Web- and Artificial Intelligence-Based Image Recognition For Sperm Motility Analysis: Verification Study.

BACKGROUND: Human sperm quality fluctuates over time. Therefore, it is crucial for couples preparing for natural pregnancy to monitor sperm motility. OBJECTIVE: This study verified the performance of an artificial intelligence-based image recognition and cloud computing sperm motility testing system (Bemaner, Createcare) composed of microscope and microfluidic modules and designed to adapt to different types of smartphones. METHODS: Sperm videos were captured and uploaded to the cloud with an app. Analysis of sperm motility was performed by an artificial intelligence-based image recognition algorithm then results were displayed. According to the number of motile sperm in the vision field, 47 (deidentified) videos of sperm were scored using 6 grades (0-5) by a male-fertility expert with 10 years of experience. Pearson product-moment correlation was calculated between the grades and the results (concentration of total sperm, concentration of motile sperm, and motility percentage) computed by the system. RESULTS: Good correlation was demonstrated between the grades and results computed by the system for concentration of total sperm (r=0.65, P<.001), concentration of motile sperm (r=0.84, P<.001), and motility percentage (r=0.90, P<.001). CONCLUSIONS: This smartphone-based sperm motility test (Bemaner) accurately measures motility-related parameters and could potentially be applied toward the following fields: male infertility detection, sperm quality test during preparation for pregnancy, and infertility treatment monitoring. With frequent at-home testing, more data can be collected to help make clinical decisions and to conduct epidemiological research.

Surgical niche for the treatment of erectile dysfunction.

Penile erection implicates arterial inflow, sinusoidal relaxation and corporoveno-occlusive function. By far the most widely recognized vascular etiologies responsible for organic erectile dysfunction can be divided into arterial insufficiency, corporoveno-occlusive dysfunction or mixed type, with corporoveno-occlusive dysfunction representing the most common finding. In arteriogenic erectile dysfunction, corpora cavernosa show lower oxygen tension, leading to a diminished volume of cavernosal smooth muscle and consequential corporoveno-occlusive dysfunction. Current studies support the contention that corporoveno-occlusive dysfunction is an effect rather than the cause of erectile dysfunction. Surgical interventions have consisted primarily of penile revascularization surgery for arterial insufficiency and penile venous surgery for corporoveno-occlusive dysfunction, whatever the mechanism. However, the surgical effectiveness remained debatable and unproven, mostly owing to the lack of consistent hemodynamic assessment, standardized select patient and validated outcome measures, as well as various surgical procedures. Penile vascular surgery has been disclaimed to be the treatment of choice based on the currently available guidelines. However, reports on penile revascularization surgery support its utility in treating arterial insufficiency in otherwise healthy patients aged <55 years with erectile dysfunction of late attributable to arterial occlusive disease. Furthermore, it is noteworthy that penile venous surgery might be beneficial for selected patients with corporoveno-occlusive dysfunction, especially with a better understanding of the innovated venous anatomy of the penis. Penile vascular surgery might remain a viable alternative for the treatment of erectile dysfunction, and could have found its niche in the possibility of obtaining spontaneous, unaided and natural erection.

Clinical and Imaging Outcomes up to 1 Year Following Balloon Angioplasty for Isolated Penile Artery Stenoses in Patients With Erectile Dysfunction: The PERFECT-2 Study.

PURPOSE: To assess the angiographic and clinical outcomes in patients with erectile dysfunction and isolated penile artery stenoses treated by balloon angioplasty. METHODS: In this prospective study, 22 patients (mean age 61.0±7.6 years, range 50-79) with erectile dysfunction and 34 isolated penile artery stenoses (mean 74.9%±9.1%) were enrolled and underwent balloon angioplasty. The mean International Index for Erectile Function-5 (IIEF-5) score at baseline was 10.3±4.5. The mean lesion length was 11.1±9.0 mm (mean reference vessel diameter 1.7±0.4 mm). The primary endpoint was in-segment restenosis ≥50% by pelvic computed tomography angiography (CTA) at 8 months. The 1-year sustained clinical success (IIEF-5 score ≥22 or a ≥4-point change in the IIEF-5 score and no later decline by ≥4) was the secondary outcome measure. RESULTS: Procedural success was achieved in 31 (91%) of 34 stenotic lesions; there was 1 flow-limiting dissection and 2 arteries with >30% residual stenosis. At 8 months, 14 of 34 lesions in 13 of 22 patients had CTA-documented binary restenosis. At 1 year, sustained clinical success was achieved in 11 of 22 patients. Of the 9 patients not developing binary restenosis, 8 achieved sustained clinical success. CONCLUSION: Our findings establish the safety and efficacy of penile artery angioplasty for patients with erectile dysfunction and isolated penile artery stenoses. They also highlight the unmet need for a more enduring treatment strategy for penile artery stenotic disease.

