Obstetric and Gynecologic Outcomes after the Transvaginal Repair of Cesarean Scar Defect in a Series of 183 Women.

STUDY OBJECTIVE: To analyze retrospectively the effect of hysteroscopy combined with transvaginal repair on the cesarean section diverticulum (CSD) and explore the clinical significance of this procedure. DESIGN: Retrospective study. SETTING: University-affiliated hospital and a gynecology hospital. PATIENTS: A total of 183 patients with scar diverticulum after cesarean section were recruited from the Southern Medical University Affiliated Maternal & Child Health Hospital of Foshan and Shenzhen In Vitro Fertilization Gynecological Hospital. INTERVENTIONS: In this study, we reported a surgical method for repairing uterine scar through uterine therapy and explored its clinical efficacy and pregnancy outcome. MEASUREMENTS AND MAIN RESULTS: The time of operation, volume of bleeding, and duration of hospitalization were recorded. The size of the scar diverticulum and the remaining myometrium were examined by B-mode ultrasonography before and after the operation. The length of the menstrual cycle and pelvic pain were recorded during follow-up to check the recovery of patients after surgery. The pregnancy of patients with pregnancy needs was recorded to check the pregnancy outcome. All 183 patients successfully completed the repair of the transvaginal uterus scar diverticulum with the help of a hysteroscopy examination. The mean (± standard deviation) operation time was 58.61 ± 18.56 minutes. The mean blood loss was 36.97 ± 22.32 mL. The mean hospital stay was 6.08 ± 1.89 days. In 57.14% of patients, the CSD completely disappeared, whereas the volume of CSD shrank by at least 50% in 88.95% of patients. The mean menstrual period of patients after surgery was 7.72 ± 2.68 days, which was significantly shorter than that recorded preoperatively (13.45 ± 3.69 days) (t = 19.62, p = .00). The pelvic pain disappeared in 81.08% of the patients. The mean postoperative thickness of the remaining muscular layer was 5.30 ± 1.27-mm, which was significantly higher than the preoperative value of 2.25 ± 0.92-mm (t = 28.21, p = .00). The mean postoperative thickness of the remaining muscular layer of patients with improved menstrual cycle was 5.40 ± 1.27-mm, which was significantly higher than the thickness of 4.88 ± 1.11-mm in patients without improved menstrual cycle (t = 2.31, p = .025). A total of 124 patients attempted to become pregnant, 83 of whom were successful. The pregnancy rate was as high as 66.95%, which included 2 scar pregnancies, 4 ectopic pregnancies, and 87 intrauterine pregnancies. No uterine rupture occurred. CONCLUSION: The transvaginal repair of the uterine diverticulum improved the symptoms and probability of a successful pregnancy effectively. This process is a surgical procedure to increase the thickness of the residual uterine muscle wall effectively.

Rce1 expression in renal cell carcinoma and its regulatory effect on 786-O cell apoptosis through endoplasmic reticulum stress.

Ras and a-factor-converting enzyme 1 (Rce1) is located in the endoplasmic reticulum (ER) and is thought to be responsible for endoproteolytic processing of the vast majority of CAAX proteins. Endoplasmic reticulum stress (ERS) plays an important role in renal cell carcinoma (RCC); however, the expression and role of Rce1 in RCC have not been extensively studied. We aimed to investigate the expression of Rce1 in RCC tissues and its molecular mechanism in ERS-induced apoptosis in RCC 786-O cells. We first used western blotting, quantitative reverse transcriptase-polymerase chain reaction, and immunohistochemistry to detect the Rce1 expression in renal carcinoma tissues and paracancerous tissues. It was found that Rce1 expression was upregulated in RCC tissues, and its positive expression level was strongly associated with clinicopathologic features. Next, we detected the expression of Rce1 in human embryonic kidney cell line HEK293 and human renal carcinoma cell lines 786-O, ACHN, and A498. Higher expression of Rce1 was found in human renal carcinoma cell lines, especially in 786-O cells. Knockdown of Rce1 in 786-O cells increased apoptosis and inhibited proliferation (P < 0.05). Moreover, downregulation of Rce1 upregulated the expression of the pro-apoptotic protein Bax, but downregulated the expression of the anti-apoptotic protein Bcl-2. Further studies showed that downregulation of Rce1 also affected the expression of ERS factors. In conclusion, our results indicated that Rce1 plays a key role in RCC. Low expression of Rce1 might indirectly increase apoptosis and inhibit proliferation of renal carcinoma cells through ERS.

