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Grape seed procyanidin B2 ameliorates mitochondrial dysfunction and inhibits apoptosis via the AMP-activated protein kinase-silent mating type information regulation 2 homologue 1-PPAR? co-activator-1? axis in rat mesangial cells under high-dose glucosamine.
Br. J. Nutr.
PUBLISHED: 11-19-2014
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Grape seed procyanidin B2 (GSPB2), an antioxidative and anti-inflammatory polyphenol in grape seed, has been found to have protective effects on diabetic nephropathy. Based on its favourable biological activities, in the present study, we aimed to investigate whether GSPB2 could inhibit apoptosis in rat mesangial cells treated with glucosamine (GlcN) under high-dose conditions. The results showed that the administration of GSPB2 (10 ?g/ml) significantly increased the viability of mesangial cells treated with GlcN at a dose of 15 mm. We found that GSPB2 inhibited apoptosis in mesangial cells using terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphates (dUTP) nick-end labelling staining and flow cytometry technique (P< 0·05 for both). GSPB2 treatment also suppressed oxidative stress by elevating the activity of glutathione peroxidase (P< 0·05) and superoxide dismutase (P< 0·01), as well as prevented cellular damage. GSPB2 enhanced the mRNA expression of nuclear respiratory factor 1, mitochondrial transcription factor A and mitochondrial DNA copy number in mesangial cells as determined by real-time PCR (P< 0·05 for each). Finally, GSPB2 treatment activated the protein expression of PPAR? co-activator-1? (PGC-1?), silent mating type information regulation 2 homologue 1 (SIRT1) and AMP-activated protein kinase (AMPK) in mesangial cells. These findings suggest that GSPB2 markedly ameliorates mitochondrial dysfunction and inhibits apoptosis in rat mesangial cells treated with high-dose GlcN. This protective effect could be, at least in part, due to the activation of the AMPK-SIRT1-PGC-1? axis.
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A Splice Variant of the Human Ion Channel TRPM2 Modulates Neuroblastoma Tumor Growth Through HIF-1/2?
J. Biol. Chem.
PUBLISHED: 11-14-2014
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The calcium permeable ion channel TRPM2 is highly expressed in a number of cancers. In neuroblastoma, full length TRPM2 (TRPM2-L) protected cells from moderate oxidative stress through increased levels of Forkhead transcription factor 3a (FOXO3a) and superoxide dismutase 2 (MnSOD). Cells expressing dominant negative short isoform (TRPM2-S) had reduced FOXO3a and MnSOD levels, reduced calcium influx in response to oxidative stress, and enhanced ROS, leading to decreased cell viability. Here, in xenografts generated with SH-SY5Y neuroblastoma cells stably expressing TRPM2 isoforms, growth of tumors expressing TRPM2-S was significantly reduced compared to tumors expressing TRPM2-L. Expression of HIF-1/2? was significantly reduced in TRPM2-S expressing tumor cells, as was expression of target proteins regulated by HIF-1/2? including those involved in glycolysis (LDHA, ENO2), oxidant stress (FOXO3a), angiogenesis (VEGF), mitophagy and mitochondrial function (BNIP3 and NDUFA4L2), and mitochondrial electron transport chain activity (complex IV, cytochrome oxidase 4.1/4.2). The reduction in HIF-1/2? was mediated both through significantly reduced HIF-1/2? mRNA levels and increased levels of von Hippel-Lindau in TRPM2-S expressing cells. Inhibition of TRPM2-L by pretreatment with clotrimazole or expression of TRPM2-S significantly increased sensitivity of cells to doxorubicin. Reduced survival of TRPM2-S expressing cells after doxorubicin was rescued by gain of HIF-1 or 2? function. These data suggest that TRPM2 activity is important for tumor growth, and cell viability and survival following doxorubicin, and that interference with TRPM2-L function may be a novel approach to reduce tumor growth through modulation of HIF-1/2?, mitochondrial function and mitophagy.
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[The expression of vascular endothelial growth factor antibody in serum of coal workers with pneumoconiosis].
Sichuan Da Xue Xue Bao Yi Xue Ban
PUBLISHED: 10-25-2014
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To determine the role of serum VEGF-Ab in pneumoconiosis of coal workers.
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15d-Prostaglandin J2 Protects Cortical Neurons Against Oxygen-Glucose Deprivation/Reoxygenation Injury: Involvement of Inhibiting Autophagy Through Upregulation of Bcl-2.
Cell. Mol. Neurobiol.
