Ruibao Chen

S-allyl cysteine restores erectile function through inhibition of reactive oxygen species generation in diabetic rats

Excessive production of reactive oxygen species (ROS) by an overactive nicotinamide adenine dinucleotide phosphate (NADPH) oxidase system in penile tissue is an important mechanism of erectile dysfunction (ED). S‐allyl cysteine (SAC), a bioactive component derived from garlic, was recently reported to exert versatile antioxidant properties. We hypothesized that SAC would be able to resolve diabetes‐related ED by reducing ROS generation, and designed this study to investigate this possibility as well as to determine the related underlying mechanisms. A streptozotocin‐induced diabetes rat model was established and used for comparative analysis of 4‐week treatment regimens with insulin or SAC. The ratio of maximal intracavernous pressure (ICP) to mean arterial blood pressure (MAP) was measured to determine erectile function. Differential levels of ROS, NADPH oxidase subunits, nitric oxide (NO)/cyclic guanosine monophosphate (cGMP) signalling pathway, and apoptosis were evaluated in cavernous tissues. Max ICP/MAP was found to be markedly decreased in untreated diabetic rats; SAC, but not insulin, treatment restored the ratio to baseline (in non‐diabetic untreated controls). The corpus cavernosum of untreated diabetic rats showed increased p47phox and p67phox expression, ROS production and penile apoptotic index, and decreased phospho‐endothelial nitric oxide synthase (phospho‐eNOS, Ser1177) expression, cGMP concentration, B‐cell lymphoma 2 (Bcl‐2)/Bcl‐2‐associated X protein (Bax) ratio and smooth muscle cell number. SAC treatment normalized all the diabetes‐induced effects, whereas insulin treatment partially normalized the alterations, but produced no effects on P47phox expression, penile ROS level, apoptotic index, Bcl‐2/Bax ratio and smooth muscle cell number. Collectively, these data indicate that SAC treatment can restore erectile function in diabetic rats by preventing ROS formation through modulation of NADPH oxidase subunit expression. Furthermore, the poor efficacy of conventional insulin treatment for diabetic ED may be associated with an elevated level of ROS in penile tissue.

Up‐regulation of VEGF by Small Activator RNA in Human Corpus Cavernosum Smooth Muscle Cells

INTRODUCTION Functional failure of smooth muscle cells and endothelial cells in corpus cavernosum contributes to erectile dysfunction (ED) in aging men. Given that vascular endothelial growth factor (VEGF) may improve the function of smooth muscle cells and endothelial cells through different mechanisms, it is thus expected that increasing the expression of VEGF may have beneficial effects on erectile function. AIM The aim of this article is to explore the possibility that VEGF can be induced by ribonucleic acid activation (RNAa) technology, and VEGF induction by RNAa has the potential of treating ED. METHODS Primary human corpus cavernosum smooth muscle cells (CCSMCs) were isolated and cultured in vitro. The expression of α-smooth muscle actin was detected by immunohistochemistry to identify CCSMCs. A previously identified VEGF promoter-targeted small activator RNA (saRNA, double-stranded [ds]VEGF-706) and a negative control dsRNA were chemically synthesized. Cultured human CCSMCs were transfected with the saRNAs. The expression of VEGF messenger RNA (mRNA) and protein in transfected CCSMCs was evaluated by real-time polymerase chain reaction (RT-PCR) and Western blotting assay, respectively. Immunofluorescent staining was also used to confirm VEGF protein expression in cultured CCSMCs. MAIN OUTCOME MEASURE The expression of VEGF was assessed by RT quantitative PCR, Western blotting, and immunofluorescence assays. RESULTS After transfection, RT quantitative PCR analysis showed that the expression of VEGF mRNA was significantly induced in dsVEGF-706 transfected cells compared with cells receiving control treatments (P < 0.05). Consistent with mRNA induction, Western blotting and immunofluorescence analysis showed that VEGF protein expression was also induced by dsVEGF-706. CONCLUSION VEGF expression can be activated by RNAa in primary human CCSMCs, suggesting a potential application of RNAa-mediated VEGF activation for the treatment of ED.

