Identification of hub genes and pathways associated with cellular senescence in diabetic foot ulcers via comprehensive transcriptome analysis.

This research aimed to find important genes and pathways related to cellular senescence (CS) in diabetic foot ulcers (DFU) and to estimate the possible pathways through which CS affects diabetic foot healing. The GSE80178 dataset was acquired from the Gene Expression Omnibus (GEO) database, containing six DFU and three diabetic foot skin (DFS) samples. The limma package was used to identify differentially expressed genes (DEGs). At the same time, DEGs associated with CS (CS-DEGs) were found using the CellAge database. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were conducted on the CS-DEGs. A protein-protein interaction (PPI) network was built using the String database, and the cytoHubba plug-in within Cytoscape helped identify hub genes. Lastly, the miRNA-TF-mRNA regulatory network for these hub genes was established. In total, 66 CS-DEGs were obtained. These genes mainly focus on CS, Kaposi sarcoma-associated herpesvirus infection and Toll-like receptor signalling pathway. Eight hub genes were identified to regulate cell senescence in DFU, including TP53, SRC, SIRT1, CCND1, EZH2, CXCL8, AR and CDK4. According to miRNA-TF-mRNA regulatory network, hsa-mir-132-3p/SIRT1/EZH2 axis is involved in senescence cell accumulation in DFU.

[Causes and management for male urethral stricture].

OBJECTIVE: To explore the etiology of male urethral stricture, analyze the therapeutic strategies of urethral stricture, and summarize the complicated cases.
 Methods: The data of 183 patients with urethral stricture were retrospectively analyzed, including etiology, obstruction site, stricture length, therapeutic strategy, and related complications.
 Results: The mean age was 49.7 years, the average course was 64.7 months, and the constituent ratio of 51 to 65 years old patients was 38.8% (71/183). The traumatic injury of patients accounted for 52.4% (96/183), in which the pelvic fracture accounted for 35.5% (65/183) and the straddle injury accounted for 16.9% (31/183). There were 54 cases of iatrogenic injury (29.5%). The posterior urethral stricture accounted for 45.9% (84/183), followed by the anterior urethral stricture (44.8%, 82/183) and the stenosis (6.6%, 12/183). A total of 99 patients (54.1%) received the end to end anastomosis, and 40 (21.9%) were treated with intracavitary surgery, such as endoscopic holmium laser, cold knife incision, endoscopic electroknife scar removal, balloon dilation, and urethral dilation. In the patients over 65-years old, the urethral stricture rate was 14.8% and the complication rate (70.4%) for transurethral resection of the prostate (TURP) was significantly higher than that of all samples (P<0.01).
 Conclusion: Both the etiology of male urethral stricture and the treatment strategy have changed and the incidence of traumatic and iatrogenic urethral stricture has increased in recent 3 years. The main treatment of urethral stricture has been transformed from endoscopic surgery into urethroplasty.

[Effect of calcium channel blockers on primary cultured human urethra scar fibroblasts].

OBJECTIVE: To investigate effects of verapamil on primary cultured human urethral scar fibroblasts (USFs) and to provide basis for protecting the formation of urethra scar.
 Methods: The cell proliferation was evaluated with the cell counting kit (CCK)-8 method after USFs were incubated various verapamil concentrations (50, 100, 150, 200, or 250 µmol/L) or solvent for 12, 24, or 48 h. The protein level of matrix metalloproteinase (MMP) was evaluated with ELISA after cells were incubated with verapamil (100 µmol/L) or solvent (control cells) for 24 h.
 Results: The proliferation of USFs was obviously suppressed after verapamil treatment, which was in a dose-dependent and time-dependent manner. Meanwhile, the protein levels of MMP-2 and MMP-9 in the verapamil treatment group increased obviously compared with those of the control groups (P<0.05).
 Conclusion: Calcium channel blockers may prevent the excessive formation of urethra scar by inhibiting the proliferation of urethral scar fibroblasts and enhancing the activity of MMP.

