shRNA constructs targeting IGFBP-3 alleviate age related erectile dysfunction in the rat.

PURPOSE: We investigated whether injecting shRNA constructs targeting IGFBP-3 in the penis of old rats would improve erectile function. MATERIALS AND METHODS: The most validated IGFBP-3 shRNA plasmid vector (pGPU6/GFP/Neo-shIGFBP-3) was prepared and injected in penile corpus cavernosum tissue. A total of 30 old (age 24 months) male Sprague Dawley® rats were randomly divided into 3 groups, including 10 each that received phosphate buffered saline only (100 µl), pGPU6/GFP/Neo-shNC (100 µg) and the most validated plasmid constructs pGPU6/GFP/Neo-shIGFBP-3 (100 µg). At 4 weeks the erectile response was measured as intracavernous pressure. The percent of smooth muscle in corpus cavernosum tissue was evaluated. Nitric oxide synthase activity and the cGMP concentration in penile tissue were also analyzed. IGFBP-3 was estimated in penile tissue by Western blot, real-time reverse transcriptase-polymerase chain reaction and immunohistochemistry. RESULTS: pGPU6/GFP/Neo-shIGFBP-3 corrected the impaired erectile response in aged rats compared with the response in those injected with phosphate buffered saline and pGPU6/GFP/Neo-shNC (each p <0.01). The percent of cavernous smooth muscle was increased in the pGPU6/GFP/Neo-shIGFBP-3 group. Nitric oxide synthase activity and the cGMP concentration were also significantly increased in rats treated with pGPU6/GFP/Neo-shIGFBP-3. IGFBP-3 shRNA effectively reduced IGFBP-3 mRNA and protein expression in penile corpus cavernosum tissue. CONCLUSIONS: Decreasing IGFBP-3 expression by plasmid expressed shRNA improved erectile function in aged rats. The therapy may modulate smooth muscle integrity and increase the cGMP concentration. This may be a new direction for treating erectile dysfunction in clinical practice.

[Influences of three surgical approaches to urethral stricture on the erectile function of the patients].

OBJECTIVE: To evaluate the impacts of three different surgical approaches to urethral stricture on the erectile function of the patients. METHODS: This study included 126 male patients with urethral stricture, 35 treated by substitution urethroplasty (group A), 52 by anastomotic urethroplasty (group B), and 39 by internal urethroplasty (group C). We evaluated the pre- and postoperative erectile function of the patients using IIEF-5 scores by telephone calls and interviews. We also monitored their nocturnal penile tumescence (NPT). RESULTS: The IIEF-5 scores in groups A, B and C were 13.5 +/- 4.5, 11.1 +/- 4.8 and 14.5 +/- 4.41 respectively after surgery, all significantly decreased as compared with 17.1 +/- 2.6, 17.1 +/- 3.0 and 17.6 +/- 2.2 preoperatively (P < 0.05). CONCLUSION: All the three surgical approaches can reduce IIEF-5 scores in patients with urethral stricture, but anastomotic urethroplasty may induce a higher incidence of erectile dysfunction than the other two approaches.

[Impact of aging on the secretion of insulin-like growth factor-1 in the corpus cavernosum smooth muscle cells of rats in vitro].

OBJECTIVE: To observe the secretion of insulin-like growth factor-1 (IGF-1) in corpus cavernosum smooth muscle cells (CCSMCs) in rats of different ages and explore the possible relationship of IGF-1 with aging-related erectile dysfunction (ED). METHODS: We primarily cultured CCSMCs of rats aged 4, 12 and 24 months, and identified them by immunohistochemistry. We quantitatively cultured the CCSMCs in 6-well culture plates, determined the levels of IGF-1 secreted from the CCSMCs by enzyme immunoassay (EIA) and analyzed the effect of age on the IGF-1 level. RESULTS: CCSMCs were successfully cultured in vitro. The level of IGF-1 secreted from the CCSMCs was decreased with the increase of age, with 7.1 ng/10(5) cells in the 4-month-old group, 2.2 ng/10(5) cells in the 12-month group, and 1.9 ng/10(5) cells in the 24-month group, with statistically significant differences among the three groups (P < 0.01). CONCLUSION: The secretion of IGF-1 is reduced with the increase of age, and the decreased expression of IGF-1 might be associated with aging-related ED.

