | ID: wpr-384267

ABSTRACT

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.

9.
Chinese Journal of Urology ; (12): 113-115, 2010.
Article in Chinese | WPRIM | ID: wpr-390955

ABSTRACT

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.

10.
Article in Chinese | WPRIM | ID: wpr-389209

ABSTRACT

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.

11.
Article in Chinese | WPRIM | ID: wpr-389128

ABSTRACT

Objective To investigate the prediction of anti-human leukocyte antigen antibodies (HLA) and anti-major histocompatibility complex class I-related chain A antibodies (MICA) to the development of acute rejection (AR) and kidney allograft function. Methods Forty-one kidney transplant patients were prospectively tested for anti-HLA and anti-MICA. Thirty-seven patients were screened using Luminex/single-antigen beads to determine the HLA and MICA-specific antibody levels at 0,30,90, 180,360,720 and 1080 days post-transplantation. The patients and donors of HLA and MICA allele typing were determined by PCR-SSOP, and donor specific antibody (DSA) and non-donor specific antibody (NDSA) were identified.Simultaneously,their serum creatinine (SCr) levels and clinical data were analyzed. Results Nine patients (21.95 % ,9/41 ) had pre-existing anti-HLA and(or) anti-MICA, including 6 cases of anti-MICA,2 cases of anti-HLA, and one case of anti-MICA and anti-HLA. Nine patients had pre-existing DSA and NDSA. In the 37 patients, 6 patients (16.2% ) developed de novo anti-HLA, and 3 (8.1%) developed de novo antiMICA. In patients positive for de novo anti-HLA, the titer of antibody was gradually increased during the follow-up of three years. Four patients out of 9 patients with pre-existing antibodies were suffered from AR (44.4%); In 6 patients positive for de novo anti-HLA,three cases (50.0%) were suffered from AR; In three patients positive for de novo anti-MICA,no AR occurred (P<0.05). In two patients positive for DSA of HLAⅡ antibody detected at the third and seventh day after transplantation, the renal grafts were renovecd due to rejection. The Scr levels in patients positive for pre-existing MICA with AR were higher than in those positive for pre-existing MICA without AR at each scheduled time point during the follow-up period (P<0.05). The Scr levels in patients negative for antibodies pre-transplantation and having AR were higher than in those having no AR at each scheduled time point during the follow-up period (P<0. 01 ). The Scr levels in patients positive for de novo HLA and MICA and having AR one month following transplantation were higher than in those negative for antibodies and having no AR (P<0.01 ). Conclusion Pre-existing and de novo anti-HLA were the irnportant factors for the development of AR, but the mismatch of HLA and MICA alleles in donors and patients was primary causes for generation of de novo antibodies.

12.
Chinese Journal of Urology ; (12): 598-600, 2010.
Article in Chinese | WPRIM | ID: wpr-387202

ABSTRACT

Objective To investigate the approach and safety of minimally invasive surgical procedures treating symptomatic caliceal diverticular calculi. Methods Clinical data of 21 cases with symptomatic caliceal diverticular calculi were retrospectively reviewed. Twelve females and 9 males aged 22 to 57 years old. The average diameter of caliceal diverticulum was 3.7 cm (2.5-7.0 cm) and average diameter of calculi was 2.3 cm (0.8-3.5 cm). The patients underwent flexible ureteroscopic lithotripsy, PCNL or mPCNL, laparoscopic techniques and laparoscopy-assisted transperitoneal PCNL, respectively. Four cases underwent flexible ureteroscopic lithotripsy. PCNL (2 cases) or mPCNL(5 cases)were performed in 7 cases. Nine cases underwent laparoscopic techniques. Laparoscopyassisted transperitoneal PCNL was performed in 1 case. Results The operations were performed successfully in 21 cases. No case need to transfer to open surgery during the operation and no major complications like perforation or organic injury were noted. One case with iatrogenic arteriovenous fistula of the kidney after 1 week postoperative was cured by delayed hemorrhage 2 days later, while clinical symptoms of 2 cases with residue calculi relieved. 19 cases without residue calculi were followed up for 6 to 12 month without recurrence. Conclusions After handling indication of treatment efficiently and creating advisable therapeutic decision-making, minimally invasive surgical procedures treating symptomatic caliceal diverticular calculi appears to be effective and safe.

13.
Chinese Journal of Urology ; (12): 163-167, 2009.
Article in Chinese | WPRIM | ID: wpr-395899

ABSTRACT

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.