[A preliminary study on mechanisms for urinary system disorders before and after ketamine withdrawal in rats].

OBJECTIVE: To explore the mechanisms for urinary system disorders before and after ketamine withdrawal in rats and to evaluate the recovery degree of the urinary system damage after ketamine withdrawal. METHODS: Fifteen male healthy Sprague-Dawley rats were randomly divided into 3 groups: A control group, an experimental group, and a withdrawal group. The rats in the control group were given normal saline. The rats in the experimental group were given ketamine 30 mg/(kg.day) for 30 days. The rats in the withdrawal group were treated as the experimental group except for drug withdrawal for 2 weeks. In the experimental period, we randomly selected 1 rat of kidney, ureter, and bladder from each group to perform HE staining. The bladder tissues in each group were used to detect mRNA expression by quantitative real-time polymerase chain reaction (qRT-PCR). RESULTS: 1) The behavior of ketamine-injected rats was obviously changed, but the weight of ketamine-induced rats was not changed. 2) As compared with the control group, the experimental and withdrawal groups showed infiltration of mononuclear inflammatory cells in the kidney tissues, the thinner epithelium of bladder and infiltration of submucosal mononuclear inflammatory cells under the optical microscope. 3) As compared with the control group, the expression of H1R mRNA was increased in the experimental group (P<0.05). As compared with the experimental group, H1R mRNA expression was significantly decreased in the withdrawal group (P<0.05). CONCLUSION: Ketamine abuse could induce behavior changes in rats. The infiltration of mononuclear inflammatory cells in kidney and bladder, the thinner bladder epithelial layer, and the increased H1R gene mRNA expression in bladder might be an important pathogenesis of KAUD. Ketamine withdrawal may effectively reverse the pathogenic process of KAUD.

[Analysis of influential factors for prostate biopsy and establishment of logistic regression model for prostate cancer].

OBJECTIVE: To establish logistic regression model for prostate cancer and provide basis for prostate biopsy.
 METHODS: A total of 117 cases of prostate biopsy were retrospectively analyzed in chronological sequence. All cases were assigned into a model group (n=78) and a validation group (n=39). Logistic regression model was established and its value was estimated by receiver operating characteristic (ROC) curve. 
 RESULTS: Digital rectal examination(DRE), transrectal ultrasound(TRUS), MRI, prostate-specific antigen density (PSAD), and free PSA/total PSA (fPSA/tPSA) were the influential factors for prostate biopsy (P<0.01). The established logistic regression model for prostate cancer by regression coefficient was: logit P=-2.362+2.561×DRE+1.747×TRUS+2.901×MRI+1.126×PSAD-2.569×fPSA/tPSA and area under curve was 0.907. When the cutoff aimed at 0.12, the sensitivity and specificity were 81.80% and 89.30%, respectively.
 CONCLUSION: Logistic regression model for prostate cancer can provide sufficient basis for prostate biopsy. Prostate biopsy should be performed when P value is more than 0.12.

Predicting prostate cancer recurrence: Introducing PCRPS, an advanced online web server.