[Application of CUA Guidelines on Prostatitis in the management of chronic pelvic pain syndrome: a nationwide survey].

OBJECTIVE: To investigate the application of the Chinese Urological Association (CUA) Guidelines on Prostatitis and its effects on the clinical practice patterns of diagnosing and treating chronic pelvic pain syndrome (CPPS) among Chinese urologists and andrologists. METHODS: We conducted a questionnaire investigation on the application of the CUA Guidelines on Prostatitis among the urologists and andrologists of 173 hospitals in 21 cities of China, and performed statistical analyses on all the eligible questionnaires collected. RESULTS: Of the 1 056 questionnaires distributed, 851 (80.6%) were eligible, of which 71.6% were from the urologists or andrologists in grade 3 hospitals, 80.7% of them with senior or intermediate professional titles and 97.5% had studied the CUA Guidelines. Most of the subjects agreed that Type III prostatitis is a clinical syndrome, whose diagnosis should exclude other conditions with similar symptoms, and whose treatment should aim at relieving pain, alleviating urination symptoms and improving the quality of life. Those who had and those who had not studied the CUA Guidelines differed in their viewpoints on CPPS as illustrated in the book. In clinical practice, the most common treatment options for CPPS were psychological therapy (80.7%), medication (80.4%) and life style adjustment (79.6%), and the most frequently used drugs were phytotherapy (80.0%), alpha-blockers (68.9%) and antibiotics (61.0%). CONCLUSION: CUA Guidelines on Prostatitis has gained a nationwide application and promoted the standardization of the management of CPPS in China.

[Insulin-like growth factor-1 gene therapy improves the levels of mRNA and protein of endothelial nitric oxide synthase in aging related erectile dysfunction in rats].

OBJECTIVE: To explore the effects of gene transfer of insulin like growth factor-1 (IGF-1) on the penis of senile rats and the altered levels of mRNA and protein of endothelial nitric oxide synthase (eNOS). METHODS: Ten young (4 months) and 20 senile (24 months) Sprague-Dawley male rats were selected. The senile rats were divided into 2 groups: phosphate buffer solution (PBS)-only (n = 10) and 100 µg IGF-1 plasmid treatment group (n = 10). After a 4-week injection of IGF-1, the responses of intracavernous pressure (ICP) with electrical stimulation to the cavernous nerve and systemic mean arterial pressure (MAP) were evaluated. In the control and transfected senile rats, the levels of eNOS mRNA and protein were examined by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot respectively. RESULTS: The ICP/MAP and total ICP were significantly higher in the young control group versus the PBS-only group at Week 4 (P < 0.05). The ICP/MAP and total ICP were significantly higher in the young control group and the 100 µg IGF-1 treatment group versus the PBS-only group at Week 4 (P < 0.05). The levels of mRNA and protein of eNOS were higher in the 100 µg IGF-1 treatment group versus the PBS-only group at Week 4 (0.62 ± 0.16 vs 0.25 ± 0.08, 0.71 ± 0.19 vs 0.27 ± 0.09, both P < 0.05, respectively). CONCLUSION: The gene therapy of IGF-1 can ameliorate erectile functions and improve the levels of mRNA and protein of eNOS in senile rats.

Higher expression of mRNA and protein of insulin-like growth factor binding protein-3 in old rat penile tissues: implications for erectile dysfunction.