PUBLISHED: 08-01-2014
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We have previously shown that PPAR-? agonist 15d-PGJ2 inhibited neuronal autophagy after cerebral ischemia/reperfusion injury. However, the underlying mechanism of its regulatory role in neuronal autophagy remains unclear. This study was designed to test the hypothesis that 15d-PGJ2 upregulated Bcl-2 which binds to Beclin 1, and thereby inhibits autophagy. We performed cell viability assay, cytotoxicity assay, western blot, and co-immunoprecipitation to analyze autophagy activities in vitro model of oxygen-glucose deprivation/reoxygenation (OGD/R). OGD/R induced autophagy in cultured cortical neurons. 15d-PGJ2 treatment significantly decreased LC3-II/LC3-I ratio and Beclin 1 expression, but increased p62 expression. Autophagic inhibitor 3-methyladenine decreased LC3-II levels, increased neuronal cell viability, and mimicked some protective effect of 15d-PGJ2 against OGD/R injury. OGD/R-induced autophagy coincided with decreases in Bcl-2 expression and increases in Beclin 1 expression. 15d-PGJ2 treatment upregulated Bcl-2 expression and decreased Beclin 1 expression, and inhibit the dissociation of Beclin1 from Bcl-2 significantly. Bcl-2 siRNA abrogated the effect of 15d-PGJ2 on Beclin 1, LC3-II and p62, and influence cell viability and LDH level, while scRNA did not. PPAR-? agonist 15d-PGJ2 exerts neuroprotection partially via inhibiting neuronal autophagy after OGD/R injury. The inhibition of autophagy by 15d-PGJ2 is mediated through upregulation of Bcl-2.
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Revealing carbon nanodots as coreactants of the anodic electrochemiluminescence of Ru(bpy)?²?.
Anal. Chem.
PUBLISHED: 07-24-2014
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Recently, research on carbon nanodots (C-dots), a new type of luminescent nanoparticles with superior optical properties, biocompatibility, and low cost, has been focused on exploring novel properties and structure-related mechanisms to extend their scope. Herein, electrochemiluminescence, a surface-sensitive tool, is used to probe the unrevealed property of carbon nanodots which is characterized by surface oxygen-containing groups. Together with chemiluminescence, carbon nanodots as the coreactants for the anodic electrochemiluminescence of Ru(bpy)3(2+) are demonstrated for the first time. During the anodic scan, the benzylic alcohol units on the C-dots surface are oxidized "homogeneously" by electrogenerated-Ru(bpy)3(3+) to form reductive radical intermediate, which further reduce Ru(bpy)3(3+) into Ru(bpy)3(2+)* that produces a strong ECL emission. This work has provided an insight into the ECL mechanism of the C-dots-involved system, which will be beneficial for in-depth understanding of some peculiar phenomena of C-dots, such as photocatalytic activity and redox properties. Moreover, because of the features of C-dots, the ECL system of Ru(bpy)3(2+)/C-dots is more promising in the bioanalysis.
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Electrical resistivity behaviors of liquid Pb-Sn binary alloy in the presence of ultrasonic field.
Ultrasonics
PUBLISHED: 07-22-2014
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Electrical resistivity behaviors of liquid Pb-Sn alloys have been investigated in the presence of ultrasonic field. The process demonstrated significantly that electrical resistivity could reveal the precise influence caused by ultrasound. Details revealed by applying the resistivity measuring approach to the liquid Pb-Sn alloy show that the short ordered structures in the liquid could be modified by ultrasonic irradiation, and the resistivity approach could have application value in the ultrasonic irradiation process on the specific liquid metals and alloys.
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High expression of CAI2, a 9p21-embedded long noncoding RNA, contributes to advanced-stage neuroblastoma.
Cancer Res.
PUBLISHED: 07-17-2014
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Neuroblastoma is a pediatric cancer with significant genomic and biologic heterogeneity. p16 and ARF, two important tumor-suppressor genes on chromosome 9p21, are inactivated commonly in most cancers, but paradoxically overexpressed in neuroblastoma. Here, we report that exon ? in p16 is also part of an undescribed long noncoding RNA (lncRNA) that we have termed CAI2 (CDKN2A/ARF Intron 2 lncRNA). CAI2 is a single-exon gene with a poly A signal located in but independent of the p16/ARF exon 3. CAI2 is expressed at very low levels in normal tissue, but is highly expressed in most tumor cell lines with an intact 9p21 locus. Concordant expression of CAI2 with p16 and ARF in normal tissue along with the ability of CAI2 to induce p16 expression suggested that CAI2 may regulate p16 and/or ARF. In neuroblastoma cells transformed by serial passage in vitro, leading to more rapid proliferation, CAI2, p16, and ARF expression all increased dramatically. A similar relationship was also observed in primary neuroblastomas where CAI2 expression was significantly higher in advanced-stage neuroblastoma, independently of MYCN amplification. Consistent with its association with high-risk disease, CAI2 expression was also significantly associated with poor clinical outcomes, although this effect was reduced when adjusted for MYCN amplification. Taken together, our findings suggested that CAI2 contributes to the paradoxical overexpression of p16 in neuroblastoma, where CAI2 may offer a useful biomarker of high-risk disease.
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Integrating novel chemical weapons and evolutionarily increased competitive ability in success of a tropical invader.
New Phytol.
