Magnitude-dependent proliferation and contractility modulation of human bladder smooth muscle cells under physiological stretch.

PURPOSE: The purpose of this study was to describe and test a kind of stretch pattern which is based on modified BOSE BioDynamic system to produce optimum physiological stretch during bladder cycle. Moreover, we aimed to emphasize the effects of physiological stretch's amplitude upon proliferation and contractility of human bladder smooth muscle cells (HBSMCs). METHODS: HBSMCs were seeded onto silicone membrane and subjected to stretch simulating bladder cycle at the range of stretches and time according to customized software on modified BOSE BioDynamic bioreactor. Morphological changes were assessed using immunofluorescence and confocal laser scanning microscope. Cell proliferation and cell viability were determined by BrdU incorporation assay and Cell Counting Kit-8, respectively. Contractility of the cells was determined using collagen gel contraction assay. RT-PCR was used to assess phenotypic and contractility markers. RESULTS: HBSMCs were found to show morphologically spindle-shaped and orientation at various elongations in the modified bioreactor. Stretch-induced proliferation and viability depended on the magnitude of stretch, and stretches also regulate contractility and contraction markers in a magnitude-dependent manner. CONCLUSION: We described and tested a kind of stretch pattern which delivers physiological stretch implemented during bladder cycle. The findings also showed that mechanical stretch can promote magnitude-dependent morphological, proliferative and contractile modulation of HBSMCs in vitro.

Intravenous Iron Sucrose versus Oral Iron in the Treatment of Pregnancy with Iron Deficiency Anaemia: A Systematic Review.

BACKGROUND: Intravenous iron sucrose and oral iron therapy are the main therapies for iron deficiency anaemia (IDA), but there is still a debate regarding their efficacy and especially as to which one is the best choice during pregnancy. METHODS: A meta-analysis of randomised controlled trials comparing patients treated with intravenous iron sucrose (intravenous group) with those treated with oral iron (oral group) for IDA during pregnancy was performed. The primary outcomes of interest were mean maternal haemoglobin and serum ferritin levels at the end of treatment. Secondary outcomes were treatment-related adverse events and foetal birth weight. RESULTS: Six randomised controlled trials, involving a total of 576 women, were included in the present review. Significant increases in haemoglobin [mean difference (MD), 0.85; 95% confidence interval (CI), 0.31-1.39; p = 0.002] and ferritin levels (MD, 63.32; 95% CI, 39.46-87.18; p < 0.00001) were observed in the intravenous group. Compared with the oral group, there were fewer adverse events in the intravenous group (risk ratio, 0.50; 95% CI, 0.34-0.73; p = 0.0003). There was no significant difference in birth weight between the two groups. CONCLUSION: For pregnant women who could not tolerate the side effects of oral treatment or required a rapid replacement of iron stores, intravenous iron sucrose was associated with fewer adverse events and was more effective than regular oral iron therapy.

Expression and proliferation profiles of PKC, JNK and p38MAPK in physiologically stretched human bladder smooth muscle cells.

OBJECTIVE: To determine protein kinase C (PKC), c-Jun NH2-Terminal Kinase (JNK) and P38 mitogen-activated protein kinases (p38MAPK) expression levels and effects of their respective inhibitors on proliferation of human bladder smooth muscle cells (HBSMCs) when physiologically stretched in vitro. MATERIALS AND METHODS: HBSMCs were grown on silicone membrane and stretch was applied under varying conditions; (equibiaxial elongation: 2.5%, 5%, 10%, 15%, 20%, 25%), (frequency: 0.05, 0.1, 0.2, 0.5, 1Hz). Optimal physiological stretch was established by assessing proliferation with 5-Bromo-2-deoxyuridine (BrdU) assay and flow cytometry. PKC, JNK and p38 expression levels were analyzed by Western blot. Specificity was maintained by employing specific inhibitors; (GF109203X for PKC, SP600125 for JNK and SB203580 for p38MAPK), in some experiments. RESULTS: Optimum proliferation was observed at 5% equibiaxial stretch (BrdU: 0.837±0.026 (control) to 1.462±0.023)%, (P<0.05) and apoptotic cell death rate decreased from 16.4±0.21% (control) to 4.5±0.13% (P<0.05) applied at 0.1Hz. Expression of PKC was upregulated with slight increase in JNK and no change in p38MAPK after application of stretch. Inhibition had effects on proliferation (1.075±0.024, P<0.05 GF109203X); (1.418±0.021, P>0.05 SP600125) and (1.461±0.01, P>0.05 SB203580). These findings show that mechanical stretch can promote magnitude-dependent proliferative modulation through PKC and possibly JNK but not via p38MAPK in hBSMCs.