Age-related changes in kallikreins–kinins system in rat corpus cavernosum

Many factors, such as nitric oxide synthase, androgen and growth factors, can regulate the tone of corpus cavernosum (CC) smooth muscle with an age-related tendency. It has been shown that the active metabolites of kallikreins-kinins system (KKS), including bradykinin, Lys-BK and Met-Lys-BK, can also relax the CC smooth muscle significantly in vitro. Our aim was to evaluate the specific association between KKS and age in rat CC. CC and thoracic aorta were isolated from rats at postnatal weeks (PW) of 2, 8, 12, 20, 30, 40 and 60, respectively. Tissue kallikrein-I (KLKI) and kinin B2 receptor (B2R) mRNA in CC and thoracic aorta were detected by real-time polymerase chain reaction (PCR). Protein expression of KLKI and B2R were determined with immunofluorescence in situ and Western blot. Real-time PCR, immunofluorescence in situ and Western blot all demonstrated that the age-related changes in expression of KLKI were similar between the CC and thoracic aorta. It significantly increased with age from PW2 to PW30, reached the peak at PW30 and then declined gradually. However, there was no statistically significant difference among PW30, PW40 and PW60. Similarly, the expression of B2R increased gradually with age reached and remained at the peak during adult stages and no significant differences were found among PW20, PW30 and PW40; then, it decreased significantly at PW60. The changes in the expression of B2R in CC and age-matched aorta were similar except that it was significantly less than that in the aorta at PW60. The expression of KLKI and B2R changed in an age-dependent pattern in rat CC and have a tendency to decline during ageing, which is of the same tendency as reported for erection capacity in ageing males and suggests why ageing is an independent predictor of ED, to some extent.

Flightless I Homolog Represses Prostate Cancer Progression through Targeting Androgen Receptor Signaling

Purpose: Flightless I (FLII), member of the gelsolin superfamily of actin-remodeling proteins, functions as a transcriptional coregulator. We aim to evaluate a tumor-suppressive function of FLII in regulating androgen receptor (AR) in prostate cancer progression. Experimental Design: We examined FLII protein and mRNA expression in clinical prostate cancer specimens by immunohistochemistry. Kaplan–Meier analysis was conducted to evaluate the difference in disease-overall survival associated with the expression levels of FLII and AR. Prostate cancer cells stably expressing FLII or shRNA knockdown were used for functional analyses. Immunoprecipitation, Luciferase reporter, and immunofluorescence staining assays were performed to examine the functional interaction between FLII and AR. Results: Our analysis of the expression levels of FLII in a clinical gene expression array dataset showed that the expression of FLII was positively correlated with the overall survival of prostate cancer patients exhibiting high levels of AR expression. Examination of protein and mRNA levels of FLII showed a significant decrease of FLII expression in human prostate cancers. AR and FLII formed a complex in a ligand-dependent manner through the ligand-binding domain (LBD) of AR. Subsequently, we observed a competitive binding to AR between FLII and the ligand. FLII inhibited AR transactivation and decreased AR nuclear localization. Furthermore, FLII contributed to castration-sensitive and castration-resistant prostate cancer cell growth through AR-dependent signaling, and reintroduction of FLII in prostate cancer cells sensitized the cells to bicalutamide and enzalutamide treatment. Conclusions: FLII plays a tumor-suppressive role and serves as a crucial determinant of resistance of prostate cancer to endocrine therapies. Clin Cancer Res; 22(6); 1531–44. ©2015 AACR.

Predictive value of FSH, testicular volume, and histopathological findings for the sperm retrieval rate of microdissection TESE in nonobstructive azoospermia: a meta-analysis

We performed this meta-analysis to evaluate the predictive value of different parameters in the sperm retrieval rate (SRR) of microdissection testicular sperm extraction (TESE) in patients with nonobstructive azoospermia (NOA). All relevant studies were searched in PubMed, Web of Science, EMBASE, Cochrane Library, and EBSCO. We chose three parameters to perform the meta-analysis: follicle-stimulating hormone (FSH), testicular volume, and testicular histopathological findings which included three patterns: hypospermatogenesis (HS), maturation arrest (MA), and Sertoli-cell-only syndrome (SCOS). If there was a threshold effect, only the area under the summary receiver operating characteristic curve (AUSROC) was calculated. Otherwise, the pooled sensitivity, specificity, positive likelihood ratio (PLR), negative likelihood ratio (NLR), and the diagnostic odds ratio (DOR) were also calculated. Twenty-one articles were included in our study finally. There was a threshold effect among studies investigating FSH and SCOS. The AUSROCs of FSH, testicular volume, HS, MA, and SCOS were 0.6119, 0.6389, 0.6758, 0.5535, and 0.2763, respectively. The DORs of testicular volume, HS, and MA were 1.98, 16.49, and 1.26, respectively. The sensitivities of them were 0.80, 0.30, and 0.27, while the specificities of them were 0.35, 0.98, and 0.76, respectively. The PLRs of them were 1.49, 10.63, and 1.15, respectively. And NLRs were 0.73, 0.72, and 0.95, respectively. All the investigated factors in our study had limited predictive value. However, the histopathological findings were helpful to some extent. Most patients with HS could get sperm by microdissection TESE.