[Expression of macrophage inflammatory protein-1alpha in EPS and its significance].

OBJECTIVE: To detect the mRNA and protein expressions of MIP-1alpha in EPS and determine their significance in the sub-typing of chronic prostatitis. METHODS: We collected samples of expressed prostatic secretion (EPS) from 50 cases of chronic prostatitis, including 16 cases of chronic bacterial prostatitis (CBP), 23 cases of chronic nonbacterial prostatitis/chronic pelvic pain syndrome (CPPS) (11 CPPS IIIA, 12 CPPS IIIB), and 11 cases of type-IV asymptomatic inflammatory prostatitis (AIP). Another 15 healthy volunteers were included as normal controls. The mRNA and protein levels of MIP-1alpha in EPS were determined by RT-PCR and ELISA, respectively, followed by statistical analysis with SPSS 15.0. RESULTS: The mRNA expression of MIP-1alpha was markedly higher in the CPPS IIIA and CPPS IIIB groups than in the others (P<0.05). The protein level of MIP-1alpha was (1174.3 +/- 89.2) pg/ml in CPPS IIIA and (842.3 +/- 76.2) pg/ml in CPPS IIIB, significantly higher than (198.0 +/- 37.8) pg/ml in the control, (347.0 +/- 61.6) pg/ml in CBP and (292.0 +/- 56.4) pg/ml in type-IV AIP (P<0.05). CONCLUSION: Determination of mRNA and protein levels of MIP-1alpha in EPS may help the sub-typing and diagnosis of chronic prostatitis.

Effects of varicocele and microsurgical varicocelectomy on the metabolites in semen.

The influence of varicocele and microsurgical varicocelectomy on semen quality remains unclear. Few studies have investigated the relationship between semen metabolism and the abnormalities in reproductive function caused by varicocele, however, there is no study on the changes of semen metabolism after microsurgical varicocelectomy. Here, we used the non-targeted and targeted metabolic analysis to investigate the different metabolites in seminal plasma within normal, varicocele, and varicocelectomy groups. We clearly showed that varicocele significantly affects semen metabolism, and microsurgical varicocelectomy can reverse this metabolic abnormality. Moreover, we characterized the landscape of three dipeptides in the seminal plasma of patients with varicocele that have not been identified previously in human tissues or biofluids. Interestingly, the levels of these three dipeptides decreased after microsurgical varicocelectomy coincident with an improvement in semen quality. Western blotting confirmed the downregulation of DPEP3 (dipeptidase 3) in the varicocele group and the upregulation of DPEP3 in the varicocelectomy group. Furthermore, we found that eight metabolites may be helpful to distinguish varicocele patients from normal subjects. Our results may be applied to earlier diagnosis or to predict the outcome of microsurgery for varicocele.

Intravesical dimethyl sulfoxide inhibits acute and chronic bladder inflammation in transgenic experimental autoimmune cystitis models.

New animal models are greatly needed in interstitial cystitis/painful bladder syndrome (IC/PBS) research. We recently developed a novel transgenic cystitis model (URO-OVA mice) that mimics certain key aspects of IC/PBS pathophysiology. This paper aimed to determine whether URO-OVA cystitis model was responsive to intravesical dimethyl sulfoxide (DMSO) and if so identify the mechanisms of DMSO action. URO-OVA mice developed acute cystitis upon adoptive transfer of OVA-specific OT-I splenocytes. Compared to PBS-treated bladders, the bladders treated with 50% DMSO exhibited markedly reduced bladder histopathology and expression of various inflammatory factor mRNAs. Intravesical DMSO treatment also effectively inhibited bladder inflammation in a spontaneous chronic cystitis model (URO-OVA/OT-I mice). Studies further revealed that DMSO could impair effector T cells in a dose-dependent manner in vitro. Taken together, our results suggest that intravesical DMSO improves the bladder histopathology of IC/PBS patients because of its ability to interfere with multiple inflammatory and bladder cell types.

[Expression and regulation of androgen receptor in the androgen-independent conversion of prostate cancer cells].