The purinergic component of human bladder smooth muscle cells' proliferation and contraction under physiological stretch.

OBJECTIVE: To investigate whether cyclic stretch induces proliferation and contraction of human smooth muscle cells (HBSMCs), mediated by P2X purinoceptor 1 and 2 and the signal transduction mechanisms of this process. METHODS: HBSMCs were seeded on silicone membrane and stretched under varying parameters; (equibiaxial elongation: 2.5%, 5%, 10%, 15%, 20%, 25%), (Frequency: 0.05Hz, 0.1Hz, 0.2Hz, 0.5Hz, 1Hz). 5-Bromo-2-deoxyuridine assay was employed for proliferative studies. Contractility of the cells was determined using collagen gel contraction assay. After optimal physiological stretch was established; P2X1 and P2X2 were analyzed by real time polymerase chain reaction and Western Blot. Specificity of purinoceptors was maintained by employing specific inhibitors; (NF023 for P2X1, and A317491for P2X2), in some experiments. RESULTS: Optimum proliferation and contractility were observed at 5% and 10% equibiaxial stretching respectively, applied at a frequency of 0.1Hz; At 5% stretch, proliferation increased from 0.837±0.026 (control) to 1.462±0.023%, p<0.05. Mean contraction at 10% stretching increased from 31.7±2.3%, (control) to 78.28 ±1.45%, p< 0.05. Expression of P2X1 and P2X2 was upregulated after application of stretch. Inhibition had effects on proliferation (1.232±0.051, p<0.05 NF023) and (1.302±0.021, p<0.05 A314791) while contractility was markedly reduced (68.24±2.31, p<0.05 NF023) and (73.2±2.87, p<0.05 A314791). These findings shows that mechanical stretch can promote magnitude-dependent proliferative and contractile modulation of HBSMCs in vitro, and P2X1 and 2 are at least partially responsible in this process.

Integrin αv mediates contractility whereas integrin α4 regulates proliferation of human bladder smooth muscle cells via FAK pathway under physiological stretch.

PURPOSE: The requirement of integrins for mechanotransduction has been recognized for some time. We investigated the role of integrin subunits and their pathway in the physiological stretch induced contractility and proliferation of human bladder smooth muscle cells. MATERIALS AND METHODS: Human bladder smooth muscle cells were seeded on silicone membrane and subjected to stretch, simulating bladder cycles of various stretches and times, as controlled by customized software on a modified BioDynamic bioreactor. Cell proliferation, viability and cycle were determined by BrdU incorporation assay, the Cell Counting Kit-8 (Beyotime Institute of Biotechnology, Haimen, People's Republic of China) and flow cytometry, respectively. Cell contractility was determined using a collagen gel contraction assay. RESULTS: Physiological stretch increased cell contractility, proliferation and viability. Knockdown of integrin αv but not α4 in the cells disrupted the enhanced contractility induced by stretch. Under physiological stretch conditions, the integrin αv level and phospho-FAK/FAK ratio correlated positively with cell stretch induced enhanced contractility. Further examination revealed that contractile marker expression was associated with integrin αv activation through the FAK pathway. At the same time integrin α4 but not integrin αv mediated stretch induced cell proliferation and viability. CONCLUSIONS: These data revealed that different integrins have different roles in the contractility and proliferation of human bladder smooth muscle cells under physiological stretch. This suggests that different integrins may become specific therapeutic targets in patients with voiding dysfunction. They may also be used to design a specific microenvironment for optimal bladder tissue regeneration.

