The effect of Heat shock protein 70 antiseuse oligonucleotides to the bladder cancer in mouse loaded with tumor / 肿瘤研究与临床

Objective To investigate The effect of Heat shock protein 70(HSP70) antisense oligonucleotides (ASO)to bladder carcinoma in mouse loaded with tumor.Methods The 40 mice loaded with tumor subcutaneously were established by cultured BIU-87 cells,and divided into 4 groups randomly when the subcutaneous neoplasms grew to about 100 mm3,namely,HSP70 mRNA ASO plus mitomycin C(MMC)group;HSP70 mRNA ASO group;MMC and blank control.HSP70 mRNA ASO were injected into neoplasms,10mmg/kg weight,twice every week,and MMC 0.1mg/kg weight,twice every week,and the above schemes were replaced with normal saline to blank.The neoplasms were peeled off,photograghed and weighed in 30 days.HSP70 expressions were examined with reverse transcription polymerase chain reaction (RT-PCR),mierovaseular density(MVD)was evaluated by immunohis to chemical staining and the tumor cells apoptosis was detected by terrainal deoxynucleotidyl transferase(TdT)-mediated dUTP-biotin nick end labeling technique (TUNEL).Results The tumor inhibition rate in ASO+MMC surpassed 50%.more than ASO or MMC respectively,and the differences were significantly(P＜0.05).The ASO and MMC exceeded blank group respectively(P＜0.05).The ASO was the same as the MMC(P＞0.05).The apoptotic index(AI)in ASO+MMC surpassed the other three groups (P＜0.05).The difference between ASO and MMC was not significant (P＞0.05),while the A1 of ASO or MMC was more than blank respectively(P＜0.05).The results of MVD were in accordance with the above results.Conclusion The injection of HSP70 mRNA ASO in tumor locally can inhibit neoplasm growth,and this effect might correlate with the inhibition of apoptosis and microvascular forming resulting from the ASO.

Effect of celecoxib on bladder cancer xenografts in nude mice and apoptosis of tumor cells of the xenografts / 中华泌尿外科杂志

Objective To investigate the effect of cyclooxygenase-2(COX-2) inhibitor celecoxib on bladder cancer xenografts in nude mice and apoptosis of tumor cells in the xenografts. Methods Models of bladder cancer xenograft in nude mice was used to observe the effect of celecoxib on the animals and the xenografts.TUNEL was used to assess apoptotic index of tumor cells in the xenografts. Results Celecoxib could effectively inhibit the growth of xenografts(P0.05). Conclusions Celecoxib maybe inhibit the growth of bladder cancer via inducing apoptosis of tumor cells and perhaps will become a choice of chemoprevention and adjuvant therapy of bladder cancer.

The inhibitive effect of siRNA on the phosphodiesterase type 5 in smooth muscle cells of human corpus cavernosum / 中华泌尿外科杂志

Objective To investigate the inhibitive effect of siRNA(small interfering RNA,siRNA) on the phosphodiesterase type 5(PDE5) in smooth muscle cells of human corpus cavernosum,and provide experimental groundwork for the gene therapy of erectile dysfunction (ED). Methods Small interfering RNAs targeting PDE5 gene were systhesized by using web design software provided by Ambion,there siRNAs and control siRNA were systhesized by Ambion. SiRNAs were transfected into smooth muscle cells of human corpus cavernosum by using siPORTTM Lipid reagent;down-regulation of PDE5 mRNA was detected by RT-PCR;the inhibitive effect of PDE5 was detected by Western Blotting. Results The results of RT-PCR indicated siRNA1、siRNA2 and siRNA3 made down-regulations of PDE5 mRNA expression in the transfected groups 58.2%、14.9% and 11.8%;the PDE5 expression decreased 70.5%、19.8% and 17.3%;however the expression did not have different in control siRNA and frank group. Conclusions The synthesized siRNAs in vitro were able to down-regulate the expression of PDE5.There were different capabilities of the specific siRNAs down-regulation.It was suggested that the siRNA technique provide not only an extremely powerful tool for the functional analysis of genome but also a new method for ED gene therapy.

