Search

Ketamine abusers develop severe lower urinary tract symptoms. The major aims of the present study were to elucidate ketamine-induced ulcerative cystitis and bladder apoptosis in association with oxidative stress mediated by mitochondria and the endoplasmic reticulum (ER). Sprague-Dawley rats were distributed into three different groups, which received normal saline or ketamine for a period of 14 or 28 days, respectively. Double-labeled immunofluorescence experiments were performed to investigate tight junction proteins for urothelial barrier functions. A TUNEL assay was performed to evaluate the distribution of apoptotic cells. Western blot analysis was carried out to examine the expressions of urothelial tight junction proteins, ER stress markers, and apoptosis-associated proteins. Antioxidant enzymes, including SOD and catalase, were investigated by real-time PCR and immunofluorescence experiments. Ketamine-treated rats were found to display bladder hyperactivity. This bladder dysfunction was accompanied by disruptions of epithelial cadherin- and tight junction-associated proteins as well as increases in the expressions of apoptosis-associated proteins, which displayed features of mitochondria-dependent apoptotic signals and ER stress markers. Meanwhile, expressions of mitochondria respiratory subunit enzymes were significantly increased in ketamine-treated bladders. Conversely, mRNA expressions of the antioxidant enzymes Mn-SOD (SOD2), Cu/Zn-SOD (SOD1), and catalase were decreased after 28 days of ketamine treatment. These results demonstrate that ketamine enhanced the generation of oxidative stress mediated by mitochondria- and ER-dependent pathways and consequently contributed to bladder apoptosis and urothelial lining defects. Such oxidative stress-enhanced bladder cell apoptosis and urothelial barrier defects are potential factors that may play a crucial role in bladder overactivity and ulceration.

Introduction The roles of testosterone and orchiectomy on male bladder subjected to ischemic/reperfusion (I/R) injuries received little attention. To fill this gap, the present study intended to examine testosterone and orchiectomy effects on male rabbits subjected to I/R damages. Aim To elucidate the effects of testosterone and orchiectomy on contractile response, bladder morphology, interstitial fibrosis, and oxidative stress in male rabbit bladder subjected to I/R surgery. Methods Male New Zealand rabbits were distributed into five groups as follows: Group 1 received sham surgical procedure. In group 2, I/R surgery was performed. In group 3, testosterone (100 μg/kg/day) was intramuscularly injected prior to I/R surgery. In group 4, orchiectomy was performed prior to I/R surgery. In group 5, orchiectomy was performed with subsequent testosterone administration, followed by I/R surgery. All the rabbits were euthanized 7 days after I/R. Comparative studies were analyzed to elucidate the effects of testosterone and orchiectomy on bladder dysfunction subjected to I/R injuries. Main Outcome Measures Bladder contractile function was evaluated. Masson's trichrome staining and immunohistochemical studies were performed to evaluate bladder morphology and intramural nerve terminals. Western blotting was examined to investigate the expressions of fibrosis and oxidative stress markers. Results I/R surgery significantly decreased bladder contractility in response to various stimulations with and without testosterone treatment. I/R damages decreased bladder nerve density with and without testosterone. The expressions of fibrosis and oxidative stress‐related proteins were increased by I/R injuries with or without testosterone treatment. Testosterone depletion significantly decreased the expressions of transforming growth factor‐β and fibronectin expressions after I/R injury. Supraphysiological testosterone treatment after orchiectomy greatly increased the expressions of these fibrosis proteins; however, orchiectomy alone ameliorated I/R injuries. Conclusions Testosterone treatment or orchiectomy affected I/R‐induced bladder damages in male rabbits. Orchiectomy decreased the level of fibrosis and oxidative stress markers and increased neurofilament densities. Supraphysiological exogenous testosterone administration after orchiectomy further exacerbated such detrimental effects of I/R.

Aims The aims of the present study were to investigate voiding patterns, tissue constituents and the expressions of cyclooxygenase-2 (COX-2) and nitric oxide synthase (NOS) involved in ketamine-induced ulcerative cystitis in rat urinary bladder. Methods Thirty Sprague–Dawley rats were distributed into three groups which received saline or ketamine (25 mg/kg/day) for a period of 14 and 28 days. In each group, cystometry was performed weekly and the concentration of ketamine and its metabolites (norketamine) was assayed. Paraffin-embedded sections were stained with Masson's trichrome stain, and ketamine-induced morphological changes were examined. Western blot analyses were carried out to examine the expressions of COX-2 and different NOS isoforms in bladder tissues. Immunofluorescence study was done to evaluate the expressions of COX-2 and macrophage infiltration (stained with ED-1 macrophage cell surface antigen) within the bladder. Results Ketamine treatment resulted in bladder hyperactivity and the non-voiding contractions were significantly increased. The urine concentrations of ketamine and norketamine were much higher in ketamine-treated group. Moreover, ulcerated urothelium and mononuclear cell infiltration were noted in ketamine-treated group. These alterations in urodynamic functions and tissue constituents were accompanied by increases in the expression of COX-2. Two NOS isoforms (iNOS and eNOS) were also overexpressed, but no significant change was observed for nNOS. COX-2 positive stained cells were significantly increased. Meanwhile, increased amounts of ED-1 positive stained macrophages were present and most of COX-2 expressed cells were co-stained with ED-1 in the early stage of ketamine treatment. Conclusions Ketamine treatment affected bladder tissues by enhancing interstitial fibrosis and accelerating macrophages infiltration. Ketamine also initiated the up-regulations of COX-2 and iNOS and eNOS expressions. These up-regulated enzymes might play an important role in contributing to ketamine-induced alterations in micturition patterns and ulcerative cystitis. Neurourol. Urodynam. 32:1137–1143, 2013. © 2013 Wiley Periodicals, Inc.