Background: Prostate cancer (PCa) is one of the leading causes of cancer death in men. About 30% of PCa will develop a biochemical recurrence (BCR) following initial treatment, which significantly contributes to prostate cancer-related deaths. In clinical practice, accurate prediction of PCa recurrence is crucial for making informed treatment decisions. However, the development of reliable models and biomarkers for predicting PCa recurrence remains a challenge. In this study, the aim is to establish an effective and reliable tool for predicting the recurrence of PCa. Methods: We systematically screened and analyzed potential datasets to predict PCa recurrence. Through quality control analysis, low-quality datasets were removed. Using meta-analysis, differential expression analysis, and feature selection, we identified key genes associated with recurrence. We also evaluated 22 previously published signatures for PCa recurrence prediction. To assess prediction performance, we employed nine machine learning algorithms. We compared the predictive capabilities of models constructed using clinical variables, expression data, and their combinations. Subsequently, we implemented these machine learning models into a user-friendly web server freely accessible to all researchers. Results: Based on transcriptomic data derived from eight multicenter studies consisting of 733 PCa patients, we screened 23 highly influential genes for predicting prostate cancer recurrence. These genes were used to construct the Prostate Cancer Recurrence Prediction Signature (PCRPS). By comparing with 22 published signatures and four important clinicopathological features, the PCRPS exhibited a robust and significantly improved predictive capability. Among the tested algorithms, Random Forest demonstrated the highest AUC value of 0.72 in predicting PCa recurrence in the testing dataset. To facilitate access and usage of these machine learning models by all researchers and clinicians, we also developed an online web server (https://urology1926.shinyapps.io/PCRPS/) where the PCRPS model can be freely utilized. The tool can also be used to (1) predict the PCa recurrence by clinical information or expression data with high accuracy. (2) provide the possibility of PCa recurrence by nine machine learning algorithms. Furthermore, using the PCRPS scores, we predicted the sensitivity of 22 drugs from GDSC2 and 95 drugs from CTRP2 to the samples. These predictions provide valuable insights into potential drug sensitivities related to the PCRPS score groups. Conclusion: Overall, our study provides an attractive tool to further guide the clinical management and individualized treatment for PCa.

LncRNA PVT1 promotes bladder cancer progression by forming a positive feedback loop with STAT5B.

BACKGROUND: Plasmacytoma Variant Translocation 1 (LncRNA PVT1) and signal transducer and activator of transcription 5B (STAT5B) play important roles in various cancers, but their interaction in bladder cancer (BC) remains unclear. PURPOSE: We aimed to explore the interaction between lncRNA PVT1 and STAT5B in BC tumorigenesis and find potential drugs for BC. METHODS: The association of the expression of lncRNA PVT1 and STAT5B to the prognosis of BC patients was evaluated via bioinformatic analysis. Loss- and gain-of-function assays were performed to determine the biological functions of lncRNA PVT1 and STAT5B. Quantitative real time polymerase chain reaction, Western blot, immunohistochemistry, and immunofluorescence were used to detect lncRNA PVT1 and STAT5B expression. Fluorescence in situ hybridization, RNA pull-down and RNA immunoprecipitation assays were conducted to determine the regulatory effect of lncRNA PVT1 on STAT5B. The transcriptional effect of STAT5B on lncRNA PVT1 gene was determined using luciferase reporter assay, chromatin immunoprecipitation and DNA-affinity precipitation assays. Connectivity Map analysis was used to screen anticancer drugs. RESULTS: LncRNA PVT1 and STAT5B enhance the expression of each other and promote the malignant phenotypes in BC, including cell viability and invasion. lncRNA PVT1 stabilizes STAT5B by decreasing ubiquitination, enhances STAT5B phosphorylation, and promotes the translocation to the nucleus of STAT5B to trigger further carcinogenesis activities. In the nucleus, STAT5B activates the transcription of lncRNA PVT1 by binding directly to its promoter region, leading to a positive feedback. Tanespimycin effectively abated the oncogenic effect. CONCLUSIONS: We first identified the lncRNA PVT1/STAT5B positive feedback loop for bladder carcinogenesis, and found a potentially effective drug for BC.

SENP3 affects the expression of PYCR1 to promote bladder cancer proliferation and EMT transformation by deSUMOylation of STAT3.

Abnormal activation of signal transducer and activator of transcription 3 (STAT3) has been found in various types of human cancers, including bladder cancer (BC). In our study, we examined the regulation of STAT3 post-translational modifications (PTMs) and found that SENP3 is high in bladder cancer. Sentrin/SUMO-specific protease3 (SENP3) and STAT3 were highly expressed in BC tissues when compared with tissue adjacent to carcinoma. SENP3 induced STAT3 protein level and p-STAT3 translocating into nuclear through deSUMOylation of STAT3. Further, nuclear STAT3, as a transcriptional activity factor, promoted pyrroline-5-carboxylate reductase 1 PYCR1 gene and protein level by interacting with the promoter of (PYCR1). Next, we found that knockdown of PYCR1 inhibited Epithelial to mesenchymal transition of bladder cancer, and simultaneously mitigated the carcinogenic effects of STAT3. In vitro, STAT3 knockdown in bladder cancer cells inhibited cell proliferation, migration, and invasion. In contrast, SENP3 overexpression reversed these effects. In all, results lend novel insights into the regulation of STAT3, which has key roles in bladder cancer progression.

