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Introduction: The development of extended-spectrum-beta-lactamase (ESBLs) Escherichia coli ( E. coli ) has become a global threat to public health. An alternative strategy to alleviate this is identifying potential natural compounds to restore antibiotic activity against ESBLs E. coli . This study aimed to find a possible compound to restore ESBLs E. coli sensitivity to ceftiofur., Methods: The synergistic effect of kaempferol and ceftiofur against ESBLs E. coli was investigated by checkerboard assays, time-kill, growth curves, and scanning electronic microscope. The impact of kaempferol with ceftiofur on the biofilm of ESBLs E. coli was evaluated by crystal violet staining and laser scanning confocal microscopy and this study also assessed the effect of kaempferol on the initial adhesion and aggregation of E. coli (SY20) by examining motility, adhesion, and surface characteristics. The RT-qPCR was used to determine the effect of kaempferol on the expression of genes related to the LuxS /AI-2 quorum sensing system in ESBLs E. coli , and the effect of kaempferol on AI-2 signaling molecules was determined by molecular docking and bioassay. The impact of kaempferol on the activity of bla CTX-M-27 protein was determined by RT-qPCR, molecular docking, and nitrofen experiments, the results were further verified by transcriptome analysis. The mouse infection model was established, and the inhibitory mechanism of kaempferol with ceftiofur on bacteria in vivo was further verified by HE staining and immunohistochemistry., Results and Discussion: Kaempferol with ceftiofur exerts synergistic antibacterial and bactericidal effects on ESBLs E. coli by influencing β-lactamase activity, biofilm formation, and LuxS /AI-2 QS system. In vivo, kaempferol protected the small intestinal villi from the damage of ESBLs E. coli . Furthermore, kaempferol fully restores the activity of ceftiofur in animal infection models by relieving the TLR 4 /NF-κb pathway. In conclusion, the sensitivity of ESBLs E. coli to ceftiofur in vitro and in vivo could be enhanced by kaempferol, which showed that kaempferol may be a kind of antibiotic adjuvant., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Li, Tong, Xiao, Fan, Ma, Liu, Zhuang, Qing and Zhang.)

Direct methanol fuel cells (DMFCs) have attracted increasing attention as a very promising and important energy source. In this paper, density functional theory (DFT) is used to study the structure and O-H fracture mechanism of methanol adsorption on Pt n Cu 4- n (111) ( n = 1, 2, 3) binary metal catalyst surfaces under different coverages. By comparing the adsorption energy and dehydrogenation energy barriers of methanol, it is found that the adsorption strength and dehydrogenation energy barriers of methanol on Pt and Cu sites decreased with increasing coverage. At the same Pt and Cu ratio, methanol is more easily adsorbed on Cu sites. When Pt/Cu = 3:1 and 1:3, the PtCu binary catalyst has a significant impact on the energy barrier of breaking the O-H bond in methanol with the increase of coverage. Especially when Pt/Cu = 1:3 and the coverage is 1/4 ML, the energy barriers of O-H bond breaking in methanol on Pt and Cu sites are 0.63 and 0.61 eV, respectively, which are lower than that on pure Pt. It means that the Cu sites played a very important role in reducing the O-H fracture energy barrier of methanol. When Pt/Cu = 1:1, the change in the dehydrogenation energy barrier of methanol on Pt sites and Cu sites is not significant, indicating that the coverage has little effect on it.

The development of drug-resistance in the opportunistic pathogen Escherichia coli has become a global public health concern. Due to the share of similar flora between pets and their owners, the detection of pet-origin antibiotic-resistant E. coli is necessary. This study aimed to detect the prevalence of feline-origin ESBL E. coli in China and to explore the resistance elimination effect of garlic oil to cefquinome on ESBL E. coli . Cat fecal samples were collected from animal hospitals. The E. coli isolates were separated and purified by indicator media and polymerase chain reaction (PCR). ESBL genes were detected by PCR and Sanger sequencing. The MICs were determined. The synergistic effect of garlic oil and cefquinome against ESBL E. coli was investigated by checkerboard assays, time-kill and growth curves, drug-resistance curves, PI and NPN staining, and a scanning electronic microscope. A total of 80 E. coli strains were isolated from 101 fecal samples. The rate of ESBL E. coli was 52.5% (42/80). The prevailing ESBL genotypes in China were CTX-M-1 , CTX-M-14 , and TEM-116 . In ESBL E. coli , garlic oil increased the susceptibility to cefquinome with FICIs from 0.2 to 0.7 and enhanced the killing effect of cefquinome with membrane destruction. Resistance to cefquinome decreased with treatment of garlic oil after 15 generations. Our study indicates that ESBL E. coli has been detected in cats kept as pets. The sensitivity of ESBL E. coli to cefquinome was enhanced by garlic oil, indicating that garlic oil may be a potential antibiotic enhancer.

Background: The development of antimicrobial resistance in the opportunistic pathogen Escherichia coli has become a global public health concern. Due to daily close contact, dogs kept as pets share the same E. coli with their owners. Therefore, the detection of antimicrobial resistance in canine E. coli is important, as the results could provide guidance for the future use of antibiotics. This study aimed to detect the prevalence of antibiotic-resistance of canine origin E. coli in Shaanxi province and to explore the inhibition effect of magnolol combined with cefquinome on MDR E. coli, so as to provide evidence for the use of antibiotics., Methods: Canine fecal samples were collected from animal hospitals. The E. coli isolates were separated and purified using various indicator media and polymerase chain reaction (PCR). Drug-resistance genes [ aacC2, ant(3')-I, aph(3')-II, aac(6')-Ib-cr, aac(3')-IIe, bla KPC , bla IMP -4 , bla OXA , bla CMY , bla TEM -1 , bla SHV , bla CTX - M -1 , bla CTX - M -9 , Qnra, Qnrb, Qnrs, TetA, TetB, TetM, Ermb ] were also detected by PCR. The minimum inhibitory concentration (MIC) was determined for 10 antibiotics using the broth-microdilution method. Synergistic activity of magnolol and cefquinome against multidrug-resistant (MDR) E. coli strains was investigated using checkerboard assays, time-kill curves, and drug-resistance curves., Results: A total of 101 E. coli strains were isolated from 158 fecal samples collected from animal hospitals. MIC determinations showed that 75.25% (76/101) of the E. coli strains were MDR. A total of 22 drug-resistance genes were detected among the 101 strains. The bla TEM -1 gene exhibited the highest detection rate (89.77%). The TetA and Sul gene also exhibited high detection rate (66.34 and 53.47%, respectively). Carbapenem-resistant E. coli strains were found in Shangluo and Yan'an. Additionally, in MDR E. coli initially resistant to cefquinome, magnolol increased the susceptibility to cefquinome, with an FICI (Fractional Inhibitory Concentration Index) between 0.125 and 0.5, indicating stable synergy. Furthermore, magnolol enhanced the killing effect of cefquinome against MDR E. coli . Resistance of MDR E. coli to cefquinome decreased markedly after treatment with magnolol for 15 generations., Conclusion: Our study indicates that antibiotic-resistance E. coli has been found in domestic dogs. After treatment with magnolol extracted from the Chinese herb Houpo ( Magnolia officinalis ), the sensitivity of MDR E. coli to cefquinome was enhanced, indicating that magnolol reverses the resistance of MDR E. coli . The results of this study thus provide reference for the control of E. coli resistance., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Tong, Zhang, Li, Cao, Ding, Yang, Lin, Gao, Sun, Fan, Liu, Qing, Ma and Zhang.)