INTRODUCTION: Previous studies have confirmed the gene transfer of insulin-like growth factor-1 (IGF-1) and the IGF-1 protein can improve the erectile function in aging rats. IGF binding protein (BP)-3 can regulates the availability of IGF-I. The higher expression of IGFBP-3 may play an important role in erectile dysfunction (ED). AIM: The study aimed to investigate the mRNA and protein expression of IGFBP-3 in young and old rat penile tissues and assess the alteration of the penile structure and the NO-guanosine 3',5'-cyclic-monophosphate (cGMP) signaling pathways-related marker in ED associated with aging. MAIN OUTCOME MEASURES: The main outcome measures for this study were the expression of IGFBP-3, morphological changes, NO-cGMP signaling pathways-related marker, erectile responses were determined. METHODS: Traditional reverse transcriptase polymerase chain reaction (RT-PCR) and real-time PCR were performed to examine the mRNA expression of the IGFBP-3. The Western blot was used to confirm the protein expression. Immunohistochemistry was also performed to identify the cellular localization of the encoded protein. The percentage of smooth muscle in corpus cavernosum tissue, the activity of nitric oxide synthase (NOS), and concentration of cGMP in penile tissue were also analyzed. RESULTS: The expression levels of IGFBP-3 of mRNA and protein were greatly increased in aging rats compared with young control rats, which is confirmed by traditional RT-PCR, real-time PCR, and Western blot (P < 0.01, respectively). Increased IGFBP-3 protein was localized to the epithelium of the urethra, penile endothelium, and smooth muscle in the corpus cavernosum. Significant depletion of the smooth muscle density relative to the connective tissue was also observed in the penis of the aged rats, and the lower activity of NOS and lower concentration of cGMP was also demonstrated accompanied with a significant reduction in the intracavernous pressure. CONCLUSIONS: Our data suggest that the increased mRNA and protein expression of IGFBP-3 in old rats may play a role in ED.

[Qianlie Beixi Capsules for unliquefiable semen: a multi-center clinical study].

OBJECTIVE: To investigate the efficacy and possible action mechanism of Qianlie Beixi Capsules in the treatment of unliquefiable semen. METHODS: A total of 190 patients with unliquefiable semen were treated with Qianlie Beixi Capsules for 1 or 2 courses (3 weeks per a course). The seminal changes were observed and recorded. RESULTS: Of the 190 patients in the 1-course treatment arm, 99 were cured and 91 failed to respond after the first course. And the effectiveness rate was 52.1%. Of the 122 patients in the 2-course treatment arm, 81 were cured and 41 failed to respond after the second course. And the effectiveness rate was 66.4%. The efficacy of 2-course regimen was obviously better than that of 1-course regiment. In the meantime, sperm density improved in the 2-course treatment arm. Sperm motility improved slightly in the effective subjects of 1-course treatment arm. All the above results had statistically significant differences (P < 0.05). CONCLUSION: Qianlie Beixi Capsules is both safe and effective for unliquefiable semen and may shorten the time of seminal liquefaction.

[Growth factors and gene therapy for erectile dysfunction].

Growth factors have a universal bioactivity. Gene therapy is a new strategy in dealing with erectile dysfunction (ED). This paper presents an overview on the value of growth factors, particularly the vascular epithelial growth factor (VEGF) and insulin-like growth factor 1 (IGF-1), in the treatment of ED.

[Induction of cell cycle arrest in bladder cancer RT4 cells by capsaicin].

OBJECTIVE: To study the effects of capsaicin on the growth of bladder cancer RT4 cell and its potential mechanism. METHODS: Cell counting kit-8 (CCK-8) assay and flow cytometry were employed to observe the effects of capsaicin (50, 100, 150, 200, 250 micromol/L) on cell growth, cell cycle and apoptosis. Capsaicin (0 micromol/L) was used as a control. The effects of mRNA and protein of transient receptor potential cation channel subfamily V 1 (TRPV1) on RT4 cells were tested by RT-PCR and immunofluorescence respectively. And the expressions of cell cycle protein P53, P21, CDK2 were detected by Western blot after the treatment of capsaicin. RESULTS: 100 micromol/L capsaicin significantly decreased the viability of RT4 cell [82.0% +/- 6.2% vs 100.0% +/- 12.4% (control), P = 0.036] while the cell viability was 7.8% +/- 2.9% at 250 micromol/L (P = 0.000). It was in a dose-dependent manner. On the other hand, capsaicin induced the cell cycle arrest of bladder cancer RT4 cells G(0)/G(1) phase in a dose-dependent way. The cell proportion of G(0)/G(1) phase in the control was 37.4% +/- 5.6%, however, it was 72.4% +/- 5.3% at 250 micromol/L (P = 0.000). It was showed that TRPV1 mRNA and protein were expressed in RT4 cells. After a 48-hour treatment with capsaicin, the expressions of P53 and P21 were up-regulated in contrary to the expression of CDK2. CONCLUSION: Capsaicin induces the cell cycle arrest of bladder cancer RT4 cells G(0)/G(1) phase and growth inhibition via TRPV1 receptor by modulating the expression of P53, P21 and CDK2.