What's known on the subject? and What does the study add? Ovary hormone deficiency and the age-related changes in post-menopausal women are subjected to a number of urological dysfunctions, including overactive bladder syndrome. Green tea is a popular healthy drink worldwide and its extract catechin has strong anti-inflammatory and antioxidant properties. EGCG, the major type of catechin, is an antioxidant polyphenol flavonoid isolated from green tea. EGCG supplement could prevent ovariectomy-induced bladder dysfunction in a dose-related manner through its anti-oxidant, anti-fibrosis and anti-apoptosis effects. OBJECTIVE • To evaluate whether green tea extract, epigallocatechin gallate (EGCG), could prevent ovariectomy-induced overactive bladder (OAB) and to investigate its antioxidant, anti-apoptotic and anti-fibrosis effects. MATERIALS AND METHODS • In all, 48 female Sprague–Dawley rats were divided into four groups. After bilateral ovariectomy, the first group served as the ovariectomy control, the second group received EGCG 1 µM/kg daily i.p. injection after ovariectomy surgery, and the third group received EGCG 10 µM/kg daily i.p. injection. The fourth group was taken as the sham without ovariectomy surgery. The rats were killed after 6 months after ovariectomy surgery. • Cystometrograms were performed for the measure of bladder overactivity. • Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labelling (TUNEL) assay was used to evaluate apoptotic cells. • Western immunoblots were performed to determine the expressions of inflammatory markers, apoptosis-associated proteins and oxidative stress markers. RESULTS • Long-term ovariectomy significantly increased non-voiding contractions and decreased bladder compliance. Treatment with EGCG significantly increased bladder compliance and diminished non-voiding contractions. • Ovariectomy significantly increased apoptotic cells and enhanced interstitial fibrosis in bladders. The expression of caspase-3 significantly increased, while that of Bcl-2 notably decreased after ovariectomy. • Inflammatory and fibrosis markers, TGF-β, fibronectin and type I collagen expressions were significantly increased after 6 months of ovariectomy surgery. Treatment with EGCG significantly decreased TGF-β and type I collagen expressions. • Oxidative stress markers, nitrotyrosine and protein carbonylation levels were significantly increased in the ovariectomy group. EGCG could attenuate this oxidative damage in dose-dependent fashion. CONCLUSIONS • Ovariectomy increased oxidative damage, enhanced voiding frequency and decreased bladder compliance. • EGCG could restore ovariectomy-induced bladder dysfunction in a dose-dependent fashion through antioxidant, anti-fibrosis and anti-apoptosis effects.

A rat model of ovariectomy-induced voiding dysfunction has been established, which mimicked the urge incontinence in postmenopausal women. Previous studies have identified strong anti-inflammatory/antioxidant properties of green tea and its associated polyphenols. The aim of this study was to evaluate whether the green tea extract, epigallocatechin gallate (EGCG), could prevent an ovariectomy-induced overactive bladder.The study included 48 female Sprague-Dawley rats, which were divided into four groups. After bilateral ovariectomy during the following 6-month period, 12 rats received an intraperitoneal injection of saline, 24 rats received either a low-dose (1 μM kg(-1) d(-1)) or a high-dose (10 μM kg(-1) d(-1)) EGCG intraperitoneal injection. The sham group consisted of twelve rats that were not ovariectomized. In vivo isovolumetric cystometrograms were performed in all groups before the animals were euthanized. The immunofluorescence study used neurofilament stains to evaluate intramural nerve damage. Western immunoblots and real-time polymerase chain reaction were performed to determine M2 and M3 muscarinic cholinergic receptors (MChRs) at both protein and messenger RNA (mRNA) expressions.Long-term ovariectomy significantly increased non-voiding contractions, whereas treatment with EGCG significantly attenuated the frequency of non-voiding contractions. Ovariectomy significantly decreased the numbers of neurofilament and increased M2 and M3 MChR protein and mRNA expressions. Treatment with EGCG restored the amount of neurofilament staining and decreased M2 and M3 MChR protein and mRNA overexpressions.This study confirmed that ovary hormone deficiency induced overactive bladder dysfunction via intramural nerve damage and muscarinic receptor overexpression. EGCG prevented ovariectomy-induced bladder dysfunction through neuroprotective effects in a dose-dependent fashion.