Epigallocatechin Gallate Attenuates Partial Bladder Outlet Obstruction-induced Bladder Injury via Suppression of Endoplasmic Reticulum Stress-related Apoptosis-In Vivo Study.

OBJECTIVES: To investigate the protective effect of epigallocatechin gallate (EGCG), a green tea extract, on partial bladder outlet obstruction (pBOO)-induced bladder injury in a rat model. METHODS: The female Sprague-Dawley rats underwent sham or BOO procedures, and were divided into several groups (sham with saline injection, sham with EGCG treatment, BOO with saline injection, and BOO with EGCG treatment). The rats in each group were randomized into 2 groups (48 hours and 30 days after the BOO procedure) for when their bladders were harvested. EGCG (4.5 mg/kg/day) and saline were administered via intraperitoneal injection after the BOO procedure during the study period. Bladder tissue was examined for inflammation, endoplasmic reticulum (ER) stress-related apoptotic markers by Western blot, and histological staining. RESULTS: BOO induced acute bladder injury (hemorrhage, edema, and neutrophil infiltration) after 48 hours. In addition, cystometry showed a decrease in micturition pressure and intercontractile interval. We also observed increased expressions of cyclooxygenase-2, poly(ADP-ribose) polymerase at 48 hours, as well as ER stress markers such as caspase-12 and CCAAT/-enhancer-binding protein homologous protein (CHOP). Treatment with EGCG significantly improved pBOO-induced histologic changes, bladder dysfunction, and the overexpression of cyclooxygenase-2, CHOP, and caspase-12 at 48 hours. Similarly, EGCG treatment for 30 days effectively recovered compliance and intercontractile interval, submucosal ER stress-related apoptosis (CHOP and caspase-12) at 30 days after pBOO. CONCLUSIONS: EGCG alleviate pBOO-induced bladder injury and dysfunction via suppression of inflammation and ER stress-related apoptosis.

Correlations of Metabolic Components with Prostate Volume in Middle-Aged Men Receiving Health Check-Up.

OBJECTIVES: To investigate the impact of metabolic components and body composition indices on prostate volume (PV) in a population of middle-aged men receiving health check-ups. METHODS: Six hundred and sixteen men receiving health assessments were stratified to large and small prostates based on the cut-off of median PV. Their demographic data, health history, and international prostate symptoms scores (IPSS) were collected. Metabolic components and body composition indices were compared between subjects with large and small prostates. Moreover, the correlations between these parameters and PV were analyzed by multivariate logistic regression. RESULTS: The median PV was 27 mL and mean age was 54.8 years. Subjects with large PV were older (56.5 vs. 52.7 years) and had higher serum prostate specific antigen (PSA) level (1.73 vs. 0.96 ng/mL), higher IPSS score (8.37 vs. 6.16), and higher body fat, body mass, and waist circumference (all p<0.05). In multivariate analysis, age (OR, 2.45; 95%CI, 1.74-3.45), serum PSA (OR, 2.75; 95%CI, 1.96-3.86), waist circumference (OR, 1.45; 95%CI, 1.02-2.07), fatness (OR, 1.47; 95%CI, 1.04-2.09), and body fat mass (OR, 1.43; 95%CI, 1.00-2.03) were significantly correlated with PV of study subjects. In subgroup analysis, raised waist circumference (OR, 1.89; 95%CI, 1.00-3.59) was the independent predictor of PV in subjects with bothersome lower urinary tract symptoms. CONCLUSIONS: Several metabolic components and body composition indices are significantly associated with PV of middle-aged men, including raised waist circumference, fatness, and body fat mass. Raised waist circumference is the only independent predictor of PV in middle-aged men with bothersome LUTS.