Aims: The role of hydrodistension in the diagnosis of interstitial cystitis/bladder pain syndrome (IC/BPS) is controversial. This study evaluated the effect of low-pressure hydrodistension on glomerulation formation in female patients diagnosed with the disease., Methods: Sixty female patients with the clinical diagnosis of IC/BPS and 30 female controls without the disease underwent cystoscopy and hydrodistension. Cold-cup biopsy was taken from bladder posterior wall at sites with normal cystoscopic appearance before hydrodistension in the IC/BPS group. The tissue samples were processed for histology study. Low-pressure (40 cmH 2 O) hydrodistension for 2 min was performed and the appearance of glomerulations was compared between the two groups. High-pressure (80 cmH 2 O) hydrodistension for 8 min was then performed as a therapeutic measure for the IC/BPS patients. Further changes to the degree of glomerulations were recorded., Results: Histology showed pathological changes in the normal-appearing IC/BPS bladder mucosa including urothelium denudation, inflammatory cell infiltration, stromal edema, fibrosis, and vascular congestion. Low-pressure hydrodistension induced significant glomerulation formation in the patient group (percentage of patients with Grades 0-4: 0%, 8.3%, 40%, 35%, 10%, respectively) while none in the controls. High-pressure hydrodistension further increased the glomerulation grading in the IC/BPS patients., Conclusions: Structural changes are present in prehydrodistension IC/BPS bladder wall, which may not be macroscopically detectable. Hydrodistension at low pressure is adequate to disrupt the integrity of such diseased mucosa and offers a more discriminative test in the diagnosis of IC/BPS., (© 2021 Wiley Periodicals LLC.)

The increasing numbers of infections caused by multidrug-resistant (MDR) pathogens highlight the urgent need for new alternatives to conventional antibiotics. Antimicrobial peptides have the potential to be promising alternatives to antibiotics because of their effective bactericidal activity and highly selective toxicity. The present study was conducted to investigate the antibacterial, antibiofilm, and anti-adhesion activities of different CTP peptides (CTP: the original hybrid peptide cathelicidin 2 (1-13)-thymopentin (TP5); CTP-NH 2 : C-terminal amidated derivative of cathelicidin 2 (1-13)-TP5; CTPQ: glutamine added at the C-terminus of cathelicidin 2 (1-13)-TP5) by determining the minimal inhibitory concentrations (MICs), minimal bactericidal concentrations (MBCs), propidium iodide uptake, and analysis by scanning electron microscopy, transmission electron microscopy, and confocal laser scanning microscopy). The results showed that CTPs had broad-spectrum antibacterial activity against different gram-positive and gram-negative bacteria, with MICs against the tested strains varying from 2 to 64 μg/mL. CTPs at the MBC (2 × MIC 64 μg/mL) showed strong bactericidal effects on a standard methicillin-resistant Staphylococcus aureus strain ATCC 43300 after co-incubation for 6 h through disruption of the bacterial membrane. In addition, CTPs at 2 × MIC also displayed effective inhibition activity of several S. aureus strains with a 40-90% decrease in biofilm formation by killing the bacteria embedded in the biofilms. CTPs had low cytotoxicity on the intestinal porcine epithelial cell line (IPEC-J2) and could significantly decrease the rate of adhesion of S. aureus ATCC 43300 on IPEC-J2 cells. The current study proved that CTPs have effective antibacterial, antibiofilm, and anti-adhesion activities. Overall, this study contributes to our understanding of the possible antibacterial and antibiofilm mechanisms of CTPs, which might be an effective anti-MDR drug candidate.

Yeast culture plus enzymatically hydrolyzed yeast cell wall (YC-EHY) contains crude protein, mannan-oligosaccharide, β-glucan and yeast culture. This study was carried out to explore the effects of dietary YC-EHY at different levels on growth performance, antioxidant capacity, and immune function of broiler chickens. A total of 320 one-day-age male broiler chicks were allocated into 4 groups and were fed with a basal diet supplemented with 0 mg/kg (the control group), 50 mg/kg, 100 mg/kg, 150 mg/kg YC-EHY for 42 d. Dietary YC-EHY improved average daily gain and feed efficiency during the starter, grower, and overall periods as well as average body weight of broiler chickens on 42 d (linear and quadratic, P < 0.05). Broiler chickens fed with YC-EHY quadratically increased jejunal sucrase activity on 21 d (quadratic, P < 0.05), and linearly and quadratically enhanced maltase activity on 21 and 42 d (linear and quadratic, P < 0.05). Supplementing YC-EHY linearly and quadratically enhanced jejunal superoxide dismutase (SOD) activity on 21 and 42 d and glutathione peroxidase (GPX) activity on 42 d whereas decreased malonaldehyde (MDA) concentration on 42 d (linear and quadratic, P < 0.05). Consistently, the jejunal genes expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and SOD1 on 21 and 42 d, heme oxygenase-1 (HO-1) and GPX1 on 42 d were enhanced by YC-EHY supplementation (linear and quadratic, P < 0.05). The concentrations of jejunal immunoglobulin G (IgG) on 21 and 42 d and secreted immunoglobulin A (SIgA) on 42 d were linearly and quadratically elevated by supplementing YC-EHY (linear and quadratic, P < 0.05). Dietary YC-EHY quadratically increased jejunal IgG and IgM genes expression on 21 d (quadratic, P < 0.05), and linearly and quadratically enhanced the genes expression of IgG and IgM on 42 d (linear and quadratic, P < 0.05). Overall, this study indicated that supplementing YC-EHY could exert beneficial effects on growth performance, intestinal antioxidant capacity and immune function in broiler chickens., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Objectives: To investigate the pathophysiological mechanism leading to lower urinary tract symptoms in prostate cancer (PCa) by using an animal model., Methods: An orthotopic PCa model in mice was established by injection of human DU145 cells into the prostate gland lateral lobe of NOD.CB17-Prkdc scid /NcrCrlBltw (NOD-SCID) mice. Cancer growth was quantified by a luciferase-based in vivo imaging system (IVIS) serially every 7 days. Comparisons were made for urodynamic parameters, bladder histology, and biological markers until the sixth week. Bladder wall structural changes were assessed by the bladder wall thickness and degree of fibrosis. Biomarker expressions in bladder tissue including muscarinic acetylcholine receptor 2 (M 2 ), transient receptor potential cation channel subfamily V member 4 (TRPV4), BCL2-associated X protein (Bax), and caspase3 were evaluated by immunohistochemical staining and immunofluorescence confocal laser scanning microscopy., Results: DU145 cell growth in the prostate was successfully monitored by a luciferase-based IVIS. after orthotopic injection. Using our injection technique, no anatomical obstruction of the bladder outlet and urethra was noted up to 6 weeks after injection. The presence of PCa induced changes in urinary bladder histology, biomarkers, and urodynamic parameters. Cystometry showed features of detrusor overactivity with increased voiding frequency and high-amplitude voiding contractions from the fourth week onward. Histological analyses 4 weeks after DU145 injection demonstrated detrusor thickening and bladder wall fibrosis. Immunohistochemistry showed increased expressions of bladder M 2 , TRPV4, Bax, and caspase3 in the PCa mice as early as in the first or second week., Conclusions: PCa can induce bladder microenvironment changes involving neural receptors and biological mediators leading to histological and functional alterations even in the absence of overt anatomical obstruction., (© 2021 John Wiley & Sons Australia, Ltd.)

A large number of experiments show that PtCu catalyst has a good catalytic effect on methanol decomposition. Therefore, density functional theory (DFT) was used to further study the dehydrogenation of methanol catalyzed by Pt n Cu m (n = 1-3, m = 0-2). The energy diagrams of O-adsorption path and H-adsorption path were drawn. By calculation, the Pt is the active site of the whole reaction process, and the barrier energy of the rate-determining step is 11.09 kcal mol -1 by Pt 2 Cu, which is lower than that of Pt 3 and PtCu 2 . However, the complete dehydrogenation product of methanol, CO, is easier to dissociate from PtCu 2 clusters than from Pt 3 and Pt 2 Cu clusters. Therefore, Cu doping can improve the catalytic activity and anti-CO toxicity of Pt to a certain extent.