OBJECTIVE: To investigate the expression and regulation of androgen receptor (AR) in prostate cancer cells from androgen dependent to androgen independent. METHODS: LNCaP cells were cultured in charcoal-stripped serum for 6 months to establish androgen-independent celline (LNCaP-AI). Proliferation of LNCaP-AI was assayed by cell viability. Expression of AR mRNA and protein was analyzed by RT-PCR and Western blot. Wnt signaling pathway inhibitor IWR-1 and proteasome inhibitor lactacystin were used to investigate effects of Wnt and proteasome pathway on AR expression in LNCaP-AI. RESULTS: LNCaP-AI exhibit enhanced proliferation and up-regulated PSA expression compared with LNCaP. During androgen deprivation, AR mRNA was up-regulated in a short early stage and then declined to a stable level in LNCaP-AI compared with LNCaP, but AR protein kept in downward trend. The mRNA and protein expression of AR was decreased by IWR-1 treatment. AR protein but not mRNA was increased by lactacystin treatment. CONCLUSION: The androgen independent prostate cancer cell line was established by androgen deprivation, in which the protein expression of AR was dramatically decreased. mRNA and protein expression of AR in LNCaP-AI was related to Wnt signaling pathway and proteasome pathway. Increased Wnt signaling or decreased proteasome pathways contribute the decreased AR protein expression.

Reproductive outcomes of intracytoplasmic sperm injection using testicular sperm and ejaculated sperm in patients with AZFc microdeletions: a systematic review and meta-analysis.

Studies have explored the assisted reproductive technology (ART) outcomes of Y-chromosome azoospermia factor c (AZFc) microdeletions, but the effect of sperm source on intracytoplasmic sperm injection (ICSI) remains unknown. To determine the ART results of ICSI using testicular sperm and ejaculated sperm from males with AZFc microdeletions, we searched Embase, Web of Science, and PubMed to conduct a systematic review and meta-analysis. The first meta-analysis results for 106 cycles in five studies showed no significant differences in the live birth rate between the testicular sperm group and the ejaculated sperm group (risk ratio: 0.97, 95% confidence interval [CI]: 0.73-1.28, P = 0.82). The second meta-analysis of 106 cycles in five studies showed no difference in the abortion rate between the testicular sperm group and ejaculated sperm group (risk ratio: 1.06, 95% CI: 0.54-2.06, P = 0.87). The third meta-analysis of 386 cycles in seven studies showed no significant difference in clinical pregnancy rates between the testicular sperm group and the ejaculated sperm group (risk ratio: 1.24, 95% CI: 0.66-2.34, P = 0.50). Inevitable heterogeneity weakened our results. However, our results indicated that testicular sperm and ejaculated sperm yield similar ART outcomes, representing a meaningful result for clinical treatment. More properly designed studies are needed to further confirm our conclusions.

Recombinant bacillus Calmette-Guérin (BCG) expressing interferon-alpha 2B enhances human mononuclear cell cytotoxicity against bladder cancer cell lines in vitro.

PURPOSE: The proper induction of cellular immunity is required for effective bacillus Calmette-Guérin (BCG) immunotherapy of bladder cancer. It has been known that BCG stimulation of human peripheral blood mononuclear cells (PBMC) leads to the generation of effector cells cytotoxic to bladder cancer cells in vitro. To improve BCG therapy, we previously developed human interferon (IFN)-alpha 2B secreting recombinant (r) BCG (rBCG-IFN-alpha). We demonstrated that rBCG-IFN-alpha augmented T helper type 1 (Th1) cytokine IFN-gamma production by PBMC. In this study, we further investigated whether rBCG-IFN-alpha could also enhance PBMC cytotoxicity toward bladder cancer cells. MATERIALS AND METHODS: PBMC were prepared from healthy individuals, left alone or stimulated with rBCG-IFN-alpha or control MV261 BCG, and used as effector cells in (51)Cr-release assays. Human bladder cancer cell lines T24, J82, 5637, TCCSUP, and UMUC-3 were used as target cells. To determine the role of secreted rIFN-alpha as well as endogenously expressed IFN-gamma and IL-2 in inducing the cytotoxicity, PBMC were stimulated with rBCG-IFN-alpha in the presence of neutralizing antibodies to IFN-alpha, IFN-gamma or IL-2. To determine the role of natural killer (NK) and CD8(+) T cells in inducing the cytotoxicity, both cell types were isolated after BCG stimulation of PBMC and used as effector cells in (51)Cr-release assays. RESULTS: Non-stimulated PBMC showed basal levels of cytotoxicity against all target cell lines tested. MV261 BCG increased the PBMC cytotoxicity by 1.8- to 4.2-fold. rBCG-IFN-alpha further increased the PBMC cytotoxicity by up to 2-fold. Elevated production of IFN-gamma and IL-2 by PBMC was observed after rBCG-IFN-alpha stimulation. Blockage of IFN-alpha, IFN-gamma or IL-2 by neutralizing antibodies during rBCG-IFN-alpha stimulation reduced or abolished the induction of PBMC cytotoxicity. Both NK and CD8(+) T cells were found to be responsible for the enhanced PBMC cytotoxicity induced by rBCG-IFN-alpha with the former cell type being more predominant. CONCLUSIONS: rBCG-IFN-alpha is an improved BCG agent that induces enhanced PBMC cytotoxicity against bladder cancer cells in vitro. This rBCG strain may serve as an alternative to BCG for the treatment of superficial bladder cancer.