PUBLISHED: 07-08-2014
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The evolution of increased competitive ability (EICA) hypothesis and the novel weapons hypothesis (NWH) are two non-mutually exclusive mechanisms for exotic plant invasions, but few studies have simultaneously tested these hypotheses. Here we aimed to integrate them in the context of Chromolaena odorata invasion. We conducted two common garden experiments in order to test the EICA hypothesis, and two laboratory experiments in order to test the NWH. In common conditions, C. odorata plants from the nonnative range were better competitors but not larger than plants from the native range, either with or without the experimental manipulation of consumers. Chromolaena odorata plants from the nonnative range were more poorly defended against aboveground herbivores but better defended against soil-borne enemies. Chromolaena odorata plants from the nonnative range produced more odoratin (Eupatorium) (a unique compound of C. odorata with both allelopathic and defensive activities) and elicited stronger allelopathic effects on species native to China, the nonnative range of the invader, than on natives of Mexico, the native range of the invader. Our results suggest that invasive plants may evolve increased competitive ability after being introduced by increasing the production of novel allelochemicals, potentially in response to naïve competitors and new enemy regimes.
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Grape seed proanthocyanidin extracts ameliorate podocyte injury by activating peroxisome proliferator-activated receptor-? coactivator 1? in low-dose streptozotocin-and high-carbohydrate/high-fat diet-induced diabetic rats.
Food Funct
PUBLISHED: 06-20-2014
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Podocytes are part of the glomerular filtration membrane in kidney and serve to prevent the filtration of protein from the blood. Several evidences suggest that mitochondrial dysfunction plays a critical role in the pathogenesis of diabetic nephropathy and it is an early event in podocyte injury. Mitochondrial dysfunction promotes oxidative stress that can favor the development of podocyte injury. Peroxisome proliferator-activated receptor-? coactivator 1? (PGC-1?) was considered to be a major regulator of metabolic homeostasis and mitochondrial function. Some studies indicated that polyphenols may improve mitochondrial dysfunction, maintain the podocyte integrity and have therapeutic effects on glomerular diseases by promoting PGC-1? expression. Our study investigated whether grape seed proanthocyanidin extracts (GSPE), a strong antioxidant, ameliorate podocyte injury by activating PGC-1? in low-dose streptozotocin-and high-carbohydrate/high-fat diet-induced diabetic rats. After 16 weeks of GSPE treatment, GSPE slightly increased the body weight and decreased plasma glucose, food intake, water intake and urine volume in diabetic rats. Further, GSPE significantly decreased 24 h albumin levels and increased the expression of nephrin and podocalyxin. The antioxidant levels were improved and the cellular damage of kidney in diabetic rats was also relieved effectively after the treatment. Moreover, GSPE increased the mRNA expression of mitochondrial biogenesis factors and mitochondrial DNA content. Finally, GSPE activated the expression of PGC-1?, silent mating type information regulation 2 homolog 1 (SIRT1) and AMP-activated protein kinase (AMPK). These results suggest that GSPE ameliorate podocyte injury in diabetic nephropathy by the activation of AMPK-SIRT1-PGC-1? signalling, which appears to inhibit oxidative stress and mitochondrial dysfunction in the kidney.
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MicroRNA-155 influences B-cell receptor signaling and associates with aggressive disease in chronic lymphocytic leukemia.
Blood
PUBLISHED: 06-09-2014
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High-level leukemia cell expression of micro-RNA 155 (miR-155) is associated with more aggressive disease in patients with chronic lymphocytic leukemia (CLL), including those cases with a low-level expression of ?-chain-associated protein of 70 kD. CLL with high-level miR-155 expressed lower levels of Src homology-2 domain-containing inositol 5-phosphatase 1 and were more responsive to B-cell receptor (BCR) ligation than CLL with low-level miR-155. Transfection with miR-155 enhanced responsiveness to BCR ligation, whereas transfection with a miR-155 inhibitor had the opposite effect. CLL in lymphoid tissue expressed higher levels of miR155HG than CLL in the blood of the same patient. Also, isolated CD5(bright)CXCR4(dim) cells, representing CLL that had been newly released from the microenvironment, expressed higher levels of miR-155 and were more responsive to BCR ligation than isolated CD5(dim)CXCR4(bright) cells of the same patient. Treatment of CLL or normal B cells with CD40-ligand or B-cell-activating factor upregulated miR-155 and enhanced sensitivity to BCR ligation, effects that could be blocked by inhibitors to miR-155. This study demonstrates that the sensitivity to BCR ligation can be enhanced by high-level expression of miR-155, which in turn can be induced by crosstalk within the tissue microenvironment, potentially contributing to its association with adverse clinical outcome in patients with CLL.
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Effect of oat intake on glycaemic control and insulin sensitivity: a meta-analysis of randomised controlled trials.
Br. J. Nutr.
PUBLISHED: 04-30-2014
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The present meta-analysis of randomised controlled trials (RCT) aimed to investigate the effect of oat intake on glycaemic control and insulin sensitivity. A literature search was carried out in PubMed, ScienceDirect Online and The Cochrane Library (up to October 2013) for RCT that assessed the effect of oat intake on glucose control and insulin sensitivity. A total of fifteen articles with 673 subjects met the inclusion criteria. A random-effects model was used when the overall pooled studies exhibited significant heterogeneity. Otherwise, a fixed-effects model was used. Compared with controls, oat intake significantly reduced the concentrations of fasting insulin by - 6·29 (95 % CI - 12·32, - 0·27) pmol/l (P= 0·04) and the values of glucose AUC (GAUC; 0-120 min) by - 30·23 (95 % CI - 43·65, - 16·81) min × mmol/l (P< 0·0001). There was a slight decrease in fasting glucose concentrations, glycated Hb concentrations and homeostatic model assessment-insulin resistance values in subjects who consumed oats, but the difference was not significant. In conclusion, oat intake significantly lowers fasting insulin concentrations and GAUC values. To further investigate the effect of oat intake on fasting glucose concentrations, additional long-term and high-quality RCT with a parallel design are required.
