Interaction of dietary carbohydrate and fat on glucose metabolism in growing pigs.

Increased consumption of fructose has been suggested to be a contributing cause of the increased rates of obesity in humans. Rodent studies have shown an increase in de novo lipogenesis and decreased insulin sensitivity in response to feeding high levels of fructose, but it is unclear if these effects occur in the same progression in humans. We aimed to develop a swine model for studying changes in glucose metabolism and insulin resistance resulting from dietary carbohydrate alone or in combination with high dietary fat. Two experiments were conducted to determine if the source of dietary carbohydrate, with or without added fat, had an effect on body weight gain, glucose metabolism, or insulin response in growing pigs. In the first experiment, pigs (24 barrows, initial body weight 28 kg) were fed one of 4 diets in which the source of carbohydrate was varied: 1) 20% starch; 2) 10% glucose + 10% starch; 3) 10% fructose + 10% starch; and 4) 20% fructose for 9 weeks. There were no differences in growth rate or glucose clearance observed. Experiment 2 was conducted as a 3 × 2 factorial with the main effects of carbohydrate source (20% starch, glucose, or fructose) and added fat level (0 vs 10%). Pigs (24 barrows, initial body weight 71 kg) were fed one of 6 experimental diets for 9 weeks. Compared to the other dietary treatments, pigs fed fructose with high fat had an elevated glucose area under the curve during the GTT (Carbohydrate x Fat interaction, P < 0.01). This same group had a lower insulin response (Carbohydrate x Fat, P < 0.05). This work demonstrates that pigs can be a viable model to assess the long-term effects of dietary carbohydrates on metabolism and body composition. Studies of longer duration are needed to determine if these changes are indicative of insulin resistance.

Kefir peptides prevent high-fructose corn syrup-induced non-alcoholic fatty liver disease in a murine model by modulation of inflammation and the JAK2 signaling pathway.

OBJECTIVE: In recent years, people have changed their eating habits, and high-fructose-containing bubble tea has become very popular. High-fructose intake has been suggested to be a key factor that induces non-alcoholic fatty liver disease (NAFLD). Kefir, a fermented milk product composed of microbial symbionts, has demonstrated numerous biological activities, including antibacterial, antioxidant and immunostimulating effects. The present study aims to evaluate the effects of kefir peptides on high-fructose-induced hepatic steatosis and the possible molecular mechanism. RESULTS: An animal model of 30% high-fructose-induced NAFLD in C57BL/6J mice was established. The experiment is divided into the following six groups: (1) normal: H2O drinking water; (2) mock: H2O+30% fructose; (3) KL: low-dose kefir peptides (50 mg kg-1)+30% fructose; (4) KM: medium-dose kefir peptides (100 mg kg-1)+30% fructose; (5) KH: high-dose kefir peptides (150 mg kg-1)+30% fructose; and (6) CFM: commercial fermented milk (100 mg kg-1)+30% fructose. The results show that kefir peptides improve fatty liver syndrome by decreasing body weight, serum alanine aminotransferase, triglycerides, insulin and hepatic triglycerides, cholesterol, and free fatty acids as well as the inflammatory cytokines (TNF-α, IL-6 and IL-1ß) that had been elevated in fructose-induced NAFLD mice. In addition, kefir peptides markedly increased phosphorylation of AMPK to downregulate its targeted enzymes, ACC (acetyl-CoA carboxylase) and SREBP-1c (sterol regulatory element-binding protein 1), and inhibited de novo lipogenesis. Furthermore, kefir peptides activated JAK2 to stimulate STAT3 phosphorylation, which can translocate to the nucleus, and upregulated several genes, including the CPT1 (carnitine palmitoyltransferase-1) involved in fatty acid oxidation. CONCLUSION: Our data have demonstrated that kefir peptides can improve the symptoms of NAFLD, including body weight, energy intake, inflammatory reaction and the formation of fatty liver by activating JAK2 signal transduction through the JAK2/STAT3 and JAK2/AMPK pathways in the high-fructose-induced fatty liver animal model. Therefore, kefir peptides may have the potential for clinical application for the prevention or treatment of clinical metabolic syndrome.

Trends and predictors of outcomes after surgery for hepatocellular carcinoma: A nationwide population-based study in Taiwan.

