Construction of an oncolytic adenovirus expressing small hairpin RNA and targeting the SATB1 gene / 中华男科学杂志

<p><b>OBJECTIVE</b>To construct an oncolytic adenovirus with the DD3 promoter regulation, expressing small hairpin RNA and targeting the SATB1 gene (SATBI-shRNA), and to evaluate its potential for inhibiting the growth of human prostatic carcinoma cells (LNCaP) in vitro.</p><p><b>METHODS</b>SATB1-shRNA expression cassettes containing the H1 promoter were produced by PCR from pSilencer3. 1-SATB1 and inserted into the pZD55 vector, and the recombinant plasmid pZD55-SATB1-shRNA was constructed, pZD55SATB1-shRNA and pZXC2-DD3-E1A were digested with EcoRV and Xba I , and the obtained expression cassettes linked each other to construct the recombinant plasmid pDD3-ZD55-SATB1, which was cotransfected with the pBHGE3 packaging plasmids mixture into 293 cells by Effectence. The recombined adenoviruses DD3-ZD55-SATB1 were identified by PCR. Viruses were propagated on HEK293 cells and purified by standard techniques, and the functional PFU titers determined by plaque assay on 293 cells. The antitumor potential of DD3-ZD55-SATB1 to LNCaP was evaluated by the crystal violet dye method. The expression level of the E1A gene was detected by Western blot, and that of the SATB1 gene in the adenovirus-infected LNCaP cells by both Western blot and RT-PCR.</p><p><b>RESULTS</b>An oncolytic adenovirus expressing SATB1-shRNA with the DD3 promoter regulation, DD3-ZD55-SATB1, was constructed successfully, whose functional PFU titer was 1.2 x 10(10) PFU/ml. DD3-ZD55-SATB1 showed an obvious cytopathic effect and a selective expression of E1A in the adenovirus-infected LNCaP cells; it exhibited a high LNCaP-targetability and specific SATB1-silencing effect.</p><p><b>CONCLUSION</b>The successful construction of the oncolytic adenovirus DD3-ZD55-SATB1 offers a new tool for researches on the gene therapy for human prostate cancer.</p>

Skip metastasis of prostate cancer: diagnosis and treatment / 中华男科学杂志

<p><b>OBJECTIVE</b>To improve the diagnosis and treatment of far advanced prostate cancer without clinically detectable bone metastasis.</p><p><b>METHODS</b>Cancer metastatic lesions were found in the liver and lungs respectively of two patients on routine medical examination, and only an abnormally elevated level of the serum prostate specific antigen (PSA) was observed in the following system examinations. The patients were diagnosed as having prostate cancer by prostate biopsy. MRI showed a discontinued prostate capsule, and ECT revealed no bone metastasis. Diagnostic treatment was conducted by giving LHRHa combined with antiandrogens. One of the patients underwent surgical castration at 12 months, and both received intensity modulated radiation therapy (80 Gy) at 15 and 18 months, respectively.</p><p><b>RESULTS</b>The metastatic lesions in the liver and lungs of the patients were either absent or significantly reduced after treated by maximal androgen blockade for 3 months, and all disappeared after 6 months'treatment, with the PSA level stabilized at less than 0.02 microg/L in one patient, and around 0.5 microg/L in the other. Antiandrogen treatment was suspended after radiotherapy. The results of liver, lung and bone scanning were normal during the 12-month follow-up, and the PSA level was below 1.0 microg/L.</p><p><b>CONCLUSION</b>Remote metastasis of prostate cancer may occur in ectosteal organs first, which deserves special attention. A combination of different treatment methods promises satisfactory results.</p>

The value of pre-operative semen analysis as a restore index of fertilizing capacity after varicocelectomy / 中华男科学杂志