Objective: To investigate whether the risk of interstitial cystitis increases among the patients with systemic lupus erythematosus., Methods: This was a nationwide population-based cohort study. Data were obtained from the National Health Insurance Research Database in Taiwan. Women aged >18 years newly diagnosed as systemic lupus erythematosus during 2001-2008 were identified as the control group. The comparison included individuals randomly selected from the National Health Insurance Research Database in the year of 2000, by matching one systemic lupus erythematosus participant with eight non-systemic lupus erythematosus participants with sex and age. These participants were followed up until being diagnosed as interstitial cystitis, or the end of 2011. Women diagnosed with lupus cystitis were excluded from this study., Results: This study included 7240 women with systemic lupus erythematosus and 57 920 women without systemic lupus erythematosus as controls. The incidence rate of interstitial cystitis was significantly higher in the systemic lupus erythematosus group, with an incidence rate ratio of 2.26 (95% confidence interval 1.57-3.27, P < 0.0001). After adjustment, the risk increased by 2.45-fold (adjusted hazard ratio 2.45, 95% confidence interval 1.57-3.27, P < 0.05). Age as a factor increases incidence rate ratios among all age groups, 2.12-, 3.32- and 4.65-fold. Age ≥45 years had an increased adjusted hazard ratio (2.07, 95% confidence interval 1.37-3.13, P < 0.05). Comorbidities, for example, hypertension, diabetes mellitus, dyslipidemia and renal disease, were insignificant., Conclusions: This is the first population-based cohort study showing a higher incidence of interstitial cystitis among patients with systemic lupus erythematosus. These findings support the concordance of interstitial cystitis with autoimmune diseases, and the temporal relationship to develop interstitial cystitis in patients with systemic lupus erythematosus., (© 2019 The Japanese Urological Association.)

Objectives: To determine the optimal surgical timing in high-risk patients with Fournier's gangrene by the Simplified Fournier's Gangrene Severity Index., Methods: From 1989 to 2018, 118 male patients diagnosed with Fournier's gangrene with complete medical records were retrospectively reviewed. Patients' demographics, laboratory parameters at initial diagnosis, Fournier's Gangrene Severity Index and Simplified Fournier's Gangrene Severity Index, and the time interval from emergency room arrival to surgical intervention were collected. The Fournier's gangrene patients were categorized into low-risk (Simplified Fournier's Gangrene Severity Index ≤2) and high-risk groups (Simplified Fournier's Gangrene Severity Index >2). Differences between the variables within the two groups were analyzed. The optimal surgical timing was analyzed with the receiver operating characteristic curve in high-risk Fournier's gangrene patients., Results: The overall mortality of 118 Fournier's gangrene patients was 14.4%. After risk stratification with the Simplified Fournier's Gangrene Severity Index scoring system, the mortality of low-risk and high-risk Fournier's gangrene patients was 1.3% and 41.0%, respectively. In the high-risk group, the time interval from emergency room arrival to surgical intervention was the only variable with a significant difference between survivors and non-survivors (P = 0.039). The optimal surgical timing was determined at 14.35 h, which allowed the highest sensitivity (0.688) and specificity (0.762) to affect mortality. The mortality was significantly lower in high-risk Fournier's gangrene patients with early surgical intervention compared with late intervention (23.8% vs 68.8%, P = 0.007)., Conclusions: The Simplified Fournier's Gangrene Severity Index is a quick and reliable screening tool for first-line physicians to identify high-risk patients with Fournier's gangrene (Simplified Fournier's Gangrene Severity Index >2) who have poor survival outcomes. We recommended early surgical intervention within 14.35 h to maximize the survival of high-risk Fournier's gangrene patients., (© 2019 The Japanese Urological Association.)

Objectives: To investigate the effect of high-fat diet (HFD) on bladder M 1,3 muscarinic receptor expression and contractile function in the rat., Methods: Eight-week-old male rats were divided into two groups including one with HFD for 8 weeks (short-term) and the other for 24 weeks (long-term). Each group was compared to age-matched rats fed with normal chow as controls. The body weight, food intake amount and blood biochemistry were monitored. Bladder muscle contractile responses to acetylcholine (0.1-10 μM), bethanechol (10 μM) and KCl (50 mM) were studied in an organ bath set-up. Bladder M 1 and M 3 muscarinic receptor protein expressions were measured by Western blotting analysis., Results: Increase in body weight as well as blood triglyceride, cholesterol and sugar levels compared to controls were noted in both 8- and 24-week HFD rats. Eating appetite change with increased food and water intakes was noted in the HFD rats. Significantly decreased bladder contractile responses to acetylcholine and bethanechol were shown in both HFD groups. On the other hand, decreased bladder contractile response to KCl was demonstrated in the 24-week group but not the 8-week group. The expressions of bladder M 1 and M 3 muscarinic receptor proteins were significantly and progressively decreased by HFD feeding from 8 to 24 weeks., Conclusions: High-fat diet induces obesity and polyphagia in rats. Short-term and long-term HFD feeding decrease rat bladder M 1 and M 3 receptor expressions as well as contractile responses to the agonistic stimulation. In addition, bladder muscle dysfunction develops after long-term HFD feeding., (© 2017 John Wiley & Sons Australia, Ltd.)

Objectives: To investigate the effect of metabolic syndrome on the bladder cannabinoid receptors 1 and 2 (CB1/CB2) expression and function in the fructose-fed rats (FR)., Methods: Adult male Sprague-Dawley rats were divided into two groups: (i) Control rats fed with normal chow; and (ii) Rats fed with high-fructose diet (FR) for 9 weeks. The body weight, blood pressure, plasma sugar, insulin, triglyceride and cholesterol were measured. Bladder muscle strips were prepared in organ bath and pre-contracted with 1 µM/L acetylcholine (ACh) or 50 mM/L KCl. The relaxation responses to CB1/CB2 agonist Bay59-3047 (0.01-1 µM/L) were recorded. The effects of CB1 antagonist AM251, CB2 antagonist AM630, protein kinase A (PKA) inhibitor H-89 and ATP-sensitive potassium channel (K ATP ) inhibitor glibenclamide on the Bay59-3047-induced response were tested. Western blotting and real-time polymerase chain reaction (RT-PCR) analyses were performed for bladder CB1/CB2 receptors., Results: Significant increases of body weight, blood pressure, plasma glucose, insulin, cholesterol and triglyceride levels were found in the FR. Bay59-3047 reduced ACh and KCl pre-contracted bladder strip tension in a dose-dependent fashion. The relaxation responses were significantly decreased in the FR. The Bay59-3047-induced relaxation was attenuated by AM251, glibenclamide and H-89. Western blotting and RT-PCR showed decreased expressions of FR bladder CB1 and CB2 receptor protein and mRNA., Conclusion: CB1/CB2 receptors mediate rat bladder relaxation through the PKA and K ATP pathway. The CB1 receptor may play a more prominent role. The response is decreased in the FR bladder due to reduced expressions of the cannabinoid receptors., (© 2017 John Wiley & Sons Australia, Ltd.)

The therapeutic action of ginsenoside Rh2 on several cancer models has been reported. This study aimed to evaluate its apoptotic effect on prostate cancer and the underlying mechanism. Cultured DU145 cells were treated with Rh2 (5 × 10 -5 to 1 × 10 -4 M), peroxisome proliferator-activated receptor-delta (PPAR-delta) antagonist GSK0660 (1 × 10 -6 to 5 × 10 -6 M); or small interfering RNA (siRNA) of PPAR-delta. The treatment effects were evaluated with cell viability assay, life/death staining and flow cytometry for apoptosis. Immunostaining was used for reactive oxygen species (ROS) and superoxide detection. Western blot analysis for PPAR-delta and signal transducer and activator of transcription 3 (STAT3) protein expression were performed. The results showed that Rh2 significantly decreased DU145 cell survival and increased cell apoptosis. ROS and superoxide induction, PPAR-delta up-regulation and phosphorylated STAT3 (p-STAT3) down-regulation by Rh2 were demonstrated. GSK0660 partially but significantly inhibited the Rh2-induced apoptosis and restored cell viability. Treatment with siRNA reversed the Rh2-induced apoptosis as well as changes in PPAR-delta and p-STAT3 expression. In conclusion, our findings have demonstrated that ginsenoside Rh2 induces prostate cancer DU145 cells apoptosis through up-regulation of PPAR-delta expression which is associated with p-STAT3 up-regulation and ROS/superoxide induction. Rh2 may be potentially useful in the treatment of prostate cancer., Competing Interests: CONFLICTS OF INTEREST The authors declare no conflicts of interest.

To investigate the chemical constituents from Barringtonia racemosa, twelve compounds were isolated by chromatography methods and identified as 3β-p-E-coumaroymaslinic acid (1), cis-careaborin (2), careaborin (3), maslinic acid (4), 2α, 3β, 19α-trihydroxyolean-12-ene-24, 28-dioic acid (5), 3β-p-Z-coumaroylcorosolic acid (6), corosolic acid (7), 1α, 2α, 3β, 19α-tetrahydroxyurs-12-en-28-oic acid (8), 19α-hydroxyl ursolic acid (9), 3α, 19α-dihydroxyurs-12-en-24, 28-dioic acid (10), tormentic acid (11), 3-hydroxy-7, 22-dien-ergosterol（12） by the NMR and MS data analysis. Among them, compounds 1-4,7-12 were obtained from the genus Barringtonia for the first time. All the compounds didn't show nocytotoxic activity against MCF-7 and A549 cell lines (IC₅₀>50 mg•L⁻¹)., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Chinese Pharmaceutical Association.)