Isoflurane inhalation anesthesia should be a new requirement in intracavernosal pressure detection-the gold standard of erectile function assessment.

Intracavernosal pressure (ICP) is gold standard for the detection of erectile function in animals, but no consensus has yet been achieved on what kind of anesthetic protocol should be applied. A total of 16 adult male Sprague-Dawley rats were randomized into two groups. In group A, chloral hydrate was injected intraperitoneally. Rats in group B were induced in 5% isoflurane for 3 min and then maintained in 1.0-1.5% isoflurane. Mean arterial pressure (MAP), respiratory rate (RR) and heart rate were monitored during all experiments. After ICP detection, tail vein and carotid artery blood were collected. The maximum ICP value, MAP and ICP/MAP ratio in group B was significantly higher than in that of group A. The RR in group A was lower than in that of group B, but the heart rate in group A was higher than in group B. There were no significant differences in both pO2 and pCO2 between groups. While the data showed that animals in group A were relatively hypoxemic. Isoflurane inhalation anesthesia in detection of erectile function could offer a relatively more stable physical state than in that under the effect of chloral hydrate intraperitoneal anesthesia. Isoflurane inhalation anesthesia is more suitable for ICP test.

Prevention of stricture recurrence following urethral internal urethrotomy: routine repeated dilations or active surveillance?

Strictures of the urethra are the most common cause of obstructed micturition in younger men and there is frequent recurrence after initial treatment. Currently, routine repeated dilations, including intermittent self-catheterisation (ISC) are prescribed by urologists to prevent urethral stricture recurrence. There is, however, no high level evidence available supporting the effectiveness of practicing these painful techniques. Balancing efficacy, adverse effects and costs, we hypothesize that active surveillance is a better option for preventing stricture recurrence as compared with routine repeated dilations. However, well designed, adequately powered multi-center trials with comprehensive evaluation are urgently needed to confirm our hypothesis. .

Water consumption and urinary tract infections: an in vitro study.

OBJECTIVES: To address to a better understanding of whether increased water consumption is associated with beneficial effects of urinary tract infections prophylaxis and treatment, and if so, the mechanism involved in this process. METHODS: Models of the catheterized bladder were infected with Escherichia coli. Artificial urine was supplied at various flow rates and various concentrations to separately assess the "flushing effect" and "dilution effect" of increased water consumption on catheter blockage time, encrustation formation, and bacterial growth. RESULTS: There were no statistical significances regarding catheter blockage time (P = 0.92), encrustation formation, and bacterial growth among bladder models supplied with various flow rates. When the flow rate was set as 1 ml/min, however, there showed significant decrease trend of the time to blockage (P = 0.0005), encrustation formation, and bacterial growth as the concentration of the artificial urine increased except the twofold-concentration urine group. CONCLUSIONS: Increased water consumption is associated with beneficial effects of urinary tract infection prophylaxis and treatment, and dilution effect of bacteria nutrition in the urine is at least partly involved in this process if not all, rather than the "flushing effect". Considering the flaws and the in vitro design of the current study, however, an in vivo study is warranted.

Cyclic stretch induces human bladder smooth muscle cell proliferation in vitro through muscarinic receptors.