Effect of surgical castration on risk factors for arteriosclerosis of patients with prostate cancer / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To analyze the effect of castration on risk factors for arteriosclerosis of patients with prostate cancer.</p><p><b>METHODS</b>Thirty patients with primary regional prostate adenocarcinoma limited to the prostate theca were selected in this study. Serum levels of testosterone (T), free testosterone (FT), dehydroepiandrosterone (DHEA), sex hormone-binding globulin (SHBG), prostatic specific antigen (PSA), triglyceride (TG), total cholesterol (TC), high density lipoprotein-cholesterol (HDL-C), low density lipoprotein-cholesterol (LDL-C), apoprotein alpha(1) (APOalpha(1)) and apoprotein beta (APObeta), insulin, plasma fibrinopeptide A (FPA), plasminogen activator inhibitor-1 (PAI-1) and fibrinogen were determined just prior to, 1 week and 1, 4 and 8 months after castration.</p><p><b>RESULTS</b>T, FT and PSA decreased significantly 1 week after castration (21.12 +/- 15.11 ng/ml vs 383.9 +/- 62.6 ng/ml, P < 0.001; 4.08 +/- 3.29 pmol/L vs 34.11 +/- 11.59 pmol/L, P < 0.001; 14.34 +/- 7.77 ng/ml vs 23.51 +/- 6.57 ng/ml, P = 0.001, respectively) and continued to decrease until reaching their lowest levels 8 months after castration. DHEA and SHBG did not undergo any changes. TG, fasting insulin and glucose, 2-hour insulin and glucose levels were significantly elevated 1 month after castration (1.84 +/- 0.61 mmol/L vs 1.30 +/- 0.40 mmol/L, P < 0.05; 18.16 +/- 5.57 mU/L vs 9.47 +/- 3.81 mU/L, P < 0.05; 4.77 +/- 0.66 mmol/L vs 3.92 +/- 0.34 mmol/L, P < 0.05; 65.52 +/- 14.78 mU/L vs 36.94 +/- 17.12 mU/L, P < 0.01; 6.98 +/- 0.79 mmol/L vs 6.01 +/- 0.23 mmol/L, P = 0.001, respectively). TC, LDL-C, FPA and PAI-1 levels were elevated 4 months after castration (6.56 +/- 0.99 mmol/L vs 5.29 +/- 0.75 mmol/L, P < 0.01; 4.09 +/- 0.86 mmol/L vs 3.04 +/- 0.15 mmol/L, P < 0.01; 3.39 +/- 1.67 nmol/L vs 1.48 +/- 0.50 nmol/L, P < 0.01; 27.02 +/- 5.98 ng/ml vs 21.78 +/- 3.16 ng/ml, P < 0.05, respectively), continuing to increase after that point. Insulin sensitive index (ISI) decreased significantly 1 month after surgery (-4.42 +/- 0.36 vs -3.50 +/- 0.39, P < 0.001), and continued to decrease from that point forward. HDL-C, APOalpha(1), APObeta and fibrinogen remained at pre-operative levels. There was a negative linear correlation between FT and TG, TC, LDL-C, PAI-1, FPA, fasting insulin and glucose, 2-hour insulin and glucose (r = -0.311, -0.384, -0.385, -0.339, -0.353, -0.381, -0.303, -0.460 and -0.395, respectively; P < 0.05). A similar phenomenon occurred with T (r = -0.308, -0.309, -0.356, -0.320, -0.430, -0.453, -0.435, -0.483 and -0.512, respectively; P < 0.05). T and FT were positively associated with ISI (r = 0.555 and 0.501; P < 0.001).</p><p><b>CONCLUSIONS</b>At 8 months follow-up of the study subjects, we found that lower androgen levels have adverse effects on lipid metabolism, coagulative function and insulin sensitivity, related to arteriosclerosis in men.</p>