Solitary fibrous tumor in bladder: a case report.

Solitary fibrous tumor (SFT) in bladder is extremely rare. In this study, we reported one case of bladder SFT and reviewed the only ten cases of the disease that had been reported so far. The patient suffered from residual urine sensation and urethral pain. Cystoscopy revealed a 7-cm protruding mass at the dome of the bladder, and bladder mucosa biopsy showed normal differentiation of the bladder mucosa with a small amount of inflammatory cells. Radical resection of the tumor was performed in this patient. Pathological examination found uniform, haphazardly arranged spindle cells, the majority of which were CD34-positive and Vimentin-positive and proved that the mass was a solitary fibrous tumor. Within a period of 9 months of follow-up, no reoccurrence was found.SummarySolitary fibrous tumor (SFT) in bladder is extremely rare. In this study, we reported one case of bladder SFT and reviewed the only ten cases of the disease that had been reported so far. The patient suffered from residual urine sensation and urethral pain. Cystoscopy revealed a 7-cm protruding mass at the dome of the bladder, and bladder mucosa biopsy showed normal differentiation of the bladder mucosa with a small amount of inflammatory cells. Radical resection of the tumor was performed in this patient. Pathological examination found uniform, haphazardly arranged spindle cells, the majority of which were CD34-positive and Vimentin-positive and proved that the mass was a solitary fibrous tumor. Within a period of 9 months of follow-up, no reoccurrence was found.

Will the kidney function be reduced in patients with renal cell carcinoma following laparoscopic partial nephrectomy? Baseline eGFR, warm ischemia time, and RENAL nephrometry score could tell

OBJECTIVES To describe the natural history of kidney function following partial nephrectomy (PN) for patients with renal cell carcinoma (RCC), and to identify independent predictors of whether patients with RCC will retain renal function unchangeable or even increased and develop functional impairment of ≧25% post-PN. PATIENTS AND METHODS We performed a retrospective analysis of 337 cases involving patients diagnosed with RCC of pT1-2N0M0 who underwent laparoscopic PN, the primary endpoints included the stabilization or increase in postoperative estimated glomerular filtration rate (eGFR) compared to the preoperative level and eGFR impairment of ≧25% following surgery. We plotted the trajectory of each patients eGFR measurement starting from their first postoperative day to the last follow-up time post-PN and used moving average method to look at trends of eGFR changing. A logistic regression model was then applied to identify associations between clinical and surgical characteristics with eGFR outcomes. RESULTS Patients were of an average age of 51.4 years and all were Chinese descent. The cohort was also primarily male (69.1%). One hundred ninety seven (58.5%) had eGFR ≧90 ml/min/1.73 m2, while 140 (41.5%) had an eGFR of 60 to 90 ml/min/1.73 m2 prior to the operation. All patients underwent minimally invasive PN with warm ischemia, with 64.1% (216/337) receiving laparoscopic surgery, and 35.9% (121/337) receiving robot-assisted laparoscopic surgery. On average, patients experienced a mean eGFR decrease of 23.8% immediately post-PN, followed by a slight increase and stabilization, with a mean 15.5% decline after 1 year. Twenty four percent (81/337) experienced GFR impairment of ≧25% over a median 10.0-month follow-up time period, while 29.1% (99/337) patients retained eGFR unchangeable or increased post-PN. And higher preoperative eGFR, longer warm ischemia time, and more complexity lesions (higher renal nephrometry score ) were found to be independently associated with higher chance of functional impairment of ≧25% and lower chance of eGFR stabilization post-PN. CONCLUSION Although, majority of patients experienced decline of renal function post-PN, functional outcomes of eGFR unchangeable and increased were also seen, and baseline total eGFR, WIT as well as RENAL nephrometry score were determined to be independent predictors of those renal functional outcomes.