What's known on the subject? and What does the study add? Protein kinase C inhibitor (PKCI) can decrease glomerular and tubular cell apoptosis and mitosis and attenuate collagen accumulation and fibronectin expression in a PUUO rat model. Although the role of PKC has been well studied in diabetic nephropathy, there is no report on its role in obstructive nephropathy. This investigation evaluated the processes that were associated with the activation of PKCα and PKCβ pathways and showed that PKCI played an important role in the protection of renal function during ureteric obstruction. OBJECTIVES • To investigate the expression of the protein kinase C (PKC) pathway after partial unilateral ureteric obstruction (PUUO). • To evaluate the therapeutic potential of a PKC inhibitor (PKCI) in obstructive nephropathy. MATERIALS AND METHODS • Thirty-six rats were divided into three groups. One sham-operated group served as the control. The other two groups received PUUO surgery, after which one group received no treatment and the other group was treated with PKCI, chelerythrine. • The severity of hydronephrosis and renal morphology were assessed: tubular and glomerularcell apoptosis, mitosis and interstitial fibrosis were examined using immunohistochemistry. • Western immunoblots were performed to determine fibronectin, transforming growth factor-β (TGF-β), and PKC isoform levels. RESULTS • Two weeks after PUUO surgery, hydronephrosis progressively developed. Tubular-interstitial fibrosis, collagen deposition and fibronectin expression were increased. • PUUO also activated the expression of PKCα and PKCβ and the translocation of PKCs from cell cytosol to cell membranes. • Treatment with PKCI significantly decreased PKCα and PKCβ expression and translocation in the renal cortex. • Treatment with PKCI also reduced the severity of hydronephrosis, decreased both glomerular and tubular cell apoptosis and mitosis, and attenuated the collagen and fibronectin accumulation in renal interstitium. CONCLUSIONS • Renal tubular apoptosis and interstitial fibrosis after obstructive nephropathy are associated with PKCα and PKCβ activation. • The PKCI, chelerythrine, is capable of decreasing PKC expression and translocation in the renal cortex, suggesting that this inhibitor may have therapeutic potential in the protection of renal function in the first few weeks after PUUO surgery.

Objectives: Ischemia/reperfusion (I/R) has been shown to be the major etiologic factor in animal models in a variety of bladder dysfunctions. Herein we investigate the direct effect of I/R on rabbit bladder contractile regulatory proteins. Methods: Male rabbit bladders were subjected to IR. The contractile responses were recorded and the protein levels of rho-kinase (ROK), caldesmon (CaD) and myosin light chain kinase (MLCK) were analyzed in both bladder muscle and mucosa. Results: For the mucosa layer, ROK was unchanged after ischemia, but increased significantly after 1 week of reperfusion. MLCK increased after ischemia and then significantly increased after 1 and 2 weeks of reperfusion. In the muscle layer, ROK increased at 2 h of reperfusion, decreased significantly following 1 week of reperfusion, then returned to control level by 2 weeks of reperfusion. MLCK expression significantly decreased as early as ischemia alone and did not recover after reperfusion. For CaD expression in the mucosa layer, both CaD isoforms significantly increased at 1 week of reperfusion, then progressively decreased at 2 weeks of reperfusion. In the muscle layer, both CaD isoforms had different expressions. In the muscle layer, smooth muscle caldesmon (h-CaD) decreased at ischemia alone and at 2 h of reperfusion, but significantly increased at 2 weeks of reperfusion; whereas non-muscle caldesmon (l-CaD) significantly decreased at 2 weeks of reperfusion. Conclusion: As bladder muscle and mucosa have markedly different purposes, it is not surprising that they respond differently to I/R. Bladder mucosa and muscle have different kinds of alternations on contractile regulatory proteins.

In a clinical skills assessment, the decision to pass or fail an examinee should be based on the test content or on the examinees' performance. The process of deciding a pass score is known as setting a standard of the examination. This requires a properly selected panel of expert judges and a suitable standard setting method, which best fits the purpose of the examination. Six standard setting methods that are often used in clinical skills assessment are described to provide an overview of the standard setting process.

Medical students are assumed to be competent to provide basic patient care independently on graduation. However, there is a gap between what students are expected to learn and what they have actually learned. This may be due to the lack of clearly defined learning objectives, well- organized curriculum, and properly administered assessment. In an attempt to tackle this problem, we conducted a three-step study. Firstly, we identified the core clinical competencies required of medical graduates in Taiwan. Secondly, we incorporated these clinical competencies into a new medical curriculum. Finally, we identified the most appropriate assessment methods for each clinical competency. In 2004, a set of minimally required clinical competencies for medical undergraduates in Taiwan was developed, which included 92 clinical skills, four communication skills, and seven kinds of attitudes. In order to prepare 3 rd and 4 th year medical students at Kaohsiung Medical University (KMU) for later clinical work, the medical curriculum committee integrated the teaching and assessment of the core clinical skills identified previously into relevant organ-system blocks of the new curriculum. To identify appropriate assessment methods for each clinical skill, a structured questionnaire of assessment methods based on the Toolbox of Assessment Methods (Accreditation Council for Graduate Medical Education) and The Scottish Doctor (Scottish Deans' Medical Curriculum Group) was developed and distributed to 40 senior clinical faculty members at KMU. Simulations and Models, Standardized Patient Examination (SP), and Objective Structured Clinical Examination (OSCE) were suggested to be most suitable to assess two-thirds of the core clinical skills. These assessment methods are commonly used in American and European medical schools. We believe that the implementation of the new curriculum at KMU accompanied by the use of Simulations and Models, SP, OSCE, and other teaching and assessment methods will help 3 rd and 4 th year students to prepare better for clinical practice in clerkships.