The effect of telmisartan on prostate cancer DU145 cell survival and the underlying mechanism of apoptosis involving peroxisome proliferator-activated receptor (PPAR) pathway were investigated. Cultured DU145 cells were treated pharmacologically with telmisartan and GSK0660 (a PPAR-delta antagonist); or by RNA interference with siRNA of PPAR-delta. The treatment effects on cell survival were evaluated with cell viability assay, life and dead cell staining and flow cytometry. Western blot analysis for PPAR-delta protein expression was also performed. The results showed that telmisartan (0-80 µm) dose-dependently reduced DU145 cell survival. Flow cytometry demonstrated cancer cell cycle arrest with increase of sub-G1 phase. GSK0660 partially but significantly restored the telmisartan-treated cell viability. Similarly, siRNA of PPAR-delta significantly reversed the telmisartan-induced apoptosis. Western blot showed that telmisartan significantly increased DU145 cell PPAR-delta protein expression. Co-incubation with siRNA of PPAR-delta inhibited the telmisartan effect of PPAR-delta up-regulation. In conclusion, telmisartan induces prostate cancer DU145 cells apoptosis through the up-regulation of PPAR-delta protein expression. Pharmacological inhibition or genetic silencing of PPAR-delta activity can both reverse the telmisartan-induced apoptotic effect. Thus the PPAR-delta pathway might be a potential target for the treatment of prostate cancer., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Purpose: To investigate the association of prostate blood flow (PBF) with lower urinary tract symptoms (LUTS) in aged males using Doppler spectral waveform (DSW) analysis., Patients and Methods: We performed a prospective analysis involving 133 aged males with clinical diagnosis of LUTS. DSW parameters (peak-systolic velocity (PSV), end-diastolic velocity (EDV), and resistive index (RI)) were measured at bilateral neurovascular bundles (NVB), periurethral, and capsular branches by Doppler transrectal ultrasound with the patient in the right lateral decubitus position. The associations of PBF parameters and the International Prostate Symptom Score (IPSS) were analyzed., Results: Overall, total IPSS scores were significantly correlated with the RI of bilateral NVB vessels (r2 = 0.03, 0.04; p = 0.04, 0.02, respectively), and PSV of left NVB vessels. PSV of bilateral NVB vessels were associated with the storage score (p = 0.022 and p = 0.016), but not with the voiding score. The sum of the frequency and urgency score was also associated with EDV of both capsular and urethral branches (p = 0.043 and p = 0.009, respectively), and PSV of NVB vessels on both sides (p = 0.045 and p = 0.019, respectively)., Conclusions: There is an association between PBF and LUTS, especially with storage symptoms. The findings may provide some insights in understanding the underlying pathophysiology of lower urinary tract dysfunction., (© 2016 S. Karger AG, Basel.)

Aims: Using the National Health Insurance (NHI) database in Taiwan, the study aimed to evaluate the rates and associated factors for reoperation of female stress incontinence., Methods: Records of female patients who had received a primary surgical treatment for stress incontinence from January 2000 to December 2006 were retrieved. Among these, patients who had reoperations during follow-up till December 2010 were identified. The data were analyzed for reoperation rates, surgery methods, patient demography, surgeon, and hospital attributes., Results: Among 14,613 patients with a mean follow-up of 86.28 ± 26.76 months, 563 (3.85%) had reoperations, an incidence rate of 54.37 per 10,000 person year (PY). Injection procedures had the highest reoperation rate of 893.30/10,000 PY. The adjusted hazard ratio (HR) of reoperation was higher for mid-urethral sling when compared to pubovaginal sling (HR 1.54, 95% CI 1.16-2.05) or retropubic urethropexy including Burch operation (HR 1.30, 95% CI 1.04-1.61). Surgeons with high service volumes tended to have fewer reoperations. No correlations were noted between the reoperation rate with patient age, surgeon age/gender, year of operation and hospital status. However, urologists had higher reoperation rates than gynecologists. For repeat surgery, the majority of patients chose the same specialty but different surgical types. Mid-urethral sling was used most commonly in 48.85% of reoperations., Conclusions: Substantial number of patients need reoperation for stress incontinence. The choice of primary surgery type and surgeon specialty may affect the reoperation rates. Mid-urethral sling is the most common reoperation choice., (© 2014 Wiley Periodicals, Inc.)

Introduction and Hypothesis: The purpose of our study was to describe the surgical trends for female stress urinary incontinence (SUI) during 2006-2010, and a time-frame comparison with 1997-2005, based upon the National Health Insurance (NHI) claims data in Taiwan., Methods: Women who underwent various primary surgeries for SUI during 2006-2010 were identified, with a total of 15,099 inpatients. The variables included surgical types, patient age, surgeon age and gender, specialty, and hospital accreditation levels. Chi-squared tests and SAS version 9.3.1 were used for statistical analysis., Results: During the follow-up study, midurethral sling (MUS) application increased significantly from 53.09 % in 2006 to 78.74 % in 2010. It was associated concomitantly with a decrease in retropubic urethropexy (RPU) from 29.68 % to 12.99 %, and pubovaginal sling treatment (PVS) from 9.33 % to 3.46 %. MUS was most commonly used among all patients' and surgeons' age groups, and different accreditation hospital levels. MUS was more commonly used by gynecologists (71.38 %) than urologists (57.91 %); while PVS and periurethral injection were more commonly performed by urologists than gynecologists. Similar surgical trends were found during time-frame comparison, 2006-2010 vs 1997-2005. SUI surgeries increased in patients aged ≥60, surgeons aged ≥ 50, and in regional hospitals., Conclusion: This follow-up study depicts the increase in popularity of MUS and offers evidence of surgical trends and a paradigm shift for female SUI surgery. More older women were willing to seek healthcare and undergo surgery. The surgical skills and knowledge spread from medical centers into regional hospitals. The time-frame shift may have a profound impact on patients, as well as the healthcare providers.

Objectives: To study the role of tumor necrosis factor-α in bladder dysfunction associated with obesity., Methods: Male 8-week-old C57BL/6J mice were divided into three groups: (i) control mice; (ii) vehicle-treated high-fat diet-fed mice; and (iii) etanercept-treated high-fat diet-fed mice. High-fat diet feeding lasted for 12 weeks, vehicle or etanercept (0.8 mg/kg/day, a tumor necrosis factor-α antagonist) treatment was given during the last 4 weeks. At the end of the treatment period, serum tumor necrosis factor-α, total cholesterol, triglyceride and blood glucose were measured. Bladder strip contractile responses to 1 μmol/L acetylcholine or 50 mmol/L KCl were studied in an organ bath. Bladder protein kinase Cζ, nuclear factor-κB and intercellular adhesion molecule-1 expressions were analyzed using western blots., Results: Serum levels of tumor necrosis factor-α total cholesterol, triglyceride and glucose were significantly elevated in high-fat diet-fed mice; and the levels were not ameliorated by etanercept treatment. High-fat diet-fed mouse bladder showed reduced contractile responses to acetylcholine and KCl stimulation accompanied by high expression levels of phospho-protein kinase Cζ, nuclear nuclear factor-κB and intercellular adhesion molecule-1. Etanercept restored normal bladder contractile responses, as well as protein kinase Cζ nuclear factor-κB and intercellular adhesion molecule-1 expressions., Conclusions: A high-fat diet induces bodyweight gain, hyperlipidemia and hyperglycemia in mice. Elevated serum tumor necrosis factor-α level associated with increased protein kinase Cζ phosphorylation, nuclear factor-κB nuclear migration, intercellular adhesion molecule-1 expression and impaired muscle contractility are shown in the high-fat diet-fed mouse bladder. Tumor necrosis factor-α antagonist treatment restores normal bladder contractility, and protein kinase Cζ nuclear factor-κB and intercellular adhesion molecule-1 levels., (© 2014 The Japanese Urological Association.)