Evidence for the Role of Mast Cells in Cystitis-Associated Lower Urinary Tract Dysfunction: A Multidisciplinary Approach to the Study of Chronic Pelvic Pain Research Network Animal Model Study.

Bladder inflammation frequently causes cystitis pain and lower urinary tract dysfunction (LUTD) such as urinary frequency and urgency. Although mast cells have been identified to play a critical role in bladder inflammation and pain, the role of mast cells in cystitis-associated LUTD has not been demonstrated. Interstitial cystitis/bladder pain syndrome (IC/BPS) is a chronic and debilitating inflammatory condition of the urinary bladder characterized by the hallmark symptoms of pelvic pain and LUTD. In this study we investigated the role of mast cells in LUTD using a transgenic autoimmune cystitis model (URO-OVA) that reproduces many clinical correlates of IC/BPS. URO-OVA mice express the membrane form of the model antigen ovalbumin (OVA) as a self-antigen on the urothelium and develop bladder inflammation upon introduction of OVA-specific T cells. To investigate the role of mast cells, we crossed URO-OVA mice with mast cell-deficient KitW-sh mice to generate URO-OVA/KitW-sh mice that retained urothelial OVA expression but lacked endogenous mast cells. We compared URO-OVA mice with URO-OVA/KitW-sh mice with and without mast cell reconstitution in response to cystitis induction. URO-OVA mice developed profound bladder inflammation with increased mast cell counts and LUTD, including increased total number of voids, decreased mean volume voided per micturition, and decreased maximum volume voided per micturition, after cystitis induction. In contrast, similarly cystitis-induced URO-OVA/KitW-sh mice developed reduced bladder inflammation with no mast cells and LUTD detected. However, after mast cell reconstitution URO-OVA/KitW-sh mice restored the ability to develop bladder inflammation and LUTD following cystitis induction. We further treated URO-OVA mice with cromolyn, a mast cell membrane stabilizer, and found that cromolyn treatment reversed bladder inflammation and LUTD in the animal model. Our results provide direct evidence for the role of mast cells in cystitis-associated LUTD, supporting the use of mast cell inhibitors for treatment of certain forms of IC/BPS.

Overexpressed Rce1 is positively correlated with tumor progression and predicts poor prognosis in prostate cancer.

Ras and a-factor-converting enzyme 1 (Rce1) have been reported to play a key role in the proteolysis processing of Ras proteins. The present study investigated the prognostic significance of Rce1 in patients with prostate cancer (PCa). The expressions of the mRNA and protein of Rce1 were analyzed in 12 pairs of PCa and benign prostatic hyperplasia (BPH) by quantitative real-time polymerase chain reaction and Western blotting, respectively. Immunohistochemistry was used to examine expression of Rce1 protein in 74 PCa tissues and 30 BPH tissues. The association between Rce1 expression and the specific clinicopathologic features was evaluated by χ(2) tests. Kaplan-Meier and Cox proportional hazards regression models were used to analyze the data. We found that expression of Rce1 mRNA and protein was markedly higher in PCa tissues than in paired BPH tissues. Expression of Rce1 in PCa was strongly associated with clinicopathologic features. It was detected in 69 (93.24%) of 74 PCa tissues by immunohistochemistry, and it was found to be associated with Gleason score (P = .013), T class (P = .015), and distant metastasis (P = .044). Patients with PCa having higher Rce1 expression had substantially shorter survival times than patients with lower Rce1 expression. Univariate and multivariate analysis revealed that Rce1 was an independent prognostic factor. In conclusion, our study suggests that expression of Rce1 can serve as an independent biomarker for the prognosis of PCa patients.