The role of nuclear matrix protein 22 combined with bladder tumor antigen stat test in surveillance of recurring bladder cancer / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate a non-invasive, effective and rapid mode of detecting the recurrence of bladder cancer during follow-up.</p><p><b>METHODS</b>Ninety patients following transurethral resection of bladder tumor (TURBt) surgery were recruited from January 1998 to March 2000. Standard ELISA was used to determine the quantity of nuclear matrix protein (NMP-22) in urine of all bladder cancer patients during their follow-up periods. Urine bladder tumor antigen (BTA) stat test was simultaneously performed and followed by cystoscopy.</p><p><b>RESULTS</b>The total positive rates of urinary NMP-22 and BTA stat test were 76.7% (33/43) and 67.4% (29/43), respectively. Comparatively, this positive rate would increase to 93.0% (40/43) when the combination of both urine NMP-22 and BTA test were adopted.</p><p><b>CONCLUSION</b>Examination of NMP-22 in urine is a rapid and effective way to detect the recurrence of bladder cancer. If combined with BTA test, NMP-22 may be used as a non-invasive method in surveillance of recurring of bladder cancer, which may reduce the frequency of patients needing to undergo conventional invasive cystoscopy.</p>

Prognostic value of matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-2 in bladder carcinoma / 中华医学杂志(英文版)

<p><b>OBJECTIVES</b>To detect the level of matrix metalloproteinase-2 (MMP-2) mRNA and the tissue inhibitor of metalloproteinase-2 (TIMP-2) mRNA in bladder transitional cell carcinoma (BTCC), and to estimate the prognosis for bladder tumor based on the quality and quantity of MMP-2 and TIMP-2 mRNA.</p><p><b>METHODS</b>Thirty-five samples of human BTCC and 15 normal fresh bladder tissues were studied by RT-PCR analysis followed by computer-assisted image analysis.</p><p><b>RESULTS</b>The level of the MMP-2 mRNA in BTCC was significantly increased compared with that in normal bladder epithelium. The positive rates of MMP-2 and TIMP-2 mRNA were 71.4% and 65.7% in BTCC, and 66.7% and 60.0% in the normal bladder wall. The expression intensity of the MMP-2 mRNA by image analysis tended to increase with tumor grading and staging, which showed statistical significance. Similarly, the MMP-2 to TIMP-2 ratio also showed statistically significant difference between normal bladder tissue and bladder carcinoma (P < 0.01).</p><p><b>CONCLUSIONS</b>A high level of the MMP-2 mRNA exists in BTCC, which may function to damage collagen IV inside the basement membrane and the extracellular basement of the bladder. The level of the MMP-2 mRNA is proportional to BTCC grading and staging, which may have prognostic value. The MMP-2 to TIMP-2 ratio may play a more significant role in the determination of the aggressiveness and prognosis of bladder tumor.</p>

Effects of transforming growth factor-β1 and transforming growth factor-α on the growth and invasive potentials of human bladder tumor cells / 中华泌尿外科杂志

Objective　To study the effects of transforming growth factor-β1 (TGF-β1) and transforming growth factor-α (TGF-α) on the growth and invasive potentials in human bladder tumor cells.　Methods　The effects of TGF-β1 and TGF-α on the growth of EJ cell line and the effects on MMPs and TIMP-2 were studied by means of MTT,Western Blot and RT-PCR methods.　Results　(1)TGF-β1 and TGF-α tended to inhibit the growth of EJ cells but statistically nonsigficant.(2)Higher levles of MMP-9 mRNA but lower levels of MMP-2,TIMP-2,MT1-MMP mRNA were found in EJ cells following the treatment with TGF-β1.The same was true for the expression of MMP-2,TIMP-2 mRNA in TGF-α groups.However,MMP-9 mRNA was not found in both TGF-α groups and the control groups. (3)TGF-β1 (0.1,1.0 ng/ml) enhanced MMP-2 protein but not the TIMP-2 protein,while TGF-β1 (5.0,10.0 ng/ml)decreased TIMP-2 protein but not on MMP-2.In TGF-α groups,when the concentration was 1.0,5.0,10.0ng/ml,TIMP-2 protein expression was decreased but MMP-2 did not.When the concentration reached 100.0 ng/ml,it increased MMP-2 protein level,not the TIMP-2 protein.　Conclusions　TGF-β1 and TGFα do not inhibit the proliferation of EJ cells whereas the enhanced-invasiveness and metastasis may be associated with regulating the expression of MMPs and TIMP-2.