Objectives: To validate the predictive value of Fournier's Gangrene Severity Index in patients with Fournier gangrene and to facilitate patient mortality risk-stratification by simplifying the Fournier's Gangrene Severity Index., Methods: From January 1989 to December 2011, 85 male patients with clinically-documented Fournier's gangrene undergoing intensive treatment and with complete medical records were recruited. The demographic information and nine parameters of Fournier's Gangrene Severity Index were compared between survivors and non-survivors. The parameters that showed a significant difference between the two groups were selected to generate a simplified scoring index., Results: Of the 85 patients recruited, 16 patients died of the disease with mortality rate of 18.8%. The Fournier's Gangrene Severity Index score at initial diagnosis was significantly higher in non-survivors than in survivors. Of the nine parameters of Fournier's Gangrene Severity Index, the scores of serum creatinine level, hematocrit level and serum potassium level were significantly different between the two groups. However, the mean body temperatures, heart rate, respiration rate, white blood cell count, serum sodium and bicarbonate levels were non-significantly different. Of the 12 patients with chronic kidney disease or end-stage renal disease, 10 died of severe sepsis. A simplified scoring index including parameters of creatinine, hematocrit and potassium was generated, which provided sensitivity and specificity of 87% and 77% in predicting patient mortality, respectively. The predictive values of this simplified Fournier's Gangrene Severity Index were shown to be non-inferior to Fournier's Gangrene Severity Index in our patients., Conclusions: The simplified Fournier's Gangrene Severity Index is easy to use at initial diagnosis, and offers a way to compare outcomes in different clinical populations., (© 2014 The Japanese Urological Association.)

Objectives: The effect of agmatine on bladder contractility and the diabetes-induced alteration of this action were studied in the rat., Methods: Bladder strips were isolated from 9-week-old streptozotocin (STZ)-diabetic rats and control Wistar rats. Strips were hung in an organ bath for measurement of isometric tension and pre-contracted with either 1 µmol/L acetylcholine (ACh) or 50 mmol/L KCl. Dose-dependent relaxation of the bladder strips was studied by cumulative administration of agmatine 1-100 µmol/L into the organ bath. Effects of specific imidazoline receptor (IR) antagonists on the agmatine-induced relaxation were studied. Western blotting analysis was used to measure bladder IR, sulphonylurea receptor (SUR) and inwardly rectifying K(+) channel subunit 6.2 (Kir 6.2) protein levels., Results: Agmatine reduced ACh and KCl pre-contracted bladder strip tension in a dose-dependent fashion. Relaxation was significantly increased in STZ-diabetic rats. The relaxation was inhibited by BU224, a selective I2 IR antagonist; but not by efaroxan (I1 IR antagonist) or KU14R (I3 IR antagonist). Moreover, the agmatine-induced relaxation was attenuated by glibenclamide (inhibitor of KATP channel) and H-89 (inhibitor of protein kinase A), but enhanced by 3-isobutyl-1-methylxanthine (IBMX, inhibitor of cyclic AMP phosphodiesterase). Western blotting showed increased expression of bladder IR but not SUR or Kir 6.2 in the STZ-diabetic rat., Conclusion: Agmatine causes rat bladder relaxation by activation of the I2 IR, which opens KATP channels through the cyclic AMP/protein kinase A pathway. Agmatine-induced bladder relaxation in STZ-diabetic rats is increased due to a higher expression of IR., (© 2013 Wiley Publishing Asia Pty Ltd.)

Aims: The use of surgical mesh for female pelvic floor reconstruction has increased in recent years. However, there is paucity of information about the biological responses of host stroma cells to different meshes. This study was aimed to establish an in vitro experimental model to study the micro-environment of extracellular matrix (ECM) with embedded mesh and the stroma cell behaviors to different synthetic meshes., Methods: Matrigel multi-cellular co-culture system with embedded mesh was used to evaluate the interaction of stroma cells and synthetic mesh in a simulated ECM environment. Human umbilical vein endothelial cells (HUVEC) and NIH3T3 fibroblasts were inoculated in the system. The established multi-cellular Matrigel co-culture system was used to detect stroma cell recruitment and tube formation ability for different synthetic meshes., Results: HUVEC and NIH3T3 cells were recruited into the mesh interstices and organized into tube-like structures in type I mesh material from Perigee, Marlex and Prolift 24 hr after cell inoculation. On the contrary, there was little recruitment of HUVEC and NIH3T3 cells into the type III mesh of intra-vaginal sling (IVS)., Conclusions: The Matrigel multi-cellular co-culture system with embedded mesh offers a useful in vitro model to study the biological behaviors of stroma cells in response to different types of synthetic meshes. The system can help to select ideal mesh candidates before actual implantation into the human body., (© 2013 Wiley Periodicals, Inc.)

In the multimodal approach to the management of postoperative pain, local infiltration and regional blocks have been increasingly utilized for pain control. One of the limitations of local anesthetics in the postoperative setting is its relatively short duration of action. Multivesicular liposomes containing bupivacaine have been increasingly utilized for their increased duration of action. Compared with bupivacaine HCl, local infiltration of liposomal bupivacaine has shown to have an increase in duration of action and causes delay in peak plasma concentration. In this article, we attempt to review the clinical literature surrounding liposomal bupivacaine and its evolving role in perioperative analgesia. This new bupivacaine formation may have promising implications in postoperative pain control, resulting in increased patient satisfaction and a decrease in both hospital stay and opioid-induced adverse events (AEs). Although more studies are needed, the preliminary clinical trials suggest that liposomal bupivacaine has predictable pharmacokinetics, a similar side effect profile compared with bupivacaine HCl, and is effective in providing increased postoperative pain control., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Objective: To investigate the change of loperamide-induced prostate relaxation in rats fed with high-fat diet (HFD)., Materials and Methods: Adult male Wistar rats were divided into 2 groups: (1) control rats fed with normal chow and (2) rats fed with HFD for 6 months. The prostate was removed for histology study. Isolated prostate strips were hung in organ bath and precontracted with 1 μmol/L phenylephrine or 50 mmol/L KCl. The relaxation responses to loperamide 0.1 to 10 μmol/L were recorded. Western blotting analyses were performed for prostate μ-opioid receptors (MOR) and ATP-sensitive potassium (K(ATP)) channel proteins: sulfonylurea receptor (SUR) and inwardly rectifying potassium channel (Kir) 6.2 subunits., Results: Body weight, prostate weight, plasma levels of glucose, insulin, triglyceride, and cholesterol, as well as systolic blood pressure, were significantly increased in the HFD rats. Histology showed prostatic hyperplasia in the HFD rat prostate. Prostatic relaxation induced by loperamide was markedly reduced in HFD when compared to the control. Protein expressions of MOR, SUR, and Kir 6.2 were decreased in HFD-fed rats., Conclusion: Loperamide-induced prostate relaxation is decreased in HFD rats due to reduced MOR and K(ATP) channel expressions.

Objectives: To study the effects of metabolic syndrome on prostate α-adrenergic contractile function using fructose-fed rats (FR)., Methods: Age-matched male Wistar rats were divided into two groups: group I, normal control rats; and group II, 9-week FR. Animal body weight, blood pressure and serum metabolic parameters were monitored. The prostate was removed 9 weeks after induction of metabolic syndrome in the FR. The contractile responses of prostatic strips to phenylephrine (10(-7) to 10(-6) M) and KCl (50 mM) were tested. Prostate α1 -adrenoceptor (α1 -AR) protein expression was studied by Western blotting analysis with a polyclonal antiserum., Results: At week 9, the FR showed significant increases in body weight, blood pressure, plasma glucose, insulin and triglyceride levels. The FR prostate weight was significantly higher than that of the controls (610.5 ± 13.2 vs 422.3 ± 7.7 mg, P < 0.05 for n = 8). FR prostate contractile responses to phenylephrine and KCl were both significantly increased. Interestingly, prostate α1 -AR protein expression level was lower in the FR. However, after in vitro 10(-6) M phenylephrine stimulation, FR prostate α1 -AR protein expression was significantly increased., Conclusion: Metabolic syndrome in FR significantly increases prostate contractile responses to KCl and α-adrenergic stimulation. Paradoxically, FR prostate α1 -AR protein expression is decreased, but significantly enhanced after in vitro phenylephrine stimulation., (© 2012 Wiley Publishing Asia Pty Ltd.)

