Phosphodiesterase type 5 siRNA increases cGMP in the smooth muscle cells of human corpus cavernosum / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the effect of phosphodiesterase type 5 (PDE5) small interfering RNA (siRNA) on the cGMP in the smooth muscle cells of human corpus cavernosum, and to provide an experimental groundwork for the gene therapy of erectile dysfunction (ED).</p><p><b>METHODS</b>Small interfering RNAs targeting PDE5 gene were synthesized by using web design software provided by Ambion, three siRNAs and a control siRNA were synthesized by Ambion. siRNAs were transfected into the smooth muscle cells of human corpus cavernosum by using siPORT Lipid reagent. cGMP was detected by ELISA at different times (24, 48, 72 and 96 h) after transfection.</p><p><b>RESULTS</b>The cGMP levels of the siRNA1, siRNA2 and siRNA3 groups were significantly higher than those of the siRNA control and blank control groups (P < 0.05), and so was it in the siRNA1 group than the siRNA2 and siRNA3 groups (P < 0.05), with significant difference between the siRNA control and the blank control group (P > 0.05).</p><p><b>CONCLUSION</b>The synthesized siRNAs in vitro are capable of increasing the level of cGMP in the smooth muscle cells of human corpus cavernosum, different siRNAs with different capabilities. The siRNA technique could provide not only an extremely powerful tool for the functional analysis of genome but also a new approach to ED gene therapy.</p>

Treatment of hyperhomocysteinemia and endothelial dysfunction in renal-transplant recipients with vitamin B / 中华外科杂志

<p><b>OBJECTIVE</b>To study the effect of vitamin B on treatment of hyperhomocysteinemia and endothelial dysfunction in renal-transplant recipients.</p><p><b>METHODS</b>Thirty-six stable hyperhomocysteinemic renal-transplant recipients were randomly assigned to vitamin treatment (group A, n = 18, folic acid 5 mg/d, vitamin B(6) 50 mg/d, B(12) 1000 microg/d) or controlled group (group B, n = 18) for 6 months. All subjects underwent assessment of levels for creatinine, creatinine clearance, average pressure, total cholesterol, triglyceride and fasting homocysteine. Endothelial function was evaluated using high-resolution vascular ultrasound.</p><p><b>RESULTS</b>The levels of homocysteine markedly decreased in group A [(13 +/- 4) micromol/L vs (20 +/- 5) micromol/L, t = 5.3, P < 0.01] after treatment, whereas no significant changes were observed in group B. In group A, endothelium dependent [(12 +/- 5)% vs (9 +/- 5)%, t = 2.9, P < 0.01] and independent [(18 +/- 4)% vs (12 +/- 5)%, t = 3.4, P < 0.01] vasodilatation responses significantly increased after treatment, no significant changes were observed in group B. Endothelium dependent [(9 +/- 6)%, t = 2.8, P < 0.01] and independent [(12 +/- 5)%, t = 3.5, P < 0.01] vasodilatation responses of group A were significantly lower than that of group B after treatment.</p><p><b>CONCLUSIONS</b>Vitamin B supplementation can reduce the levels of homocysteine and improve the endothelial function in hyperhomocysteinemic renal-transplant recipients.</p>

Analysis of eleven cases of severe pneumonia in kidney transplant recipients / 中华外科杂志

<p><b>OBJECTIVE</b>To analyse retrospectively the diagnosis and treatment of severe pneumonia in kidney transplantation recipients.</p><p><b>METHODS</b>Between January 1999 and December 2003, 172 adult patients underwent kidney transplantation at our department. In all severe pneumonia cases, empirical therapy was initiated with aztreonam, erythromycin and ganciclovir. And the therapy was switched to proper antibiotics according to the results of sensitivity testing. Responsible pathogen was detected by study of BAL (bronco-alveolar-lavage), sputum and blood specimen. Analyses included cell differential count, cytopathologic examination and cultures for bacteria, fungi and viruses. The immunosuppressive therapy was drastically reduced. Hypoxia was relieved by BiPAP (Bi-level Positive Airway Pressure) or mechanical ventilation if necessary.</p><p><b>RESULTS</b>Seventeen cases (9.9%) of pneumonia were observed in the 172 recipients, only 11 (65%) patients experienced severe pneumonia, 1 (9%) of them died. Fever was the most common symptom on presentation (82%). On presentation 46% of the patients presented with classical clinical syndrome of fever accompanied by cough and dyspnea. Positive rate of BAL and blood culture were 100% and 46% respectively. BiPAP and mechanical ventilation were required in 6 and 2 cases respectively.</p><p><b>CONCLUSION</b>BAL is preferred for early detection of responsible pathogen. A combination of drastic reduction of the immunosuppressive regimen, implementation of appropriate empirical antibiotics, proper BiPAP or mechanical ventilation are important.</p>

