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.

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.

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.

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.

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.

Transrectal sonographic cystourethrography: studies in stress urinary incontinence.

Transrectal ultrasonic sagittal transducer is an excellent modality to image the bladder and urethra in dynamic change. In female patients, we found it is also helpful for the diagnosis of urinary stress incontinence. The posterior urethrovesical (PUV) angle is measured with the transrectal sonoprobe under strain and non-strain conditions. We compared the results of sonographic cystourethrogram with the radiographic chain cystourethrogram. The sonographic cystourethrography is superior to the radiography. The former may estimate the PUV angle accurately and differentiate between the patients with and without stress urinary incontinence. Furthermore, we also use the transrectal sagittal probe intraoperatively to adjust the suspension force as well as PUV angle in patients who underwent vesical neck suspension for stress urinary incontinence (SUI).

Fertilization and embryo cleavage after intracytoplasmic spermatid injection in an obstructive azoospermic patient with defective spermiogenesis.

OBJECTIVE: To achieve fertilization and cleavage by spermatids without tails from testicular biopsy. DESIGN: Clinical trial. SETTING: Reproductive unit of a university teaching hospital. PATIENT: A patient of obstructive azoospermia with defective spermiogenesis. INTERVENTION: Testicular biopsy after scrotal exploration and spermatid injection into the cytoplasm of oocyte. MAIN OUTCOME MEASURES: Fertilization and cleavage. RESULTS: Four of 13 spermatid injections achieved normal fertilization and 2 of them cleaved. CONCLUSION: Intracytoplasmic spermatid injection may be a possible treatment for patients of defective spermiogenesis who have no viable spermatozoa available.

Abnormal transcripts of FHIT gene in thyroid cancer.

Abnormalities of the FHIT gene were found in many carcinoma cell lines and human tumors as reported which suggest an etiological function in tumorigenesis. To investigate whether the FHIT gene is a target of thyroid tumor specific 3p alterations, we screened the FHIT gene for alteration in thyroid tumors and found that the tumors exhibited aberrant FHIT gene expression. The complete sequence of the FHIT gene in seven cases was determined and deletions between exon 2-9 in different regions were found. Goiter samples as control had normal FHIT transcripts while both normal and aberrant transcripts of FHIT were found not only in the tumor samples but also in the adjacent non-tumorous portion of the thyroid tumor. Our results support the hypothesis that FHIT gene alteration is involved in tumorigenetic development of human neoplasms in thyroid glands.

Abnormal transcripts of FHIT gene in Chinese brain tumors.

FHIT located at chromosome 3p14.2 was discovered and proposed as a candidate tumor suppressor gene in several cancers. To determine whether the FHIT gene at 3p14.2 is altered in Chinese brain tumors, we examined 13 brain tumors for deletions within FHIT locus. Evaluation of the FHIT gene in the panel of brain tumors led to a comprehensive mutation analysis. The complete sequence of the FHIT gene was determined and deletions between exon 5-8 were found in all 13 cases. In addition, single point mutation of amino acid from two glioblastoma and one atypical meningioma cases and multiple amino acid mutations from one pituitary tumor were observed. Our results support the hypothesis that FHIT gene alteration is involved in tumorigenic development of human neoplasms.

A pilot study of AFL-T (doxorubicin, 5-fluorouracil, leucovorin, and tamoxifen) combination chemotherapy for hormone-refractory prostate cancer.

BACKGROUND: Tamoxifen had been used to treat advanced prostate cancer with limited success. In vitro data suggested that tamoxifen could enhance the cytotoxic effect of chemotherapeutic agents, including doxorubicin, on prostate cancer cell lines. We applied this observation into a phase II trial for patients with hormone refractory prostate cancer (HRPC). PATIENTS AND METHODS: The AFL-T regimen consisted of doxorubicin 30 mg/m2/day on day 1; 5-FU 2,000 mg/m2/day 24-hour infusion and leucovorin 200 mg/m2/day 24-hour infusion on days 15 and 29; tamoxifen 50 mg/m2 four times daily on days 1, 2, 16, 17, 30, and 31. The protocol was designed to be of low dose-intensity and tolerable to most HRPC patients who may have reduced bone marrow reserve and poor renal function. Between Feb. 1994 and April 1999, 17 patients (median age 67, range 60-81) with HRPC were enrolled. Extensive hormonal manipulations had been done prior to the chemotherapy. Three patients had measurable diseases, 14 had only bone metastases, and all had elevated PSA levels (median 498 ng/ml, range 7.4-3,970 ng/ml). RESULTS: All 17 patients were eligible for analysis of toxicity. ECOG Grade III/IV leukopenia and thrombocytopenia occurred in 1 and 3 patients, respectively. There was no febrile neutropenia; there was no treatment-related mortality. Grade III/IV nausea, vomiting, mucositis, and diarrhea were noted in 0, 0, 1 and 0 patient, respectively. There was no venous thrombosis. One partial response, 1 stable disease, and 1 progressive disease were found in the three patients with measurable lesions. Eleven of the 17 patients (64.7%, 95% confidence interval: 41-88%) who were eligible for the evaluation of PSA response (PSA decrease > 50% for at least 6 weeks) were responders. The median overall and progression-free survivals were 13 and 7 months, respectively. Seventy-six percent of patients showed decreased analgesic usage or enhanced performance status. CONCLUSION: AFL-T, that has a low toxicity profile, is comparable to most other active regimens in terms of the PSA response rate. Randomized trials are needed to determine if there exists a survival benefit for patients with HRPC.

Intractable hiccup due to multiple sclerosis: MR imaging of medullary plaque.