The present study aimed to investigate whether the cyclic stretch­induced proliferation of human bladder smooth muscle cells (HBSMCs) is mediated by muscarinic (M) receptors, together with the signal transduction mechanisms involved in this process. HBSMCs seeded onto silicone membranes were subjected to different cyclic stretches (5, 10, 15 and 20%) for 6 and 12 h. As the effect of cyclic stretch on M2 and M3 mRNA expression levels was maximal at 6 h 10% stretch, all subsequent experiments were performed at this stretch. Western blot analysis was used to quantify M2, M3, protein kinase C (PKC) and phosphorylated (p)­PKC protein expression levels, flow cytometry was employed to examine cell cycle distribution and a 5-bromo­2-deoxyuridine (BrdU) incorporation assay was used to assess cell proliferation at this stretch. Subsequently, HBSMCs were exposed to different acetylcholine concentrations and/or cyclic stretch, M receptor antagonists [AF-DX16, an M2 receptor antagonist; 1,1-dimethyl-4-diphenylacetoxypiperidinium iodide (4-DAMP), an M3 receptor antagonist and atropine, a non­selective antagonist] and GF 109203X, a PKC antagonist, to assess the possible underlying signaling mechanisms. Cyclic stretch was found to increase the proliferation of HBSMCs and the expression levels of M2, M3, PKC and p­PKC proteins. M receptor and PKC antagonists exerted no apparent effect on nonstretched cells, but reduced the incorporation of BrdU into stretched cells; the most pronounced effects were observed when non­selective M receptor and PKC antagonists were applied. Notably, 4­DAMP did not inhibit stretch­induced PKC activation. These results indicate that the activation of the M3 receptor signaling pathway in stretch­induced HBSMC proliferation occurs via PKC-independent mechanisms.

Combined Dorsal Plus Ventral Double-Graft Urethroplasty in Anterior Urethral Reconstruction.

This study aims to investigate the effect of combined dorsal plus ventral double-graft urethroplasty in anterior urethral reconstruction. Patients who underwent graft urethroplasty for anterior urethral strictures at West China Hospital from 2005 to 2010 were followed up with clinical evaluation. According to the site of graft fixed, patients were divided into single-onlay group (dorsal or ventral) and double-onlay group (dorsal plus ventral). Success rate and complications were compared between the two groups and were analyzed using t test and chi-square. A total of 77 patients completed the follow-up, 51 in single-onlay group and 26 in double group. There was no statistical difference in terms of age, length, site, stricture reason, and the type of graft used between the two groups. The mean follow-up time was 15.6 months (range from 4 to 33 months) in double group and 39.5 months (range from 15 to 59 months) in single group. The total success rate was 72.5 % in single-onlay group and 88.5 % in double-onlay group; no statistical difference existed (p > 0.05). Subgroup analysis shows the success rate was higher for double-onlay urethroplasty for the stricture of penoscrotal junction (88.9 vs 60.9 %, p < 0.05). The main postoperative complications including infection and urethrocutaneous fistula were not significant between the two groups (p > 0.05). Combined dorsal plus ventral double-graft urethroplasty showed a high success and low complication rate for anterior urethral strictures, especially for the penoscrotal junction.

Prevention of stricture recurrence following urethral endoscopic management: what do we have?

OBJECTIVE: Strictures of the urethra are the most common cause of obstructed micturition in younger men and there is frequent recurrence after initial treatment. This review was performed to determine the best strategy for stricture recurrence prevention following urethral endoscopic management. METHODS: We reviewed the published literature in PubMed, the Cochrane Library, and Google Scholar focusing on this intractable problem regardless of language restrictions. Outcomes of interest included the study methods and the applied strategy's efficacy. The level of evidence and grade of recommendations of included studies were appraised with an Oxford Centre for Evidence-Based Medicine Scale. RESULTS: Currently, numerous techniques, including catheterization, repeated dilation, brachytherapy, and intraurethral use of various antifibrosis agents, have been employed to oppose the process of wound contraction or regulate the extracellular matrix. But unfortunately, none of these techniques or agents have demonstrated efficacy with enough evidence. CONCLUSIONS: Although lots of strategies are available, still, we do not have a suitable, single optimum solution for all the conditions. The clinical decision of stricture-recurrence-prevention techniques should be carefully tailored to every individual patient. As the studies are not sufficient, more efforts are warranted to address this interesting but challenging issue.

The management of urethral stricture after kidney transplantation.