<p><b>OBJECTIVE</b>To discuss the value of pre-operative semen analysis of patients with varicocele as a predictive restore index of sperm motility and fertilizing capacity after varicocelectomy.</p><p><b>METHODS</b>Semen analysis was carried out with computer-aided sperm analyzer in 107 patients with varicocele and all patients were referred to the clinic with diagnosis of male infertility. Stratification of patients as group A (n = 32), B ( n = 36) and C (n = 39) was based on pre-operative total motile sperm count (TMSC). Follow-up included semen analysis and pregnancy data after three months following left or bilateral varicocelectomy.</p><p><b>RESULTS</b>The average post-operative TMSC increased significantly when compared with the pre-operative. However, a mean absolute increase in group A and B was better than that in group C (P < 0.05). Of the 68 patients in groups A and B based on pre-operative TMSC, 56 patients' TMSC (82.4%) was > or =20 x 10(6) after varicocelectomy, and that of only 8 (20.5%) patients in group C was > or =20 x 10(6) following varicocelectomy. Of the 98 patients wives, 36 had natural conception. Pregnancy rates in groups A and B were higher than that in group C (P < 0.05).</p><p><b>CONCLUSION</b>Varicocelectomy may be the most effective method to patients with varicocele with pre-operative TMSC > or = 5 x 10(6), but it may be not the best method for patients with severe oligoasthenospermia (pre-operative TMSC < 5 x 10(6)).</p>

Association between polymorphism of CYP17 gene and serum hormone concentrations in aged men / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the association between polymorphism of CYP17 gene and serum hormone concentrations in aged men.</p><p><b>METHODS</b>Eighty-three healthy men at the average age of 66.7 were divided into a < 66.7 group (n = 36) and a > 66.7 group (n = 47), and the polymorphism of CYP17 gene in the 5' promoter region was investigated by PCR using DNA from the men's peripheral blood lymphocytes. A new recognition site was created for the restriction enzyme MspA1 I by transition (T --> C) in the risk allele (A2). Three genotypes A1/A1, A1/A2, A2/A2 were established, serum sex-hormone levels measured, and mean hormone concentration evaluated in each genotype and age group.</p><p><b>RESULTS</b>No evidence was found that the testosterone (T) level, estrogen (E2) level and T/E2 ratio were associated with the genotype of CYP17 gene. There was no significant difference in T and E2 levels between the two groups, but there was a significant increase in the T/E2 ratio (P < 0.05).</p><p><b>CONCLUSION</b>A2 allele does not increase sex hormone levels in aged men, but the T/E, ratio was higher in the > 66.7 group than in the < 66.7 group. This may be closely associated with the mechanism of benign prostate hyperplasia and prostate cancer in aged men.</p>

Effects of siRNA against Ki67 gene on the proliferation and apoptosis of human renal carcinoma cells / 中华泌尿外科杂志

Objective To evaluate the effects of small interfering RNA(siRNA)against Ki67 gene on the proliferation and apoptosis of human renal carcinoma cell line 786-0 cells.Methods The human renal carcinoma 786-0 cells were treated with Ki67-siRNA(100 nmol/L).The mRNA expression of Ki67 was detected by RT-PCR.The protein expression of Ki67 was detected by Western blot and immunohisto- chemical technique,respectively.The proliferation of 786-0 cells was detected by MTT assay.The apoptosis of 786-0 cells was detected by TUNEL assay.Results RT-PCR and Western blot analysis showed that the Ki67 mRNA and Ki67 protein expression levels of the 786-0 cells treated with Ki67-siRNA were(37.6?1.9)% and(46.4?0.9)% ,respectively,which were significantly lower than those of controls [(97.3?0.9)% and(95.3?0.9)%,P＜0.01],The Ki67 positive expression rate of 786-0 cells treated with Ki67-siRNA by immunohistochemical technique was 52.5?2.3,which was significantly lower than that of controls(114.5?4.9 ,P＜0.01).The proliferation-inhibiting rate and apoptosis rate of the 786-0 cells trea- ted with Ki67-siRNA were( 63.6?1.6)% and(41.7?0.6)% ,respectively,which were significantly higher than those of controls [(2.8?0.2)% and(10.3?1.4)%,P＜0.01].Conclusions siRNA against Ki67 gene can inhibit the proliferation and induce the apoptosis by blocking Ki67 expression of hu- man renal carcinoma 786-0 cells.The inhibition of Ki67 expression by siRNA may be a promising approach in gene therapy for renal cancer.