Oral Diseases (2012) Objective:  The aim of this study was to determine the expression of N-Glycolyl GM3 (NeuGcGM3) ganglioside in oral mucosal melanomas. Materials and Methods:  To assess the presence of cytidine monophospho-N-acetylneuraminic acid hydroxylase (CMAH) mRNA, an RT-PCR assay was performed in melanoma cell line (ME), an oral mucosal ME, and two fresh oral mucosal melanoma tissues. Expression of NeuGcGM3 ganglioside was evaluated by immunohistochemistry in 44 primary oral mucosal melanomas and 10 oral melanotic nevi. Also, the expression of NeuGcGM3 was examined in ME by immunocytochemistry. Results:  We first checked the expression of CMAH in ME and two fresh oral mucosal melanoma tissues. Presence of NeuGcGM3 ganglioside was evident in 37 of 44 cases (84.1%), showing a diffuse cytoplasmic and membranous staining. Patients with primary occurrence showed high levels of NeuGcGM3 ganglioside compared to patients with secondary occurrence. On the other hand, negative immunoreaction was evidenced in oral melanotic nevi. ME also presented the expression of NeuGcGM3 by immunocytochemistry. Conclusions:  In this work, we for the first time evaluated the expression of 14F7 MAb immunorecognition in oral mucosal melanomas. Our results were in agreement with the assumption that NeuGcGM3 ganglioside may be considered as target for passive and active immunotherapy in oral mucosal melanomas expressing this molecule and indicate less risk of recurrence and a better prognosis. Moreover, ME provides a platform for more studies on the specific function of NeuGcGM3 in oral mucosal melanomas.

This study was conducted to evaluate the effects of dietary factors that alter ruminal fermentability on intake, duodenal flows and intestinal digestibility of individual amino acids (AA) in lactating dairy cows. The experiment was designed as a 4×4 Latin square using 4 ruminally and duodenally cannulated lactating dairy cows. Treatments were arranged in a 2×2 factorial design; 2 forage particle lengths (FPL) of alfalfa silage (short and long) were combined with low (35:65) and high (60:40) forage-to-concentrate ratio (F:C; dry matter basis). Four diets were formulated using 2 cuts of alfalfa silage [short (7.9mm) and long (19.1mm)], combined with 2 ratios of forage to barley grain concentrate (35:65 and 60:40). Overall, the interactions between dietary F:C and FPL on intake, duodenal flows, and intestinal digestibility of AA were marginal. Intakes of total AA and nonessential AA were not different between low- and high-F:C diets, whereas that of essential AA tended to be less with high-F:C diet as a result of lower intakes of Met, Phe, Arg, and His. The flows of total AA and microbial AA were reduced by 22 and 19%, respectively, with increasing F:C ratio in the diets due to consistently reduced flows of individual AA, whereas AA profiles (% of AA-N) of the duodenal protein were not different. Altering F:C from 35:65 to 60:40 decreased the intestinal digestibility of Ile, Leu, Thr, Val, Ala, Cys, and Ser, and consequently, tended to decrease the digestibility of total AA, essential AA, and nonessential AA. Intakes of total AA, essential AA, and nonessential AA were overall not affected by dietary FPL so FPL did not affect the flows or intestinal digestibility of AA. These results indicate that increasing dietary F:C ratio decreased overall AA supply because flow to the duodenum and intestinal digestibility of AA were decreased. However, increasing FPL had no effect on AA supply. The measured duodenal flows of AA were consistent with the predictions of the Cornell Net Carbohydrate and Protein System model for the low-forage diet, and were consistent with the National Research Council model for the high-forage diet. Furthermore, the digestibility of individual AA in the intestine varied considerably, regardless of dietary treatment. The results revealed the necessity to consider the both flows and digestibility of individual AA when optimizing ration formulation to meet AA requirements of dairy cows.