[Bipolar transurethral resection of the prostate versus monopolar transurethral prostatectomy: a pathological study in a canine model].

OBJECTIVE: To compare the postoperative depths of the coagulation zones and pathological changes between bipolar transurethral resection of the prostate with plasmakinetic energy (PKRP) and monopolar transurethral prostatectomy (TURP) in canines. METHODS: Twenty-five male dogs were randomly divided into a PKRP group (n = 12), a TURP group (n = 12) and a sham-operation control group (n = 1). The dogs were sacrificed, their prostates harvested at 0 week (immediately after surgery), 1 week, 2 weeks and 8 weeks postoperatively and sectioned for pathologic analysis and measurement of the coagulation zones. RESULTS: At 0, 1 and 2 weeks after the operation, the coagulation depths were (237.73 +/- 20.12) microm, (113.03 +/- 16.65) microm and (106.01 +/- 16.36) microm in the PKRP group, and (200.75 +/-19.34) microm, (129.46 +/- 17.81) microm and (116.04 +/- 25.67) microm in the TURP group (P < 0.01). At 8 weeks, the coagulation zones completely peeled off and the wounds were covered by regenerated urothelial in both of the groups. At 0, 1, 2 and 8 weeks, different inflammatory reactions were observed in the prostates of the PKRP and TURP groups, with some glandular lumens beneath the coagulation zones expanded and epithelia damaged. However, none of these phenomena occurred in the sham-operation control group. CONCLUSION: Pathologically, PKRP and TURP inflicted basically similar effects on the prostate of the canine. However, the coagulation zone was deeper intraoperatively and became thinner postoperatively with the former than with the latter, which suggests that PKRP causes less bleeding and less penetrative thermal damage than TURP.

Effects of TRPM8 on the proliferation and motility of prostate cancer PC-3 cells.

We investigated the effects of transient receptor potential M8 (TRPM8) channel on the proliferation and motility of androgen-independent prostate cancer PC-3 cells. After being permanently transfected with an empty vector and cDNA encoding the TRPM8 protein, cells were analysed for cell cycle distribution and motility using flow cytometry and scratch assay. Immunocytochemistry and Ca2+ imaging analysis revealed the overexpression of functional TRPM8 channel on both endoplasmic reticulum and plasma membrane of PC-3-TRPM8 cells. Cell cycle distribution and scratch assay analysis revealed that TRPM8 induced cell cycle arrest at the G0/G1 stage (P < 0.05) and facilitated the cell apoptosis induced by starvation (P < 0.05). Furthermore, TRPM8 inhibited the migration of PC-3-TRPM8 cells (P < 0.01) through the inactivation of focal-adhesion kinase. It appears that TRPM8 was not essential for the survival of PC-3 cells; however, the overexpression of TRPM8 had negative effects on the proliferation and migration of PC-3 cells. Thus, TRPM8 and its agonists may serve as important targets for the treatment of prostate cancer.

The value of combined use of survivin, cytokeratin 20 and mucin 7 mRNA for bladder cancer detection in voided urine.

OBJECTIVES: To evaluate the value of combined use of survivin, cytokeratin (CK) 20 and mucin (MUC) 7 mRNA in comparison with voided urine cytology in the detection of bladder cancer patients. METHODS: One hundred and fifty three patients and 20 healthy volunteers were evaluated by RT-PCR for detecting survivin, CK-20 and MUC7 mRNA in voided urine before cystoscopy. The three markers and cytology were evaluated independently or in combinations. RESULTS: The overall sensitivity and specificity were 90.4 and 94.7% for survivin, 82.6 and 97.4% for CK-20, 62.6 and 94.7% for MUC7 and 46.0 and 100% for voided urine cytology. Combined sensitivity of voided urine cytology with the three biomarkers together was higher than either combined sensitivity of voided urine cytology with one of the biomarkers or than that of the biomarker alone. CONCLUSIONS: Combined use of the three markers can improve the sensitivity for detecting bladder cancer.

Insulin-like growth factor-1 restores erectile function in aged rats: modulation the integrity of smooth muscle and nitric oxide-cyclic guanosine monophosphate signaling activity.