Effects of icariin on erectile function and expression of nitric oxide synthase isoforms in castrated rats.

AIM: To investigate the effect of icariin on erectile function and the expression of nitric oxide synthase (NOS) isoforms in castrated rats. METHODS: Thirty-two adult male Wistar rats were randomly divided into one sham-operated group (A) and three castrated groups (B, C and D). One week after surgery, rats were treated with normal saline (groups A and B) or oral icariin (1 mg/[kg.day] for group C and 5 mg/[kg.day] for group D) for 4 weeks. One week after treatment, the erectile function of the rats was assessed by measuring intracavernosal pressure (ICP) during electrostimulation of the cavernosal nerve. The serum testosterone (ST) levels, the percent of smooth muscle (PSM) in trabecular tissue, and the expression of mRNA and proteins of neuronal nitric oxide synthase (nNOS), inducible nitric oxide synthase (iNOS), endothelial nitric oxide synthase (eNOS) and phosphodiesterase V (PDE5) in corpus cavernosum (CC) were also evaluated. RESULTS: ICP, PSM, ST and the expression of nNOS, iNOS, eNOS and PDE5 were significantly decreased in group B compared with those in group A (P 0.01). However, ICP, PSM and the expression of nNOS and iNOS were increased in groups C and D compared with those in group B (P 0.05). Changes in ST and the expression of eNOS and PDE5 were not significant (P 0.05) in groups C and D compared with those in group B. CONCLUSION: Oral treatment with icariin ( 98.6 % purity) for 4 weeks potentially improves erectile function. This effect is correlated with an increase in PSM and the expression of certain NOS in the CC of castrated rats. These results suggest that icariin may have a therapeutic effect on erectile dysfunction.

YAP is closely correlated with castration-resistant prostate cancer, and downregulation of YAP reduces proliferation and induces apoptosis of PC-3 cells.

Yes-associated protein 65 (YAP65) has been implicated as an oncogene, and its expression is increased in human cancer. Previous studies have demonstrated that alterations in YAP activity may result in tumourigenesis of the prostate. With androgen deprivation therapies becoming progressively ineffective, often leading to life­threatening androgen­resistant prostate cancer (CRPC). The present study aimed to analyse the role of YAP in prostate cancer (PCa), particularly in CRPC. YAP protein was detected using immunohistochemistry and western blot analysis in different prostatic tissues. In addition, three specific RNA interference vectors targeting the human YAP gene were synthesised, and PC­3 cells with a stable inhibition of YAP were obtained by transfection. MTT, flow cytometry, reverse transcription­quantitative polymerase chain reaction and western blot assays were used to analyse the effects of YAP inhibition on the proliferation and apoptosis of PC­3 cells. The frequency of cells that were positive for YAP protein in PCa (78.13%) was significantly higher, compared with para­PCa (26.67%; P=0.007) and benign prostatic hyperplasia (0%; P=0.002). The frequency of cells, which were positive for the expression of YAP exhibited a positive correlation (P=0.008) with the Gleason score, the tumour­node­metastasis staging (P=0.033) and the level of prostate specific antigens (P=0.0032) in PCa. The proliferative capacity of the transfected group was significantly lower, compared with the negative control group (P=0.022). The cell­cycle of the transfected group was arrested in the G1 stage, which was detected using flow cytometry, and there was a significant increase in the apoptosis of cells in the transfected group (P=0.002). The mRNA and protein levels of TEA domain family member 1 were inhibited in the transfected group (P=0.001 and P=0.00, respectively). Therefore, it was concluded that gene transcription and protein expression of YAP may be involved in the development of PCa, particularly CRPC, and may be a novel biomarker for investigation of the occurrence and progression of CRPC. However, the mechanism underlying the modulation of YAP in CRPC remains to be fully elucidated.

Effect of renewed SS-cream on spinal somatosensory evoked potential in rabbits.