Interleukin-15 (IL15) is a potential immunotherapeutic treatment for cancer. Caspy2 is an active zebra caspase for inducing apoptosis and immune response in murine tumors. In this study, we aim to evaluate the potential of gene therapy using IL15 and Caspy2 against the murine tumors. Plasmid expressing both Caspy2 and IL15 genes was constructed, encapsulated in DOTAP/cholesterol cationic liposome and injected intratumorally into the mice bearing CT26, B16-F10 and 4T1 carcinoma. We found that coexpression of IL15 and Caspy2 could significant inhibit tumor growth and prolong survival of the mice bearing CT26 or B16F10 tumor. A significant reduction in spontaneous lung metastasis was observed in the 4T1 tumor model. In CT26 model, the mice treated with IL15 and Caspy2 acquired a long-time protective immunity against the parental tumor cell rechallenge. Cytotoxic T lymphocytes and terminal deoxynucleotidyltransferase-mediated nick end labelling assays showed that the combination of capsy2 and IL15 could enhance both the apoptosis and immune response induction, which may account for its extraordinary antitumor effect. Furthermore, we showed that the observed tumor suppression by IL15 and Caspy2 concurred with the Caspy2-mediated downregulation of IL10 and upregulation of interferon-γ and tumor necrosis factor-α. Our results therefore suggested that the combination regimen might be a novel and effective strategy for cancer treatment.Cancer Gene Therapy advance online publication, 27 April 2012; doi:10.1038/cgt.2012.17.

The purpose of this study is to investigate the anti-osteoporotic effects of Radix Dipsaci total saponins (RTS). We showed that RTS was able to improve bone properties by either an increase of osteoblastic activity or a decrease in osteoclastic activity. INTRODUCTION: Radix Dipsaci has long been used as an anti-osteoporotic drug. The present study investigates the anti-osteoporotic effects of RTS. METHODS: Three-month-old female rats were randomly assigned into a sham-operated group (sham) and five ovariectomy (OVX) subgroups, namely, OVX with vehicle (OVX), OVX with 17β-ethinylestradiol (E(2)), and OVX with graded doses of RTS (50, 100, or 200 mg/kg/d). RTS and E(2) were administered orally, daily from 1 week after OVX treatment for 4 months. Bone mass, turnover, and strength were evaluated by dual-energy X-ray absorptiometry, biochemical markers, and the three-point bending test. The trabecular bone microarchitecture was assessed by microCT. In vitro experiments were performed to determine the potential molecular mechanisms of the anti-osteoporotic effect of RTS. RESULTS: RTS prevented the loss of bone mass induced by OVX. The preventive effect on bone loss was primarily indicated by decreasing levels of bone turnover markers and confirmed by the changes in urinary calcium and phosphorus excretion. The treatment also enhanced the biomechanical strength of bone and prevented the deterioration of trabecular bone microarchitecture. RTS induced MC3T3-E1 and primary osteoblastic cell maturation and differentiation and increased bone formation by increasing BMP-2 synthesis. In addition, RTS inhibited osteoclastogenesis through an increase in osteoprotegrin and a decrease in NF-kB ligand expression in vitro. CONCLUSIONS: RTS treatment can effectively suppress the loss of bone mass induced by OVX and in vitro evidence suggests this could be through actions on both osteoblasts and osteoclasts.

Background and study aims: The use of natural orifice transluminal endoscopic surgery (NOTES) for gastroenterostomy has been previously reported, but it remains technically challenging and additional assistance is often needed. The aim of this study was to develop and evaluate a novel method for the creation of a gastroenterostomy using NOTES with an occluder.Methods: Transgastric endoscopic gastroenterostomy was performed in 12 healthy female dogs using a therapeutic upper gastrointestinal endoscope and a partially covered occluder. The occluder was removed with a snare 1 week later. The patency of the gastroenterostomy was confirmed by endoscopy, contrast radiological study, necropsy, and histological examination after 2 weeks.Results: NOTES gastroenterostomy with an occluder was successful in all 12 dogs. The mean operative time was 32.3 ± 10.3 min (range 20.3 - 53.5). One dog (the first; 8.3 %) died 4 days after the operation of severe intra-abdominal infection due to incorrect deployment of the occluder and poor bowel preparation. Minor bleeding occurred at the anastomosis after removal of the occluder in two of the remaining dogs (18.2 %). Necropsy revealed postoperative adhesions that had developed at the anastomotic site in one dog (9.1 %). No anastomotic leakage or intestinal obstruction was observed. Complete healing of the anastomosis was confirmed on histological evaluation.Conclusion: Gastroenterostomy performed entirely by NOTES using an occluder was technically feasible in this survival animal model.