Purpose: Extracorporeal shock wave lithotripsy has been popular since the 1980s. Only 30% to 50% of the shock waves of all conventional lithotripters are focused on stones. We developed an ultrasound based, real-time stone tracking system (version 1) to improve accuracy and treatment efficiency. However, some problems remained. We revised the existing system (version 2) and tested its reliability and performance., Materials and Methods: We revised the system by adding more algorithms to decrease renal stone misidentification. We verified the advanced system by 2 tests using no tracking and tracking with 13 stone trajectories for versions 1 and 2. We performed the coincidence test to evaluate the accuracy of targeting the stone within the effective focal area and the stone fragmentation efficiency test to clarify the decrease in the number of shocks needed for stone fragmentation., Results: In the coincidence test the mean ± SD results of the nontracking system, and tracking versions 1 and 2 were 68.8% ± 18.8%, 89.9% ± 5.2% and 94.3% ± 4.5%, respectively. Version 2 was statistically significantly better than version 1 (p = 1.5 × 10(-4)). In the stone fragmentation efficiency test the mean results of the nontracking system, and versions 1 and 2 were 49.5% ± 14.2%, 85.1% ± 4.5% and 89.5% ± 4.2%, respectively. Version 2 was statistically significantly better than version 1 (p = 1.9 × 10(-8))., Conclusions: The revised tracking system is better than version 1. It improves treatment efficiency, decreases stone misidentification and can shorten treatment time., (Copyright © 2013 American Urological Association Education and Research, Inc. Published by Elsevier Inc. All rights reserved.)

Objectives: The effect of agmatine on prostate contractility as well as the roles of imidazoline receptors and potassium channels in this action were studied using isolated Wistar rat prostate tissue., Methods: Rat prostate strips were pre-contracted with 1 µmol/L phenylephrine or 50 mmol/L KCl. The relaxation response to agmatine (1-100 µmol/L) was measured. The effects of imidazoline receptor blockers: efaroxan, BU224, KU14R; ATP-sensitive K(+) channels (KATP ) channel inhibitor: glibenclamide; cyclic AMP (cAMP) phosphodiesterase inhibitor: IBMX; or protein kinase A (PKA) inhibitor: H-89 on the agmatine-induced relaxation were studied., Results: Agmatine produced relaxation in prostate strips pre-contracted with phenylephrine or KCl in a dose-dependent manner. This relaxation was significantly reduced by BU224, a selective I2 imidazoline receptor (IR) blocker, but not by I1 or I3 IR blockers (efaroxan, KU14R respectively). Moreover, the agmatine-induced relaxation was attenuated by glibenclamide and H-89, but enhanced by IBMX., Conclusion: The results suggest that agmatine causes rat prostate relaxation by activation of the I2 IR, which opens KATP channels through cAMP/PKA pathway., (© 2012 Blackwell Publishing Asia Pty Ltd.)

Objective: To investigate the healthcare-seeking prevalence of those with lower urinary tract symptoms (LUTS) among National Health Insurance enrollees in Taiwan., Materials and Methods: Data from the National Health Insurance Research Database were obtained from a random population sample of about one million as a representative cohort from 2000 to 2009. Subjects aged≥18 years with ≥1 outpatient service claim and coding for a LUTS-related diagnosis were identified and analyzed annually., Results: The healthcare-seeking prevalence for LUTS increased from 23.11‰ in 2000 to 38.37‰ in 2009. Similar trends were found for storage symptoms and voiding symptoms (P<.0001). In 2009, the age distribution of LUTS was 4.32%, 5.82%, 10.92%, 19.23%, 21.25%, and 38.47% in those aged 18-29, 30-39, 40-49, 50-59, 60-69, and ≥70 years, respectively. The overall LUTS prevalence was more common in men (54.45‰) than in women (21.73‰). For LUTS subtypes, storage symptoms were more common in women (19.46‰) than in men (13.13‰); however, the trend was reversed in the population>60 years old. Voiding and mixed symptoms were more common in men (4.69‰, and 0.83‰, respectively) than in women (2.80‰ and 0.58‰, respectively)., Conclusion: In the present cohort study, patients with LUTS seeking medical help increased annually from 2000 to 2009 in Taiwan. The occurrence of LUTS subtypes varied according to age and sex. LUTS is becoming a substantial healthcare burden as the number of aged people who want to maintain a good quality of life increases., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Unlabelled: What's known on the subject? and What does the study add? Neurotransmitters are known to control prostate contractility. Agmatine is one of them and induces relaxation through imidazoline receptors. The paper shows that the action of agmatine is reduced in hypertensive rats, and that this change is related to the decrease of ATP-sensitive potassium channels in the prostate. The findings can increase our understanding of the possible underlying mechanism for the development of clinical benign prostatic hyperplasia., Objectives: To compare agmatine-induced prostatic relaxation in hypertensive and control rats. To investigate the responsible mechanism(s) and the role of the ATP-sensitive potassium channel., Methods: Prostate strips were isolated from male spontaneously hypertensive (SH) rats and normal Wistar-Kyoto (WKY) rats for measurement of isometric tension. The strips were precontracted with 1 µmol/L phenylephrine or 50 mmol/L KCl. Dose-dependent relaxation of the prostatic strips was studied by cumulative administration of agmatine, 1 to 100 µmol/L, into the organ bath. Effects of specific antagonists on agmatine-induced relaxation were studied. Western blotting analysis was used to measure the gene expression of the ATP-sensitive potassium channel in the rat prostate., Results: Prostatic relaxation induced by agmatine was markedly reduced in SH rats compared with WKY rats. The relaxation caused by agmatine was abolished by BU224, a selective imidazoline I(2)-receptor antagonist, but was not modified by efaroxan at a dose sufficient to block imidazoline I(1)-receptors. The relaxation induced by diazoxide at a concentration sufficient to activate ATP-sensitive potassium channels was markedly reduced in the SH rat prostate. Expressions of ATP-sensitive potassium channel sulphonylurea receptor and inwardly rectifying potassium channel (Kir) 6.2 subunits were both decreased in the prostate of SH rats., Conclusion: The decrease of agmatine-induced prostatic relaxation in SH rats is related to the change in ATP-sensitive potassium channels., (© 2012 THE AUTHORS. BJU INTERNATIONAL © 2012 BJU INTERNATIONAL.)

Aims: The effects of metabolic syndrome on the prostate blood perfusion and histological structure were studied using fructose-fed rats (FR)., Methods: Age-matched male Wistar rats were divided into two groups: group I, normal control rats and group II, 9-week FR. Animal body weight, blood pressure, and serum metabolic parameters were monitored. With the rats under anesthesia, prostatic blood flow volume and flow velocity were measured by laser Doppler flowmetry. The prostate was then removed for histological examination and morphometric study., Results: The 9-week FR showed significant increases in body weight, blood pressure, plasma glucose, insulin, and triglyceride levels. The blood flow volume (345.8 ± 20.57 ml/min/100 g vs. 440.4 ± 35.57 ml/min/100 g of tissue, P < 0.05 for n = 8) and flow rate (29.4 ± 1.25 units vs. 40.9 ± 2.65 units, P < 0.05 for n = 8) in the FR ventral prostate were significantly decreased when compared to controls. Structurally, the FR prostate weight was significantly higher than that of the controls (612.4 ± 16.9 mg vs. 427.3 ± 6.5 mg, P < 0.05 for n = 8). Prostate histology showed glandular hyperplasia while morphometry showed increased stromal component in the FR., Conclusions: Metabolic syndrome in the FR decreases prostatic perfusion and induces prostatic glandular hyperplasia. The findings suggest that chronic ischemia may be a link between metabolic syndrome and prostatic enlargement., (Copyright © 2012 Wiley Periodicals, Inc.)

