Search

Opioids are widely prescribed analgesics, but their use is limited due to development of tolerance and addiction, as well as high variability in individual response. The development of improved opioid analgesics requires high-throughput functional assays to assess large numbers of potential opioid ligands. In this study, we assessed the ability of a proprietary "no-wash" fluorescent membrane potential dye to act as a reporter of µ-opioid receptor (MOR) activation and desensitization via activation of G-protein-coupled inwardly rectifying potassium channels. AtT-20 cells stably expressing mouse MOR were assayed in 96-well plates using the Molecular Devices FLIPR membrane potential dye. Dye emission intensity decreased upon membrane hyperpolarization. Fluorescence decreased in a concentration-dependent manner upon application of a range of opioid ligands to the cells, with high-efficacy agonists producing a decrease of 35% to 40% in total fluorescence. The maximum effect of morphine faded in the continued presence of agonist, reflecting receptor desensitization. The effects of opioids were prevented by prior treatment with pertussis toxin and blocked by naloxone. We have demonstrated this assay to be an effective method for assessing ligand signaling at MOR, which may potentially be scaled up as an additional high-throughput screening technique for characterizing novel opioid ligands. NHMRC Grant Numbers: 1011979 & 1045964

To investigate the nutritional status, and provide evidence for nutritional treatment option.A total of 452 patients with gastrointestinal cancer were selected, including 156 gastric cancer,117 colon cancer, and 180 rectal cancer. The nutritional risk screening 2002(NRS2002) was applied to grade the nutritional risk. A multi-frequency bioelectrical impedance analysis was used to measure the patients' body composition. Albumin (Alb), prealbumin(PA), transferring(Tf), retinol binding protein(RBP), red blood cell(RBC), hemoglobin (Hb), haematocrit(Hct) were measured after fasting.The rate of patients with NRS2002 score more than 3 was 70.5%(110/156) for gastric cancer, 53.8%(63/117) for colon cancer, and 46.7%(86/180) for rectal cancer. The score for impaired nutritional status more than 1 for gastric cancer was higher than that for colorectal cancer(P0.05), while patients with disease score more than 2 was less for gastric cancer(P0.05). Body mass index(BMI), obesity degree, fat content, fat percentage, and arm circumference were lower in gastric cancer patients as compared to colorectal cancer patients(P0.05); but protein percentage, muscle percentage, ratio of muscles of arm, and cell mass percentage were higher in gastric cancer patients(P0.05). The proportions of patients with low Alb, PA, Tf, BC, Hb, Hct were higher for gastric cancer and colon cancer(P0.05).Patients with gastric cancer are prone to fat loss and therefore have a higher nutritional risk and malnutrition than those with colorectal cancer. Combination of body composition analysis and laboratory examination may achieve comprehensive evaluation of the nutritional status of patients, and provide the evidence of nutritional therapy by being combined with NRS2002 score.

Gastric cancer (GC) is one of the most common types of cancer in the world. A change in the metabolism of lipids in tumor cells could lead to the pathogenesis of cancer. In this study, we investigated fatty acid and fatty acid amide metabolic perturbations associated with GC morbidity.Gas chromatography/mass spectrometry (GC/MS) was utilized to analyze fatty acids (FAs) and fatty acid amides (FAAs) of GC tissues and matched normal mucosae from 30 GC patients. Acquired lipid data was analyzed using non parametric Wilcoxon rank sum test to find the differential biomarkers for GC and diagnostic models for GC were established by using orthogonal partial least squares discriminant analysis (OPLS-DA).A total of 13 FAs and 4 FAAs were detected using GC/MS and 5 differential FAs as well as oleamide were identified with significant difference (P0.05). The OPLS-DA model generated from lipid profile showed adequate discrimination of GC tissues from normal mucosae while the OPLS-DA model failed to separate GC specimens of different TNM stages. A total of 8 variables were obtained for their most contribution in the discriminating model (Variable importance in the projection (VIP) value1.0), five of which were detected with significant difference (P0.05).FA and FAA metabolic profiles have great potential in detecting GC and helping understand perturbations of lipid metabolism associated with GC morbidity.