Recent attempts by other investigators to enhance repair processes in the spinal cord have involved the administration of X rays to spinal cord injury sites. Although some functional improvement has been reported, the underlying cellular changes within the irradiated spinal cords are not clear. Studies initiated recently in this laboratory examined the potential of X rays to modulate nonneuronal cell populations associated with an injury site in adult mammalian spinal cords. These studies revealed a unique and previously unreported radiosensitivity of the microglial cell population. Administration of X radiation to a unilateral dorsal lesion cavity in the cervical spinal cord revealed a significant decrease (approximately half) in numbers of microglia associated with the cavity. Even more unexpected were the significant decreases in microglial cells observed on the nonlesioned side of the spinal cord or in sham-operated spinal cords in irradiated rats. In contrast to reports of others, densitometric quantification of GFAP immunoreactive cells and processes indicated no differences in the astrocytic reactions associated with the lesion cavities between nonirradiated and irradiated groups in our studies. The demonstration that exposure of a spinal cord injury site to radiation modifies the responses of certain components of the glial environment to injury may offer a noninvasive approach for direct treatment of that site. Studies are in progress to determine if this altered glial environment enhances the extension of regrowing axons from a peripheral nerve graft across the interface with the irradiated lesion cavity and into the spinal cord parenchyma.

Background Measuring clinical learning environment is crucial for the quality improvement of medical education, especially for medical trainees at transition stages. Medical education in Taiwan is shaped by multiple socio-cultural influences. The aims of this study were to construct an instrument for measuring clinical learning environment in alignment with the local socio-cultural factors and medical training settings, and further investigate the relationship between the quality of the clinical learning environment and the status of mental distress among medical trainees. Methods Participants consisted of 189 medical trainees (62 interns, 52 postgraduate year one (PGY1) residents, and 75 senior residents). Instruments included a designed 40-item Clinical Learning Environment Questionnaire (CLENQ) and a five-item Brief Symptoms Rating Scale (BSRS-5) for evaluating mental distress. Constructs of CLENQ were examined using factor analysis. Correlations were calculated between BSRS-5 and CLENQ across the three levels of medical trainees. Results Factor analysis of CLENQ yielded five factors- I: Teaching (13 items), II: Workload (7 items), III: Relationship pressure (9 items), IV: Organisational support (4 items) and V: Mutual trust (6 items). Intern trainees reported the lower total CLENQ scores in comparison to PGY1 and senior resident trainees. Mental distress using BSRS-5 was negatively correlated with total CLENQ scores and several key factors in all three groups. Conclusions Our study using CLENQ has identified five major factors of clinical learning environment that are closely linked with our local socio-cultural factors and medical training settings. Medical trainee’s mental distress status was negatively correlated with the quality of CLENQ. These findings have socio-cultural relevance and medical contextual significance and might be applicable to other countries. It warrants further study to investigate the impact of clinical learning environment improvement on the medical trainee’s mental distress and performance.

This study sought to establish a culture model of cardiac ganglia (CG) neurons of the Sprague-Dawley (SD) rat which could by used to study the distinct characteristics of CG neurons. After culturing, the morphology and immunocytochemistry of CG neurons obtained on different days after birth were compared. Samples of CG neurons were taken from the posterior atrial wall of rats aged 7, 14, 21 and 40 postnatal days (designated as P7, P14, P21 and P40, respectively). During 3-6 days of culture, the morphological changes of the cultured neurons were monitored using a light microscope. Immunocytochemical staining of the neurofilaments (NF-L, -M and -H) was performed to identify the CG neurons and the changes in morphology. The differences in size of the CG soma of each culture were compared by morphometry. Frozen sections of CG neurons were used as the in vivo control of the above experiments. The results showed that the rate of growth in size of the CG soma was highest in the P7 group, and was slower after weaning (21 days after birth). Cultured neurons were categorized into unipolar-like (Type I), multipolar-like (Type II), and bipolar-like (Type III) based on their morphological characteristics. In NF immuocytochemical staining, there were strong responses to NF-H and NF-M in all cultures, but not to NF-L. More specifically, responses to NF-H were mainly observed in perikaryons and neurites, whereas the responses to NF-M were mainly in perikaryons. The present study has established a culture system for cardiac ganglia neurons of SD rats. Our results show that the intracardiac neurons were still developing in their somata and the processes and that various responses to different antibodies of NF for CG neurons occurred in different postnatal stages in rats.