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Rescue of defective MC4R cell-surface expression and signaling by a novel pharmacoperone Ipsen 17.
J. Mol. Endocrinol.
PUBLISHED: 04-29-2014
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Melanocortin 4 receptor (MC4R) is a key factor in regulating energy homeostasis, and null mutations occurring in the gene encoding MC4R cause severe early-onset morbid obesity in humans. Many obesity-causing mutations affecting MC4R clinically identified so far lead to failure of mutant receptors to shuttle to the plasma membrane. In this study, we show that a novel human MC4R antagonist, Ipsen 17, acted as an pharmacological chaperone of human MCR4. As tested with 12 obesity-causing human MC4R variants including S58C, E61K, N62S, I69T, P78L, C84R, G98R, T162I, R165W, W174C, C271Y, and P299H, Ipsen 17 was found to be the most universal pharmacological chaperone of MC4R reported so far because it can completely rescue nearly all mutant receptors (except P299H) with the highest potency (an EC50 value of approximately 10(-8)?M) and efficiency when compared with results for other tested pharmacological chaperones of MC4R including ML00253764, PBA, MTHP, PPPone, MPCI, DCPMP, and NBP described in the literature. Once restored to the plasma membrane, defective human MC4R variants responded to ?-MSH stimulation with an EC50 value of approximately 10(-8)?M and displayed dramatically enhanced signaling ability (except for G98R) in a mutant-specific efficacy and potency profile. Taken together, these results indicate that Ipsen 17 represents a candidate for the development of a targeted treatment of severe early-onset morbid obesity caused by a large subset of inherited mutations in the human MC4R gene.
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Effects of analog P165 of amyloid precursor protein 5-mer peptide on learning, memory and brain insulin receptors in the rat model of cognitive decline.
Neurol. Sci.
PUBLISHED: 03-28-2014
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We aim to study the therapeutic efficacy of analog P165 of amyloid precursor protein 5-mer peptide in streptozotocin (STZ)-induced cognitive decline model. Rats were divided into four groups: control, STZ, STZ+P165, and STZ+rosiglitazone (RSG). STZ model was established by intracerebroventricular injection of STZ. Three weeks following surgery, rats received daily gavage administration of distilled water (control and STZ groups), P165 (STZ+P165), or RSG (STZ+RSG) for four consecutive weeks. Learning and memory abilities were assessed with the Morris water maze test. Insulin-like growth factor-1 (IGF-1) was detected by ELISA. Expressions of insulin receptor-? (IR-?), insulin receptor substrate-1 (IRS-1), serine/threonine kinase (Akt), and phosphorylation of CREB (p-CREB) were observed by immunohistochemistry. Both P165 and RSG significantly reduced the escape latency relative to the STZ group (P165, P < 0.05; RSG, P < 0.01). STZ model rats had reduced levels of IGF-1 relative to control, and this deficit was attenuated in the STZ+P165 group (P < 0.01). IR and IRS-1 were elevated in STZ rats, and these levels were restored to near control in the STZ+P165 and STZ+RSG groups (P < 0.01). Our findings demonstrate that P165 and RSG improved hippocampus-dependent spatial learning and memory in STZ rats by regulating the insulin signaling pathway.
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Effects of grape seed proanthocyanidin extract on renal injury in type 2 diabetic rats.
Mol Med Rep
PUBLISHED: 03-23-2014
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Grape seed proanthocyanidin extract (GSPE) is known to be an effective natural polyphenol capable of removing free radicals in vivo. It has been reported that GSPE has biological functions including antioxidant, anti?cancer, anti?hyperglycemic, anti?radiation, and prevention and treatment of cardiovascular diseases. This study aims to investigate the effects of GSPE on renal injury in type 2 diabetic rats induced with low?dose streptozotocin and a high?carbohydrate/high?fat diet. Rats (n=12 per group) were administered GSPE at either a low (125 mg/kg·bw), medium (250 mg/kg·bw) or high (500 mg/kg·bw) dose, while control rats and diabetes mellitus group rats received no specific treatment. After 16 weeks, GSPE slightly increased body weight and decreased food consumption, water intake and urine volume in rats. Diabetic rats treated with GSPE demonstrated decreased fasting blood glucose, serum insulin, HbA1c and systolic blood pressure (P<0.05). GSPE significantly improved renal function parameters, reduced the expression of tissue inhibitor of metalloproteinase?1 and also increased the activity of matrix metalloproteinase?9. Moreover, GSPE (particularly at a dose of 500 mg/kg·bw) increased the activity of antioxidant enzymes and reduced the levels of c?reactive proteins (P<0.01) in serum and the expression of tumor necrosis factor??, monocyte chemoattractant protein?1 and intercellular adhesion molecule?1 (P<0.05) in the kidney. These findings suggest that GSPE ameliorates renal injury in type 2 diabetic rats through its antioxidative activity and anti?inflammatory effects.