Purpose:To evaluate the prevalence of glaucoma in adults of the Bai Nationality populations in rural China.Methods:A population-based survey of Chinese Bai Nationality aged ≥50 years from randomly selected block groups in southwestern China was conducted. Eligible persons were invited to local examination sites for a complete ophthalmic examination. Glaucoma was diagnosed using the International Society of Geographical and Epidemiological Ophthalmology Classification scheme.Results:In the study, 2133 subjects (77.8% participation rate) were examined, with a crude prevalence of all glaucoma of 2.2%(95% confidence interval [CI], 1.6%-2.9%). Primary open angle glaucoma (POAG) was found in 1.0%(95%CI, 0.6%-1.6%) and primary angle-closure glaucoma (PACG) in 0.9%(95%CI, 0.6%-1.4%). The prevalence of all glaucoma was significantly higher in older people and women. Mean IOP was 16.17±3.74 mmHg (97.5th and 99.5th percentiles, 24 mmHg and 30 mmHg). The mean vertical cup-to-disc ratio (VCDR) was 0.43±0.17 (97.5th and 99.5th percentiles, 0.7 and 0.8). Unilateral blindness was found in 80% of PACG while only in 36.3% of POAG cases.Conclusions:Prevalence of POAG is similar to PACG in ethnic Bai population living in rural south-western China; PACG has a worse visual impairment and prognosis compared to POAG.

The jet-edge system using a vibrating element (liquid whistle) was investigated experimental using high-speed camera. The vibrating element vibrated periodically with a few millimeters displacement and cavitaion bubbles were observed near the tip of the vibrating element. The acoustic emission of the system was measured with hydrophones in some place. The low frequency and shock signals were observed in the pressure signal, and whistle can be heard at the same time. In frequency field, the line and broadband spectrum exist. The foundation frequency f0 of the vibrating element and harmonics compose the line spectrum. The former 10 order harmonics can be seen obviously. As the flow velocity increasing, the frequency f0 and the harmonic do not change, but the intensity of the high order harmonics and broadband spectrum become strong. Base the experimental result, we consider that the f0 is the foundation frequency of the vibrating element, and the harmonics and the continuous spectrum is the bubble cavitation emission. Acknowledgement: This work was supported by the National Natural Sciences Foundation of China (10804118)and he National Science and Technology Major Project of China (No. 2011ZX05032-003).

AIMS/HYPOTHESIS: Sirtuin-1 (SIRT1) is a potential therapeutic target to combat insulin resistance and type 2 diabetes. This study aims to identify a microRNA (miRNA) targeting SIRT1 to regulate hepatic insulin sensitivity. METHODS: Luciferase assay combined with mutation and immunoblotting was used to screen and verify the bioinformatically predicted miRNAs. miRNA and mRNA levels were measured by real-time PCR. Insulin signalling was detected by immunoblotting and glycogen synthesis. Involvement of SIRT1 was studied with adenovirus, inhibitor and SIRT1-deficient hepatocytes. The role of miR-181a in vivo was explored with adenovirus and locked nucleic acid antisense oligonucleotides. RESULTS: miR-181a targets the 3' untranslated region (3'UTR) of Sirt1 mRNA through a miR-181a binding site, and downregulates SIRT1 protein abundance at the translational level. miR-181a is increased in insulin-resistant cultured hepatocytes and liver, and in the serum of diabetic patients. Overexpression of miR-181a decreases SIRT1 protein levels and activity, and causes insulin resistance in hepatic cells. Inhibition of miR-181a by antisense oligonucleotides increases SIRT1 protein levels and activity, and improves insulin sensitivity in hepatocytes. Ectopic expression of SIRT1 abrogates the effect of miR-181a on insulin sensitivity, and inhibition of SIRT1 activity or SIRT1 deficiency markedly attenuated the improvement in insulin sensitivity induced by antisense miR-181a. In addition, overexpression of miR-181a by adenovirus impairs hepatic insulin signalling, and intraperitoneal injection of locked nucleic acid antisense oligonucleotides for miR-181a improves glucose homeostasis in diet-induced obesity mice. CONCLUSIONS/INTERPRETATION: miR-181a regulates SIRT1 and improves hepatic insulin sensitivity. Inhibition of miR-181a might be a potential new strategy for treating insulin resistance and type 2 diabetes.