Verapamil inhibited the development of ureteral stricture by blocking CaMK II-mediated STAT3 and Smad3/JunD pathways.

BACKGROUND: Ureteral stricture (US) is a fibrotic process that leads to urinary tract obstruction and even kidney damage, with the characteristic of reduced extracellular matrix (ECM) degradation and increased collagen synthesis. Verapamil, as a calcium channel blocker, was reported to prevent scar formation. Our work aimed to investigate the biological effects and mechanism of verapamil in US. METHODS: Fibroblasts were subjected to transforming growth factor-beta 1 (TGF-ß1) to stimulate collagen synthesis, and the messenger ribonucleic acid (mRNA) and protein expressions in fibroblasts were assessed using quantitative real-time polymerase chain reaction (qRT-PCR) and western blot. The location of phosphorylation-signal transducer and activator of transcription 3 (p-STAT3) and Jund proto-oncogene subunit (JunD) in fibroblasts were determined by immunofluorescence (IF). The binding relationship between signal transducer and activator of transcription 3 (STAT3) and collagen type I alpha1 (COL1A1)/collagen type III alpha 1 chain (COL3A1) and the binding relationship between JunD and tissue inhibitor of metalloproteinases-1 (TIMP-1) were verified by dual luciferase reporter gene and chromatin Immunoprecipitation (ChIP) assays. RESULTS: Herein, we found that verapamil could inhibit TGF-ß1/Ca2 + /calmodulin-dependent protein kinase II (CaMK II)-mediated STAT3 activation in fibroblasts, and STAT3 inhibition repressed collagen production. In addition, verapamil could inhibit TGF-ß1/CaMK II-mediated Mothers against DPP homolog 3 (Smad3)/JunD pathway activation in fibroblasts, and JunD silencing inhibited TIMP1 (a matrix metalloproteinase inhibitor) expression. Our subsequent experiments revealed that STAT3 bound with COL1A1 promoter and COL3A1 promoter and activated their transcription, and JunD bound with TIMP1 promoter and activated its transcription. Moreover, as expected, STAT3 activation could eliminate the inhibitory effect of verapamil treatment on TGF-ß1-induced collagen production in fibroblasts, and JunD overexpression reversed the inhibitory effect of verapamil treatment on TGF-ß1-induced TIMP1 expression in fibroblasts. CONCLUSION: Verapamil inhibited collagen production and TIMP-1 expression in US by blocking CaMK II-mediated STAT3 and Smad3/JunD pathways.

Verapamil inhibits ureteral scar formation by regulating CaMK II-mediated Smad pathway.

Verapamil is reported to prevent scar formation. However, whether verapamil is involved in the ureteral stricture scar and the underlying mechanism need further investigation. Fibroblasts were isolated from ureteral scar tissues. TGF-ß1 stimulation was used to induce fibrosis of fibroblasts. Inhibition of CaMK II was achieved by shRNA transfection. CCK-8 was performed to evaluate cell viability. qRT-PCR was applied to determine the level of mRNA while western blotting was used to determine the level of proteins. Immunofluorescence was used to detect the level of vimentin, collagen I and collagen III. Primary fibroblasts was successfully isolated from ureteral scar tissues. TGF-ß1 stimulation was capable to induce collagen production and fibrosis in primary fibroblasts while inhibition of CaMK II attenuate collagen production. Overexpression of wild type CaMK II lead to further increase of collagen production upon TGF-ß1 stimulation while the mutated CaMK II did not exert this promotion. Treatment of verapamil inhibits TGF-ß1 induced collagen production via inhibiting CaMK II. In present study, we revealed a vital role of Verapamil and CaMK II in the formation of ureteral scar. Verapamil inhibited TGF-ß1 induced collagen fiber formation by regulating CaMK II. Our finding might provide new insight into mechanism of prevention and treatment of ureteral scar.