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Photosensitizing effectiveness of a novel chlorin-based photosensitizer for photodynamic therapy in vitro and in vivo.
J. Cancer Res. Clin. Oncol.
PUBLISHED: 03-10-2014
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Photodynamic therapy (PDT) is a promising noninvasive treatment, which has been approved by the US Food and Drug Administration for the treatment of localized tumors. With the aim to select an appropriate photosensitizer for tumor treatment in PDT, the antitumor effect of a novel chlorin-based photosensitizer, meso-tetra (3-morphlinomethyl-4-methoxyphenyl) chlorin (TMMC) (Fig. 1a) on two types of human malignant tumor cells in vitro and a esophageal cancer model in nude mice, was evaluated in the present paper. Fig. 1 Chemical structure and spectrum properties of TMMC in DMF. a Chemical structure of TMMC in DMF. b UV-Vis absorption spectrum of TMMC in DMF. Its maximum absorbance is at 423 nm, and at 527, 555, 600, 655 nm and 712 nm, also it has absorption. c Emission spectrum of TMMC, which was excited at 514 nm, and its peaks were at 656 and 720 nm. d The matrix of excitation and emission spectra (Ex: 300-550 nm, Em: 600-780 nm)
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Influencial factors of the performance of interferon-? release assays in the diagnosis of childhood tuberculosis.
Clin. Exp. Med.
PUBLISHED: 03-03-2014
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Diagnosis of active tuberculosis (TB) in children remains difficult. This study aimed at evaluating the ability of interferon-gamma release assays (IGRAs) in the detection of active TB in human immunodeficiency virus-negative children vaccinated with Bacille Calmette-Guérin and investigating the effect of prednisolone treatment on the IGRAs performance. Among the 162 children with suspected TB disease recruited in China, 60 were tested with QuantiFERON-TB Gold In Tube (QFT-GIT) and 102 were tested with T-SPOT.TB. QFT-GIT presented a sensitivity of 83.9 % (95 % CI 66.9-93.4 %) and a specificity of 88.5 % (95 % CI 70.2-96.8 %), while T-SPOT.TB had a sensitivity of 75.9 % (95 % CI 63.4-85.2 %) and a specificity of 94.7 % (95 % CI 81.8-99.5 %). The positive predictive value was high in both assays, 92.9 % for QFT-GIT and 95.7 % for T-SPOT.TB. In total of these two kinds of IGRAs, false negative rate was significantly higher in children receiving systemic prednisolone (1 mg/kg/day) therapy for >1 week (two tested with T-SPOT.TB and five tested with QFT-GIT) than in those with ?1 week of prednisolone therapy and without prednisolone therapy (57.1 vs. 18.3 %, p = 0.035). There was no significant difference of the positive rate of both tests in children <5 years old compared with those ?5 years old. Both types of IGRAs showed good diagnostic values in detecting childhood TB before microbiological evidence was available. Glucocorticoids had a significant negative influence on IGRAs if treated for >1 week. Age made no difference on the performance of these tests in children.
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Histone H2B monoubiquitination is involved in regulating the dynamics of microtubules during the defense response to Verticillium dahliae toxins in Arabidopsis.
Plant Physiol.
PUBLISHED: 02-24-2014
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Histone H2B monoubiquitination (H2Bub) is being recognized as a regulatory mechanism that controls a range of cellular processes in plants, but the molecular mechanisms of H2Bub that are involved in responses to biotic stress are largely unknown. In this study, we used wild-type and H2Bub loss-of-function mutations of Arabidopsis (Arabidopsis thaliana) to elucidate which of its mechanisms are involved in the regulation of the plant's defense response to Verticillium dahliae (Vd) toxins. We demonstrate that the depolymerization of the cortical microtubules (MTs) was different in the wild type and the mutants in the response to Vd toxins. The loss-of-function alleles of HISTONE MONOUBIQUITINATION1 and HISTONE MONOUBIQUITINATION2 mutations present a weaker depolymerization of the MTs, and protein tyrosine phosphorylation plays a critical role in the regulation of the dynamics of MTs. Moreover, H2Bub is a positive regulator of the gene expression of protein tyrosine phosphatases. These findings provide direct evidence for H2Bub as an important modification with regulatory roles in the defense against Vd toxins and demonstrate that H2Bub is involved in modulating the dynamics of MTs, likely through the protein tyrosine phosphatase-mediated signaling pathway.
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Decitabine facilitates immune recognition of sarcoma cells by upregulating CT antigens, MHC molecules, and ICAM-1.
Tumour Biol.