INTRODUCTION: Insulin-like growth factor-1 (IGF-1) is one of the growth factors that have a wide range of biologic effects. We have confirmed that gene transfer of IGF-1 to the penis could improve erectile capacity. However, there are some limitations in gene therapies, such as toxicity or a risk of insertional mutagenesis. Protein treatment may be another choice for decreasing these risks. AIM: To investigate whether intracavernosal injection of IGF-1 protein can restore erectile function in the aging rat. MAIN OUTCOME MEASURES: Erectile responses, morphological changes, and nitric oxide-cyclic guanosine monophosphate (NO-cGMP) signaling pathways-related marker were determined. METHODS: Ten young (4 months) and 30 old (24 months) Sprague-Dawley male rats were enrolled in this study. The old rats were divided into three groups: vehicle-only (N = 10), IGF-1 1 microg/kg (N = 10) and IGF-1 10 microg/kg treatment group (N = 10). After 4 and 8 weeks of single IGF-1 injection treatment, intracavernous pressure (ICP) responses with electrical stimulation to the cavernous nerve were evaluated. The percent of smooth muscle in corpus cavernosum tissue, the expression of mRNA and protein of endothelial nitric oxide synthase (eNOS) were also evaluated. The activity of nitric oxide synthase (NOS) and concentration of guanosine 3',5'-cyclic-monophosphate (cGMP) that act upon the major NO-cGMP signaling pathways in penile tissue were also analyzed. RESULTS: After IGF-1 treatment, the ICP responses was significantly increased as the young control group in both the IGF-1 1 microg/kg and the IGF-1 10 microg/kg group compared with the vehicle-only group at 4 and 8 weeks (P < 0.05). Masson's trichrom staining showed the percentage of cavernosal smooth muscle was increased in IGF-1 treatment group. IGF-1 increased e-NOS expression. NOS activities and cGMP concentrations were also significantly increased in IGF-1 treatment rats. CONCLUSIONS: IGF-1 improved erectile function in aged rats via restoration the integrity of smooth muscle of corpus cavernosum and modulation of NO-cGMP pathways.

Use of bipolar energy for transurethral resection of superficial bladder tumors: long-term results.

PURPOSE: To evaluate the long-term efficacy and safety of transurethral resection of bladder tumor (TURBT) with bipolar plasmakinetic energy. PATIENTS AND METHODS: We reviewed the records of 121 patients with superficial transitional cell carcinoma of the bladder treated at our institute. Bipolar TURBT with plasmakinetic energy was performed for diagnostic and therapeutic purposes in all patients. Resected tissue was examined by a pathologist who recorded the number of tumors, tumor size, tumor shape, location, grade, invasion of the muscularis propria, and presence of muscular invasion. The operating time, length of hospital stay, blood loss, and intraoperative and postoperative complications were recorded by a urologist. Follow-up was 3 to 5.5 years after operation. RESULTS: The median age of the patients was 61 years; 41 patients had multiple tumors and 80 had single tumors. The mean tumor size was 1.9 cm in diameter. The tumor was located in the lateral wall of the bladder in 67 patients. The mean operative time was (25 +/- 16) minutes and the mean postoperative hospitalization period was 3 days. Three (2.5%) patients had hematuria requiring blood transfusion and 2 (1.7%) patients had bladder perforation. Adductor contraction was noted in 6 patients (4.9%), and urethral strictures occurred in 5 patients (4.1%). CONCLUSION: Transurethral resection of bladder tumors with bipolar plasmakinetic energy is safe and effective in the treatment of superficial bladder tumors.

[Yiqigushen capsule combined with levofloxacin highly effective for type III A prostatitis].

OBJECTIVE: To evaluate the clinical efficacy of the Chinese medicine Yiqigushen Capsule combined with levofloxacin on type III A prostatitis. METHODS: A total of 180 patients with type III A prostatitis were equally randomized into a treatment group, which received Yiqigushen Capsule combined with levofloxacin, and a control group, which was given Yiqigushen Capsule only, both for 4 weeks. NIH-CPSI scores and leukocyte counts in the expressed prostatic secretions of all the patients were analyzed before and after the treatment. RESULTS: After the treatment, NIH-CPSI scores and leukocyte counts in the expressed prostatic secretions of both the groups were significantly decreased compared with pre-treatment (P < 0.05 or 0.01), with statistic differences in NIH-CPSI scores but not in leukocyte counts (P < 0.05) between the two groups. CONCLUSION: Yiqigushen Capsule combined with levofloxacin is highly effective for type III A prostatitis.