OBJECTIVE: To investigate the incidence and causes of urethral stricture after kidney transplantation, as well as analyze its diagnosis, treatment and prevention. METHODS: Clinical data of patients who developed urethral stricture after living-donor kidney transplantation in our center between January 2007 and June 2012 were retrospectively analyzed. RESULTS: Urethral stricture occurred in 8 of the 677 eligible kidney recipients (1.18 %) during the study period; the complication occurred at a mean of 4.4 months (range 2-7 months) after transplantation. Cystoscope-related iatrogenic injury and urinary tract infection seemed to be the most likely causes. In addition to frequency and dysuria, three patients had hydronephrosis and four had elevated serum creatinine levels. Urethrography showed that the urethral stricture was anterior in two patients and posterior in the remaining six. Two patients were treated by urethral dilation, four by internal urethrotomy and two by urethra reconstruction surgery. All patients urinated readily after treatment and four patients with impaired renal function recovered. CONCLUSION: Urethral strictures after kidney transplantation are rare, and they can be safely and effectively treated by urethral dilation, internal urethrotomy or urethra reconstruction. Avoiding iatrogenic injury and shortening catheterization time may help reduce the risk of this complication.

Maximizing the donor pool: left versus right laparoscopic live donor nephrectomy--systematic review and meta-analysis.

OBJECTIVE: To assess the current evidence regarding the efficiency, safety, and potential advantages of right-laparoscopic live donor nephrectomy (LLDN) compared with left-LLDN. MATERIALS AND METHODS: We comprehensively searched PubMed, Embase, the Cochrane Library, and Web of Science to perform a systematic review and cumulative meta-analysis of a randomized controlled trial (RCT) and prospective and retrospective comparative studies assessing the two techniques. RESULTS: A single RCT and 28 comparative studies including a total of 32,426 cases were identified. Although left-LLDN was associated with lower delayed renal function [odds ratio (OR) 0.73; 95 % confidence interval (CI) 0.62, 0.87, p = 0.0003] and less thrombosis (OR 0.35; 95 % CI 0.13, 0.96, p = 0.04) in our meta-analysis, sensitivity analysis did not show any significant difference between the two groups. There were no other considerable differences between the two groups. CONCLUSIONS: With enough surgical experience, right-LLDN can be performed with equivalent safety and efficacy. Although large-volume centers are routinely performing right-LLDN, there is under appreciation of the right side in smaller centers given the associated fear of poor outcomes and technically demanding procedure. Due to the inherent limitations of the included studies, future well-designed RCTs are awaited to confirm and update the findings of this analysis.

Primary adenocarcinoma of the rete testis: A case report and review of the literature.

Primary adenocarcinoma of the rete testis is an extremely rare extratesticular neoplasm. Due to its low occurrence and the scarcity of data, sonographic characteristics of adenocarcinoma of the rete testis are still poorly defined. A 46-year-old male complained of swelling and pain in the right side of the scrotum. No associated symptoms were observed. Anti-tuberculosis chemotherapy yielded no response. Postoperative pathology revealed a diagnosis of poorly-differentiated adenocarcinoma of the rete testis. Using the ultrasonography features observed, combined with a review of current literature, the diagnosis and differential diagnosis of this neoplasm are presented.

Antimuscarinics and α-blockers or α-blockers monotherapy on lower urinary tract symptoms--a meta-analysis.

OBJECTIVE: To assess the clinical efficiency and safety of combination pharmacotherapy of antimuscarinics and α-blockers vs α-blockers monotherapy on patients with moderate to severe lower urinary tract symptoms (LUTS). METHODS: We searched the Cochrane Central Register of Controlled Trials, PubMed, EMBASE, the Cochrane Database of Systematic Review, and Web of Science from their inception until June 2013 to identify all eligible studies that compare the 2 pharmacotherapy strategies on LUTS. The Cochrane Collaboration's RevMan 5.2 software was used for data analysis, and the fixed or the random effect model was selected depending on the proportion of heterogeneity. RESULTS: Eighteen eligible randomized controlled trials were included in this systematic review, including 2106 (51.57%) in cotherapy group and 1978 (48.43%) in monotherapy group. Synthetic data showed that there were significant improvements on Storage International Prostate Symptom Score (mean difference [MD] = -1.51; 95% confidence interval [CI] -2.10 to -0.91, P <.00001), quality of life score (MD = -0.53; 95% CI -0.89 to -0.17, P = .004), micturitions per 24 hours (MD = -1.14; 95% CI -1.84 to -0.45, P = .001), and urgency episodes per 24 hours (MD = -0.99; 95% CI -1.46 to -0.51, P <.0001) in the cotherapy group. There were no significant difference regarding maximum flow rate (MD = -0.05; 95% CI -0.27 to 0.17, P = .64), Total International Prostate Symptom Score (TIPSS) (MD = -0.88; 95% CI -1.64 to -0.12, P = .02), and Voiding International Prostate Symptom Score (VIPSS) (MD = 0.40; 95% CI -0.34 to 1.15, P = .29). As to postvoid residual volume, however, a worse condition was showed in the cotherapy group (MD = -6.53; 95% CI 3.06-10.00, P <.0002). CONCLUSION: Antimuscarinics could and should be added to the drug regimen for patients with LUTS attributed to benign prostatic hyperplasia/bladder outlet obstruction, particularly are dominated by storage symptoms. For patients with increased acute urinary retention risk, they should be carefully monitored.

