[Psychological factors and erectile function in men with refractory chronic prostatitis].

OBJECTIVE: To study the psychological factors and erectile function in patients with refractory chronic prostatitis. METHODS: We obtained and compared the scores on the NIH scales of chronic prostatitis symptoms, anxiety, depression and erectile function among 232 refractory and medical chronic prostatitis patients who had never received any psychotherapy. RESULTS: No significant differences were observed in the scores on chronic prostatitis symptoms between the refractory and the medical chronic prostatitis groups, while the scores on anxiety and depression were significantly higher and that on erectile function significantly lower in the refractory than in the medical group (P < 0.01), with a negative correlation between the scores on the former two items and that on the latter. CONCLUSION: Obvious psychological factors exist in patients with refractory chronic prostatitis, which may affect their erectile function.

[Effects of Omi/HtrA2 on expression of anti-apoptotic protein PED/PEA-15 and apoptosis of prostate cancer cell line PC-3].

BACKGROUND & OBJECTIVE: The interaction between pro-apoptotic factors and anti-apoptotic factors is closely related to the genesis and development of tumors. Omi/HtrA2 is a novel gene involved in the regulation of apoptosis. PED/PEA-15 is a widely expressed anti-apoptotic protein. This study was to explore the effects of Omi/HtrA2 on PED/PEA-15 expression and apoptosis of prostate cancer cell line PC-3. METHODS: Omi/HtrA2 expression and specific siRNA vectors were constructed and transiently transfected into PC-3 cells. The effect of Omi/HtrA2 on PED/PEA-15 expression was assayed by Western blot, and its effect on apoptosis of PC-3 cells was analyzed by ELISA. Caspase-8 activity was assayed using Caspase-8 colorimetric assay kit. The effects of Omi/HtrA2-specific siRNA sequence on its transcription and translation were analyzed by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. The sensitivity of PC-3 cells to cisplatin (DDP) after Omi/HtrA2 gene silencing was determined by flow cytometry. RESULTS: Enzyme digestion analysis and DNA sequencing confirmed Omi/HtrA2 expression, and specific siRNA vectors were successfully constructed. After transfection of Omi/HtrA2 expression vector, PED/PEA-15 expression was inhibited, Caspase-8 activity was promoted, and the apoptosis of PC-3 cells was enhanced. The sensitivity of PC-3 cells to DDP was suppressed after Omi/HtrA2 gene silencing. CONCLUSION: Omi/HtrA2 can promote the apoptosis of PC-3 cells through inhibiting PED/PEA-15 expression.

[The changes of IGF-I in testis and epididymis on a rat model with oligozoospermia/azoospermia induced by cyclophosphamide].

OBJECTIVE: To evaluate the effect of the levels of IGF-I in the epididymis and the expression of IGF-I in the testis of adult male rat after the administration of cyclophosphamide. METHODS: Ninety-six male adult rats (8 weeks age) were divided into 6 groups. The doses given to the rats of the groups 1 to 5 were 10, 20, 40, 80 and 100 mg/(kg x d), respectively. The remaining group was served as control. All those rats were sacrificed and IGF-I were quantitatively determined by ELISA techniques 2 and 4 weeks after the administration of the drug (by gastric fudge). Immunohistochemical SP technique was used to examine expression of IGF-I in rat testis. RESULTS: The levels of cell factors (IGF-I) in the epididymis of the rats were gradually reduced with the increasing time and dose after administration of the drug. In the mean time the expression of IGF-I in the tissues of the testis of those rats were also gradually reduced. CONCLUSION: In the time of oligozoospermia/azoospermia induced by the administration of cyclophosphamide, the expression levels of IGF-I in the genetic system were significantly reduced. The possible mechanism of these changes could be attributed to the lower spermatogenesis function of the testis caused by the administration of cyclophosphamide.

Downregulation of XIAP expression induces apoptosis and enhances chemotherapeutic sensitivity in human gastric cancer cells.