Expression of HSP 70 mRNA and its association with cell apoptosis in bladder transitional cell carcinoma / 中华泌尿外科杂志

Objective To investigate the expression of HSP 70 mRNA in transitional cell carcinoma of bladder and association of HSP 70 mRNA with cell apoptosis. Methods In situ hybridization histochemistry(ISHH) was applied to detect HSP 70 mRNA expression in 60 patients with transitional cell carcinoma of bladder,and cell apoptosis was evaluated by means of terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick end labeling technique(TUNEL). Results Of 60 cases of bladder transitional cell carcinoma,HSP 70 mRNA expression was 56.7%(34/60),and it increased with grade and stage progressing( P

Endostatin expression in tumor tissues and serum levels of bladder cancer patients / 中华泌尿外科杂志

Objective To study endostatin expression in tumor tissues and serum levels of bladder cancer patients. Methods Expression of endostatin in 45 cases of bladder cancer and 12 subjects with normal bladder tissues was examined immunohistochemically.Serum levels of endostatin in 58 patients with bladder cancers and 43 healthy controls were analyzed by competitive enzyme immunoassay. Results Positive expression of endostatin was found in 61.5% patients with superficial bladder cancer,90.6% with invasive bladder cancer and 33.3% in normal bladder tissues.Serum level of endostatin was significantly higher in patients with bladder cancer than that in healthy controls (P

IL-6 and IL-8 levels in expressed prostate secretion of chronic prostatitis / 中华泌尿外科杂志

ObjectiveTo investigate the levels of anti-inflammatory cytokine IL-6 and pro-inflammatory cytokine IL-8 in expressed prostatic secretion(EPS)of chronic prostatitis.To study the relationship of cytokines to the mechanism of chronic prostatitis.MethodsIL-6 and IL-8 levels were measured in 102 chronic prostatitis EPS and in 28 normal EPS by radioimmunoassay.Urine was cultured and EPS was studied according to 2-glass test.NIH-CPSI(NIH-Chronic prostatitis symptom index) was performed by every patient.ResultsIL-6 and IL-8 levels were higher in Ⅱ,ⅢA and Ⅳ prostatitis (IL-6 0.51?0.57 ng/ml,IL-8 10.75?7.96 ng/ml,n=90) than those in controls (IL-6 0.32?0.51 ng/ml,IL-8 4.56?5.68 ng/ml,n=28),P0.05.There was a correlation between WBC count and IL-8 level,r=0.529,P0.05.ConclusionsIL-6 and IL-8 are elevated in EPS of Ⅱ,ⅢA and IV prostatitis, and might be considered as mediators of inflammatory response in prostate.The results indicate that IL-6 and IL-8 can be the objective parameters in the diagnosis of chronic prostatitis.

The inhibitive effect of antisenes oligodeoxynucleotide on the phosphodiesterase type 5 in smooth muscle cells of human corpus cavernosum / 中华泌尿外科杂志

Objective To investigate the inhibitive effect of antisenes oligodeoxynucleotide(ASON) on the phosphodiesterase type 5(PDE5) in smooth muscle cells of human corpus cavernosum,and provide experimental groundwork for the gene therapy of erectile dysfunction. Methods PDE5 gene ASON (containing exon 1) was transfected into the corpus cavernosum smooth muscle cells with the presence of liposome DOTAP.Another sense oligodeoxynucleotide(SON) and 1% of bovine serum were also transducted into the cells as controls.PDE5 was probed and measured by Western blot at 1,2,4,6,10,24 and 48 h after transfection. Results After transfection(1～6 h), PDE5 in human corpus cavernosum smooth muscle cells was significantly lower than it was in controls( P = 0.000 ～0.014). Conclusions The results indicate that PDE5 gene antisense oligodeoxynucleotide has inhibitive effect on PDE5 in smooth muscle cells of human corpus cavernosum in vitro,and the present study provides experimental groundwork for the gene therapy of erectile dysfunction.