To investigate bone health conditions in 1637 aged women.From May 2004 to October 2008, Bone mineral density (BMD) of 1637 women at age of more than 60 years old were measured by Hologic DephiA dual energy X-ray absorptiometry (DXEA) in Huadong hospital affiliated to Fudan University. All data were compared and analyzed among each group which will be divided by every ten years. Those women were divided into groups on 10 years range. BMD of lumbar vertebral and hip bone, fracture incidence and bone turnover marker were compared and analyzed.(1) BMD: at age of ≥90, 80-89, 70-79, 60-69, BMD of the lumbar vertebral 2-4 (L2-4) values were (0.96±0.18), (0.90±0.20), (0.81±0.16), (0.83±0.14) g/cm2, respectively. There were significantly increased BMD of lumbar of women at the age of 80-89 and ≥90 year-old compared with those of 60-69 year-old (P0.05). At age of ≥90, 80-89, 70-79, 60-69 BMD of femur neck, Total, Torch, Ward's trianger were (0.60±0.11), (0.65±0.11), (0.47±0.09), (0.37±0.09) g/cm2; at age of 80-89 BMD of FN, Total, Torch, Ward's trianger were (0.57±0.10), (0.68±0.13), (0.48±0.11), (0.35±0.10) g/cm2; at age of 70-79 BMD of FN, Total, Torch, Ward's trianger were (0.57±0.10), (0.69±0.12), (0.49±0.10), (0.36±0.11) g/cm2; at age of 60-69 BMD of FN, Total, Torch, Ward's trianger were (0.63±0.10), (0.76±0.12), (0.54±0.10), (0.45±0.15) g/cm2; There were significantly decreased in BMD of hip at the age of 70-79, 80-89, ≥90 year-old compared with those of 60-69 year-old (P0.05). (2) Fracture incidence:one time fracture incidence at age of 60-69, 70-79, 80-89, ≥90 were 34.8% (242/695), 45.0% (296/658), 51.3% (137/267), 5/17. There were increasing trend of fracture in aged women. (3) Bone turnover marker of bone Gla protein (BGP) N-mid (N-midBGP) in serum and C-terminal telopeptide of type I collagen/Cr (CTX/Cr) in urine values were (17±5) µg/L, (106±56) µg/mmol at age of more than 90 years, (17±7) µg/L, (128±99) µg/mmol at age of 80-89 years, (21±14) µg/L, (182±173) µg/mmol at age of 70-79 years, (25±13) µg/L, (190±168) µg/mmol at age of 60-69 years. There were significant decreased trends of N-midBGP at age of 70-79, 80-89 compared with that of 60-year (P0.05). There were significant decreased trends of CTX/Cr 80-89 compared with that of 60-year (P0.05).There were significant decreased bone metabolism in aged women. The risk of hip fracture is significantly increased in aged women. Diagnosis of osteoporosis based on BMD of hip in aged women is more reliable.

To evaluate the influence of mutations in epidermal growth factor receptor (EGFR) gene on the prognosis of non-small cell lung cancer (NSCLC).A total of 107 NSCLC patients undergoing tumor resection from December 2005 to August 2009 at our hospital were recruited. Mutations of exons 18 - 21 of EGFR gene in patients were detected by polymerase chain reaction (PCR) amplification and gene sequencing. The influence of mutations in EGFR gene on the growth, metastasis and survival rate of NSCLC was evaluated.EGFR mutations were detected in 30 (28.0%) of 107 patients. The mutation distribution was as follows: exon 18 (n = 1), exon 19 (n = 8), exon 21 (n = 20), and exons 18 and 19 multiple (triple mutations, n = 1). NSCLC with EGFR mutations had a higher growth velocity than that without EGFR mutations [M(Q(1-3)): 0.42 (0.17 - 1.04) mm/d vs 0.21 (0.19 - 1.00) mm/d, P0.05]. EGFR mutations had no significant impaction on neither the lymph node metastasis of NSCLC (P0.05) nor on the tumor metastasis to other organs (P0.05). The mean survival time of patients with and without EGFR mutations were (18.2 ± 8.9) and (25.5 ± 7.8) months respectively. The 1, 2 and 3-year survival rate were 62.2% vs 72.2%, 47.7% vs 57.3% and 46.9% vs 56.3% respectively between patients with and without EGFR mutations. Log-rank test didn't show a significant difference among them (χ(2) = 0.59, P0.05).The EGFR mutations promote the growth of NSCLC. But it may not be a factor of predicting prognosis.