X-linked inhibitor of apoptosis (XIAP) is the most potent member of the inhibitor of apoptosis protein (IAP) gene family in terms of its ability to inhibit caspases and suppress apoptosis. Recent evidence has suggested that XIAP is a key determinant in chemoresistance of cancer cells. To explore a novel approach for ameliorating chemotherapy of gastric cancer, the antisense expression vector for the XIAP gene was constructed and transferred into gastric cancer cell lines, MKN-45 (wild-type p53) and MKN-28 (mutant-type p53). This transfer resulted in significant downregulation of XIAP expression, decreased in vitro cell viabilities, and induced apoptosis. In transferred cells, inactive caspase-3 precursors were cleaved into the active subunits (p20 and p17) during apoptosis induced by downregulation of XIAP. The inhibitory effects of cisplatin and mitomycin C on the growth of XIAP downregulated cancer cells were significantly enhanced. In addition, this process occurred only in wild-type p53 (MKN-45), but not in mutant-type p53 (MKN-28) gastric cancer cells. The data presented suggest that downregulation of XIAP via antisense RNA can lead to apoptosis of gastric cancer cells in vitro, correlating with cellular p53 status and activation of caspase-3. This finding could lead to a potential strategy for improving the efficiency of therapies for gastric cancer.

Selection of optimal antisense accessible sites of survivin and its application in treatment of gastric cancer.

AIM: To select the optimal antisense accessible sites of survivin, a highly expressed gene in tumor tissues, in order to explore a novel approach to improve biological therapy of gastric cancer. METHODS: The 20 mer random oligonucleotide library was synthesized, hybridized with in vitro transcribed total survivin cRNA, then digested by RNase H. After primer extension and autoradiography, the antisense accessible sites (AAS) of survivin were selected. Then RNADraw software was used to analyze and choose the AAS with obvious stem-loop structures, according to which the complementary antisense oligonucleotides (AS-ODNs) were synthesized and transferred into survivin highly- expressing gastric cancer cell line MKN-45. Survivin expression was detected by RT-PCR and Western Blotting. Cellular growth activities were assayed by tetrazolium bromide (MTT) colorimetry. Cellular ultrastructure was observed by electronic microscopy, while apoptosis was detected by annexin V-FITC and propidium iodide staining flow cytometry. RESULTS: Thirteen AAS of survivin were selected in vitro. Four AAS with stem-loop structures were chosen, locating at 207-226 bp, 187-206 bp, 126-145 bp and 44-63 bp of survivin cDNA respectively. When compared with non-tranfection controls, their corresponding AS-ODNs (AS-ODN(1), AS-ODN(2), AS-ODN(3) and AS-ODN(4)) could reduce Survivin mRNA levels in MKN-45 cells by 54.3+/-1.1% (t = 6.12, P<0.01), 86.1+/-1.0% (t = 5.27, P<0.01), 32.2+/-1.3% (t = 7.34, P<0.01) and 56.2+/-0.9% (t = 6.45, P<0.01) respectively, while survivin protein levels were decreased by 42.2+/-2.5% (t = 6.26, P<0.01), 75.4+/-3.1% (t = 7.11, P<0.01), 28.3+/-2.0% (t = 6.04, P<0.01) and 45.8+/-1.2% (t = 6.38, P<0.01) respectively. After transfection with 600 nmol/L AS-ODN1-AS-ODN(4) for 24 h, cell growth was inhibited by 28.12+/-1.54% (t = 7.62, P<0.01), 38.42+/-3.12% (t = 7.75, P<0.01), 21.46+/-2.63% (t = 5.94, P<0.01) and 32.12+/-1.77% (t = 6.17, P<0.01) respectively. Partial cancer cells presented the characteristic morphological changes of apoptosis, with apoptotic rates being 19.31+/-1.16% (t = 7.16, P<0.01), 29.24+/-1.94% (t = 8.15, P<0.01), 11.87+/-0.68% (t = 6.68, P<0.01) and 21.68+/-2.14% (t = 7.53, P<0.01) respectively. CONCLUSION: The AAS of survivin could be effectively selected in vitro by random oligonucleotide library/RNase H cleavage method combined with computer software analysis, this has important reference values for further studying survivin-targeted therapy strategies for gastric cancer.

[Ureteroscopic holmium: YAG laser lithotripsy for managing urinary tract calculi].