AIM: The effect of a renewed SS-cream (RSSC) on the treatment of premature ejaculation (PE) was evaluated and compared with the original SS-cream (OSSC). METHODS: Sixty male white New Zealand rabbits, weighing 2.5 kg-3.0 kg, were divided at random into 3 groups: the RSSC, OSSC and placebo groups. The spinal somatosensory evoked potential (SSEP) elicited by electric stimulation of the glans penis with disk electrode was investigated with an electrophysiograph (Poseidomn, Shanghai, China) before and 10, 30 and 60 min after drug or placebo application on the glans. The Onset and the N1 latencies and the amplitude of SSEP were recorded and analyzed. RESULTS: There was no significant difference (P>0.05) in the mean Onset and N1 latency of SSEP among the 3 groups before drug application. Compared with the pre-application value, the mean Onset and N1 latencies in the RSSC and OSSC groups were significantly prolonged at 10, 30 and 60 min after treatment (P<0.05), while they were not significantly changed (P>0.05) in the placebo group. The mean Onset latency of RSSC at 10 and 30 min and that of OSSC at 30 min were significantly delayed (P<0.05) compared with the placebo group. The mean N1 latency of RSSC at 30 and 60 min and that of OSSC group at 30 min were also significantly delayed (P<0.05). CONCLUSION: RSSC delays the latencies of SSEP, suggesting a local desensitizing effect on the sensory receptor of the glans penis dorsal nerve, which provides the potential for PE treatment. The desensitizing effect of RSSC is higher than that of OSSC.

Sexual function of premature ejaculation patients assayed with Chinese Index of Premature Ejaculation.

AIM: To assess the psychometric properties of the Chinese Index of Premature Ejaculation (CIPE). METHODS: The sexual function of 167 patients with and 114 normal controls without premature ejaculation (PE) were evaluated with CIPE. All subjects were married and had regular sexual activity. The CIPE has 10 questions, focusing on libido, erectile function, ejaculatory latency, sexual satisfaction and difficulty in delaying ejaculation, self-confidence and depression. Each question was responded to on a 5 point Likert-type scale. The individual question score and the total scale score were analyzed between the two groups. RESULTS: There were no significant differences between the age, duration of marriage and educational level (P> 0.05) of patients with and without PE and normal controls. The mean latency of patients with PE and normal controls were 1.6 +/- 1.2 and 10.2 +/- 9.5 minutes, respectively. Significant differences between patients with (26.7 +/- 4.6) PE and normal controls (41.9 +/- 4.0) were observed on the total score of CIPE (P< 0.01). Using binary logistic regression analysis, PE was significantly related to five questions of the original measure. They are the so-called the CIPE-5 and include: ejaculatory latency, sexual satisfaction of patients and sexual partner, difficulty in delaying ejaculation, anxiety and depression. Receiver Operating Characteristic (ROC) curve analysis of CIPE-5 questionnaire indicated that the sensitivity and specificity of CIPE were 97.60 % and 94.74 %, respectively. Employing the total score of CIPE-5, patients with PE could be divided into three groups: mild (>15 point) 19.8 %, moderate (10-14 point) 62.8 % and severe (< 9 point) 16.7 %. CONCLUSION: The CIPE-5 is a useful method for the evaluation of sexual function of patients with PE and can be used as a clinical endpoint for clinical trials studying the efficacy of pharmacological intervention.

[Peroxisome proliferator-actived receptor-gamma ligand troglitazone induces apoptosis in renal cell carcinoma].

UNLABELLED: OBJECTIVE To investigate the expression of peroxisome proliferator-actived receptor-gamma (PPAR-gamma)and the inducement of apoptosis by PPAR-gamma ligand in renal cell carcinoma(RCC)-derived cell lines. METHODS: RT-PCR and Western blot analysis were performed to determined the expression of PPAR-gamma mRNA and protein in two RCC derived cell lines(786-O and A498) and two normal kidney(NK)-derived cell lines(HK-2 and HMCC). Two RCC cell lines were treated with 50 micromol/L troglitazoned for and evaluated for the effects of antidiabetic thiazolidinediones (TZDs) on the cells apoptosis by fluorescence microscopy and DNA ladder assay. The mutative expressions of Bcl-2 and Bax before and after TZDs treatment were also performed by western blot analysis. RESULTS: The expression of PPAR-gamma was observed to be stronger in 786-O and A498 cells than in HK-2 and HMCC cells by RT- PCR and Western blot analysis. Treated with 50 micromol/L troglitazone (for 48 h) it induced typical apoatosis in 786-O and A498 cells. After treatment, a decrease in Bcl-2 expression in RCC cells was observed by Western blot analysis,and the expression of Bax,however,was up-regulated. CONCLUSION: The results reveal that troglitazone has the tumor-suppressive effect on RCC cells. High-affinity PPAR-gamma ligands (TZDs) may be the candidates for a novel approach to the treatment of this refractory neoplasm.