Recent genome-wide association studies reported that GCKR rs780094 polymorphism is associated with elevated fasting serum triglyceride levels and elevated levels of C-reactive protein (CRP). There are a ample of data on the association between circulating triglyceride, CRP concentrations and risk of non-alcoholic fatty liver (NAFLD). To determine whether the GCKR rs780094 polymorphism contributes to the development of non-alcoholic fatty liver, a case-control study was performed in 903 Chinese subjects. Among study population, 436 patients with B-mode ultrasound-proven NAFLD (318 with steatosis hepatis I°, 90 with steatosis hepatis II° and 28 with steatosis hepatis III°) and 467 controls were genotyped by using TaqMan allelic discrimination assays. We confirmed the association of GCKR rs780094 with NAFLD in Chinese people (OR = 1.607, 95% CI 1.139-2.271, P[dom] = 7.2 × 10(-3)). In this study, polymorphism in GCKR rs780094 was not significantly associated with the degree of fatty infiltration of the liver. In addition, the T-allele of GCKR rs780094 was significantly associated with increasing fasting triglyceride (P[add] = 3.8 × 10(-4)) and CRP (P[add] = 2.9 × 10(-4)) concentrations after adjusting for age, gender, and BMI. The association with NAFLD remained significant after adjustment for triglyceride, while adjustment for CRP abolished the association. Genetic variation in GCKR gene rs780094 polymorphism contributes to the risk of NAFLD in Chinese people. The effect of genotype on NAFLD is probably mediated through chronic low-grade systemic inflammation rather than through dislipidemia.

Techniques for the fast and accurate detection of bacterial infection are critical for early diagnosis, prevention and treatment of bacterial translocation in clinical severe acute pancreatitis (SAP). In this study, the availability of a real-time PCR method in detection of bacterial colonization in SAP rat models was investigated.Samples of blood, mesenteric lymph nodes (MLN), pancreas and liver from 24 specific pathogen-free rats (8 in a control group, 16 in a SAP group) were detected for bacterial infection rates both by agar plate culture and a real-time PCR method, and the results were made contrast.Bacterial infection rates of the blood, MLN, pancreas and liver in the SAP group and the control group by the two different methods were almost the same, which were 5/16, 12/16, 15/16, 12/16 in the SAP group compared with 0/8, 1/8, 0/8, 0/8 in the control group by agar plate culture, while 5/16, 10/16, 13/16, 12/16 and 0/8, 1/8, 0/8, 0/8 respectively by a real-time PCR method. Bacterial number was estimated by real-time PCR, which showed that in the same mass of tissues, the pancreas contained more bacteria than the other three kinds of organs in SAP rats (P0.01), that may be due to the edema, necrosis and hemorrhage existing in the pancreas, making it easier for bacteria to invade and breed.Fast and accurate detection of bacterial translocation in SAP rat models could be carried out by a real-time PCR procedure.

Background The development and progression of liver cancer may involve abnormal changes in DNA methylation, which lead to the activation of certain proto-oncogenes, such as c-myc, as well as the inactivation of certain tumor suppressors, such as p16. Betaine, as an active methyl-donor, maintains normal DNA methylation patterns. However, there are few investigations on the protective effect of betaine in hepatocarcinogenesis. Methods Four groups of rats were given diethylinitrosamine (DEN) and fed with AIN-93G diets supplemented with 0, 10, 20 or 40 g betaine/kg (model, 1%, 2%, and 4% betaine, respectively), while the control group, received no DEN, fed with AIN-93G diet. Eight or 15 weeks later, the expression of p16 and c-myc mRNA was examined by Real-time PCR (Q-PCR). The DNA methylation status within the p16 and c-myc promoter was analyzed using methylation-specific PCR. Results Compared with the model group, numbers and areas of glutathione S-transferase placental form (GST-p)-positive foci were decreased in the livers of the rats treated with betaine (P < 0.05). Although the frequency of p16 promoter methylation in livers of the four DEN-fed groups appeared to increase, there is no difference among these groups after 8 or 15 weeks (P > 0.05). Betaine supplementation attenuated the down-regulation of p16 and inhibited the up-regulation of c-myc induced by DEN in a dose-dependent manner (P < 0.01). Meanwhile, increases in levels of malondialdehyde (MDA) and glutathione S-transferase (GST) in model, 2% and 4% betaine groups were observed (P < 0.05). Finally, enhanced antioxidative capacity (T-AOC) was observed in both the 2% and 4% betaine groups. Conclusion Our data suggest that betaine attenuates DEN-induced damage in rat liver and reverses DEN-induced changes in mRNA levels.