We describe a 31-year-old man with clinically definite multiple sclerosis who presented with intractable hiccup for one month prior to admission. The responsible lesion was confirmed by magnetic resonance imaging to be a plaque in the ventral paramedian portion of the medulla oblongata.

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.

Emergency separation of omphaloischiopagus tetrapus conjoined twins in the newborn period.

This article reports an experience on subemergent separation of xyphoomphaloischiopagus tetrapus conjoined twins in the newborn period when survival of one of the twins was threatened by a severely malformed counterpart.

A new sperm collection method for treatment of retrograde ejaculation.

Retrograde ejaculation is an uncommon but treatable form of male infertility. Successful recovery of motile spermatozoa from a post-ejaculatory urine for artificial insemination is dependent on careful regulation of osmolarity and acidity of the bladder contents into which the ejaculate emanates, and the separation of the motile spermatozoa from the debris and cells which are found in these specimens. A pregnancy established by artificial insemination of spermatozoa recovered from bladder contents of a patient suffering from retrograde ejaculation is presented. The techniques for preparing the urinary bladder for spermatozoal survival and the removal of debris and cells by delicate centrifugation are discussed.

Measurement of viscosity of human semen with a rotational viscometer.

The viscosity of 130 human seminal plasma samples was studied with a rotational viscometer instead of by the traditional subjective rating method. The average seminal viscosity one hour after ejaculation was 9.35 +/- 0.99 centipoise (cps), which was statistically identical to that in the third hour (8.63 +/- 0.77 cps). Seminal viscosity showed a significant negative correlation with the percentage of motile sperm (p < 0.05); however, no significant correlation between seminal viscosity and sperm concentration could be found (r = -0.15, p = 0.098). The seminal viscosity of the oligoasthenospermic group was significantly higher than that of the normospermic group (p < 0.01); there was also a trend towards higher viscosity in the semen of asthenospermia and oligospermia when compared with the normospermic group. It is concluded that seminal viscosity may be higher in cases of poor-quality semen; however, sperm motility and concentration are not the sole determinants of viscosity. Determining the seminal viscosity with this rapid, objective and quantitative method is valuable in identifying and treating the subgroup of infertile men with viscid semen.

Toxic megacolon secondary to infective colitis in children.

BACKGROUND AND PURPOSE: Toxic megacolon is a fulminating and potentially lethal complication of severe colitis. Toxic megacolon secondary to infective colitis in children is rare. We analyzed the clinical course, pathology, treatment, and outcome of toxic megacolon secondary to infective colitis in children. METHODS: The medical records of all 20 children treated for infective colitis complicated with toxic megacolon during a 12-month (October 1997-October 1998) period were retrospectively reviewed. RESULTS: There were 10 boys and 10 girls, with a mean (+/- standard deviation, SD) age of 26.2 +/- 12.9 months (range, 6-57 mo). With an initial presentation of nonspecific gastroenteritis syndrome lasting several days, the disease progressed rapidly. In the acute stage, most patients developed toxic signs such as mental change, ranging from irritability to stupor (20, 100%), fever (19, 95%), tachycardia (20, 100%), abdominal distension (20, 100%), and abnormal stool pattern (19, 95%). Initial investigations revealed anemia (11, 55%), leukocytosis (11, 55%), and elevated levels of C-reactive protein ranging from 25.0 mg/L to 483.0 mg/L with a mean +/- SD of 185.7 +/- 129.1 mg/L (normal range, < 8 mg/L) (20, 100%). Salmonella enteritidis (12 patients, 60%) and Clostridium difficile (1, 5%) were isolated from stool samples in some cases. Plain abdominal x-rays revealed severe colonic dilatation. Prolonged hospitalization (mean, 33.6 d) and intensive therapy including a combination of broad-spectrum antibiotics, physical decompression, and total parenteral nutrition were necessary. Three patients (15%) underwent surgical management; the pathologic findings in these patients demonstrated severe transmural inflammation. We believe that bacterial and/or endotoxin translocation played an important role in gut failure. Three patients (15%) in the study died. CONCLUSION: Toxic megacolon in infective colitis is a fulminating illness that has a high mortality rate. The disease course can be divided into three stages: the acute toxic stage, the gut failure stage, and the convalescence or deterioration stage. Early diagnosis and aggressive management are important.

Ileal neobladder reconstruction after radical cystoprostatectomy.

To improve the outcome and patient acceptance of bladder substitution, five male patients underwent O-shaped ileal neobladder reconstruction, after radical cystoprostatectomy for invasive bladder cancer, with a mean follow-up of 11 months. With only 40 cm of ileal segment, an O-shaped neobladder was constructed after complete detubularization. Bilateral ureters were implanted using the Le Duc-Camey method. Six months after operation, all patients were totally continent during the day time, and one patient suffered from mild incontinence at night, which could be overcome by waking to void once or twice. The satisfactory continence levels are in agreement with a urodynamic study of the neobladder which showed a low pressure, high-capacity (mean, 456 mL) reservoir in the cystometric tracings. The mean maximal flow rate was 22.2 mL/sec, the mean residual urine was minimal (10 to 20 mL), the mean maximal urethral closing pressure was 74.4 cm H2O and the mean functional profile length was 2.9 cm. All renal units do not have neovesico-ureteral reflux. Two patients showed unilateral hydronephrosis which subsided later, one patient sustained bilateral hydronephrosis and died of jejunal perforation five months postoperatively. There were few perioperative complications and no patient expressed regret at having undergone the procedure. We consider bladder substitution to be the treatment of choice in male patients requiring radical cystoprostatectomy.