The Biochemical and Pathological Correlates of Successful Semen Cryopreservation From Patients With Testicular Cancer: A Single Tertiary Center Experience.

OBJECTIVE: To determine the predictors for success with regard to semen cryopreservation and good semen quality of patients with testicular cancer. MATERIALS AND METHODS: A total of 50 men (aged 16-36 years) with testicular cancer, referred for semen cryopreservation prior to gonadotoxic treatment, were included. Age, alpha fetal protein (α-FP), beta human chorionic gonadotropin, lactate dehydrogenase, clinical staging, tumor volume, and pathological reports were evaluated as correlates of successful semen cryopreservation and good semen quality. RESULTS: The overall success rate was 52%. α-FP (4113.1 ng/mL vs 81.2 ng/mL) and tumor volume (77.7 mL vs 25.5 mL) were significantly higher in the failure group as compared to the success group. The seminoma to nonseminomatous germ cell tumor ratio was lower in the failure group as compared to the success group (9/17 vs 3/21). There was nearly a significant difference (P = .066). The optimal cutoff value for α-FP > 1000 ng/mL showed the highest Youden index (0.689) and resulted in a sensitivity of 0.625 and specificity of 1.0 for predicting poor outcome. In terms of multivariate analysis, the α-FP (P = .013), tumor volume (P = .047), and α-FP > 1000 ng/mL (P = .010) were significantly associated with poor semen quality and failure to preserve semen. Sperm quality was found to be higher in the seminoma versus the nonseminomatous germ cell tumor patients in: sperm concentration (21.5 million/mL vs 11.8 million/mL, P < .027). Furthermore, tumor volume is correlated to α-FP (P = .018) and is weakly correlated to lactate dehydrogenase (P = .067) CONCLUSION: Elevated α-FP and tumor volume are independently poor factors for semen quality and semen cryopreservation. In clinical use, α-FP is a noninvasive tool to predict the success of semen cryopreservation and patients with α-FP > 1000 ng/mL should be informed of the higher risk of poor semen quality and semen cryopreservation concerns.

MLN4924 Synergistically Enhances Cisplatin-induced Cytotoxicity via JNK and Bcl-xL Pathways in Human Urothelial Carcinoma.

Cisplatin-based chemotherapy is the primary treatment for metastatic bladder urothelial carcinoma. However, the response rate is only 40-65%. This study investigated the anti-tumor effect and underlying mechanisms of the combination of cisplatin and the NEDD8-activating enzyme inhibitor MLN4924 in human bladder urothelial carcinoma. The combination of cisplatin and MLN4924 exerted synergistic cytotoxicity on two high-grade bladder urothelial carcinoma cell lines, NTUB1 and T24 (combination index <1). MLN4924 also potentiated the cisplatin-induced apoptosis and activation of caspase-3 and -7, phospho-histone H2A.X and PARP. c-Jun N-terminal kinase (JNK) activation and a down-regulation of B-cell lymphoma-extra large (Bcl-xL) were also observed during cisplatin and MLN4924 treatment. Inhibition of JNK activation partially restored cell viability and Bcl-xL expression. Bcl-xL overexpression also rescued cell viability. MLN4924 significantly potentiated cisplatin-induced tumor suppression in urothelial carcinoma xenograft mice. In summary, MLN4924 synergistically enhanced the anti-tumor effect of cisplatin via an increase in DNA damage, JNK activation and down-regulation of Bcl-xL in urothelial carcinoma cells. These findings provide a new therapeutic strategy for the treatment of bladder cancer.

MLN4924, a novel protein neddylation inhibitor, suppresses proliferation and migration of human urothelial carcinoma: In vitro and in vivo studies.