[Dose and long-term effect of hIGF-1 injection for erectile dysfunction in aged rats].

OBJECTIVE: To investigate the best dose and the long-term effect of the human insulin-like growth factor-1 (hIGF-1) gene injection into the penis of aged rats. METHODS: Included in this study were 10 young (4 months old) and 40 aged (24 months old) Sprague-Dawley male rats, the latter equally divided into a PBS control and a 10 microg, a 100 microg and a 1 000 microg hIGF-1 injection group. Electrical stimulation was conducted 4 and 8 weeks after hIGF-1 injection into the penile corpus cavernous of the rats to detect the intracavernous pressure (ICP) and mean arterial pressure (MAP). Dose - and time -associated therapeutic results were analyzed and the mRNA expression of hIGF-1 determined by RT - PCR. RESULTS: ICP, MAP and total ICP were significant decreased by electrical stimulation in the aged rats as compared with the young ones (P < 0.05), statistically increased in the three hIGF-1 dose groups in comparison with the PBS controls (P < 0.05), and showed no obvious difference between the young rats and the latter two dose groups at 4 and 8 weeks. Although less obvious effect was achieved in the 10 microg group than in the young rats, the therapeutic result was still of significance. The mRNA expression of the hIGF-1 gene was confirmed in all the hIGF-1 treated rats. CONCLUSION: The hIGF-1 therapy can improve erectile function in aged rats, 100 microg suffices for effective erection and the effect may last at least 8 weeks for a single dose.

Comparative study of the impact of 3- versus 8-month neoadjuvant hormonal therapy on outcome of laparoscopic radical prostatectomy.

PURPOSE: To comparatively evaluate the effect of 3- versus 8-month neoadjuvant hormonal therapy (NHT) on laparoscopic radical prostatectomy (LRP) operative and postoperative parameters. METHODS: We evaluated 55 patients with clinically localized prostate cancer treated by 3-month (25 patients, group-1), 8-month NHT (19 patients, group-2) and non-neoadjuvant therapy (11 patients, group-3) before LRP. Serum PSA levels and prostate volume were measured and evaluated monthly before operation. The operative and postoperative parameters were recorded and compared in the three different groups. RESULTS: Transrectal ultrasound determined that prostate volume decreased 39.6% in group 1 and 35.4% in group 2 after 3-month NHT and a further 34.4% in group 2 after 8-month NHT. The mean prostate volume was significantly smaller after 8-month than 3-month NHT in group 2 (P < 0.05). Mean serum PSA decreased 97.8% in group 1 and 98.1% in group 2 after 3-month NHT. A further 72.9% PSA decrease was determined after 8-month NHT in group 2. There were no significant differences in the three groups with respect to mean operative time, mean blood loss, transfusion rate, operative difficulty, catheterization time, hospital time and complication rate (P > 0.05, respectively). Positive margin rate was significantly lower in the 3- or 8-month NHT group than in the non-adjuvant group (P < 0.05, respectively). However, there was no significant difference between the 3- and 8-month groups with respect to positive margin rate (P > 0.05). CONCLUSIONS: The prolonged NHT (8-month) did not affect the operative or postoperative parameters. The LRP can be safely performed in patients after prolonged NHT. Prospective randomized studies are required to determine whether prolonged NHT reduces the risk of biochemical recurrence after LRP.

Distribution profiles of transient receptor potential melastatin-related and vanilloid-related channels in prostatic tissue in rat.

