SUMMARYExcessive 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 p47 phox and p67 phox 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 P47 phox 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.

Adipose-derived stem cells (ADSCs) have recently been considered as a promising therapy for erectile dysfunction (ED). However, the mechanism of ADSC-based therapy is unclear. Insulin-like growth factor-1 (IGF-1), basic fibroblast growth factor (bFGF), and vascular endothelial growth factor (VEGF) secreted by ADSCs were assessed in vitro. Sixteen 24-month-old male Sprague-Dawley rats were used for comparative analysis of 2-week treatment with labeled ADSCs or PBS. Eight additional 5-month-old rats were used as a young rat group. At 2 weeks post-transplantation, all rats were analyzed for erectile function, cavernous IGF-1, bFGF and VEGF levels, and penile histology. Conditioned medium and co-culture systems were used in cell experiments to detect how growth factors act on corpus cavernosum smooth muscle cells (CCSMCs) under oxidative stress conditions via crystal violet staining and immunofluorescence staining. We found that ADSCs secreted significantly higher IGF-1, bFGF, and VEGF levels in culture medium compared with basal medium. Compared with young rats, untreated aged rats had significantly lower Max ICP/MAP and ADSC treatment significantly increased the ratio. Immunofluorescence staining demonstrated a small number of labeled ADSCs in the corpus cavernosum. The untreated aged rats showed significantly decreased cavernous IGF-1, bFGF, and VEGF levels and significantly decreased contents of cavernous smooth muscle and endothelium compared with young rats. ADSC treatment partially normalized these alterations. In cell experiments, the groups receiving growth factor neutralizing antibody separately or combined had significantly decreased numbers of CCSMCs compared with control groups. These results indicated that ADSC treatment may improve aging-related ED partially through the secretion of IGF-1, bFGF, and VEGF.

The aim of the present study was to evaluate the diagnostic value of the ImmunoCyt test compared with urine cytology in detecting bladder cancer. A systematic literature search was performed to locate all publications reporting on the diagnostic accuracy of the ImmunoCyt test for bladder cancer. Data were extracted from 2×2 tables or calculated from reported accuracy data. Collected data were meta-analyzed for sensitivity, specificity, positive likelihood ratio (LR), negative LR, diagnostic odds ratio (DOR), and summary receiver operator characteristic (sROC) curve analysis. We applied the Meta-DiSc 1.4 and STATA 13.0 software to the meta-analysis. Seven separate studies consisting of 1,602 patients with bladder cancer were considered in the meta-analysis. We found that the ImmunoCyt test had a higher sensitivity than the urine cytology test [0.725, 95% confidence interval (CI) 0.683–0.765 vs. 0.566, 95% CI, 0.521–0.611], but the specificity, positive LR, negative LR, DOR, area under the curve (AUC) and Q index of the ImmunoCyt test were lower compared with the urine cytology test. In addition, the pooled sensitivity, specificity, positive LR, negative LR, DOR, AUC, and Q index of the combined method (combination of ImmunoCyt and cytology) were 0.833, 0.644, 2.804, 0.228, 13.50, 0.8554 and 0.7863, respectively. The results of the Eggers test showed no publication bias (P>0.05). In conclusion, specificity, positive LR, negative LR, DOR, the AUC, and the Q index of the urine cytology test may be superior to the ImmunoCyt test, but the ImmunoCyt test has greater sensitivity than the urine cytology test. Use of ImmunoCyt and cytology in combination has the potential to improve the sensitivity and promises to be an alternative in the detection of bladder cancer.

BackgroundThis study aimed to determine whether the enhancer of the rudimentary homolog (ERH) gene regulates cell migration and invasion in human bladder urothelial carcinoma (BUC) T24 cells and the underlying mechanism.MethodsFirst, we knocked down ERH in BUC T24 and 5637 cells by shRNA and then used wound healing cell scratch migration assays, transwell cell migration assays, transwell cell invasion chamber experiments and nude mouse tail vein transfer assays to determine the migration and invasion ability after ERH was knocked down. Moreover, we used gene expression profiling chip analysis and further functional experiments to explore the possible mechanism through which ERH knockdown downregulated metastasis ability in T24 cells.ResultsWound healing cell scratch migration assays, transwell cell migration assays, transwell cell invasion chamber experiments and nude mouse tail vein transfer assays all showed that the metastasis ability was significantly inhibited in human BUC T24 and 5637 cells with ERH knockdown. A gene expression profiling chip analysis in T24 cells showed that the MYC gene may be an important downstream target of the ERH gene, and the functional experiments showed that MYC is a functional target of ERH in BUC T24 cells.ConclusionERH knockdown could inhibit the metastasis of BUC T24 cells in vitro and in vivo. This study further explored the mechanism of the ERH gene in the metastasis of the T24 human bladder cancer cell line and found that ERH may regulate MYC gene expression. The results of this research provide a basis for the clinical application of ERH as a potential target for BUC treatment.Electronic supplementary materialThe online version of this article (10.1186/s12885-019-5423-9) contains supplementary material, which is available to authorized users.