PUBLISHED: 02-14-2014
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Rhabdomyosarcoma, osteosarcoma, and Ewing's sarcoma are the most common types of sarcoma in children. Despite standard therapy, nearly one third of the patients with Ewing's sarcoma relapse, and there are limited options with curative potential. Immunotherapy is a promising approach as it can target tumor-specific antigens that are specifically expressed on tumors while sparing non-malignant cells. We have demonstrated that a demethylating chemotherapeutic drug, 5-aza-2'-deoxycytidine (decitabine, DAC) can upregulate the expression of cancer-testis (CT) antigens, MHC molecules, and intracellular cell adhesion molecule-1 on pediatric sarcoma cell lines, resulting in enhanced killing of tumor cells by CT antigen-specific cytotoxic T lymphocytes derived from pediatric sarcoma patients. A significant increase in the mRNA expression levels of MAGE-A1 and MAGE-A3 were found in 70 %, and NY-ESO-1 in 80 % of the sarcoma lines following exposure to pharmacological levels of DAC. The high expression levels of MAGE-A1, MAGE-A3, and NY-ESO-1 were sustained in sarcoma lines and primary tumor lines over 30 days after the cessation of DAC. Furthermore, DAC treatment induced upregulation of MAGE-A1, MAGE-A3, or NY-ESO-1 protein expression in seven of nine lines studied. These studies show that demethylating chemotherapy could be combined with CT antigen-directed immunotherapy for treating pediatric sarcoma.
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Survivin mRNA expression in blood as a predictor of the response to EGFR-tyrosine kinase inhibitors and prognosis in patients with non-small cell lung cancer.
Med. Oncol.
PUBLISHED: 02-14-2014
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The epidermal growth factor receptor (EGFR) mutations have been proven to be a reliable predictive marker for the response to EGFR-tyrosine kinase inhibitors (TKIs) in patients with non-small cell lung cancer (NSCLC). However, the responses to EGFR-TKIs vary even among patients with EGFR mutation. Recent study showed that survivin overexpression attenuated EGFR-TKI-induced apoptosis and inhibited the antitumor effect of EGFR-TKIs on EGFR mutation NSCLC cells. We investigated the role of survivin mRNA expression in peripheral blood on predicting response and prognosis in NSCLC patients treated with EGFR-TKIs. Survivin mRNA expression levels in blood was detected using quantitative real-time-PCR assay in 62 patients with advanced NSCLC before (D0) and 4 weeks after treatment of EGFR-TKIs (D4w). The associations between survivin mRNA expression in blood and tumor response to treatment, time to progression (TTP), and overall survival (OS) were analyzed. Blood survivin mRNA levels at D0 and D4w were significantly higher in patients with progressive disease than in those with partial response and stable disease. The detections of blood survivin mRNA positivity at D0 and D4w were associated with unfavorable response to EGFR-TKIs treatment and shorter TTP and OS. Multivariate Cox analysis showed that blood survivin mRNA positivity before and 4 weeks after EGFR-TKIs treatment were independent predictor for worse TTP and OS. In conclusion, Blood survivin mRNA positivity was strongly related to a poor treatment outcome of EGFR-TKIs and may be a potential non-invasive biomarker for NSCLC treatment.
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Design and construction of a cryogenic distillation device for removal of krypton for liquid xenon dark matter detectors.
Rev Sci Instrum
PUBLISHED: 02-13-2014
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Liquid xenon (Xe) is one of the commendable detecting media for the dark matter detections. However, the small content of radioactive krypton-85 ((85)Kr) always exists in the commercial xenon products. An efficient cryogenic distillation system to remove this krypton (Kr) from commercial xenon products has been specifically designed, developed, and constructed in order to meet the requirements of the dark matter experiments with high- sensitivity and low-background. The content of krypton in regular commercial xenon products can be reduced from 10(-9) to 10(-12), with 99% xenon collection efficiency at maximum flow rate of 5 kg/h (15SLPM). The purified xenon gases produced by this distillation system can be used as the detecting media in the project of Panda X, which is the first dark matter detector developed in China.
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Quantum dot-functionalized porous ZnO nanosheets as a visible light induced photoelectrochemical platform for DNA detection.
Nanoscale
PUBLISHED: 01-24-2014
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This work reports the synthesis of novel CdTe quantum dot (QD)-functionalized porous ZnO nanosheets via a covalent binding method with (3-aminopropyl)triethoxysilane as a linker. The functional nanosheets showed an excellent visible-light absorbency and much higher photoelectrochemical activity than both CdTe QDs and ZnO nanosheets due to the porous structure and appropriate band alignment between the CdTe QDs and ZnO nanosheets. Using hydrogen peroxide as an electron acceptor the nanosheet-modified electrode showed a sensitive photocurrent response. This speciality led to a novel methodology for the design of hydrogen peroxide-related biosensors by the formation or consumption of hydrogen peroxide. Using biotin-labeled DNA as capture probe, a model biosensor was proposed by immobilizing the probe on a nanosheet-modified electrode to recognize target DNA in the presence of an assistant DNA, which produced a "Y" junction structure to trigger a restriction endonuclease-aided target recycling. The target recycling resulted in the release of biotin labeled to the immobilized DNA from the nanosheet-modified electrode, thus decreased the consumption of hydrogen peroxide by horseradish peroxidase-mediated electrochemical reduction after binding the left biotin with horseradish peroxidase-labeled streptavidin, which produced an increasing photoelectrochemical response. The 'signal on' strategy for photoelectrochemical detection of DNA showed a low detection limit down to the subfemtomole level and good specificity to single-base mismatched oligonucleotides. The sensitized porous ZnO nanosheets are promising for applications in both photovoltaic devices and photoelectrochemical biosensing.