BACKGROUND: Despite the huge and growing global burden of hepatocellular carcinoma (HCC), high-quality population-based studies of HCC prevalence and outcomes are scarce. PURPOSE: To analyze trends and predictors of hospital resource utilization and mortality rates in a population of patients who had received HCC surgery. PATIENTS AND MATERIALS: This population-based patient cohort study retrospectively analyzed 23,107 patients who had received surgical treatment for HCC from 1998 to 2009. RESULTS: The prevalence rate of surgical treatment in HCC patients significantly increased by 167.4% from 4.857 per 100,000 persons in 1998 to 12.989 per 100,000 persons in 2009 (P < 0.001). Age, gender, Deyo-Charlson co-morbidity index score, hospital volume, surgeon volume, digestive system disease, hepatitis type and liver cirrhosis were significantly associated with HCC surgical outcomes (P < 0.05). Over the 12-year period analyzed, the estimated mean hospital treatment cost increased 9.4% whereas mean length of stay (LOS) decreased 25.3%. The estimated mean overall survival time after HCC surgery was 40.9 months (SD 1.2 months), and the overall in-hospital 1-, 3-, and 5-year survival rates were 97.2%, 79.9%, 61.1%, and 54.6%, respectively. CONCLUSIONS: These population-based data reveal that the prevalence of HCC has increased, especially in older patients. Additionally, hospital treatment costs for HCC have increased despite decreases in LOS. These analytical results should be applicable to most countries with relatively small populations. Additionally, healthcare providers and patients should recognize that attributes of both the patient and the hospital may affect outcomes.

Kefir improves bone mass and microarchitecture in an ovariectomized rat model of postmenopausal osteoporosis.

SUMMARY: Kefir treatment in ovariectomized (OVX) rats could significantly decrease the levels of bone turnover markers and prevent OVX-induced bone loss, deterioration of trabecular microarchitecture, and biomechanical dysfunction that may be due to increase intracellular calcium uptake through the TRPV6 calcium channel. INTRODUCTION: Osteoporosis is a disease characterized by low bone mass and structural deterioration of bone tissue, leading to an increased fracture risk. The incidence of osteoporosis increases with age and occurs most frequently in postmenopausal women due to estrogen deficiency, as the balance between bone resorption and bone formation shifts towards increased levels of bone resorption. Among various methods of prevention and treatment for osteoporosis, an increase in calcium intake is the most commonly recommended preventive measure. Kefir is a fermented milk product made with kefir grains that degrade milk proteins into various peptides with health-promoting effects, including immunomodulating-, antithrombotic-, antimicrobial-, and calcium-absorption-enhancing bioactivities. METHODS: The aim of this study is to investigate the effect of kefir on osteoporosis prophylaxis in an ovariectomized rat model. A total of 56 16-week-old female Sprague-Dawley (SD) rats were divided into 7 experimental groups: sham (normal), OVX/Mock, OVX/1X kefir (164 mg/kg BW/day), OVX/2X kefir (328 mg/kg BW/day), OVX/4X kefir (656 mg/kg BW/day), OVX/ALN (2.5 mg/kg BW/day), and OVX/REBONE (800 mg/kg BW/day). After 12-week treatment with kefir, the bone physiology in the OVX rat model was investigated. Accordingly, the aim of this study was to investigate the possible transport mechanism involved in calcium absorption using the Caco-2 human cell line. RESULTS: A 12-week treatment with kefir on the OVX-induced osteoporosis model reduced the levels of C-terminal telopeptides of type I collagen (CTx), bone turnover markers, and trabecular separation (Tb. Sp.). Additionally, treatment with kefir increased trabecular bone mineral density (BMD), bone volume (BV/TV), trabecular thickness (Tb. Th), trabecular number (Tb. N), and the biomechanical properties (hardness and modulus) of the distal femur with a dose-dependent efficacy. In addition, in in vitro assay, we found that kefir increased intracellular calcium uptake in Caco-2 cell through TRPV6 calcium channels and not through L-type voltage-operated calcium channels. CONCLUSION: The protective effect of kefir in the OVX rat model may occur through increasing intracellular calcium uptake through the TRPV6 calcium channel.

Prevalence of food allergy in Taiwan: a questionnaire-based survey.