Objective: To investigate the relationships among the cardiovascular risk factors, prostate blood flow, and prostate volume in patients with clinical benign prostatic hyperplasia (BPH)., Methods: A total of 130 patients with the clinical diagnosis of BPH were recruited. The presence of 5 cardiovascular risk factors, including obesity, diabetes, hypertension, hyperlipidemia, and a history of cardiovascular events, was recorded. The urologic evaluation included digital rectal examination, serum prostate-specific antigen, International Prostate Symptom Score, and transrectal ultrasonography. Doppler spectrum analysis was performed with the patient in the right lateral decubitus position to measure the blood flow in the prostate capsular arteries, periurethral arteries, and neurovascular bundles. The correlations were analyzed between the resistive indexes of the prostatic branches and the cardiovascular risk factors, as well as the transrectal ultrasound findings., Results: The resistive indexes of the periurethral arteries and right neurovascular bundles showed positive correlations with the number of cardiovascular risk factors in the patients (r=.228, P=.01 and r=.225, P=.011, respectively). The periurethral artery resistive index also correlated positively with both prostate and transitional zone volumes, with the capsular artery correlating positively only with the latter. No significant correlations were noted between the resistive indexes and the International Prostate Symptom Score., Conclusion: Prostate vascular resistance in patients with BPH has positive correlations with cardiovascular risk factors and prostate size. These findings suggest that prostate hypoxia might play a role in the pathogenesis of BPH., (Copyright © 2012 Elsevier Inc. All rights reserved.)

This study attempts to investigate the transdermal permeability, the bioavailability and gene expression of plasmid formulated with nonionic poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) (PEO-PPO-PEO) polymeric micelles (PM). Dynamic light scattering (DLS) and atomic force microscopy (AFM) were used to analyze the PM formulated pCMV-Lac Z (P/PM) containing the gene for β-galactosidase (β-Gal) driven by cytomegalovirus early promoter. Franz diffusion cell was used for in vitro transdermal permeability analysis. Real-time PCR was used to quantify the permeated plasmid in vitro and in vivo. β-Gal activity assay was performed to evaluate transgene expression in vivo. The size of P/PM was ~50 nm with round shape. PM significantly enhanced the in vitro transdermal permeability of plasmid in a direction- and temperature-dependent manner. Following transdermal application of P/PM, higher area under the curve (AUC(P/PM): 98.34 h·ng/mL) and longer half-life of plasmid were detected compared with that of plasmid alone (AUC(P): 10.12 h·ng/mL). Additionally, the β-Gal activity was significantly increased in skin, stomach, brain and spinal cord at both 48 and 72 h after P/PM application and in testis and spleen at 72 h postapplication. In conclusion, PM formulation enhanced the permeation of plasmid through skin into blood circulation, increasing its absorption and the transgene expression in various tissues.

Aims: The effect of µ-opioid receptor (MOR) agonist, loperamide on prostate relaxation and the role of potassium channel in this action were studied in isolated Wistar rat prostate., Methods: Tissue strips from rat prostate ventral lobe were hung in organ bath containing: group 1: standard Tyrode's solution (TS); group 2: TS with 1 µM naloxone; group 3: TS with 0.1 µM naloxonazine; and group 4: TS with 0.01-1 µM glybenclamide. The strips were pre-contracted with either 50 mM KCl or 1 µM phenylephrine. Dose-response study on the prostate strip was performed by cumulative administration of loperamide 0.1-10 µM into the organ bath. Western blot study was performed to detect the presence of MOR protein and adenosine triphosphate (ATP)-sensitive potassium channel (K(ATP) ) subunit Kir6.2 protein expressions in the prostate tissue., Results: Loperamide induced relaxation of the pre-contracted prostate strips in a dose-dependent fashion. Pre-treatment with 1 µM naloxone significantly inhibited the relaxation, thus suggesting activation of MOR in the loperamide effect. Pre-treatment with 0.1 µM naloxonazine inhibited relaxation only in the phenylephrine-contracted strips. The K(ATP) channel blocker glybenclamide significantly inhibited the loperamide-induced relaxation, indicating involvement of K(ATP) channels in mediating the prostate relaxation. Western blots showed the expression of MOR and Kir6.2 protein in the rat prostate., Conclusions: MOR and Kir6.2 are expressed in the rat prostate and loperamide induces rat prostate relaxation through activation of peripheral MOR. K(ATP) channels are involved in mediating the effect of loperamide on the relaxation of prostate., (Copyright © 2011 Wiley-Liss, Inc.)

Background: Prior clinical studies have used injection port lumen culture as a marker of intravenous (IV) fluid system contamination. We hypothesized that culturing injected saline (effluent) is a more sensitive method of detecting IV fluid system bacterial contamination than lumen culture. To test this hypothesis, we compared the incidence of lumen contamination with effluent contamination in a simulated setting. We also measured the effect of a novel injection port protective device (Port Guide; Matrix Tooling, Inc, Wood Dale, IL) on contamination., Methods: In this ex vivo study, 33 providers performed 5 injections of 1 mL sterile saline into each of 4 injection port designs: (1) stopcock, (2) stopcock with Port Guide, (3) stopcock with disinfectable needleless closed connector (DNCC), and (4) stopcock with DNCC and Port Guide. The primary outcome was the rate of effluent contamination with simultaneously contaminated injection port lumen., Results: Bacterial organisms were recovered from the effluent in 17 of the 132 injection ports evaluated. Of those 17 injection ports with contaminated effluent, 4 injection port lumens were simultaneously contaminated (24%). Additionally, use of the stopcock with Port Guide significantly reduced effluent contamination., Conclusion: Effluent culture is a more sensitive marker of IV fluid system contamination than injection port lumen culture. A novel protective device on the stopcock (Port Guide) significantly reduced IV fluid system bacterial contamination., (Copyright © 2010 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Mosby, Inc. All rights reserved.)

Stromal keratocyte apoptosis triggered by epithelial injury is one mechanism of corneal disorders. A model of epithelial injury by epithelial debridement is established, and keratocyte apoptosis is evidenced by DNA fragmentation and cellular morphological changes in the anterior stroma underlying the injured epithelium. Delivery of plasmid (pCMV-bcl-x(L)-eGFP) encoding an anti-apoptotic gene, the bcl-x(L) with a nano-carrier, polymeric micelles (PM) via eye drop to cornea after epithelial debridement, the mRNA level of bcl-x(L) was significantly increased (2.2-fold, P<0.05) at 48 h and the eGFP mRNA was detected (4571.7 ± 1194.5 copies/μg total RNA). The bcl-x(L)-eGFP fusion protein was also detected in wounded cornea at 48 h after delivery, accompanying with decreased DNA fragmentation and lower caspase-3 activity (P<0.05). In conclusion, eye drop of pCMV-bcl-x(L)-eGFP/PM reduced corneal apoptosis following epithelial debridement., (Copyright © 2010 Elsevier B.V. All rights reserved.)

The study investigated the effects of metabolic syndrome on urinary bladder nerve-growth factor (NGF) and p75(NTR) expression in fructose-fed obese rats. Age-matched male Wistar rats were divided into four groups; group I: normal control rats; group II: 6-week fructose-fed rats (FR); group III: 9-week FR; and group IV: 12-week FR. In vivo cystometry under anesthesia was performed. In vitro bladder NGF levels were measured by enzyme-linked immunosorbent assay (ELISA). Expressions of the mRNAs that encoded NGF and NGF receptor p75(NTR) in control and 9-week FR bladder were studied using the method of reverse transcription combined with polymerase chain reaction (RT-PCR). The three groups of FR showed significant increases in body weight, blood pressure, plasma glucose, insulin and triglyceride levels when compared to control rats. The bladder NGF levels in the 9-week and 12-week FR were significantly lower than the control (66.8+/-5.4, 49.4+/-7.1 and 95.2+/-6.5 pg/microg protein; p<0.05 and p<0.01 respectively, for n=8 in each group). The bladder emptying function was deteriorated in these two groups of FR as shown by the significantly increased residual volume and decreased emptying fraction. The mRNA expressions of bladder NGF and p75(NTR) in the 9-week FR were significantly decreased when compared to the control group (p<0.05 and p<0.001 respectively, n=8 in each group). In conclusion, long-term metabolic syndrome in FR significantly decreases bladder NGF and p75(NTR) expression. These alterations are associated with deterioration in bladder emptying function.