MLN4924, a small molecule inhibitor of NEDD8 activating enzyme (NAE), has been reported to elicit an anti-tumor effect on various malignancies. In this study, we investigated the anti-tumor effect of MLN4924 in human urothelial carcinoma (UC) in vitro and in vivo by using three human UC cell lines of various grading (T24, NTUB1 and RT4). The impact of MLN4924 on UC cells was determined by measuring viability (MTT), proliferation (BrdU incorporation), cell cycle progression (flow cytometry with propidium iodide staining) and apoptosis (flow cytometry with annexin V-FITC labeling). The cell cycle regulatory molecules, apoptosis-related molecules, and cell stress-related proteins were examined by Western blotting. The influence of tumor cell migration and invasion was analyzed by Transwell and wound healing assays. We also evaluated the effects of MLN4924 on tumor growth by a SCID xenograft mouse model. The data show that MLN4924 induced dose-dependent cytotoxicity, anti-proliferation, anti-migration, anti-invasion and apoptosis in human UC cells, accompanied by activations of Bad, phospho-histone H2A.X, caspase-3, 7 and PARP, decreased level of phospho-Bcl2, and caused cell cycle retardation at the G2M phase. Moreover, MLN4924 activated endoplasmic reticulum stress-related molecules (caspase-4, phospho-eIF2α, ATF-4 and CHOP) and other stress responses (JNK and c-Jun activations). Finally, we confirmed MLN4924 inhibited tumor growth in a UC xenograft mouse model with minimal general toxicity. We concluded that MLN4924 induces apoptosis and cell cycle arrest, as well as activation of cell stress responses in human UC. These findings imply MLN4924 provides a novel strategy for the treatment of UC.

MLN4924, a Novel NEDD8-activating enzyme inhibitor, exhibits antitumor activity and enhances cisplatin-induced cytotoxicity in human cervical carcinoma: in vitro and in vivo study.

MLN4924, an inhibitor of NEDD8 activating enzyme (NAE), has been reported to have activity against various malignancies. Here, we investigated the antitumor properties of MLN4924 and MLN4924 in combination with cisplatin on human cervical carcinoma (CC) in vitro and in vivo. Two human CC cell lines, ME-180 and HeLa, were used in this study. The cytotoxic effects of MLN4924 and/or cisplatin were measured by cell viability (MTT), proliferation (BrdU incorporation), apoptosis (flow cytometry with annexin V-FITC labeling), and the expression of cell apoptosis-related proteins (Western blotting). In vivo efficacy was determined in Nu/Nu nude mice with ME-180 and HeLa xenografts. The results showed that MLN4924 elicited viability inhibition, anti-proliferation and apoptosis in human CC cells, accompanied by activations of apoptosis-related molecules and Bid, Bcl-2 phosphorylation interruption, and interference with cell cycle regulators. Moreover, MLN4924 caused an endoplasmic reticulum stress response (caspase-4, ATF-4 and CHOP activations) and expression of other cellular stress molecules (JNK and c-Jun activations). Additionally, MLN4924 suppressed growth of CC xenografts in nude mice. Furthermore, we demonstrated that MLN4924 potentiated cisplatin-induced cytotoxicity in CC cells with activation of caspases. Consistently with this, MLN4924 significantly enhanced cisplatin-induced growth inhibition of CC xenografts. Together, these findings suggest that MLN4924 alone or in combination with cisplatin is of value in treating human CCs.

The prevalence and the risk factors of testosterone deficiency in newly diagnosed and previously known type 2 diabetic men.

INTRODUCTION: While the epidemiology of testosterone deficiency has been well described in men with previously known type 2 diabetes mellitus (T2DM), it was less reported in those with untreated, newly diagnosed T2DM. AIM: The aim of this study was to investigate the prevalence and the risk factors of testosterone deficiency of men with newly diagnosed T2DM. METHODS: The cross-sectional study included 105 men (mean age: 61.2 ± 6.8 years) with previously known T2DM and another 81 (57.8 ± 8.8 years) with newly diagnosed T2DM. All received health checkup and sex hormone measurement at our institute in 2009. MAIN OUTCOME MEASURES: We calculated the prevalence and explored the risk factors of low total (<300 ng/dL) and free (<6 ng/dL) testosterone in men with newly diagnosed and previously known T2DM. RESULTS: Men with previously known T2DM were older and had higher diastolic pressure and greater fasting glucose. There was no significant difference in total (358.0 [155.0] ng/dL vs. 363.0 [154.0] ng/dL, P=0.68) and free (7.2 [2.5] ng/dL vs. 7.4 [2.4]ng/dL, P=0.84) testosterone and sex-hormone binding globulin (SHBG) (27.3 [22.3]nmol/L vs. 28.7 [14.9]nmol/L, P=0.46). The prevalence of low total and free testosterone was 28.4% and 21.0%, respectively, in men with newly diagnosed T2DM, and was 26.7% and 19.0% in those with previously known T2DM. In men with previously known T2DM, better glycemic control (HbA1c <7%) was associated with a higher level of total testosterone and a lower risk of low total testosterone. Men with newly diagnosed and previously known T2DM shared similar risk factors of low total testosterone, including high HbA1c (≥ 7%), low SHBG (<20 nmol/L), obesity, hyperuricemia, hypertriglycemia, and metabolic syndrome. Elevated prostate-specific antigen was a protective factor of low total testosterone. However, none of these factors was associated with low free testosterone. CONCLUSIONS: The prevalence and the risk factors of testosterone deficiency are similar between newly diagnosed and previously known type 2 diabetic men.