[Studies on the testis regression related gene profile in aged male].

OBJECTIVE: To understand the mechanism of testis regression in aging male. METHODS: Human testis tissues were obtained during related operation with informed consent (normal young male 3 cases and aged male 3 cases). Total RNA was isolated by QIAGEN RNAeasy kit. Differentiations of gene expression were studied by Clon-Tech cDNA microarray methods and the differential expression gene in aged male were classified by Venter's classify system and the candidate genes were investigated by RT-PCR analysis. RESULTS: In the results of cDNA microarray we found 117(1.46%) gene differentiations in aged male at least more than 1.0 fold. Among them, the 83 genes were down-regulated and the 34 genes up-regulated. The down expressed genes related to metabolism were 16(19.3%), gene or protein expression 18(21.7%), cell signaling or cell communication 16(19.3%), cell division 19(22.9%), cell structure or motility 6(7.2%) and unknown function 4 genes (4.8%). The up expressed genes related to cell division were 11 (32.4%), gene or protein expression 10(29.4%) and metabolism 3 (8.8%). It is interesting to find that respiratory chain related gene cox7a2 was up-regulated and atp50 down-regulated significantly which as further confirmed by RT-PCR analysis with sequence analysis in the products of the RT-PCR by T-A cloning. CONCLUSION: The gene expression profile in aged male testis was changed significantly as compared with that in normal young controls; testis regression in aging male may relate multi-gene differentiations, especially the differentiations of respiratory chain related gene cox7a2, atp50, which may be an important candidate gene in the study of the mechanism of testis regression in aging male.

[Effects of icariin on intracavernosal pressure and systematic arterial blood pressure of rat].

OBJECTIVE: To realize the effect of icariin on erectile function of penis. METHODS: After the cavernous nerve (CN) of rat was isolated unilaterally, the corpora cavernosa (CC) and right carotid artery were exposed. A 26G needle catheter was inserted into the right CC to monitor the intracavernous pressure (ICP), and another 26G needle catheter was inserted into the left CC for drug administration. Another catheter was placed into the carotid artery to monitor mean systematic arterial blood press (MBp). Icariin of different concentrations was administrated intracavernosally, and the ICP and MBp were recorded during electric stimulation on CN. Sildenafil and papaveine were used as controls. The effects of nitric oxide syntheses (NOS) inhibitor N(omega)-nitro-L-arginine (LNNA), and soluble guanylate cyclase inhibitor H-[1,2,4] oxadiazolo [4,3,-a] quinoxalin-1-one (ODQ) on icariin (10(-4)mol/L) induced ICP changes were investigated also. RESULTS: Icariin, sidenafil and papaverine increased the ICP in a dose-depended manner (P < 0.01). Icariin and sildenafil did not influence the MBp (P > 0.05), however, papaverine significant influenced MBp (P < 0.01). EC(50) of Icariin, sildenafil and papaveine on ICP/MBp were 2.23 micro mol/L, 0.24 micro mol/L and 9.73 micro mol/L respectively. ODQ and LNNA significantly decreased ICP induced by icariin (10(-4)mol/L). CONCLUSION: Icariin increases ICP without influence on MBp and such effect is inhibited by LNNA and ODQ significantly. Icariin regulates the activity of NO-cGMP signal pathway on CC to enhance erectile function by oral therapy.

[Effects of icariin on the erectile function and expression of nitrogen oxide synthase isoforms in corpus cavernosum of arterigenic erectile dysfunction rat model].