The influences of endostatin on bladder tumor growth and its mechanism / 中华泌尿外科杂志

Objective To evaluate the inhibitory effect of endostatin on bladder tumor and to investigate the possible mechanism of the inhibition. Methods (1)By means of MTT method,the effects of endostatin on ECV304 and EJ cells proliferation were studied.(2)Ten nude mice were subcutaneously implanted with EJ bladder tumor cells (1?10 7/ml). After the tumor volume exceeded 100～200 mm 3,the mice were randomized into two groups.One group received recombinant human endostatin (2 mg?kg -1 ?d -1 ) whereas the other group received PBS as control.All the treatment lasted 21 days.(3) After that,the mice were sacrificed and then MMPs and TIMPs mRNA expression were measured in implanted tumor by RT PCR and Western blot method. Results (1)Endostatin inhibits ECV304 proliferation stimulated with bFGF (5 ng/ml).In addition,we report for the first time that endostatin also inhibits EJ cells proliferation.(2)The mean tumor weight of endostatin treated group (0.70?0.16 g) was significantly lower than that of control group (1.14?0.21)g, P

Expression of MMP-2, MMP-9, TIMP-2 in bladder carcinoma as a prognostic marker / 中华泌尿外科杂志

Objective To detect both the protein and gene expression of matrix metalloproteinase (MMP 2,MMP 9) and its tissue inhibitor (TIMP 2) in tissues of bladder transitional cell carcinoma (BTCC), and to study the correlation with the grading and staging of the neoplasm and with the prognosis. Methods 41 human BTCC ,15 normal bladder tissue were studied by Western Blot and RT PCR analysis followed by computer assisted image analysis in order to detect the A values of their expression. Results In BTCC, the A value of MMP 2, MMP 9 were increased significantly as compared to normal bladder epithelium. There was no statistical significant difference of A value of TIMP 2 between normal bladder tissue and bladder cancer tissue. The ratio of MMP 2/TIMP 2 in bladder cancer showed statistical significantly difference as compared with normal bladder tissue. Conclusions This study demonstrated there was a high expression of MMP 2 and MMP 9 in BTCC,by which collagen Ⅳ inside basement membrane of bladder was damaged.However, the ratio of MMP 2/TIMP 2 has more prognostic value than the expression of MMP 2 and MMP 9 alone.

Expression of TNF receptor in human spermatozoa / 中华泌尿外科杂志

Objective To verify the presence of tumor necrosis factor receptor (TNF R 2) in spermatozoa of fertile men,and to provide experimental evidence for the direct action of TNF on spermatoza. Methods Using Western blotting the expressions of TNF R 2 in spermatozoa of 15 fertile men were assessed. Results TNF R 2 was expressed in spermatozoa of all the 15 fertile men. Conclusions The spermatozoa of fertile men have TNF R 2,which may transmit the cytokines signal into cytoplasm and modulate the spermatozoa function.

Effects of transforming growth factor-?_1 and transforming growth factor-? on the growth and invasive potentials of human bladder tumor cells / 中华泌尿外科杂志

Objective To study the effects of transforming growth factor ?1 (TGF ?1) and transforming growth factor ? (TGF ?) on the growth and invasive potentials in human bladder tumor cells. Methods The effects of TGF ?1 and TGF ? on the growth of EJ cell line and the effects on MMPs and TIMP 2 were studied by means of MTT,Western Blot and RT PCR methods. Results (1)TGF ?1 and TGF ? tended to inhibit the growth of EJ cells but statistically nonsigficant.(2)Higher levles of MMP 9 mRNA but lower levels of MMP 2,TIMP 2,MT1 MMP mRNA were found in EJ cells following the treatment with TGF ?1.The same was true for the expression of MMP 2,TIMP 2 mRNA in TGF ? groups.However,MMP 9 mRNA was not found in both TGF ? groups and the control groups. (3)TGF ?1 ( 0.1 , 1.0 ng/ml) enhanced MMP 2 protein but not the TIMP 2 protein,while TGF ?1 ( 5.0 , 10.0 ng/ml)decreased TIMP 2 protein but not on MMP 2.In TGF ? groups,when the concentration was 1.0 , 5.0 , 10.0 ng/ml,TIMP 2 protein expression was decreased but MMP 2 did not.When the concentration reached 100.0 ng/ml,it increased MMP 2 protein level,not the TIMP 2 protein. Conclusions TGF ?1 and TGF? do not inhibit the proliferation of EJ cells whereas the enhanced invasiveness and metastasis may be associated with regulating the expression of MMPs and TIMP 2.