Enhanced sensitivity to mitomycin C by abating heat shock protein 70 expression in human bladder cancer cell line of BIU-87 / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Bladder cancer is a relatively common tumor in the urinary system, in which mitomycin C (MMC)-based chemotherapy or combination chemotherapy has been mainly used to treat patients with advanced bladder cancer. The prognosis of patients with advanced bladder cancer is still extremely poor in spite of recent therapeutic advances. To improve the prognosis, the sensitivity of tumor cells to mitomycin C by the induction of apoptosis with the abating heat shock protein 70 (HSP70) expression in human bladder cancer cell lines of BIU-87 was investigated.</p><p><b>METHODS</b>HSP70 expression was abated in BIU-87 cells by HSP mRNA antisense oligomers. MTT assay and the clone-forming test were used for evaluating the sensitivity of cells to MMC. Apoptosis was assessed using both fluorescent microscopy after staining the cells with Hoechst 33258 and DNA fragment ladder agarose electrophoresis. Thirty-two male six-week-old BALB/c nude mice, at the beginning of the experiment, were used to evaluate the effect of antisense oligomers (ASO) on the tumor formation in vivo.</p><p><b>RESULTS</b>HSP70 expression in BIU-87 was effectively abated by HSP70 mRNA antisense oligomers. The percentage of apoptotic cells in ASO group was greater than in sense oligomers (SO) [P < 0.05, (18.31 +/- 2.89)% vs (1.89 +/- 0.74)%], nonsense oligomers (NO) [P < 0.05, (18.31 +/- 2.89)% vs (1.78 +/- 0.92)%] and blank groups [P < 0.05, (18.31 +/- 2.89)% vs (1.87 +/- 0.84)%], while the sensitivity of tumor cells to mitomycin C was enhanced. The in vivo tumor inhibition rate of ASO plus MMC (> 50%) was more than that of ASO or MMC group alone (all P < 0.05).</p><p><b>CONCLUSIONS</b>The abating level of HSP70 expression can strengthen the sensitivity of BIU-87 to MMC. One of this effect might be related to the induction of apoptosis by abating HSP70 expression.</p>

Diagnosis and treatment of bilateral testicular intratubular seminoma: a case report and review of literature / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the diagnosis and treatment of testicular intratubular seminoma.</p><p><b>METHODS</b>One case of testicular intratubular seminoma was diagnosed with testicular biopsy. Epididymal sperm was aspirated and intracytoplasmic sperm injection ( ICSI) was performed. And local radiotherapy was conducted on the bilateral testes after fertilization.</p><p><b>RESULTS</b>The result of testicular biopsy revealed bilateral testicular intratubular seminoma. Large numbers of sperms were found in the epididymal aspirate. ICSI did not succeed for the first time. The second ICSI succeeded. The local radiotherapy by 60Co had been conducted on the bilateral testes, and testicular tumor didn't develop further.</p><p><b>CONCLUSION</b>Testicular intratubular seminoma is a type of intratubular germ cell neoplasia, with no clinical manifestations, and usually found in testicular biopsy. Earlier management promises better prognosis.</p>

The growth inhibition effect of alpha-adrenoceptor antagonists on androgen- independent prostate cancer cell line / 中华外科杂志