Free bladder mucosa graft harvested by water‑jet: A novel, minimally invasive technique for urethral reconstruction

The aim of the current study was to describe a novel approach of urethral reconstruction through minimally invasive harvesting of the bladder graft via endoscopic sub-mucosal dissection of water-jet. The records of two patients were reviewed, who underwent transurethral endoscopic surgical bladder mucosa graft harvest by water-jet and urethral reconstruction with informed consent. Case 1 was a 35-year-old male with anterior urethral stricture; case 2 was a 22-year-old male with secondary anterior urethral stricture and hypospadias following a failed hypospadias surgery. The two male patients successfully underwent urethral reconstruction using bladder mucosa graft harvested via endoscopic assisted by water-jet; no perforation, cysthemorrhagia or any other postoperative bladder-related complication was observed. Voiding cystourethrogram of case 1 indicated that the reconstructed urethra was unobstructed, and no recrudescence was observed within 4 months of follow-up. In case 2, dysuria had disappeared completely within 1 month of follow-up, and the urethra plate was successfully reconstructed by first-stage. To the best of our knowledge, this is the first report to demonstrate urethral reconstruction using a bladder mucosa graft harvested by transurethral endoscopic sub-mucosal dissection, assisted by water-jet. Transurethral endoscopic surgery may provide a minimally invasive approach instead of the traditional open surgery for harvesting bladder mucosa graft. Urethral reconstruction conducted with bladder mucosa graft harvested via endoscopic sub-mucosal dissection assisted by water-jet is a feasible and safe method, and the short-term follow-up results are encouraging.

AB061. Clinical analysis of small cell carcinoma of the bladder: 9 cases report and literature review

Objective To present our experience with nine patients with small cell carcinoma of the bladder (SCCB) who were treated with different modalities and review the literature for patients with SCCB who have been reported in 56 literatures. SCCB is a rare, highly aggressive tumor that presents in an advanced stage and has a propensity for early metastasis. Hematuria is the main clinical manifestations. Surgery, chemotherapy, and radiotherapy, either alone or as part of combined therapy have been used for treatment. Methods We retrospectively evaluated nine patients with SCCB by medical record review between February 1980 and January 2014 at Tongji Hospital of Huazhong University of Science and Technology. In order to better understand the clinical features of SCCB, 56 literatures were concerned (from January 1979 to March 2014). The general characteristics, clinical manifestation, the pathological and immunohistochemical characteristics, treatment options and prognostication in those eligible manuscripts are analyzed, a retrospective analysis is performed. Results All the nine cases in Tongji hospital were successfully operated and tissue samples were carried on pathological examination. All the tumor tissue contained small cell carcinoma components, there were four cases coexisting with other histologic types of bladder cancers, and 2 of the 9 cases have three different cell components. All patients had muscle-invasive disease at presentation, 4 cases showed lymph nodes metastasis, 3 cases showed invasion of the surrounding seminal vesicle or uterus, and 1 case greatly suspected the liver metastasis. Specimens of all cases underwent immunohistochemistry examination showed that NSE, PCK, Syn, and CD56 were all positive, but LCA was negative. After operation, 3 patients underwent chemotherapy and only one patient received post operational radiotherapy. Patients were followed up range between 3 to 84 months and the median survival time was 33 months. The main reasons of the patients die are tumor recurrence and metastasis, but two patients are still alive. Conclusions SCCB is different from transitional cell carcinoma (TCC) of the bladder. It has its unique cytology, immunohistochemistry and ultrastructural features. Diagnosis depends on pathological examination and immunohistochemistry. Surgery followed by chemotherapy is the main treatment method currently. In view of the disease is easily early metastasis, the overall prognosis for this cancer is poor. Further research is required to understand the molecular pathogenesis so that novel targeted therapies can be developed for this rare cancer.

Peripheral Primitive Neuroectodermal Tumor (pPNET) of the Penis: A Case Report and Literature Review

Primitive neuroectodermal tumors are derived from primitive neuroectodermal cells and belong to a highly malignant subgroup of round-cell tumors. Peripheral primitive neuroectodermal tumors of the penis are an extremely rare malignant form among penile neoplasms. These tumors are often difficult to diagnose due to atypical symptoms. Here, we report a case of a 24-year-old patient in China with a neoplasm localized at the base of his penis. The initial symptom was dysuria without any inducement. The results of blood and urine examinations indicated no abnormalities. The imaging examination results indicated a firm mass near the base of the penis. The hematoxylin and eosin (H&E) staining revealed round, small tumor cells with heterotypical darkened nuclei. In addition, immunohistochemistry (IHC) revealed strong and diffuse positive staining for CD99 (mic-2), VIM (vimentin), and NCAM1/CD56 (neural cell adhesion molecule 1). In addition, 60% of cells were positive for the cell proliferation marker Ki-67. Base...