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Generation of GGTA1 biallelic knockout pigs via zinc-finger nucleases and somatic cell nuclear transfer.
Sci China Life Sci
PUBLISHED: 01-15-2014
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Genetically modified pigs are valuable models of human disease and donors of xenotransplanted organs. Conventional gene targeting in pig somatic cells is extremely inefficient. Zinc-finger nuclease (ZFN) technology has been shown to be a powerful tool for efficiently inducing mutations in the genome. However, ZFN-mediated targeting in pigs has rarely been achieved. Here, we used ZFNs to knock out the porcine ?-1, 3-galactosyl-transferase (GGTA1) gene, which generates Gal epitopes that trigger hyperacute immune rejection in pig-to-human transplantation. Primary pig fibroblasts were transfected with ZFNs targeting the coding region of GGTA1. Eighteen mono-allelic and four biallelic knockout cell clones were obtained after drug selection with efficiencies of 23.4% and 5.2%, respectively. The biallelic cells were used to produce cloned pigs via somatic cell nuclear transfer (SCNT). Three GGTA1 null piglets were born, and one knockout primary fibroblast cell line was established from a cloned fetus. Gal epitopes on GGTA1 null pig cells were completely eliminated from the cell membrane. Functionally, GGTA1 knockout cells were protected from complement-mediated immune attacks when incubated with human serum. This study demonstrated that ZFN is an efficient tool in creating gene-modified pigs. GGTA1 null pigs and GGTA1 null fetal fibroblasts would benefit research and pig-to-human transplantation.
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AbsCN-seq: a statistical method to estimate tumor purity, ploidy and absolute copy numbers from next-generation sequencing data.
Bioinformatics
PUBLISHED: 01-02-2014
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Detection and quantification of the absolute DNA copy number alterations in tumor cells is challenging because the DNA specimen is extracted from a mixture of tumor and normal stromal cells. Estimates of tumor purity and ploidy are necessary to correctly infer copy number, and ploidy may itself be a prognostic factor in cancer progression. As deep sequencing of the exome or genome has become routine for characterization of tumor samples, in this work, we aim to develop a simple and robust algorithm to infer purity, ploidy and absolute copy numbers in whole numbers for tumor cells from sequencing data.
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Rac2 controls tumor growth, metastasis and M1-M2 macrophage differentiation in vivo.
PLoS ONE
PUBLISHED: 01-01-2014
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Although it is well-established that the macrophage M1 to M2 transition plays a role in tumor progression, the molecular basis for this process remains incompletely understood. Herein, we demonstrate that the small GTPase, Rac2 controls macrophage M1 to M2 differentiation and the metastatic phenotype in vivo. Using a genetic approach, combined with syngeneic and orthotopic tumor models we demonstrate that Rac2-/- mice display a marked defect in tumor growth, angiogenesis and metastasis. Microarray, RT-PCR and metabolomic analysis on bone marrow derived macrophages isolated from the Rac2-/- mice identify an important role for Rac2 in M2 macrophage differentiation. Furthermore, we define a novel molecular mechanism by which signals transmitted from the extracellular matrix via the ?4?1 integrin and MCSF receptor lead to the activation of Rac2 and potentially regulate macrophage M2 differentiation. Collectively, our findings demonstrate a macrophage autonomous process by which the Rac2 GTPase is activated downstream of the ?4?1 integrin and the MCSF receptor to control tumor growth, metastasis and macrophage differentiation into the M2 phenotype. Finally, using gene expression and metabolomic data from our Rac2-/- model, and information related to M1-M2 macrophage differentiation curated from the literature we executed a systems biologic analysis of hierarchical protein-protein interaction networks in an effort to develop an iterative interactome map which will predict additional mechanisms by which Rac2 may coordinately control macrophage M1 to M2 differentiation and metastasis.
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Cell-Specific and pH-Activatable Rubyrin-Loaded Nanoparticles for Highly Selective Near-Infrared Photodynamic Therapy against Cancer.
J. Am. Chem. Soc.
PUBLISHED: 12-09-2013
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Spatiotemporal control of singlet oxygen ((1)O2) release is a major challenge for photodynamic therapy (PDT) against cancer with high therapeutic efficacy and minimum side effects. Here a selenium-rubyrin (NMe2Se4N2)-loaded nanoparticle functionalized with folate (FA) was designed and synthesized as an acidic pH-activatable targeted photosensitizer. The nanoparticles could specifically recognize cancer cells via the FA-FA receptor binding and were selectively taken up by cancer cells via receptor-mediated endocytosis to enter lysosomes, in which NMe2Se4N2 was activated to produce (1)O2. The pH-controllable release of (1)O2 specially damaged the lysosomes and thus killed cancer cells in a lysosome-associated pathway. The introduction of selenium into the rubyrin core enhanced the (1)O2 generation efficiency due to the heavy atom effect, and the substitution of dimethylaminophenyl moiety at meso-position led to the pH-controllable activation of NMe2Se4N2. Under near-infrared (NIR) irradiation, NMe2Se4N2 possessed high singlet oxygen quantum yield (??) at an acidic pH (?? = 0.69 at pH 5.0 at 635 nm) and could be deactivated at physiological pH (?? = 0.06 at pH 7.4 at 635 nm). The subcellular location-confined pH-activatable photosensitization at NIR region and the cancer cell-targeting feature led to excellent capability to selectively kill cancer cells and prevent the damage to normal cells, which greatly lowered the side effects. Through intravenous injection of FA-NMe2Se4N2 nanoparticles in tumor-bearing mice, tumor elimination was observed after NIR irradiation. This work presents a new paradigm for specific PDT against cancer and provides a new avenue for preparation of highly efficient photosensitizers.