The objective, structured clinical examination (OSCE) is a method to assess clinical competency based on objective testing, through direct observation in a formal setting. The Kaohsiung Medical University (KMU) has pioneered OSCEs in Taiwan. In KMU, three groups of examinees---medical students in years 3 and 4, medical students in years 5 and 6, and medical students in year 7---were assessed using different OSCEs. Each OSCE was set up using the following five steps: (1) create cases; (2) decide on the items or clinical skills to be evaluated; (3) train standardized patients; (4) run the OSCE and (5) review videos to improve the curriculum. We expect that KMU will become the premier OSCE center in Taiwan.

One layer of attenuated endothelia of continuous capillaries provides a partially selective diffusion barrier between the blood and the interstitium. Ultrastructures of membrane specialization without the known physiologic functions have been found in blood vessel endothelia. The vacuolar profiles or vacuole-like, membrane-bound structures, which are larger than plasmalemmal vesicles, have been observed routinely in normal endothelial cytoplasm or in blood vessels challenged by insults in electron microscopic studies. Three-dimensional information from serial sections is required to understand the organization and functions of vacuole-like structures in capillary endothelium. The capillaries in eel retia mirabile were perfused with electron-dense tracers, glutaraldehyde in buffer, and were processed for transmission electron microscopy. Ribbons of serial thin sections without counterstaining were examined under a transmission electron microscope. The vacuolar profiles inside endothelial cytoplasm were investigated with the techniques of serial section analysis and three-dimensional reconstruction from serial sections. The vacuole-like structures inside endothelial cytoplasm either were connected to extracellular (luminal, abluminal) compartments or existed as isolated vacuoles from serial section analysis. In the eight series examined in this study, six of ten vacuole-like structures were classified as isolated vacuoles inside endothelia, and their diameters ranged between 186 nm and 266 nm. Two of ten vacuole-like structures were found to extend to the luminal surface of capillaries as luminal, pocket-like invaginations. One of ten vacuole-like structures was found to be connected to the albuminal compartment, and another one existed as an extracellular compartment surrounded by endothelia. Three-dimensional projection of the vacuolar compartments from serial sections showed that endothelial cytoplasm of sheet shape protruded and folded over adjacent endothelium. Three-dimensional information from serial sections reveals the organization of vacuolar profiles and pocket-like invaginations from the cell surfaces in capillary endothelium. The vacuolar profiles in capillary endothelia in two-dimensional electron photomicrographs may represent the extracellular compartments surrounded by the endothelial finger-like extensions. The results indicate that the luminal and abluminal surfaces of the capillary lumen are not smooth or static, and endothelia may change their shape in three dimensions through cytoplasmic protrusions when the tissue environment changes.

Objective To investigate the early genetic and cellular responses in the smooth muscle layer of completely obstructed ureters, and to determine whether myocytes proliferate (hyperplasia) in the ureters during the early stage of obstructive uropathy. Materials and methods The study comprised 35 female Sprague-Dawley rats which had undergone unilateral ligation of their ureters. After ureteric ligation, five rats each were killed and examined at 0.5, 1, 2, 4, 8, 16 and 24 h after ligation. The proximal portion of the ureters was prepared for light and electron microscopy. The expression of c-Fos, c-Jun, c-Myc and Ki-67 antigen was assessed immunohistochemically. Results c-Fos and c-Jun were detected 2 h after ligation and the expression of these two proteins reached a maximum after 4 h, becoming undetectable 16 h after ligation. The expressions of c-Fos and c-Jun were strongly correlated (r=0.9854, P

Furosemide has been used in the diuretic renography and diuretic radionuclide scan to evaluate the severity of hydroureter and hydronephrosis. To elucidate the influence of furosemide on obstructed ureters, unilateral ligation of ureter was performed in 45 rats. Twenty-four of the rats received intramuscular injections of furosemide (6 mg/kg/day) after the third day postligation. Eight rats were sacrificed for examination on days 7, 10 and 14 after ligation, respectively. The remaining 21 untreated rats were also sacrificed for comparison. The severity of hydroureter and hydronephrosis in the ligated side of the furosemide-treated rats was significantly higher than that of the untreated rats. However, the histological changes in the treated and untreated rats showed no significant difference. The ultrastructural alterations aggravated along the course of ureteric obstruction. Intriguingly, the ultrastructural changes were significantly milder in the treated rats. We conclude that the administration of furosemide might increase the severity of hydroureter, but it does not accelerate the ureteric damage of the obstructed ureters.