Two-stage laparoscopic repair of two-level ureteral strictures: our experience of 8 patients.

PURPOSE: To evaluate the feasibility and effectiveness of two-stage laparoscopic repair for two-level ureteral strictures. MATERIALS AND METHODS: From October 2010 to January 2017, 8 patients with two-level ureteral strictures, which were located in upper and lower ureter, received two-stage laparoscopic repair in our institution. Laparoscopic ureteroureterostomy was conducted for the upper ureteral strictures in first stage and 8 weeks later laparoscopicureterovesical reimplantation was performed for lower stricture after the patency of upper lesion was confirmed by antegrade ureteropyelography. The kidney was drained by a nephrostomy tube during the interval period of two operations. RESULT: All the operations were performed successfully without intraoperative complications except one patient converted to open surgery during second-stage operation. For first-stage surgery, mean operating time was 120.88 ± 16.88 min, mean blood loss was 89.38 ± 13.74 mL, and mean duration of postoperative hospitalization was 3.63± 0.74 days. While in second-stage surgery, mean operating time took 125.25 ± 17.00 min, mean blood loss was 65.63 ± 10.16 mL, and mean duration of postoperative hospitalization was 3.62 ± 1.41 days.. On ureteropyelography 10 weeks after second-stage surgery, the contrast medium flowed from kidney down into bladder unrestrictedly and the patency of entire ureter was restored in all patients. During the follow-up, one female was observed kidney atrophy with ureteral calculus formed on the lesion side, and was successfully treated by ureteroscopiclithotripsy. No sign of stricture recurrence was found on other patients. CONCLUSION: Two-stage laparoscopic repair is a feasible and effective treatment for two-level ureteral strictures.But its indication is relatively narrow and confined to ureteral strictures located in two sites with sufficient intervaldistance and minor stricture length.

Urethral calculi with a urethral fistula: a case report and review of the literature.

BACKGROUND: To explore and summarize the reasons why urethral calculi cause a urethral fistula. CASE PRESENTATION: We retrospectively studied 1 patient in Xiangya hospital and all relevant literature published in English between 1989 and 2015. The patients (including those reported in the literature) were characterized by age, origin, location of calculus, size of calculus, fistulous track, and etiological factors. Most of urethral calculi associated with a urethral fistula were native generated. Urethral calculi can be formed in various locations of the urethra, and the size of the calculus ranged from small (multiple) calculi to giant stones. The fistula external orifice located at the root of the penis was relatively common, and there were various etiological factors, such as urethral strictures, urethral trauma induced by long-term catheterization, lumbar fractures, and congenital anomaly factors. They were managed by the excision of the fistulous tract, retrieval of the urethral stones, and/or debridement and pus drainage operations. CONCLUSION: Some elements, such as trauma, recurrent urinary tract infections, abscess formation induced by long-term catheterization, and urethral calculus, may be the risk factors for a urethral fistula.

Factors Associated with Knowledge of and Willingness for Adult Male Circumcision in Changsha, China.