The role of hyperglycaemia in the pathogenesis of hypotension in diabetic disorders was investigated using the changes in cardiac M(2)-muscarinic receptor (M(2)-mAChR) gene expression in type-1-like diabetic rats and cultured cardiomyocytes. Blood pressure was markedly decreased in diabetic rats following the intravenous injection of streptozotocin (STZ) for 8 weeks. Also, the baroreflex sensitivity (Delta HR/Delta BP), as measured by the changes in heart rate (Delta HR) and mean blood pressure (Delta BP) 1 min after the intravenous injection of phenylephrine (10 microg/kg), was significantly increased. Arecaidine propargyl ester (APE), a M(2)-mAChR agonist produced a marked reduction in heart rate in these diabetic rats. Normalization of plasma glucose in diabetic rats using insulin (0.5 IU) or phlorizin (1 mg/kg) injection attenuated the blood pressure reduction and reversed the mRNA and protein levels of cardiac M(2)-mAChR. A high concentration of glucose (20 mmol/l) directly influenced the increase in gene expression of M(2)-mAChR in the H9c2 cardiac cell line. Hyperglycaemia induced an increase in cardiac M(2)-mAChR gene expression, suggesting a role in the pathogenesis of hypotension in diabetic disorders.

Purpose: We assessed the treatment effect of the aldose reductase inhibitor ONO-2235 (China Chemical and Pharmaceutical, Taipei, Taiwan) on diabetes associated alterations in bladder nerve growth factor and the nerve growth factor neurotrophin receptor p75 mRNA expressions using the streptozotocin (Sigma) induced diabetic rat model., Materials and Methods: Male Wistar rats were divided into 3 groups, including group 1--vehicle treated normal rats, group 2--vehicle treated 9-week streptozotocin diabetic rats and group 3--ONO-2235 treated 9-week streptozotocin diabetic rats. In vivo cystometry was performed using anesthesia. Bladder nerve growth factor levels were measured by enzyme linked immunosorbent assay. Expression of the mRNA encoding nerve growth factor and neurotrophin receptor p75 in the rat bladder was studied using reverse transcriptase-polymerase chain reaction., Results: Cystometry showed increased bladder capacity and decreased emptying function in diabetic rats. ONO-2235 treatment improved voiding volume, voiding fraction and residual volume. The nerve growth factor concentration in streptozotocin induced diabetic rat bladders was significantly lower than the control level in 8 experiments each (mean +/- SEM 45.78 +/- 4.36 and 96.44 +/- 8.73 pg/microg protein, respectively, p <0.01). The mRNA expression of bladder nerve growth factor and neurotrophin receptor p75 in diabetic rats was significantly decreased compared to that in controls in 8 experiments each (p <0.01 and <0.001, respectively). Treatment with ONO-2235 did not significantly change the blood sugar level in diabetic rats. However, administration of the drug significantly increased the bladder nerve growth factor concentration as well as nerve growth factor mRNA and neurotrophin receptor p75 mRNA expression to normal levels in 8 experiments each (p <0.01, <0.01 and <0.001, respectively)., Conclusions: ONO-2235 improved bladder emptying function and restored the decreased genetic expression of bladder nerve growth factor and neurotrophin receptor p75 in 9-week streptozotocin induced diabetic rats, indicating involvement of the sorbitol pathway in the genetic down-regulations of nerve growth factor and p75(NTR) during diabetic cystopathy.

Background: This study evaluates the eye drop delivery of genes with cornea-specific promoters, i.e., keratin 12 (K12) and keratocan (Kera3.2) promoters, by non-ionic poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) (PEO-PPO-PEO) polymeric micelles (PM) to mouse and rabbit eyes, and investigates the underlying mechanisms., Methods: Three PM-formulated plasmids (pCMV-Lac Z, pK12-Lac Z and pKera3.2-Lac Z) containing the Lac Z gene for beta-galactosidase (beta-Gal) whose expression was driven by the promoter of either the cytomegalovirus early gene, the keratin 12 gene or the keratocan gene, were characterized by critical micelle concentration (CMC), dynamic light scattering (DLS), and atomic force microscopy (AFM). Transgene expression in ocular tissue after gene delivery was analyzed by 5-bromo-4-chloro-3-indolyl-beta-D-galactoside (X-Gal) color staining, 1,2-dioxetane beta-Gal enzymatic activity measurement, and real-time polymerase chain reaction (PCR) analysis. The delivery mechanisms of plasmid-PM on mouse and rabbit corneas were evaluated by EDTA and RGD (arginine-glycine-aspartic acid) peptide., Results: The sizes of the three plasmid-PM complexes were around 150-200 nm with unimodal distribution. Enhanced stability was found for three plasmid-PM formulations after DNase I treatment. After six doses of eye drop delivery of pK12-Lac Z-PM three times a day, beta-Gal activity was significantly increased in both mouse and rabbit corneas. Stroma-specific Lac Z expression was only found in pKera3.2-Lac Z-PM-treated animals with pretreatment by 5 mM EDTA, an opener of junctions. Lac Z gene expression in both pK12-Lac Z-PM and pKera3.2-Lac Z-PM delivery groups was decreased by RGD peptide pretreatment., Conclusions: Cornea epithelium- and stroma-specific gene expression could be achieved using cornea-specific promoters of keratin 12 and keratocan genes, and the gene was delivered with PM formulation through non-invasive, eye drop in mice and rabbits. The transfection mechanism of plasmid-PM may involve endocytosis and particle size dependent paracellular transport., (2007 John Wiley & Sons, Ltd)

Purpose: The effects of metabolic syndrome on bladder function and M2,3-muscarinic receptor expression were studied using the fructose fed obese rat., Materials and Methods: Male Wistar rats were divided into 2 groups, including group 1-normal control rats and group 2-6-week fructose fed rats. In vivo cystometry using anesthesia was performed. In vitro the bladder was divided into the urothelium and muscle layer by microdissection. Tissue M2,3-muscarinic receptor protein levels were measured by Western blotting. Expression of the mRNAs that encode M2,3-muscarinic receptors was estimated using reverse transcriptase-polymerase chain reaction., Results: Premicturition unstable bladder contractions suggestive of detrusor overactivity were noted in 62.5% of fructose fed rats but in no controls. M2,3-muscarinic receptor protein and mRNA expression was noted in the urothelium and muscle layer of the rat bladder. In control rats the M2-to-M3-muscarinic receptor protein expression ratio was 1:0.68 in urothelium and 1:0.28 in the muscle layer, and that for mRNA expression was 1:0.32 and 1:0.36, respectively. Compared to controls bladder M2,3-muscarinic receptor protein expression in the urothelium and muscle layer of 8 preparations each was significantly increased in fructose fed rats by 89% and 90% for M2-muscarinic receptor (each p <0.001), and by 35% and 93% for M3-muscarinic receptor (p <0.01 and <0.001, respectively). Correspondingly M2,3-muscarinic receptor mRNA expression in the fructose fed rat bladder urothelium and muscle layer of 8 preparations each was also significantly increased by 31% and 49% for M2-muscarinic receptor (each p <0.01), and by 121% and 117% for M3-muscarinic receptor (each p <0.001, respectively)., Conclusions: Metabolic syndrome induces increased expression of M2,3-muscarinic receptor mRNA and protein in the urothelium as well as the muscle layer of the bladder in 6-week fructose fed rats. The receptor alterations are associated with functional evidence of detrusor overactivity.

Objective: To assess the selenium status of Southern Tasmanians., Design: Cross-sectional., Settings: One thousand and five hundred adults randomly selected from the electoral roll living in the Greater Hobart region of Southern Tasmania, Australia, were invited to participate., Subjects: The overall response rate was 22% (335/1500)., Interventions: A venous blood sample was collected and a questionnaire administered (consisting of brief demographic details and health questions) to subjects who granted informed consent. A previously validated assay using magnetic sector ICP-MS was employed for plasma analysis., Results: Total plasma selenium levels for this sample population were normally distributed with a mean level of 110 microg/l (range 67-268 microg/l) indicating that the majority of the subjects were not selenium-depleted (71% with levels greater than 100 microg/l). Adjustment for differential age/gender response rates produced similar values. More women under 50 (42%) and men over 50 (32%) had selenium levels <100 microg/l with the potential for sub-optimal selenoprotein activity. Low education attainment was associated with low total selenium level (P<0.02)., Conclusions: The majority of participants were not deficient in selenium. Given the narrow therapeutic window of supplementation, dietary advice to increase foods rich in selenium, particularly to higher risk groups, may be an effective means of increasing plasma selenium toward target levels.