14.
Chinese Journal of Urology ; (12): 755-758, 2008.
Article in Chinese | WPRIM | ID: wpr-397763

ABSTRACT

Objective To study the influence of human leucocyte antigen(HLA) and major his-tocompatibility complex class Ⅰ chain-related gene A (MICA) specific antibodies on renal allograft function and graft rejective reaction by monitoring their changes from preoperative to postoperative pe-riods. Methods Twenty-seven patients with renal aliografts were tested with the specificity of anti-HLA antibodies (anti-HLA class Ⅰ and anti-HLA class Ⅱ) and anti-MICA antibodies and their posi-tive value changes by flow PRATM beads. The HLA genotype was integrated to distinguish donor specific antibody(DSA) and non-donor specific antibody(NDSA). Their serum creatinine levels and clinical data were analyzed simultaneously. Results Of the 27 patients, 22 cases accepted renal transplantation from dead bodies and 5 eases accepted from live donors. Except 1 failed patient, the other 26 patients had good functional renal allografts. Twenty-four survival patients were followed up on month 1, 3, 6 and 12 after transplantation. Seven out of 27 patients had pre-exist antibody before transplantation. Among them, 2 patients had anti-HLA antibody; 3 patients had anti-MICA antibody; 2 patients had both anti-HLA and anti-MICA antibody. Three patients with no anti-HLA and anti-MICA antibodies before transplantation created antibodies after transplantation from 3 to 6 months. One patient created NDSA after transplantation and appeared chronic rejection. There were 3 patients who had anti-MICA antibodies before transplantation. The expression levels of antibodies had changed from high to low, but the specific anti-MICA antibody had not changed during the follow-up on month 1, 3, 6 and 12 after transplantation. The patient with pre-transplantation low level of anti-HLA class Ⅱ antibody appeared acute rejection with fever and his CMV was positive as well. The patient's SCr levels changed from 171 μmol/L to 236 μmol/L after I to 3 months post-transplantation. Twenty-four patients were divided into positive and negative groups according to the specific antibody. There was significant difference of SCr levels between the 2 groups 1 month and 1 year after transplantation(P= 0.03, 0.05). Conclusions It is important to detect the specificity and positive value of anti-HLA antibodies and anti-MICA antibody regularly during the post transplantation follow-up. This will make an effective therapy for decreasing the occurrenee and development of acute or chronic rejection and hy-pofunction on renal allograft.

15.
Article in Chinese | WPRIM | ID: wpr-544101

ABSTRACT

Objective To investigate the relationship between the ADAM gene expression and unknown reason infertile patients. Methods With RT-PCR mehtod, we checked from normal group semen 30 cases and infertile group semen 30 cases in order to know the ADAM1,2,3,32 mRNA expression. Results there are the ADAM1,2,3,32 gene expression in all 30 cases of normal group whereas in 30 cases infertile patients there exists 1 case ADAM1,2,32 lacking expression, 1 case ADAM2,32 lacking expression, 1 case ADAM1 and 1 case ADAM3 lacking expression. Conclusions The lacking of ADAM1,2,3,32 may be one of the most reasons which cause the infertility.

16.
Article in Chinese | WPRIM | ID: wpr-588170

ABSTRACT

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.

17.
Article in Chinese | WPRIM | ID: wpr-541910

ABSTRACT

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.

18.
National Journal of Andrology ; (12): 595-597, 2004.
Article in Chinese | WPRIM | ID: wpr-308292

ABSTRACT

<p><b>OBJECTIVE</b>To elucidate the regulation of epidermal growth factor receptor (EGFR) expression by transforming growth factor (TGFalpha) and epidermal growth factor (EGF) in human prostate androgen-unresponsive cancer cells.</p><p><b>METHODS</b>EGFR mRNA expression and its protein level were measured by means of RT-PCR and Western blot respectively in human prostate cancer androgen-unresponsive cell lines, ARCaP and PC3, all treated with exogenous TGFalpha.</p><p><b>RESULTS</b>In the TGFalpha group, the levels of EGFR mRNA were 5.01 0.45 and 9.05 0.63 in PC3 and ARCaP respectively, significantly higher than in the control group (P < 0.05). The level of EGFR protein in PC3 treated with TGFalpha was 2.28 0.53, higher than in the control group (P < 0.05); however, the level of EGFR protein in ARCaP treated with TGFalpha was only 1.24 0.22, not different from the control (P > 0.05).</p><p><b>CONCLUSION</b>TGFalpha/EGF-EGFR pathway serves as a key growth regulator in prostate cancer. TGFalpha, but not EGF, preferentially maintains an autocrine loop in human androgen-unresponsive prostate cancer.</p>


Subject(s)
Blotting, Western , Cell Line, Tumor , Humans , Male , Prostatic Neoplasms , Metabolism , Pathology , RNA, Messenger , Genetics , ErbB Receptors , Genetics , Reverse Transcriptase Polymerase Chain Reaction , Transforming Growth Factor alpha , Pharmacology
19.
Article in Chinese | WPRIM | ID: wpr-675636

ABSTRACT

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.

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