To study the effect of betaine on the formation of atherosclerotic plaque in apolipoprotein E (ApoE)-deficient mice and explore its anti-inflammatory mechanism.Seven-week-old ApoE-deficient mice (C57BL/6J background) were divided into four groups randomly based on body weight: model group and three betaine groups. Wild-type mice with the same age and genetic background were used as control group. The control group and model group were fed AIN-93G diet. Three betaine groups were fed AIN-93G diet supplemented with 1, 2, 4 g betaine/100 g diet, respectively. Serum tumor necrosis factor-alpha (TNF-alpha), monocyte chemoattractant protein-1, lipid levels and methylation status of TNF-alpha promotor in aorta were determined at 0, 7 and 14 weeks. The percentage of aorta sinus plaque to lumen area was measured at 14-week.The percentage of aorta sinus plaque to lumen area of 1% and 2% betaine groups were (11.43+/-2.65)% and (12.09+/-3.07)%, respectively, which were 41% and 33% smaller than that of the model group (t=3.117, 3.010, respectively, and P0.01). Serum TNF-alpha level of three betaine groups were (56.33+/-3.86), (63.04+/-4.67) and (65.52+/-3.97) pg/ml, respectively, which were lower than that of the model group (79.40+/-4.68) pg/ml (t=9.270, 6.571 and 5.576, respectively, P0.001), but there was no significant difference in the methylation status of TNF-alpha promotor among all five groups.Betaine could inhibit the development of atherosclerosis via anti-inflammation.

AIMS/HYPOTHESIS: Recently, three groups independently reported that variation in MTNR1B, the gene encoding melatonin receptor 1B, was associated with an increased risk of type 2 diabetes, increased fasting plasma glucose and impaired insulin secretion in populations of European ancestry. In the present study, we investigated whether a single MTNR1B polymorphism was associated with type 2 diabetes in Han Chinese individuals, to elucidate whether this is a cross-populational effect. METHODS: The MTNR1B variant rs10830963 was genotyped in 1,165 type 2 diabetic patients and 1,105 normoglycaemic control individuals of southern Han Chinese ancestry who were residents of the metropolitan area of Shanghai. The risk of developing type 2 diabetes was calculated using a logistic regression model adjusted for age, sex and BMI. A possible association with fasting plasma glucose was analysed in the normoglycaemic control individuals using a multiple linear regression analysis with adjustments for age, sex and BMI. RESULTS: The genetic variant rs10830963 was associated with an increased risk of type 2 diabetes in our Han Chinese cohort (OR 1.16, 95% CI 1.03-1.31, p = 0.015). As previously described, the risk variant was also associated with increased fasting plasma glucose, showing an increase of 0.068 mmol/l (95% CI 0.036-0.100, p = 4 x 10(-5)) per risk allele. CONCLUSIONS/INTERPRETATION: A common variant in the MTNR1B gene is associated with an increased risk of type 2 diabetes and increased fasting plasma glucose in Han Chinese, suggesting an important role for this polymorphism in populations of different ethnic and environmental backgrounds.