Primary squamous cell carcinoma of seminal vesicle: an extremely rare case report with literature review.

Primary squamous cell carcinoma of seminal vesicle is extremely rare, and most cases regarding seminal vesicle tumors failed to address this kind of tumor. A 54-year-old male patient presented with intermittent painless visual hematuria for 6 months was hospitalized. Ultrasonography, computerized tomography and magnetic resonance imaging demonstrated a 4.4 cm × 3.6 cm × 3.0 cm mixed tumorous lesion in the left seminal vesicle. A transrectal needle biopsy revealed severe chronic inflammation. The mass was completely resected in a laparoscopic approach and was verified as a moderately differentiated squamous cell carcinoma in the seminal vesicle by post-surgical histopathological examination. The patient received totally 5 cycles of chemotherapy. A rectal metastasis was detected 7 months after the surgery.

Effects of circumcision on male sexual functions: a systematic review and meta-analysis.

This meta-analysis was performed to assess sexual functions following adult male circumcision. We searched the Cochrane Central Register of Controlled Trials, PUBMED, EMBASE, the Cochrane Database of Systematic Review and Web of Science from their inception until January 2013 to identify all eligible studies that reported on men's sexual function after circumcision. The Cochrane Collaboration's RevMan 5.2 software was employed for data analysis, and the fixed or the random effect model was selected depending on the proportion of heterogeneity. We identified 10 studies, which described a total of 9317 circumcised and 9423 uncircumcised men who were evaluated for the association of circumcision with male sexual function. There were no significant differences in sexual desire (odds ratio (OR): 0.99; 95% confidence interval (CI): 0.92-1.06), dyspareunia (OR: 1.12; 95% CI: 0.52-2.44), premature ejaculation (OR: 1.13; 95% CI: 0.83-1.54), ejaculation latency time (OR: 1.33; 95% CI: 0.69-1.97), erectile dysfunctions (OR: 0.90; 95% CI: 0.65-1.25) and orgasm difficulties (OR: 0.97; 95% CI: 0.83-1.13). These findings suggest that circumcision is unlikely to adversely affect male sexual functions. However, these results should be evaluated in light of the low quality of the existing evidence and the significant heterogeneity across the various studies. Well-designed and prospective studies are required for a further understanding of this topic.

Active targeted therapy for metastatic collecting duct carcinoma of the kidney: a case report and review of the literature.

Collecting duct carcinoma (CDC) is a rare and aggressive renal cell carcinoma (RCC) with extremely poor prognosis, which has been shown to have a poor response to several kinds of systemic therapy. Targeted agents have greatly changed the therapeutic landscape in advanced RCC. Nonetheless, patients with CDC are always excluded from the prospective trials with targeted therapies due to its rarity. We present a case of metastatic CDC that responded favorably to the multiple tyrosine kinase inhibitor, sorafenib, achieving a partial response in both lungs and retroperitoneal lymph nodes metastases. We also reviewed the limited number of reports of metastatic CDC treated with targeted agents and found that 33.33 % (4/12) of patients had favorable clinical activity. These suggest that targeted therapy should be considered for the treatment of metastatic CDC and its prospective evaluation is encouraged.