OBJECTIVE: To evaluate the effectiveness and safety of ureteroscopic holmium: YAG laser lithotripsy for managing ureteral calculi. METHODS: Ureteroscopic holmium: YAG laser lithotripsy was used in 168 ureteral calculi (proximal 27 cases, middle 33 cases, distal 108 cases). Transurethral cystoscopic holmium: YAG laser lithotripsy in 12 bladder calculi. RESULTS: Four to six weeks after operation, The stone-free rate was 93% (25/27) in the proximal ureteral calculi, 94% (31/33) in the middle ureteral calculi, 94% (102/108) in the distal ureteral calculi, respectively. The complication rate was 5% (8 cases). the stone-free rate of bladder calculi was 100% (12/12), no complication. CONCLUSION: Ureteroscopic holmium: YAG laser lithotripsy is a highly effective and safe treatment modality for managing ureteral calculi.

[Effects of mTR antisense oligodeoxynucleotide on telomerase activity and the expression of telomerase mTR in type A spermatogonia].

OBJECTIVES: To study the inhibitory effects of mouse telomerase RNA (mTR) antisense oligodeoxynucleotide(ASODN) on telomerase activity in rat spermatogonia. METHODS: 9-mer phosphorothioate mTR-ASODN was encapsulated by Lipofect AMINE 2000 (LF 2000) and transfected to type A spermatogonia in Snrague Dawley (SD) rat. Telomerase activity was detected by aid of TRAP-SYBR-Green staining and Bioluminescence technique in type A spermatogonia treated or untreated with ASODN. RESULTS: mTR-ASODN conjugated with LF 2000 could significantly inhibit telomerase activity of spermatogonia(P < 0.01). mTR mRNA level also decreased while the spermatogonia were treated with ASODN for 24 h. No change of telomerase activity and apoptosis were observed when SODN, RODN or single LF 2000 was used. CONCLUSIONS: Antisense oligodeoxynucleotide of mTR conjugated with LF 2000 could significantly inhibit telomerase activity of spermatogonia. mTR-ASODN might inhibit telomerase activity of spermatogonia at transcription level.

[Influence of suramin on the proliferation of prostatic carcinoma cells PC-3M].

OBJECTIVES: To examine the effects of suramin on the growth, cell cycle and apoptosis of a hormone refractory prostate cancer cell line PC-3M, and to explore the possible mechanisms. METHODS: The roles of diverse concentrations (10, 50, 100 and 200 mumol/L) of suramin on PC-3M cell proliferation at different ratios of fetal calf serum (FCS) (2%, 5%, 10%) were assayed respectively by trypan blue exclusion and tetrazolium (MTT) assay. The effect of suramin on cell cycle distribution and apoptosis induction of PC-3M cells was evaluated with flow cytometry (FCM). RESULTS: A higher dosage of suramin (200 mumol/L) had a cytotoxic effect on PC-3M cells, while lower dosages from 10 to 100 mumol/L produced a predominant inhibiting effect. Suramin could also play a growth suppressive role in the culture media containing 10% FCS, but to a much less extent than in the media containing lower concentrations(5%, 2%) of FCS. FCM analysis exhibited that suramin at a high dosage of 200 mumol/L could induce apoptosis, and at the other concentrations, G0/G1 cell cycle arrest. CONCLUSION: Suramin's proliferative suppression on PC-3M cells might result from several mechanisms including antagonistic action on growth stimulation via growth factor, arrest of cell cycle and induction of apoptosis.

[Expression of telomerase gene hTERT in testes of infertile male and its significance].

OBJECTIVES: To study the expression and the significance of telomerase gene hTERT in testes of infertile male. METHODS: By using in situ hybridization(ISH) techniques, the expression of telomerase gene hTERT mRNA in testes of 47 infertile male and 10 normal testicular tissues were observed. RESULTS: In male testes, there was a positive correlation between the expression of hTERT and the quantity and density of germ cells(spermatogonia, spermatocyte, spermatid). The expression of hTERT in some germinal cell of maturation arrest patients were not significantly different with those of normal. CONCLUSIONS: Our results suggest that the deficiency of telomerase might be a factor for germinal cell maturation arrest and there might be some other etiological factors in these patients. Our study provides experimental groundwork for the gene therapy of male infertility.