AIM: Food allergy is common in children and adults, and could be potentially fatal in minor groups. It is important for physicians to identify the prevalence of food allergies and to recognise common food allergens to make precise diagnosis and choose correct therapeutic approaches. METHODS: We used a nationwide, cross-sectional, random questionnaire-based survey to estimate the self-reported and expert-screened prevalence of food allergies and to identify the common food allergens in Taiwan. In this study, the perceptional diagnosis of food allergies was screened by physicians according to descriptions of convincing symptoms and medical recordings; in the meantime, non-allergic adverse reactions to foods, including food intolerance or food avoidance, were clarified. RESULTS: A total of 30 018 individuals who met the inclusion criteria was evaluated, and 6.95% of them were diagnosed as victims of food allergies. The prevalence was 3.44% in children under 3 years of age, 7.65% in children aged 4-18 years and 6.40% in adults respectively. About 77.33% of the food allergy population had experienced recurrent allergic attacks. Systemic reactions happened about 4.89% in food allergies group. The most commonly reported food allergen in Taiwan is seafood, including shrimp, crab, fish and mollusc. In addition, mango, milk, peanuts and eggs were also important food allergens in the general population; while milk, shellfish, peanuts and eggs were common in children. CONCLUSIONS: Less than 10% of the Taiwan population suffers from food allergy with different allergic symptoms to variable food allergens in different age groups.

Application of high-frequency ultrasound for the detection of surgical anatomy in the rodent abdomen.

Rats are used extensively in abdominal disease research. To monitor disease progress in vivo, high-frequency ultrasound (HFU) can be a powerful tool for obtaining high-resolution images of biological tissues. However, there is a paucity of data regarding the correlation between rat anatomy and corresponding HFU images. Twenty-four adult male Sprague-Dawley (SD) rats underwent abdominal scans using HFU (40 MHz) surgical procedures to identify abdominal organs and major vessels as well as in situ scanning to confirm the imaging results. The results were compared with those of human abdominal organs in ultrasonographic scans. The rat liver, paired kidneys, stomach, intestines, and major blood vessels were identified by HFU and the ultrasonic morphologies of the liver and kidneys showed clear differences between rats and humans. Clinically relevant anatomical structures were identified using HFU imaging of the rat abdomen, and these structures were compared with the corresponding structures in humans. Increased knowledge with regard to identifying the anatomy of rat abdominal organs by ultrasound will allow scientists to conduct more detailed intra-abdominal research in rodents.

Identification of sex-specific polymorphic sequences in the goat amelogenin gene for embryo sexing.

Amelogenin (AMEL) is a conserved gene located on the sex chromosomes of mammals. It is involved in the formation of enamel, which is the hard, white material that forms the protective outer layer of each tooth. In this study, we first cloned and determined the intron sequences of the goat AMELX and AMELY genes from female and male ear tissues. The polymorphic AMEL alleles were further analyzed by PCR-based RFLP and Southern blot hybridization analyses. Results showed that intron 5 nucleotide sequences of the goat AMELY gene contains multiple deletions/insertions and shares only 48.5% identity to intron 5 of the goat AMELX gene. Based on the polymorphic AMEL intron sequences, a set of sex-specific triplex primers was designed to PCR amplify a single fragment of 264 bp from the X chromosome of female goats and 2 fragments of 264 and 206 bp from the X and Y chromosomes, respectively, of male goats. An increased sensitivity for sex determination was reached with a single blastomere at the blastula stage isolated from goat embryos. A total of 43 goat embryos were used to estimate a 100% accuracy rate of this method confirmed by chromosomal karyotyping and live births. The embryo sexing technique has been successfully applied in different strains of goats including Alpine, Saanen, Nubian, and Taiwan goats.

A longitudinal study of the impact of interviews on medical school admissions in Taiwan.

Medical schools in Taiwan have recently adopted the U.S. medical school admissions model by incorporating interviews into the selection process. The objective of this study was to investigate factors that contribute to successful medical school applications through the national entrance examination and interview admission routes. The sample consisted of survey data from five entry cohorts of medical students admitted to the National Yang-Ming University Faculty of Medicine from 2003 to 2007. Of the 513 students, 62% were admitted through the traditional national entrance examination route and 38% were admitted early after achieving a threshold score on the composite national exam followed by a structured interview. Students admitted through the interview route were more likely to be female, with an odds ratio (OR) of 2.17 (1.20-3.93). Maternal education level was an independent predictor of both early admission through a successful interview and higher medical school grade point average (GPA). Students admitted through the interview route had a 3.20 point higher first-year medical school GPA (p < .001) as determined by regression analyses. Those students who were admitted via interview did not have significantly different personality traits than those admitted through the traditional route. This study calls into question the ability of an admissions interview to select for noncognitive character traits.