Predictive clinical indicators of biochemical progression in advanced prostate cancer patients receiving Leuplin depot as androgen deprivation therapy.

Therapeutic planning and counseling for advanced prostate cancer patients receiving androgen deprivation therapy (ADT) is complicated because the prognoses are highly variable. The purpose of this study is to identify predictive clinical indicators of biochemical progression (BCP). In this retrospective analysis, data from 107 newly diagnosed patients (from November 1995 to April 2008) with advanced prostate adenocarcinoma receiving Leuprorelin acetate depot were analyzed. Data was collected from the computerized registry of two collaborating medical centers in Taiwan. Cox regression and Kaplan-Meier analyses were used to evaluate the relationship between potential predictive parameters and BCP. Univariate analysis revealed that predictors of BCP included (1) initial serum prostate-specific antigen (PSA) (hazard ratio [HR], 1.00; 95% confidence interval [CI] 1.00-1.00); (2) log of initial PSA (HR, 1.35; 95% CI 1.17-1.56); (3) PSA density at diagnosis (HR, 1.00; 95% CI 1.00-1.01), and (4) pathological bone fracture (HR, 2.22; 95% CI 1.20-4.11). Age (HR, 0.94; 95% CI 0.91-0.98) and hemoglobin levels (HR, 0.86; 95% CI 0.76-0.97) were also associated with greater risk of BCP. After adjusting for age, pathologic fracture, and hemoglobin level, the initial PSA and PSA density were no longer significantly associated with BCP. However, age and hemoglobin levels continued to be associated with greater risk of BCP (P ≤ 0.007). Using Kaplan-Meier analysis, patients with higher initial PSA concentration, pathological bone fracture, and low hemoglobin had a greater probability of BCP. Thus, low hemoglobin and age are predictive indicators of BCP and therefore early indicators of BCP despite ADT therapy.

Safety and six-month durability of angioplasty for isolated penile artery stenoses in patients with erectile dysfunction: a first-in-man study.

AIMS: Obstructive pelvic arterial lesions are highly prevalent in patients with erectile dysfunction and commonly located in penile artery segments. In this first-in-man study, we intended to assess the safety and feasibility of balloon angioplasty for isolated penile artery stenoses in patients with erectile dysfunction. METHODS AND RESULTS: Twenty-five patients with erectile dysfunction and isolated penile artery stenoses (unilateral stenosis ≥70% or bilateral stenoses ≥50%) identified by pelvic computed tomographic angiography were enrolled. A total of 20 patients (mean age 61 years [range, 48-79 years]) underwent balloon angioplasty. Three patients had bilateral penile artery stenoses. Procedural success was achieved in all 23 penile arteries, with an average balloon size of 1.6 mm (range, 1.00-2.25 mm). The average International Index for Erectile Function-5 (IIEF-5) score improved from 10.0±5.2 at baseline to 15.2±6.7 (p<0.001) at one month and 15.2±6.3 (p<0.001) at six months. Clinical success (change in the IIEF-5 score ≥4 or normalisation of erectile function [IIEF-5 ≥22]) was achieved in 15 (75%), 13 (65%), and 12 (60%) patients at one, three, and six months, respectively. There were no adverse events through follow-up. CONCLUSIONS: For the first time we demonstrated that penile artery angioplasty is safe and can achieve clinically significant improvement in erectile function in 60% of patients with erectile dysfunction and isolated penile artery stenoses.