Background The Objective Structured Clinical Examination (OSCE) has been widely applied as a high-stakes examination for assessing physicians’ clinical competency. In 1992, OSCE was first introduced in Taiwan, and the authorities announced that passing the OSCE would be a prerequisite for step-2 medical licensure examination in 2013. This study aimed to investigate the impacts of the announced national OSCE policy on implementation of OSCE at the institutional level. Further, the readiness and the recognition of barriers toward a high-stakes examination were explored. Methods In 2007 and 2010, the year before and after the announcement of high-stakes OSCE policy in 2008, respectively, questionnaires on the status of OSCE implementation were distributed to all hospitals with active OSCE programs in Taiwan. Information on OSCE facilities, equipment, station length, number of administrations per year, and the recognition of barriers to the success of implementing an OSCE were collected. The missing data were completed by telephone interviews. The OSCE format, administration, and facilities before and after the announcement of the nationwide OSCE policy were compared. Results The data were collected from 17 hospitals in 2007 and 21 in 2010. Comparing the OSCE formats between 2007 and 2010, the number of stations increased and the station length decreased. The designated space and the equipment for OSCE were also found to have been improved. As for the awareness of OSCE implementation barriers, the hospital representatives concerned mostly about the availability and quality of standardized patients in 2007, as well as space and facilities in 2010. Conclusions The results of this study underscored an overall increase in the number of OSCE hospitals and changes in facilities and formats. While recruitment and training of standardized patients were the major concerns before the official disclosure of the policy, space and facilities became the focus of attention after the announcement. The study results highlighted the influence of government policy on different aspects of OSCE implementation in Taiwanese training institutes that showed high level of support as reflected in the improved hardware and the change in OSCE format to serve the summative purpose.

Dear SirBoursicot et al. (2011) stated “The current situation in relation to performance assessment and national regulatory standards are that Canada, China and Japan have established national lice...

Objective The present study investigated the effects of ovarian hormone depletion and estrogen administration on ischemia/reperfusion (I/R)-induced bladder damage in female rabbits. Methods Female New Zealand white rabbits were divided into five groups. A sham surgical procedure was performed on rabbits in group 1. In group 2, both vesical arteries were clamped for 2 hours and then released (I/R surgical procedure). In group 3, 17β-estradiol (100 μg/kg/d) was injected intramuscularly before I/R surgical procedure. In group 4, ovariectomies were performed before I/R surgical procedure. Group 5 had ovariectomy, recovered for 2 weeks, and then received 17β-estradiol for 2 weeks. I/R surgical procedure was performed thereafter. Rabbits were killed 7 days after I/R surgical procedure. Masson's trichrome stain was used, and immunohistochemical experiments were performed to evaluate interstitial fibrosis and intramural nerve changes. Western immunoblots were examined to determine the expressions of markers for inflammation, fibrosis, and oxidative stress. Results I/R surgical procedure decreased bladder contractile responses by 30% to 50%. Ovarian hormone depletion further reduced bladder contractile function by 45% to 55% compared with the I/R group members that retained their ovaries. Moreover, I/R surgical procedure significantly decreased intramural neurofilament staining by two thirds compared with the control group. Estrogen replacement after ovariectomy significantly increased the density of nerve terminals. In addition, the expression of transforming growth factor-β and fibronectin increased twofold and fivefold after I/R, respectively. Ovarian hormone depletion further increased the expression of these inflammatory and fibrosis markers. Ovariectomy significantly exacerbated oxidative damage, whereas estrogen replacement diminished oxidative stress to a level approaching that of the control group. Conclusions I/R surgical procedure increases oxidative damage, enhances interstitial fibrosis, and results in bladder denervation. Ovarian hormone deficiency exacerbates this I/R-induced bladder damage, whereas estrogen therapy after ovariectomy attenuates this injury. These results reveal estrogen's protective effects on bladders subjected to I/R injury and the potential benefits of estrogen therapy on I/R-induced bladder damage.

The aims of the present study were to investigate voiding patterns, tissue constituents and the expressions of cyclooxygenase-2 (COX-2) and nitric oxide synthase (NOS) involved in ketamine-induced ulcerative cystitis in rat urinary bladder.Thirty Sprague-Dawley rats were distributed into three groups which received saline or ketamine (25 mg/kg/day) for a period of 14 and 28 days. In each group, cystometry was performed weekly and the concentration of ketamine and its metabolites (norketamine) was assayed. Paraffin-embedded sections were stained with Masson's trichrome stain, and ketamine-induced morphological changes were examined. Western blot analyses were carried out to examine the expressions of COX-2 and different NOS isoforms in bladder tissues. Immunofluorescence study was done to evaluate the expressions of COX-2 and macrophage infiltration (stained with ED-1 macrophage cell surface antigen) within the bladder.Ketamine treatment resulted in bladder hyperactivity and the non-voiding contractions were significantly increased. The urine concentrations of ketamine and norketamine were much higher in ketamine-treated group. Moreover, ulcerated urothelium and mononuclear cell infiltration were noted in ketamine-treated group. These alterations in urodynamic functions and tissue constituents were accompanied by increases in the expression of COX-2. Two NOS isoforms (iNOS and eNOS) were also overexpressed, but no significant change was observed for nNOS. COX-2 positive stained cells were significantly increased. Meanwhile, increased amounts of ED-1 positive stained macrophages were present and most of COX-2 expressed cells were co-stained with ED-1 in the early stage of ketamine treatment.Ketamine treatment affected bladder tissues by enhancing interstitial fibrosis and accelerating macrophages infiltration. Ketamine also initiated the up-regulations of COX-2 and iNOS and eNOS expressions. These up-regulated enzymes might play an important role in contributing to ketamine-induced alterations in micturition patterns and ulcerative cystitis.