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Background Erectile dysfunction is a major complication of diabetes mellitus. Adipose-derived stem cells (ADSCs) have attracted much attention as a promising tool for the treatment of diabetes mellitus-induced erectile dysfunction (DMED). Inducible nitric oxide synthase (iNOS) plays an important role in protecting penile tissues from fibrosis. The aim of this study was to determine the efficacy of ADSCs overexpressing iNOS on DMED in rats. Methods ADSCs were isolated and infected with adenovirus overexpressing iNOS (named as ADSCs-iNOS). The expression of iNOS was detected using western blot analysis and real-time PCR. Rats were randomly assigned into five groups: control group, DMED group, ADSCs group, ADSCs-EGFP group and ADSCs-iNOS group. 5 × 105 cells were given once via the intracorporal route. Two weeks after treatment, erectile function was assessed by electrical stimulation of the cavernous nerve. Penile tissues were obtained and evaluated at histology level. Results We found that ADSCs-iNOS had significantly higher expression of iNOS at mRNA and protein levels and generated more nitric oxide (NO). ADSCs-iNOS reduced collagen I and collagen IV expression of corpus cavernosum smooth muscle cells (CCSMCs) in cell co-culture model. Transforming growth factor-β1 expression in CCSMCs reduced following co-culture with ADSCs-iNOS. Injection of ADSCs-iNOS significantly ameliorated DMED in rats and decreased collagen/smooth muscle cell ratio of penile tissues. Moreover, elevated NO and cyclic guanosine monophosphate concentrations were detected in penile tissues of ADSCs-iNOS group. Conclusion Taken together, ADSCs-iNOS significantly improved erectile function of DMED rats. The therapeutic effect may be achieved by increased NO generation and the suppression of collagen I and collagen IV expression in the CCSMCs to decrease penile fibrosis.

Solasonine, a steroidal alkaloid extracted from Solanum nigrum L., has been found to exert inhibitory effect on cancers. However, the underlying anticancer mechanisms of solasonine, particularly in urinary bladder cancer (BC), remain unclear. In this study, we identified the potential targets and biological functions associated with solasonine activity using a bioinformatics approach. Ingenuity pathway analysis revealed that neuropilin-1 (NRP1) and other signaling pathways, such as PI3K/AKT and ERK/MAPK pathways, were potentially involved in the therapeutic effects of solasonine. The ability of solasonine in inducing apoptosis and inhibiting proliferation in BC cells was confirmed experimentally, and the inhibition of ERK/MAPK, P38/MAPK, and PI3K/AKT pathways was validated by Western blot. Mechanistically, solasonine suppressed the expression of NRP1 protein, but not that of mRNA. Further results of molecular docking and molecular dynamics simulation analysis indicated that solasonine could directly bind to the b1 domain of NRP1 protein with a reasonable and stable docking conformation. We previously found that targeting NRP1 is a potential antitumor strategy. Combined with these findings, it can be speculated that the binding of solasonine with NRP1 on the cell membrane could prevent the formation of NRP1/VEGFA/VEGFR2 and NRP1/EGFR complexes, resulting in the inhibition of downstream signaling, including ERK/MAPK, P38/MAPK, and PI3K/AKT pathways. Additionally, intracellular solasonine could inhibit the membrane localization of NRP1 and provoke its cytoplasmic retention, facilitating the degradation of NRP1 protein in the cytoplasm. The dual effects induced by the binding of solasonine to NRP1 extracellularly and intracellularly could account for the antiproliferative and proapoptotic effects of solasonine on BC.

BackgroundAdipose‐derived stem cells have been considered as a promising therapy for erectile dysfunction. However, the therapeutic efficacy of adipose‐derived stem cell‐based therapy requires improvement.ObjectiveTo determine whether the inhibition of phosphodiesterase type 5 in adipose‐derived stem cells would improve stem cell therapy for rats with diabetes‐induced erectile dysfunction.Materials and MethodsA phosphodiesterase type 5 siRNA was incorporated into lentiviral vectors and transduced into adipose‐derived stem cells. The mRNA and protein levels of phosphodiesterase type 5 were evaluated. Three days after transduction, the adipose‐derived stem cell supernatant was collected to determinate the levels of insulin‐like growth factor 1 and vascular endothelial growth factor. Streptozotocin‐induced diabetic rat models were established and used for comparative analysis of 1‐ and 2‐week treatment regimens with intracavernosal injection of adipose‐derived stem cells or Lv‐siPDE5‐modified adipose‐derived stem cells.ResultsLv‐siPDE5‐ADSCs secreted more insulin‐like growth factor 1 and vascular endothelial growth factor in supernatants than unmodified adipose‐derived stem cells. Preconditioned Adipose‐derived stem cells‐treated diabetic rats showed consistently superior erectile function when compared with non‐preconditioned adipose‐derived stem cells after 2 weeks of treatment. Lv‐siPDE5‐ADSCs provided additional benefits in recovery of cavernous structures with rapid effects (1 week) when compared to plain adipose‐derived stem cells. These features were associated with the significantly increased levels of insulin‐like growth factor 1 and vascular endothelial growth factor in Lv‐siPDE5‐ADSC‐treated diabetic rats.ConclusionsAdipose‐derived stem cell therapy could serve as an alternate approach for diabetes‐induced erectile dysfunction, albeit with a long onset period. In vitro preconditioning of adipose‐derived stem cells could accelerate the functional and structural recovery in vivo, indicating that preconditioning by inhibition of phosphodiesterase type 5 may improve adipose‐derived stem cells therapy following diabetes‐induced erectile dysfunction.