Surgeon volume for percutaneous nephrolithotomy is associated with medical costs and length of hospital stay: a nationwide population-based study in Taiwan.

BACKGROUND AND PURPOSE: To investigate the factors associated with outcomes and medical costs for percutaneous nephrolithotomy (PCNL). METHODS: The present study uses a subset of the National Health Insurance Research Database (NHIRD), known as the Longitudinal Health Insurance Database 2005 (LHID 2005), which contains the data of all medical benefit claims from 1997 to 2010 for a subset of 1 million enrollees randomly drawn from the population of 22.72 million persons who were enrolled in 2005. The claims data for all subjects with a diagnosis of urolithiasis who underwent PCNL were analyzed. Hospital and surgeon case volume were classified by quartile. The correlations of all patient, surgeon, and hospital variables with the outcomes and medical costs of PCNL were analyzed by generalized estimating equations. RESULTS: A total of 995 subjects received PCNL. In univariate analysis, PCNL performed by high-volume surgeons (≥12) cost 26% less ($2684 vs $1986) and resulted in a 34.3% shorter hospital stay (6.5 vs 9.9 days) compared with low-volume surgeons (≤3). In multivariate analysis, surgeon volume was a significant predictor for medical cost, length of stay, and intensive care unit transfer but not complications and mortality. CONCLUSIONS: Surgeon volume was associated with lower medical costs and shorter length of stay after PCNL. Surgeon volume, however, was not an independent predictor of complications and mortality. Our findings have important implications for urologists and policymakers with regard to the cost and effectiveness of PCNL.

Prediabetes is associated with an increased risk of testosterone deficiency, independent of obesity and metabolic syndrome.

OBJECTIVE: The association between type 2 diabetes and low testosterone has been well recognized. However, testosterone levels in men with prediabetes have been rarely reported. We aimed to investigate whether prediabetes was associated with an increased risk of testosterone deficiency. METHODS: This study included 1,306 men whose sex hormones was measured during a medical examination. Serum total testosterone and sex hormone-binding globulin were measured; free and bioavailable testosterone concentrations were calculated by Vermeulen's formula. Prediabetes was defined by impaired fasting glucose (IFG), impaired postprandial glucose (IPG), or glycated hemoglobin (HbA1c) 5.7%-6.4%. Logistic regression was performed to obtain the odds ratios (OR) for subnormal total testosterone (<300 ng/dL) or free testosterone (<6 ng/dL) in prediabetic and diabetic men compared with normoglycemic individuals, while adjusting for age, BMI, waist circumference, and metabolic syndrome (MetS). RESULTS: Normoglycemia, prediabetes, and diabetes were diagnosed in 577 (44.2%), 543 (41.6%), and 186 (14.2%) men, respectively. Prediabetes was associated with an increased risk of subnormal total testosterone compared to normoglycemic individuals (age-adjusted OR=1.87; 95%CI=1.38-2.54). The risk remained significant in all multivariate analyses. After adjusting for MetS, the OR in prediabetic men equals that of diabetic patients (1.49 versus 1.50). IFG, IPG, and HbA1c 5.7%-6.4% were all associated with an increased risk of testosterone deficiency, with different levels of significance in multivariate analyses. However, neither prediabetes nor diabetes was associated with subnormal free testosterone in multivariate analyses. CONCLUSIONS: Prediabetes is associated with an increased risk of testosterone deficiency, independent of obesity and MetS. After adjusting for MetS, the risk equals that of diabetes. Our data suggest that testosterone should be measured routinely in men with prediabetes.

2-methoxyestradiol induces mitotic arrest, apoptosis, and synergistic cytotoxicity with arsenic trioxide in human urothelial carcinoma cells.