What's known on the subject? and What does the study add? Ovary hormone deficiency and the age-related changes in post-menopausal women are subjected to a number of urological dysfunctions, including overactive bladder syndrome. Green tea is a popular healthy drink worldwide and its extract catechin has strong anti-inflammatory and antioxidant properties. EGCG, the major type of catechin, is an antioxidant polyphenol flavonoid isolated from green tea. EGCG supplement could prevent ovariectomy-induced bladder dysfunction in a dose-related manner through its anti-oxidant, anti-fibrosis and anti-apoptosis effects.To evaluate whether green tea extract, epigallocatechin gallate (EGCG), could prevent ovariectomy-induced overactive bladder (OAB) and to investigate its antioxidant, anti-apoptotic and anti-fibrosis effects.In all, 48 female Sprague-Dawley rats were divided into four groups. After bilateral ovariectomy, the first group served as the ovariectomy control, the second group received EGCG 1 µM/kg daily i.p. injection after ovariectomy surgery, and the third group received EGCG 10 µM/kg daily i.p. injection. The fourth group was taken as the sham without ovariectomy surgery. The rats were killed after 6 months after ovariectomy surgery. Cystometrograms were performed for the measure of bladder overactivity. Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labelling (TUNEL) assay was used to evaluate apoptotic cells. Western immunoblots were performed to determine the expressions of inflammatory markers, apoptosis-associated proteins and oxidative stress markers.Long-term ovariectomy significantly increased non-voiding contractions and decreased bladder compliance. Treatment with EGCG significantly increased bladder compliance and diminished non-voiding contractions. Ovariectomy significantly increased apoptotic cells and enhanced interstitial fibrosis in bladders. The expression of caspase-3 significantly increased, while that of Bcl-2 notably decreased after ovariectomy. Inflammatory and fibrosis markers, TGF-β, fibronectin and type I collagen expressions were significantly increased after 6 months of ovariectomy surgery. Treatment with EGCG significantly decreased TGF-β and type I collagen expressions. Oxidative stress markers, nitrotyrosine and protein carbonylation levels were significantly increased in the ovariectomy group. EGCG could attenuate this oxidative damage in dose-dependent fashion.Ovariectomy increased oxidative damage, enhanced voiding frequency and decreased bladder compliance. EGCG could restore ovariectomy-induced bladder dysfunction in a dose-dependent fashion through antioxidant, anti-fibrosis and anti-apoptosis effects.

What's known on the subject? and What does the study add? Protein kinase C inhibitor (PKCI) can decrease glomerular and tubular cell apoptosis and mitosis and attenuate collagen accumulation and fibronectin expression in a PUUO rat model. Although the role of PKC has been well studied in diabetic nephropathy, there is no report on its role in obstructive nephropathy. This investigation evaluated the processes that were associated with the activation of PKCα and PKCβ pathways and showed that PKCI played an important role in the protection of renal function during ureteric obstruction.• To investigate the expression of the protein kinase C (PKC) pathway after partial unilateral ureteric obstruction (PUUO). • To evaluate the therapeutic potential of a PKC inhibitor (PKCI) in obstructive nephropathy.• Thirty-six rats were divided into three groups. One sham-operated group served as the control. The other two groups received PUUO surgery, after which one group received no treatment and the other group was treated with PKCI, chelerythrine. • The severity of hydronephrosis and renal morphology were assessed: tubular and glomerularcell apoptosis, mitosis and interstitial fibrosis were examined using immunohistochemistry. • Western immunoblots were performed to determine fibronectin, transforming growth factor-β (TGF-β), and PKC isoform levels.• Two weeks after PUUO surgery, hydronephrosis progressively developed. Tubular-interstitial fibrosis, collagen deposition and fibronectin expression were increased. • PUUO also activated the expression of PKCα and PKCβ and the translocation of PKCs from cell cytosol to cell membranes. • Treatment with PKCI significantly decreased PKCα and PKCβ expression and translocation in the renal cortex. • Treatment with PKCI also reduced the severity of hydronephrosis, decreased both glomerular and tubular cell apoptosis and mitosis, and attenuated the collagen and fibronectin accumulation in renal interstitium.• Renal tubular apoptosis and interstitial fibrosis after obstructive nephropathy are associated with PKCα and PKCβ activation. • The PKCI, chelerythrine, is capable of decreasing PKC expression and translocation in the renal cortex, suggesting that this inhibitor may have therapeutic potential in the protection of renal function in the first few weeks after PUUO surgery.