As there are conflicting reports regarding the association between obstructive sleep apnoea (OSA) and cancer incidence and mortality, a meta-analysis was performed to evaluate whether OSA is independently associated with cancer incidence and mortality. Pubmed, EMBASE and Web of Science were searched up until November 2014. Studies that assessed OSA and the future risk of cancer incidence or mortality were included. Pooled hazard ratios (HR) and corresponding 95% confidence intervals (CI) were calculated. Subgroup analysis was conducted based on the polysomnographic variable, apnoea-hypopnoea index. Six studies, which involved 114 105 participants, were pooled in this meta-analysis. Fixed-effects analysis showed the pooled adjusted HR of cancer incidence as 0.91 (95% CI, 0.74-1.13; P = 0.408) for mild OSA, 1.07 (95% CI, 0.86-1.33; P = 0.552) for moderate OSA and 1.03 (95% CI, 0.85-1.26; P = 0.743) for severe OSA. Random-effects analysis demonstrated neither mild OSA (adjusted HR, 0.79; 95% CI, 0.46-1.34; P = 0.381), moderate OSA (adjusted HR, 1.92; 95% CI, 0.63-5.88; P = 0.251) nor severe OSA (adjusted HR, 2.09; 95% CI, 0.45-9.81; P = 0.349) correlated with cancer mortality. This meta-analysis indicates that OSA is not independently associated with cancer incidence and mortality according to currently available data. Additional experimental and human research is required to determine the exact association between OSA and cancer.

To investigate the direct electrochemical behaviors of matrine (MT), oxymatrine (OMT), sophoridine (SR) and oxysophoridine (OSR) at a glassy carbon electrode in a physiological medium, and to determine the contents of commercial MT and SR pharmaceutical products were determined.Their electrochemical behaviors and contents in 0.15 mol x L(-1) NaCl aqueous solutions were obtained by cyclic voltammetry (CV) and square wave voltammetry (SWV) methods.The electrochemical experimental results show that the two totally irreversible oxidation peaks for both MT and SR were observed, which potential values are 0.81 V and 0.92 V, and the electrode reaction processes were diffusion-controlled. But for both OMT and OSR, they have no oxidation peaks observed in the same condition as well. The linear range of the concentration of MT and SR and the contents in their products were obtained by SWV.The research results can be applied in determination of the contents of commercial MT and SR pharmaceutical products.

To evaluate the efficacy and safety of cefdinir in the treatment of patients with mild to moderate bacterial community acquired pneumonia (CAP).A prospective single-blind randomized controlled clinical study was performed comparing cefdinir with cefaclor in the treatment of sixty-four patients with CAP. The clinical and bacteriological efficacy and safety were compared between cefdinir and cefaclor in treating mild to moderate CAP. Thirty-three patients were treated with cefdinir 100 mg, orally, three times a day (cefdinir group), thirty-one patients were treated with cefaclor 500 mg, orally, three times a day (cefaclor group). In both groups 7-14 d was a treatment course.The cure rate of cefdinir and cefaclor was 84.8% and 77.4% respectively and the overall efficacy rate was 93.9% and 87.1% respectively. The bacterial positive rates and bacterial eradication rates of the two groups were 81.8%, 80.7% and 96.3%, 88%, respectively. The adverse drug reaction rate were 3% in cefdinir group and 6.5% in cefaclor group. There was no statistical significant difference between the two groups for the above results (P0.05). The time of given medicine of cefdinir and cefaclor was (10.8 +/- 1.6) d and (12.1 +/- 1.7) d (P0.01) respectively.cefdinir is safe and effective, shorten the course of treatment in the treatment of mild to moderate bacterial community acquired pneumonia.

The title compound, K+·C7H3N2O6−, crystallized from aqueous solution, is not isostructural with sodium 3,5-dinitro­benzoate. Both the anion and the cation lie on a twofold axis. The K+ cation is surrounded by eight O atoms and the crystal structure is stabilized by π–π inter­actions.

Background The objective of this study was to systematically evaluate the contribution of the insulin-like growth factor 2 mRNA-binding protein 2 (IGF2BP2) to type 2 diabetes mellitus (T2DM) and its interaction with obesity to T2DM susceptibility. Methods To clarify whether IGF2BP2 is an independent risk factor for T2DM in Chinese population, we conducted a study with a total of 2,301 Chinese Han subjects, including 1,166 T2DM patients and 1,135 controls, for the genotype of a most common and widely studied polymorphism—rs4402960 of IGF2BP2. Genotyping was performed by iPLEX technology. Gene and environment interaction analysis was performed by using multiple logistic regression models. Results The repeatedly confirmed association between IGF2BP2 (rs4402960) and T2DM had not been replicated in this cohort (P = 0.182). Interestingly, we found that obese subjects (body mass index (BMI) ≥ 28.0 kg/m2) bearing the minor A allele had an increased risk to develop T2DM (P = 0.008 for allele analysis and P