2-Methoxyestradiol (2-ME), an endogenous derivative of 17ß-estradiol, has been reported to elicit antiproliferative responses in various tumors. In this study, we investigated the effects of 2-ME on cell viability, proliferation, cell cycle, and apoptosis in human urothelial carcinoma (UC) cell lines. We used two high-grade human bladder UC cell lines (NTUB1 and T24). After treatment with 2-ME, the cell viability and apoptosis were measured by MTT assay and flow cytometry (fluorescence-activated cell sorting), with annexin V-FITC staining and propidium iodide (PI) labeling. DNA fragmentation was analyzed by agarose gel electrophoresis. Flow cytometry with PI labeling was used for the cell cycle analyses. The protein levels of caspase activations, poly (ADP-ribose) polymerase (PARP) cleavage, phospho-histone H2A.X, phospho-Bad, and cell cycle regulatory molecules were measured by Western blot. The effects of the drug combinations were analyzed using the computer software, CalcuSyn. We demonstrated that 2-ME effectively induces dose-dependent cytotoxicity and apoptosis in human UC cells after 24 h exposure. DNA fragmentation, PARP cleavage, and caspase-3, 7, 8, 9 activations can be observed with 2-ME-induced apoptosis. The decreased phospho-Bad (Ser136 and Ser155) and mitotic arrest of the cell cycle in the process of apoptosis after 2-ME treatment was remarkable. In response to mitotic arrest, the mitotic forms of cdc25C, phospho-cdc2, cyclin B1, and phospho-histone H3 (Ser10) were activated. In combination with arsenic trioxide (As2O3), 2-ME elicited synergistic cytotoxicity (combination index <1) in UC cells. We concluded that 2-ME significantly induces apoptosis through decreased phospho-Bad and arrests bladder UC cells at the mitotic phase. The synergistic antitumor effect with As2O3 provides a novel implication in clinical treatment of UC.

Sperm quality assessment via separation and sedimentation in a microfluidic device.

A major reason for infertility is due to male factors, including the quality of spermatozoa, which is a primary factor and often difficult to assess, particularly the total sperm concentration and its motile percentage. This work presents a simple microfluidic device to assess sperm quality by quantifying both total and motile sperm counts. The key design feature of the microfluidic device is two channels separated by a permeative phase-guide structure, where one channel is filled with raw semen and the other with pure buffer. The semen sample was allowed to reach equilibrium in both chambers, whereas non-motile sperms remained in the original channel, and roughly half of the motile sperms would swim across the phase-guide barrier into the buffer channel. Sperms in each channel agglomerated into pellets after centrifugation, with the corresponding area representing total and motile sperm concentrations. Total sperm concentration up to 10(8) sperms per ml and motile percentage in the range of 10-70% were tested, encompassing the cutoff value of 40% stated by World Health Organization standards. Results from patient samples show compact and robust pellets after centrifugation. Comparison of total sperm concentration between the microfluidic device and the Makler chamber reveal they agree within 5% and show strong correlation, with a coefficient of determination of R(2) = 0.97. Motile sperm count between the microfluidic device and the Makler chamber agrees within 5%, with a coefficient of determination of R(2) = 0.84. Comparison of results from the Makler Chamber, sperm quality analyzer, and the microfluidic device revealed that results from the microfluidic device agree well with the Makler chamber. The sperm microfluidic chip analyzes both total and motile sperm concentrations in one spin, is accurate and easy to use, and should enable sperm quality analysis with ease.

The frequency of vesicoureteral reflux diagnosis is correlated with urbanization level of residence in pediatric population: a nationwide study in Taiwan.

OBJECTIVE: To investigate the frequency and characteristics of newly diagnosed vesicoureteral reflux (VUR) in children younger than 18 years based on a nationwide database in Taiwan. METHODS: The present study utilizes a subset of the Taiwan's National Health Insurance Research Database, known as the Longitudinal Health Insurance Database 2005, which contains the data of all paid medical benefit claims over 1997-2007 for a subset of 1,000,000 beneficiaries randomly drawn from the population of 22.72 million individuals during any part of the 2005 calendar year. Our analysis includes the data of all pediatric patients with the diagnosis of VUR. RESULTS: A total of 738 subjects with VUR diagnosis were identified, including 412 (55.8%) boys and 326 (44.2%) girls. The peak age of VUR occurrence was the first year for males and 1-4 years for females. Approximately 49.7% of all subjects presented with urinary tract infection (UTI); moreover, there were significant differences between genders concerning the presence of UTI (RR = 0.8; p = 0.002). The occurrence rate of VUR in the pediatric population ranged from 2.63 in 1998 to 3.94 in 2003 per 10,000 children during 1998-2005. The frequency of newly-diagnosed VUR in the pediatric population was significantly correlated with urbanization levels of residence. CONCLUSION: The nationwide, population-based study of pediatric VUR shows there were gender differences in age distribution and presence of UTI. Further studies are warranted to clarify the correlations between urbanization level of residence and occurrence of VUR.