Mammalian cortical neurogenesis occurs on a precise time schedule during development. The earliest born neurons form the preplate and later separate into layer 1, which includes Cajal-Retzius (C-R) neurons, and the subplate. The preplate and its derivatives play a critical role in regulating subsequent neuron migration and cortical lamination. Using an early B cell factor 2 (Ebf2)-enhanced green fluorescent protein (EGFP) transgenic mouse line, we show that Ebf2-EGFP is expressed in the preplate and persists in C-R neurons, allowing us to follow the development of these early born neurons. Therefore, Ebf2 is a genetic marker for preplate neurons from the earliest stage of their differentiation and C-R cells after the preplate is split. Additionally, we examined the function of Ebf2 in the development of C-R neurons using both lentiviral-mediated knock-down overexpression approaches to perturb Ebf2 expression. Our data show that Ebf2 overexpression increases the generation of early born neurons including C-R cells, while Ebf2 knock-down decreases it, without affecting the generation of other layer-specific neurons in vitro. These results indicate that Ebf2 is important for early cortical neurogenesis and regulates the generation of C-R neurons in the developing cerebral cortex.

The present study was designed to investigate the effect of nitric oxide precursor, L: -arginine, on bladder function following ovariectomy.Twenty-eight New Zealand white female rabbits were separated into seven groups. Groups 1 to 6 underwent ovariectomy surgery. Among them, groups 1 and 2 received ovariectomy without treating with L-arginine. Groups 3, 4, 5, and 6 were given high L-arginine diet and were sacrificed 1, 3, 7, and 14 days after ovariectomy, respectively. Group 7 served as the control group. The effects of L: -arginine on the contractility of bladder tissues were determined in response to various stimulations. In addition, L-arginine effects on the expression of Rho kinase (ROK), protein kinase C potentiated inhibitor (CPI-17), caldesmon (CaD), and calponin (CaP) were studied by immunoblotting.Ovariectomy significantly decreases contractile response to all forms of stimulation. Feeding rabbits L: -arginine significantly increases contractile response at 1 day following ovariectomy, but the response decreases to the control level by 14 days. Ovariectomy increases the expressions of both isoforms of CaD, CaP, and CPI-17; L-arginine treatment induces ROK underexpression, while CaP is overexpressed in the early few days of ovariectomy but returns to the control level at 2 weeks after ovariectomy.Ovariectomy appreciably reduced bladder contractility. Treatment with L-arginine reversed the ovariectomy-induced bladder dysfunction. Decreased bladder contractile response was observed in the early days following ovariectomy.

Neuron density, volume of the area and total neuron number were measured in the medial preoptic area, anterior hypothalamic area and arcuate nucleus of the hypothalamus of young (6-month-old), middle age (14-month-old) and old (22-month-old) male and female rats. Intact male rats did not show neuron loss even in old age, while intact female rats manifested neuron loss in the medial preoptic area, anterior hypothalamic area and arcuate nucleus in old age. Long-term administration of estradiol benzoate to castrated male rats induced neuron loss in the anterior hypothalamic area and arcuate nucleus of the middle age group and in all three areas of the old age group. However, long-term ovariectomy could not prevent neuron loss in these hypothalamic areas in old age. The results suggested that estradiol can have a cumulative impact on the degree of neuron damage and simulate female-type age-related neuron loss in male rats and that there may be the possibility of an intrinsic aging process inducing neuron loss in female rats.

Two sperm motility inhibtors (SMI1 and SMI2) were purified from porcine seminal plasma with high performance liquid chromatography (HPLC). Their molecular weights are about 15,000 as estimated by SDS-polyacrylamide gel electrophoresis (SDS-PAGE). Both of them decreased the percentage of motile spermatozoa in a dose-dependent manner. The inhibitory effect can be abolished by addition of the porcine follicular fluid. Both SMI1 and SMI2 have similar amino acid composition, suggesting that they may be structurally related. They also have inhibitory effect on chymotrypsin.

The present work is an attempt to develop a new method to determine sex from the skull with lateral radiographic cephalometry and discriminant function analysis. The superciliary ridges, frontal sinuses, external occipital protuberance, and mastoid processes were adopted as objects of lateral radiographic cephalometric measurements. With discriminant functions created from 18 established cephalometric variables, a total of 100 cases were classified into two sexual groups with 100% accuracy in a random sample of Taiwanese adults. Therefore, we may obtain a much greater reliability of sex determination from skulls according to this newly developed technique.

Adult albino rats were subjected to unilateral surgical removal of the eyeball. After survival times of 7-140 days, the numerical response of the neuroglial cells, and the progressive disintegration of the myelin sheaths in the optic nerves, were studied qualitatively and quantitatively in electron-microscopic montages. The distribution density of microglia and astroglia in degenerating optic nerve increased to peaks after 35 and 56 days respectively, whereas, the oligodendroglia gradually decreased. During the early stage of degeneration, microglial cells appeared and invaded the sheath at the intraperiod line, peeling off the outer lamellae, which were then engulfed by phagocytosis. Within the microglia, myelin sheath fragments were surrounded by a membrane curled to form a myelin ring. In the intermediate stage of degeneration, the paired electron-dense lines of the ring, made up of myelin basic protein, decomposed and formed a homogeneous or heterogeneous osmiophilic layered structure, the myelin body, which, in the final stages, disintegrated and transformed into globoid lipid droplets and needle shaped cholesterol crystals.