<p><b>OBJECTIVE</b>The aim of the present study is to compare the effects of two alpha1-adrenoceptor antagonist terazosin and alfuzosin together with one alpha-adrenoceptor antagonist phenoxybenzamine on androgen-independent prostate cancer cell lines PC-3 and DU145.</p><p><b>METHODS</b>Two androgen- independent cell lines, PC-3 and DU145, were used to determine the cell viability, colony-forming ability as well as cell cycle characteristics after exposure to these three drugs.</p><p><b>RESULTS</b>This study showed that terazosin inhibited not only prostate cancer cell growth but also colony-forming ability, which is the main target of clinical treatment. On the other hand, alfuzosin and phenoxybenzamine have no effect on cell viability and colony forming ability of PC-3 and DU145. In addition, the terazosin inhibits cell growth through G(1) phase cell cycle arrest.</p><p><b>CONCLUSION</b>This study provided the evidence that alpha1-adrenoceptor antagonist terazosin may have a therapeutic potential in the treatment of advanced androgen-independent prostate cancer.</p>

Electron paramagnetic resonance in monitoring of nitric oxide production after kidney transplantation in rats / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Much research has been focused on ischemia/reperfusion injury (IRI) to the transplanted organs. As a free radical, nitric oxide (NO) plays an important role in IRI. In this study, the production of NO and its functions during IRI were monitored in rat models after allotransplantation of kidney grafts.</p><p><b>METHODS</b>Of 75 male LEW rats, 30 served as donors, and the remaining 45 rats were divided into three groups (15 rats in each group): controls (group 1), kidney allotransplantation followed by bilateral nephrectomy during reperfusion (group 2), 2 hours before operation, donors and recipients were treated with N(G)-nitro L-arginine methyl ester (L-NAME), a NO synthase inhibitor, at a dose of 30 mg/kg (group 3). Bilateral nephrectomies were performed while kidney grafts were reperfused. The kidney grafts were hypothemically stored for 24 hours. The production of NO before and after reperfusion was measured by electron paramagnetic resonance (EPR). The creatinine level, the glomerular filtration rate (GFR) and the protein carbonyl content in tissue samples were recorded on the first and the fifth day after operation. The data were evaluated by one-way analysis of variance. Differences were considered to be statistically significant when a P value was less than 0.05.</p><p><b>RESULTS</b>After reperfusion for 15 minutes, the production of NO increased remarkably and kept increasing till 120 minutes, after which the level returned to normal. In group 3, which was pretreated with L-NAME, creatinine levels were higher than those in group 2 at the 24th hour (4.10 +/- 0.50 mg/dl vs. 3.77 +/- 0.42 mg/dl, P < 0.05) and the 120th hour (3.19 +/- 0.79 mg/dl vs. 2.22 +/- 0.53 mg/dl, P < 0.05). GFR levels in group 3 were lower than those in group 2 at the 24th hour (0.50 +/- 0.12 ml/min vs. 0.71 +/- 0.19 ml/min, P < 0.05) and the 120th hour (0.59 +/- 0.38 ml/min vs. 1.27 +/- 0.23 ml/min, P < 0.01). The content of protein carbonyl in tissue samples of group 3 was lower than that in group 2 at the 24th hour (29.01 +/- 7.02 nmol/mg protein vs. 49.39 +/- 13.13 nmol/mg protein, P < 0.05), but was higher than that at the 120th hour (75.71 +/- 16.74 nmol/mg protein vs. 57.93 +/- 15.32 nmol/mg protein, P < 0.05).</p><p><b>CONCLUSIONS</b>After transplantation of hypothemically stored kidney grafts, the increased NO production in the early stage has protective effects on the transplanted kidney. Application of L-NAME to inhibit NO production is harmful to the recovery of the renal functions of kidney grafts.</p>