The role of TURP in the management of prostate carcinoma / 中华泌尿外科杂志

ObjectiveTo evaluate the role of transurethral resection of the prostate (TURP) in the management of prostate carcinoma.MethodsTURP were randomly performed on 19 suspected patients with prostate carcinoma, based on their mean serum PSA level (14.5 ng/ml) and negative result of anal touch,so as to take more biopsy samples and to make histopathologic diagnosis.Meanwhile,another 8 patients with prostate carcinoma underwent palliative TURP so as to resolve bladder outlet obstruction.ResultsOf the 19 suspected patients, 6 (33%) were pathologically diagnosed to have prostate carcinoma.These 6 patients were all alive during a follow-up for 3 months to 6 years.Obstructive symptom was relieved in 4 patients who underwent palliative TURP,but repeat palliative TURP was needed in 2 patients 3 months after initial operations.ConclusionsIn certain circumstances,TURP may play a role in the diagnosis of prostate carcinoma. In addition, it relieves obstructive symptom caused by enlarged prostate. However, the long-term effects are unpredictable.

Primary culture and identification of human corpus cavernosum smooth muscle cells / 中华泌尿外科杂志

Objective To provide an ideal material for the investigation of penile erection. Methods Human corpus cavernosum smooth muscle cells were cultured with DMEM ( 20% of it was human serum)in vitro and the bred cells were identified by immunohistochemistry. Results After 10～14 days, spindle cells overspreaded in 6 culture bottles and they were validated as human corpus cavernosum smooth muscle cells immunohistochemically. Conclusions Human corpus cavernosum smooth muscle cells can be cultivated in feasible medium and can also become a cell line.These bred cells can serve as a material for the study of penile erection.

Risk factors of the occurrence bladder tumor after resection of upper urinary tract urothelial carcinoma / 中华泌尿外科杂志

0) and the correlation coefficient were low.The absence of intravesical occurrence of the cancer in solitary primary tumor cases was significantly less than that in the multiple ones ( P =0).The intravesical occurrence rate of the cancer elevated with the pathological stage of the primary cancer ( P =0.0039).92.5% of cases with initial concurrent involvement of the bladder have had the occurrence of bladder cancer within 2 years after surgical treatment. Conclusions The number and the stage of the primary tumor,with concurrent bladder involvement or not and the internal between the occurrence of the bladder cancer and the upper urinary tract cancer are the significant factors for prognosis,the coefficient correlation between the 4 factors being low.

Study of test-retest reliability for pressure-flow parameters for bladder outlet obstruction / 中华泌尿外科杂志

Objective To study the test-retest reliability for parameters of repeated pressure flowexaminations in patients with benign prostatic hyperplasia (BPH) and bladder outlet obstruction (BOO). Methods A total of 24 patients with BPHwere included.Their mean age was 65.3 years and the prostatevolume measured by transrectal ultrasound was (62.7?30.4)ml.Pressure flowexaminations were performedat baseline,10 min and 4 weeks.Data of parameters were studied,including Pdet.open,Pdet.max,Pdet.min.void,PdetQmax,and Qmax.In addition, Abrams-Griffiths (AG) number,Linearized passive urethral resistance relationcategory (LPURR),urethral resistance algorithm (URA) were calculated. Results The relative coeffi-cients of repeatability of Pdet.open,Pdet.max,Pdet.min.void,Pdet.Qmax,Qmax,AG,LPURR,URA were 17.1%,16.3%,24.0%,12.5%,16.5%,18.7%,21.4%,23.7%,respectively,at the second time compared with those atbaseline;and they were 19.7%,20.5%,32.7%,14.8%,24.0%,23.9%,27.3%,27.2%,respectively,atthe third time compared with those at baseline.The mean coefficients of variation were 22.9%,16.6%,24.5%,12.5%,19.9%,17.9%,20.7%,31.5%,respectively,at the second time compared with those atbaseline;and they were 20.6%,18.8%,20.6%,13.2%,21.0%,16.7%,21.7%,29.7%,respectively,atthe third time compared with those at baseline.Among all the parameters,the lowest 3 of the mean coeffi-cients of variation were PdetQmax,AG number and Pdet.maxin turn; and the lowest3 of the relative coefficients ofrepeatability were PdetQmax,Pdet.openand Pdet.max. Conclusions In patients with BPHand BOO,of all the pa-rameters of repeated pressure flow examinations,PdetQmaxhas the best test-retest reliability of both short andlong term.