ErbB4 (JM-b/CYT-1)-induced expression and phosphorylation of c-Jun is abrogated by human papillomavirus type 16 E5 protein.

Human papillomavirus type 16 E5 (HPV-16 E5) is a highly hydrophobic membrane protein with weak-transforming activity, which is associated with ErbB4 receptor in HPV-16-infected cervical lesions. Presently, we investigated the transforming mechanisms of E5 involving ErbB4 signaling. Firstly, we report a role for ErbB4 (JM-b/CYT-1) receptor that activates c-jun gene expression and phosphorylating at Ser63 and Ser73 of the c-Jun protein in ligand-independent and Ras-c-jun NH(2)-terminal kinase-dependent pathway. Secondly, we show that HPV-16 E5 protein can form a complex with ErbB4 via binding to the extracellular and transmembrane domains of ErbB4 (JM-b/CYT-1). When co-expressing HPV-16 E5 and ErbB4 in cells, E5 can abrogate ErbB4-induced c-Jun protein expression and phosphorylation resulted in increasing cell proliferation compared to ErbB4-expressing cells. The interaction between of HPV-16 E5 and ErbB4 provides more insight into the mechanisms of HPV-16 E5 transformation induction.

Psychosocial effects on caregivers for children on chronic peritoneal dialysis.

The study was designed to explore the psychosocial effects on caretakers of children in Taiwan on chronic peritoneal dialysis (CPD). This is a case-control study, performed with subjects drawn from eight medical centers. The study group consisted of caretakers of 32 children with renal failure being treated with CPD. For comparison, a control group of caretakers of 64 healthy children as well as the regional Taiwanese studies were used. Two instruments were used to explore the presence of probable depression and quality of life (QOL) of the caretakers: the Taiwanese Depression Questionnaire, and the World Health Organization QOL BRIEF-Taiwan Version. In the study group, only 25% of caregivers had full-time jobs, and 66% of families had an annual income of less than US dollar 15,000. Of the 32 families in the study group, 16% had only a single parent. The prevalence of probable depression was significantly more common in the study group compared with control and referent group (28% vs 5% and 9.44%; P = 0.001). QOL scores in four domains were also significantly lower in the study group. In conclusion, even with the advances of peritoneal dialysis techniques, caring for children on CPD in Taiwan has significant adverse psychosocial effects on the primary caregivers. Attention should be paid to the psycho-social status of the caregivers.

Using children as standardised patients for assessing clinical competence in paediatrics.

BACKGROUND: Standardised patients (SPs) have been widely used to assess physicians' clinical competence. However, in paediatrics, the use of children in such a way has long been questioned with regard to ethics and the examination quality (in terms of validity, reliability, and feasibility). AIMS: To summarise the current state of the use of child SPs, and to highlight the difficulties inherent in the use of children for this purpose. METHODS: Nineteen articles dealing with the use of child SPs for clinical assessment were reviewed. RESULTS: Child SPs, ranging in age from infancy to adolescence, were present in varied proportions of paediatric objective structured clinical examination stations (12-27%). In most of these reports, there were several children with cases who could substitute for one another. Child SPs successfully portrayed various roles, although only older children had to learn a scenario. In general, clinical examinations using child SPs were found to be valid and generated reliable scores. Child SPs also provided effective feedback. The experience tended to be considered negative for younger children but was quite positive for a number of older children. The use of young SPs should be avoided for ethical reasons, and the use of child SPs should be limited to assessments that cannot be satisfactorily measured by other methods. CONCLUSION: Through meticulous attention to detail and careful planning, a clinical examination using children as SPs can be practical, valid, and reliable.

The quality of a simulation examination using a high-fidelity child manikin.

PURPOSE: Developing quality examinations that measure physicians' clinical performance in simulations is difficult. The goal of this study was to develop a quality simulation examination using a high-fidelity child manikin in evaluating paediatric residents' competence about managing critical cases in a simulated emergency room. Quality was determined by evidence of the reliability, validity and feasibility of the examination. In addition, the participants' responses regarding its realism, effectiveness and value are presented. METHOD: Scenario scripts and rating instruments were carefully developed in this study. Experts were used to validate the case scenarios and provide evidence of construct validity. Eighteen paediatric residents, 'working' as pairs, participated in a manikin-based simulation pre-test, a training session and a post-test. Three independent raters rated the participants' performance on task-specific technical skills, medications used and behaviours displayed. At the end of the simulation, the participants completed an evaluation questionnaire. RESULTS: The manikin-based simulation examination was found to be a realistic, valid and reliable tool. Validity (i.e. face, content and construct) of the test instrument was evident. The level of inter-rater concordance of participants' clinical performance was good to excellent. The item analysis showed good to excellent internal consistency on all the performance scores except the post-test technical score. CONCLUSIONS: With a carefully designed rating instrument and simulation operation, the manikin-based simulation examination was shown to be reliable and valid. However, a further refinement of the test instrument will be required for higher stake examinations.