OBJECTIVE: The study the effects of oral administered icariin on intracavernosal pressure (ICP) and on expression of the nitrogen oxide synthase (NOS) isoforms in corpus cavernosum (CC) of arteriogenic erectile dysfunction (A-ED) rat model. METHODS: Forty adult male Wistar rats were randomly divided into 4 groups of 10 rats: shame operated group (group A) and three A-ED model groups (group B, C and D). The internal pudendal arteries were isolated and ligated with 7-O nylon thread at both the main trunk and the penile branches to establish the A-ED model. ICP were tested after the operation to make sure the successful model establishment. The groups A and B were treated with saline: and the groups C and D were treated with icariin (5 mg/kg/day and 10mg/kg/day respectively) orally for 30 days. Then the ICP was measured again. The tissues of corpus cavernosum were taken and RT-PCR was used to detect the mRNA expression of nNOS, iNOS and eNOS in CC, and Western-blot was used to detect the protein expression of these NOS isoforms. RESULTS: The ICP in the group B was significantly decreased compared to the group A (P < 0.01), but the ICP values in the groups C and D were both increased compared to those in the group B (both P < 0.01). The expressions of the mRNA and protein of nNOS, iNOS, and eNOS were all decreased in the group B, however, the mRNA and protein expressions of eNOS were increased a in the groups C and D. In the group C, iNOS also increased. The expression of nNOS showed no obvious changes in the group C and group D. CONCLUSION: Chronic oral treatment with Icariin increases the erectile function (ICP) and restores the eNOS expression in CC of A-ED rats. Icariin may have a long-term therapeutic effect on ischemia/hypoxia induced ED.

[Male reproduction related genes and their function].

With the proceeding of the human genome program(HGP), new genes related to male reproduction have been cloned continuously by different methods. However, investigation of the gene function is still at the rudimentary stage. In spite of the advances in the common methods of studying gene function, some special methods have yet to be developed concerning the study of the genes related to male reproduction. This paper summarizes the current methods of study on gene function relevant to male reproduction.

[Seminiferous tubule scores used for quantitative assessment of spermatogenic function of patients with azoospermia].

OBJECTIVE: To investigate the clinical reliability of quantitative evaluation by seminiferous tubule scores on spermatogenesis dysfunction, using the testis tissues of azoospermia patients for analysis of histological changes. METHODS: One hundred and twelve Chinese patients with azoospermia underwent open testicular biopsy and their testicular biopsy specimens were evaluated by 10-score (on testicular biopsy) and 5-Grade (on seminiferous tubule spermatogenesis) scale. The 112 patient, 22 to 46 years old [(29.0 +/- 4.4) years old] included 105 cases of obstructive and 7 cases non-obstructive azoospermia. Of the total number, there were 96 primary infertile cases and 16 secondary infertile cases with infertile marriage of 2-12 years [(4.0 +/- 2.8) years]. Various seminiferous tubule characteristics were categorized by 10-score as follows: [1] degenerating Sertoli cells and no germinal epithelium; [2] no germ cells and only Sertoli cells; [3] no spermatids and primary spermatocytes and only spermatogonia; [4] no spermatids and few primary spermatocytes; [5] no spermatids and numerous primary spermatocytes; [6] no mature spermatids and few round immature spermatids; [7] no mature spermatids and numerous round immature spermatids; [8] < 20 mature spermatids/tubules, germinal epithelium height < 80 microns and spermiation absent; [9] > 20 mature spermatids/tubules, germinal epithelium height < 80 microns and spermiation rarely < 80 microns; [10] > 20 mature spermatids/tubule and germinal epithelium height 80 microns and spermiation common. Seminiferous tubule spermatogenesis was catagorized by 5-Grade scale as follows: [1] tubular sclerosis; [2] sertoli cell only; [3] arrested spermatogenesis; [4] reduced spermatogenesis; [5] intact spermatogenesis. RESULTS: In terms of the 10-score scale on testicular biopsy, scores of 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10 corresponded with total patient numbers of 5 (4.5%), 38(33.9%), 2(1.8%), 6(5.4%), 2(1.8%), 17(15.2%), 6(5.4%), 19(17%), 10(8.9%) and 7(6.3%), respectively. According to the 5-Grade scale on the seminiferous tubule spermatogenesis, Grades 1, 2, 3, 4 and 5 corresponded with 5(4.5%), 38(33.9%), 33(29.5%), 29(25.9%) and 7 (6.3%), respectively. Tubular diameter, the thickness of the lamina propria, the height of the germinal epithelium and serum FSH correlated with the average seminiferous tubule scores (P < 0.01). CONCLUSION: The seminiferous tubule scores obtained through testicular biopsy may provide important quantitative information concerning the etiology and pathogenesis and of azoospermia may serve as a helpful guide to the fundamental, clinical and therapeutical study of element, clinic and therapy.