The significance of the expression of cyclooxygenase-2 in bladder transitional cell carcinoma tissues / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the expression of cyclooxygenase-2 (COX-2) in bladder transitional cell carcinoma tissues, and understand its clinical significance.</p><p><b>METHODS</b>Reversal transcription polymerase chain reaction (RT-PCR) was used to assess the expression of COX-2 mRNA in 52 cases of bladder transitional cell carcinoma tissues and 17 cases of normal bladder tissues far from neoplasm; Western blot was used to assess the expression of COX-2 protein in 49 cases of bladder cancerous tissues and 17 cases of normal tissues.</p><p><b>RESULTS</b>Positive expression of COX-2 mRNA was detected in 83% (43/52) of bladder cancer tissues and in 29% (5/17) of normal tissues by RT-PCR and there was significant difference in expression of COX-2 mRNA between cancer tissues and normal tissues. Western blot analysis showed that expression of COX-2 protein was correlation with the stage and grade of cancer.</p><p><b>CONCLUSION</b>COX-2 is overexpressed in bladder transitional cell carcinoma. COX-2 maybe play a certain role in carcinogenesis and progression of bladder cancer and turn into a useful target of chemoprevention of bladder cancer.</p>

The heat shock protein 70 antisense oligomers enhance the sensitivity of bladder cancer cell EJ to mitomycin C / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate whether the heat shock protein (HSP) 70 antisense oligomers can enhance the sensitivity of bladder cancer cell EJ to mitomycin C.</p><p><b>METHODS</b>The HSP70 mRNA of EJ cells was blocked by the 10 micromol/L HSP70 antisense oligomers, while its effect on cell growth was evaluated by methyl thiazolyl tetrazolium (MTT) and colony forming ability test.</p><p><b>RESULTS</b>The HSP70 expressions in HSP70 antisense treated group were lower than the corresponding sense and nonsense treated groups (P < 0.01). While, the increased sensitivity of EJ to mitomycin C was found in antisense treated group, compared with the corresponding sense and nonsense treated groups (P < 0.01).</p><p><b>CONCLUSION</b>The sensitivity of bladder cancer cell EJ to mitomycin C was enhanced by the blockage of the HSP70 expression.</p>

Detection of serum levels of MMP-9 and VEGF in patients with bladder cancer / 中华外科杂志

<p><b>OBJECTIVE</b>To detect serum levels of MMP-9 and VEGF in patients with bladder cancer.</p><p><b>METHODS</b>Serum levels of MMP-9 and VEGF in 58 patients with bladder cancer and 45 healthy controls were measured by sandwich-ELISA.</p><p><b>RESULTS</b>Serum levels of MMP-9 and VEGF (737.12 micro g/L and 1148.88 ng/L) were significantly higher in the cancer patients than those of controls (423.51 micro g/L and 846.96 ng/L, P < 0.01). The serum levels were associated with tumor stage and grade. In patients with invasive cancer, the levels were significantly higher than those of superficial cancer (P < 0.01). Patients with distant metastasis had significantly higher levels of MMP-9 and VEGF than those with localized invasion (P < 0.01). But there was no significant difference between patients with superficial cancer and controls. Patients with G(3) tumors had significantly higher levels of MMP-9 and VEGF than those of patients with G(1) and G(2) tumors (P < 0.01).</p><p><b>CONCLUSIONS</b>Elevated MMP-9 and VEGF levels are associated with a high stage and grade of bladder cancer and they may serve as markers of tumor progression in the future.</p>

The effects of antisenes oligodeoxynucleotide on the cyclic nucleotide monophosphates in smooth muscle cells of human corpus cavernosum / 中华男科学杂志

<p><b>OBJECTIVES</b>To investigate the effects of antisense oligodeoxynucleotide(ASON) on the cyclic nucleotide monophosphates (cNMP) in smooth muscle cells of human corpus cavernosum, and provide experimental groundwork for the gene therapy of erectile dysfunction.</p><p><b>METHODS</b>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. Two of cNMP, cAMP and cGMP, were probed and measured by ELISA at 1, 2, 4, 6, 10, 24 and 48 h after transfection.</p><p><b>RESULTS</b>After transfection, the level of cGMP(1-6 h) in human corpus cavernosum smooth muscle cells was significantly higher than that in controls(P < 0.01).</p><p><b>CONCLUSIONS</b>The PDE5 gene ASON had been showed to manifest stimulative effect on the cGMP in smooth muscle cells of human corpus cavernosum in vitro, and it provides experimental groundwork for the gene therapy of erectile dysfunction.</p>