BACKGROUND: Male circumcision (MC) has been shown to reduce the risk of male genital diseases. MC is not commonly practiced among Chinese males and little is known about the factors associated with their knowledge of and willingness for MC. This study was to explore the knowledge regarding the foreskin among Chinese males and to identify factors associated with their willingness to undergo circumcision. METHODS: A total of 237 patients with redundant prepuce/phimosis were interviewed through face-to-face interviews. The items on the questionnaire included: demographics, an objective scale assessing knowledge about the foreskin, willingness to have MC, the attitudes of sexual partners and doctors toward redundant prepuce/phimosis, and the approaches that patients used to acquire knowledge regarding the prepuce. Univariate analysis and multiple logistic regression analysis were performed to identify factors that are associated with willingness to be circumcised (WTC). RESULTS: A total of 212 patients completed the interview. Multivariable logistic regression showed that three factors were significantly associated with WTC: being married (OR = 0.43), perceiving redundant prepuce/phimosis as a disease (OR = 1.93), and if a patient's partner supported MC (OR = 1.39). 58% (n = 122) had received information about the foreskin from another party: 18% (n = 37) from school, 8% (n = 17) from family, 17% (n = 36) from friends, 27% (n = 57) from health care providers. About 4% (n = 8) believed that their partners disliked their redundant prepuce/phimosis. 20% (n = 42) had received doctors' advice to undergo circumcision. CONCLUSION: Knowledge about the foreskin was low among Chinese males. Our study elucidates the factors associated with WTC and suggests that more education of the population about the foreskin can help improve the recognition of a correctible abnormality and help patients assess the potential role of MC in their health.

Downregulation of VEGFA inhibits proliferation, promotes apoptosis, and suppresses migration and invasion of renal clear cell carcinoma.

OBJECTIVE: The aim of this study was to investigate the effects of vascular endothelial growth factor A (VEGFA) on cell proliferation, apoptosis, migration, and invasion in renal clear cell carcinoma (RCCC). METHODS: Between June 2012 and June 2015, RCCC tissues were obtained for the experimental group, and RCCC adjacent tumor-free kidney parenchyma tissues were obtained for the control group. VEGFA mRNA and protein expressions and phosphoinositide 3-kinase, serine/threonine-specific protein kinase (AKT), and phosphorylated-AKT protein expressions were detected. The chemically synthesized specific siRNA using RNA interference technology was used to inhibit VEGFA gene expression in human RCCC 786-O cells. The negative control (NC) group was transfected with NC sequence, and the blank group was transfected with no sequence. Flow cytometry, scratch test, and cell-penetrating experiment were used to detect cell proliferation, apoptosis, migration, and invasion of 786-O cells. RESULTS: Positive expression of VEGFA protein was 60.62% in RCCC tissue and 18.34% in adjacent tissue with statistically significant difference (P<0.001). VEGFA protein and mRNA expressions were higher in RCCC tissue than those in adjacent tissue (both P<0.01). VEGF expression in RCCC tissue was associated with Fuhrman grading and American Joint Committee on Cancer staging (both P<0.05). After RCCC 786-O cells transfecting the VEGFA siRNA, the VEGFA mRNA and protein expressions and phosphoinositide 3-kinase and phosphorylated-AKT protein expressions were significantly decreased, cell proliferation was remarkably inhibited, cell apoptotic ratio was obviously increased, and migration distance and invasive cell number were markedly decreased compared to those in the NC group and the blank group (all P<0.05). CONCLUSION: Inhibition of VEGFA inhibited proliferation, promoted apoptosis, and suppressed migration and invasion of RCCC 786-O cells. VEGF has a potential role in diagnosis and therapy of RCCC.

Diagnosis and management of testicular rupture after blunt scrotal trauma: a literature review.

Testicular rupture, one of the most common complications in blunt scrotal trauma, is the rupture of tunica albuginea and extrusion of seminiferous tubules. Testicular rupture is more inclined to young men, and injury mechanisms are associated with sports and motor accidents. After history taking and essential physical examination, scrotal ultrasound is the first-line auxiliary examination. MRI is also one of the vital complementary examinations to evaluate testicular rupture after blunt scrotal trauma. Surgical exploration and repair may be necessary when the diagnosis of testicular rupture is definite or suspicious. Postoperative follow-up is to monitor the relief of local symptoms and changes of testicular functions. This review sums up the literatures about testicular rupture after blunt scrotal trauma in recent 16 years and also refers some new advantages and perspectives on diagnosis and management of testicular rupture.