AIM: To investigate the expression and distribution of the members of the transient receptor potential (TRP) channel members of TRP melastatin (TRPM) and TRP vanilloid (TRPV) subfamilies in rat prostatic tissue. METHODS: Prostate tissue was obtained from male Sprague-Dawley rats. Reverse transcription polymerase chain reaction (RT-PCR) and quantitative real-time polymerase chain reaction (PCR) were used to check the expression of all TRPM and TRPV channel members with specific primers. Immunohistochemistry staining for TRPM8 and TRPV1 were also performed in rat tissues. RESULTS: TRPM2, TRPM3, TRPM4, TRPM6, TRPM7, TRPM8, TRPV2 and TRPV4 mRNA were detected in all rat prostatic tissues. Very weak signals for TRPM1, TRPV1 and TRPV3 were also detected. The mRNA of TRPM5, TRPV5 and TRPV6 were not detected in all RT-PCR experiments. Quantitative real-time RT-PCR showed that TRPM2, TRPM3, TRPM4, TRPM8, TRPV2 and TRPV4 were the most abundantly expressed TRPM and TRPV subtypes, respectively. Fluorescence immunohistochemistry indicated that TRPM8 and TRPV1 are highly expressed in both epithelial and smooth muscle cells. CONCLUSION: Our results demonstrate that mRNA or protein for TRPM1, TRPM2, TRPM3, TRPM4, TRPM6, TRPM7, TRPM8, TRPV1, TRPV2, TRPV3 and TRPV4 exist in rat prostatic tissue. The data presented here assists in elucidating the physiological function of TRPM and TRPV channels.

Improvement in erectile dysfunction after insulin-like growth factor-1 gene therapy in diabetic rats.

AIM: To determine whether adenoviral gene transfer of insulin like growth factor-1 (IGF-1) to the penis of streptozotocin (STZ)-induced diabetic rats could improve erectile capacity. METHODS: THE STZ diabetic rats were transfected with AdCMV-betagal or AdCMV-IGF-1. These rats underwent cavernous nerve stimulation to assess erectile function and their responses were compared with those of age-matched control rats 1 to 2 days after transfection. In control and transfected STZ diabetic rats, IGF-1 expression were examined by reverse transcription polymerase chain reaction (RT-PCR), Western blot and histology. The penis beta-galactosidase activity and localization of the STZ diabetic rats were also determined. RESULTS: One to two days after transfection, the beta-galactosidase was found in the smooth muscle cells of the diabetic rat penis transfected with AdCMV-betagal. One to 2 days after administration of AdCMV-IGF-1, the cavernosal pressure, as determined by the ratio of maximal intracavernous pressure-to-mean arterial pressure (ICP/MAP) and total intracavernous pressure (ICP), was increased in response to cavernous nerve stimulation. Transgene expression was confirmed by RT-PCR, Western blot and histology. CONCLUSION: Gene transfer of IGF-1 significantly increased erectile function in the STZ diabetic rats. These results suggest that in vivo gene transfer of IGF-1 might be a new therapeutic intervention for the treatment of erectile dysfunction (ED) in the STZ diabetic rats.

[Antifertility effect of 30% ethanol retro-injection into rat vas deferens].

OBJECTIVE: To explore the antifertility effect and safety of 30% ethanol retro-injection into the vas deferens of the rat. METHODS: Thirty Sprague-Dawley male rats, 3 m of age and (200 +/- 20) g in weight, were equally randomized into an experimental group and a control group. The former received 30% ethanol (0.5 ml) and the latter 0.9% sodium chloride (0.5 ml), both retro-injected into the vas deferens. Pregnancy rates were obtained through pregnancy tests with 60 Sprague-Dawley female adult rats 1.5 m and 3 m after the injection. All the male rats were sacrificed three months later, and tests were done for the rates of sperm motility and deformity as well as for the apoptosis of spermatogenic cells with TUNEL. RESULTS: The 1.5 m pregnancy rate was 0 and the 3 m sperm motility and pregnancy rates were (0.32 +/- 1.12)% and (0.58 +/- 1.27)%, significantly decreased (P < 0.05) as compared with those of the control group, which were (80.62 +/- 2.68)%, (70.68 +/- 1.62)% and (86.62 +/- 1.68)%, respectively. While the 3 m sperm deformity rate in the experimental group was (78.26 +/- 1.08)%, increased significantly (P < 0.05), and the apoptosis index (AI) of spermatogenic cells was (7.63 +/- 1.16)% as compared with (5.62 +/- 1.32)% of the control group, with no significant difference between the two groups (P > 0.05). CONCLUSION: Retro-injection of 30% ethanol into the vas deferens of the rat produces significant antifertility effect on rats, but has no significant influence on their spermatogenic cells.