The incidence rate and characteristics in patients with testicular torsion: a nationwide, population-based study.

AIM: To determine the incidence rate and characteristics in patients with testicular torsion in Taiwan using a nationwide insurance database. METHODS: This study utilizes a subset of national health insurance research database, which contains the data of all paid medical benefit claims over the period 1997-2010 for in 1 000 000 beneficiaries in 2005. We analysed claims data for all male subjects younger than 25 years with the diagnosis of testicular torsion. RESULTS: A total of 86 subjects younger than 25 years with the diagnosis of testicular torsion were included. Among them, 22 (25.6%) underwent orchiectomies and 64 (74.4%) underwent orchiopexies. The estimated incidence of testicular torsion was 3.5 per 100 000 person-years. There are two peaks in the age-specific incidence rates: the first in boys aged 10-14 years (at 7.7 per 100 000 person-years) and the second in male infants aged <1 year (at 7.6 per 100 000 person-years). There was significant difference between orchiectomy and orchiopexy groups in the Insurance fee (p = 0.032). CONCLUSION: The incidence of testicular torsion in Taiwan was similar with previous report in the United States. It is important to improve the medical access to achieve better outcomes of testicular torsion.

Direct characterization of motion-dependent parameters of sperm in a microfluidic device: proof of principle.

BACKGROUND: Semen analysis is essential for evaluating male infertility. Besides sperm concentration, other properties, such as motility and morphology, are critical indicators in assessing sperm quality. Nevertheless, rapid and complete assessment of these measures still presents considerable difficulty and involves a range of complex issues. Here we present a microfluidic device capable of quantifying a range of properties of human sperm via the resistive pulse technique (RPT). METHODS: An aperture, designed as a long channel, was used to allow the quantification of various properties as sperm swam through. RESULTS: The time trace of the voltage drop across the aperture during sperm passage contained a wealth of information: the sperm volume was presented by the amplitude of the induced pulse, the swim velocity was evaluated via the duration, and the beat frequency was calculated from the voltage undulation superposed on the pulse signal. The RPT measurement of swim velocity and beat frequency showed a correlation with the same observation in a microscope (R(2) = 0.94 and 0.70, respectively). CONCLUSIONS: The proposed proof of principle enables substantial quantification of the motion-dependent properties of sperm. Because this approach requires only a current/voltage source and data analysis, it is economically advantageous compared with optical methods for characterizing sperm motion. Furthermore, this approach may be used to characterize sperm morphology.

The activation of peripheral 5-HT1A receptors can inhibit seminal vesicle contraction: an in vivo animal study.

OBJECTIVES: To elucidate the differential effects of stimulating various peripheral 5-HT receptor subtypes on the contractile response of seminal vesicles (SVs) induced by electrical stimulation (ES). METHODS: Male Wistar rats (aged 12-14 weeks) were prepared as our previously established model, which allows an intraarterial injection of test agents to directly act on SV. Four selective 5-HT agonists-8-OH-DPAT (5-HT1A), 5-nonyloxytryptamine (5-HT1B), BW723C86 (5-HT2B), and MK-212 (5-HT2C)-were injected at various concentrations (from 10(-8) to 10(-4) mmol/kg). After an injection, the SV contractile response was recorded after ES of lesser splanchnic nerve was applied. Relationships between the concentration of an agonist and its effect on SV contraction were plotted and analyzed. RESULTS: The peripheral injection of 5-HT1A agonist had a dose-dependent inhibitory effect on SV contraction and could achieve an inhibition of >50%; the IC50 was 3.16x10(-6) mmol/kg. No significant effects were observed with the peripheral injection of 5-HT1B, 5-HT2B, or 5-HT2C agonist. CONCLUSIONS: Our in vivo animal study shows that the activation of peripheral 5-HT1A receptors can inhibit ES-induced SV contraction, whereas the activation of peripheral 5-HT1B, 5-HT2B, or 5-HT2C receptors has no significant effect. The results suggest that the peripheral 5-HT pathway is a potential therapeutic target of the treatment for premature ejaculation.