The biochemical and biological functions of human papillomavirus type 16 E5 protein.

Human papillomavirus type 16 (HPV-16) E5 protein, along with the more publicized E6 and E7 proteins of this virus, has been found to be oncogenic. E5 is a highly hydrophobic membrane-bound protein of 83 amino acids associated with the Golgi apparatus, endoplasmic reticulum, and nuclear membrane in infected cells. E5 can activate epidermal growth factor receptor (EGFR) through binding to the 16 kD subunit of protein pump ATPase leading to a reduced downregulation of EGFR receptors. The activation of EGFR can initiate biochemical cascades that lead to overexpression of a variety of protooncogenes and stimulate rapid cell growth. Moreover, E5 can inhibit the expression of tumor suppressor gene p21((WafI/SdiI/CipI)) and impair the control of cell cycle checkpoint. E5 protein has been identified as a potential tumor vaccine target antigen.

Primary cerebral anaplastic large cell lymphoma containing abundant reactive histiocytes and eosinophils. A case report and literature review.

Primary cerebral anaplastic large cell lymphoma (ALCL) is very rare. We report on our experience with such a case and review the literature. A 46-year-old Taiwanese woman presented with headache, weakness of her right extremity, and limited eye movement. A solid mass (5 cm x 4 cm) at the left occipital lobe was almost completely removed. The neoplastic cells, some of which had reniform or embryo-like nuclei, were large and were admixed with abundant eosinophils, histiocytes, and some small lymphocytes. These neoplastic cells expressed CD30, CD43, granzyme B and T-cell intracellular antigen-1, but not ALK1, CD3, CD20, CD45, CD79a, cytokeratin, and EMA. They were positive for Epstein-Barr virus-encoded mRNA by in situ hybridization. Polymerase chain reaction study of formalin-fixed tissue showed a clonal gene arrangement of the T-cell receptor-gamma chain. ALCL of T-cell lineage with cytotoxic phenotype was diagnosed. The patient received cranial irradiation and has remained with no evidence of disease for 25 months of follow-up.

Suppression of choroidal neovascularization by adeno-associated virus vector expressing angiostatin.

PURPOSE: To test the efficacy of a recombinant adeno-associated virus (rAAV) vector that expresses mouse angiostatin in suppressing experimental choroidal neovascularization (CNV) in a rat model. METHODS: An rAAV vector, rAAV-angiostatin, was constructed to deliver the mouse angiostatin gene. rAAV-angiostatin and a control virus, rAAV-lacZ, were delivered in vivo by subretinal injection in Brown Norway rats, and the delivery was confirmed by reverse-transcriptase polymerase chain reaction (RT-PCR). For a CNV suppression experiment, CNV was generated by fundus krypton laser photocoagulation 7 days after the viral vector injection and was evaluated by fluorescein angiography (FA) and histology. Apoptosis in retina was analyzed using the TUNEL assay. Inflammation in the retina was investigated by immunohistochemistry, using antibodies that recognize lymphocytes. RESULTS: rAAV-angiostatin injection led to sustained expression of the angiostatin gene in chorioretinal tissue for up to150 days. FA analysis revealed significant reduction of the average sizes of CNV lesions in rAAV-angiostatin-injected eyes when compared with rAAV-lacZ-injected eyes at both 14 (P = 0.019) and 150 (P = 0.010) days after injection. Moreover, histologic analysis of CNV lesions also revealed significantly smaller lesions in rAAV-angiostatin-injected eyes (P = 0.004). As for adverse effects, rAAV-angiostatin injection did not cause inflammation or apoptosis of cells in retina and choroid. CONCLUSIONS: This is the first report that subretinal injection of rAAV-angiostatin can significantly reduce the sizes of CNV lesions. This and the absence of apoptosis and inflammation in chorioretinal tissue indicate the feasibility of a gene therapy approach for treatment of CNV disease.