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[Endovascular repair for isolated iliac artery aneurysm].
Zhonghua Yi Xue Za Zhi
PUBLISHED: 11-29-2013
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To evaluate the short and middle-term efficacies of endovascular repair for isolated iliac artery aneurysms (IIAAs).
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Determination of aflatoxins B1, B2, G1, and G2 in olive oil, peanut oil, and sesame oil using immunoaffinity column cleanup, postcolumn derivatization, and liquid chromatography with fluorescence detection: first action 2013.05.
J AOAC Int
PUBLISHED: 11-29-2013
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A collaborative study of a method for determination of aflatoxins (AFs) B1, B2, G1, and G2 in olive oil, peanut oil, and sesame oil using immunoaffinity column cleanup, postcolumn derivatization, and LC with fluorescence detection, previously published in J. AOAC Int. 95, 1689-1700 (2012), was approved as First Action 2013.05 on March 29, 2013 by the Method-Centric Committee for Aflatoxins in Edible Oils. The method uses methanol for extraction followed by filtration. The extract is applied to an immunoaffinity column with antibodies specific for AFs, which are then eluted from the column with a methanol solution. Determination and quantification occur using RP-LC with fluorescence detection after postcolumn derivatization. The average recovery of AFs in olive, peanut, and sesame oils in spiked samples (levels between 2.0 and 20.0 microg/kg) ranged from 84 to 92%. The recoveries for AFs B1, B2, G1, and G2 were 86-93, 89-95, 85-97, and 76-85%, respectively. Within-laboratory RSD (RSDr) values for AFs ranged from 3.4 to 10.2%. RSDr values forAF B1, B2, G1, and G2 were 3.5-10.9, 3.2-9.5, 6.5-14.9, and 4.8-14.2%, respectively. Between-laboratory RSD (RSDR) values for AFs were 6.1-14.5%. RSD, values for AFs B1, B2, G1, and G2 were 7.5-15.4, 7.1-14.6, 10.8-18.1, and 7.6-23.7%, respectively. Horwitz ratio values were < or =2 for the analytes in the three matrixes.
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Associations between Fas/FasL polymorphisms and susceptibility to cervical cancer: a meta-analysis.
Tumour Biol.
PUBLISHED: 09-02-2013
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Genetic polymorphisms in the Fas/Fas ligand (FasL) gene were proposed to be associated with susceptibility to cervical cancer, but previous studies reported controversial findings. We performed a meta-analysis to assess the associations between Fas/FasL polymorphisms and susceptibility to cervical cancer. We carried out a literature search in PubMed and Embase databases for studies on the associations between Fas/FasL polymorphisms and susceptibility to cervical cancer. The associations were assessed by odds ratio (OR) together with its 95 % confidence intervals (CIs). Eleven individual studies with a total of 6,919 subjects were finally included into the meta-analysis. Overall, there was no association between Fas 1377G?>?A polymorphism and susceptibility to cervical cancer (A vs. G: OR?=?0.99, 95 % CI 0.88-1.12, P?=?0.91; AA vs. GG: OR?=?1.00, 95 % CI 0.76-1.32, P?=?0.99; AA/GA vs. GG: OR?=?0.95, 95 % CI 0.81-1.12, P?=?0.54; AA vs.
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Clinical progression and predictors of death in patients with severe fever with thrombocytopenia syndrome in China.
J. Clin. Virol.
PUBLISHED: 08-26-2013
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Severe fever with thrombocytopenia syndrome (SFTS) is an emerging infectious disease of which the clinical progression and factors related to death are still unclear.
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Generating hypoimmunogenic human embryonic stem cells by the disruption of Beta 2-microglobulin.
Stem Cell Rev
PUBLISHED: 08-13-2013
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Immune rejection hinders the application of human embryonic stem cells (hESCs) in transplantation therapy. Human leukocyte antigens (HLAs) on the cell surface are the major cause of graft rejection. In this study, we generated HLA class I-deficient hESCs via disruption of beta 2-microglobulin (?2m), the light chain of HLA Class I. We found that HLA class I proteins were not present on the cell surface of ?2m-null hESCs. These cells showed the same pluripotency as wildtype hESCs and demonstrated hypoimmunogenicity. Thus, HLA class I-deficient hESCs might serve as an unlimited cell source for the generation of universally compatible "off-the-shelf" cell grafts, tissues or organs in the future.
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[Significance of carbonic anhydrase IX protein expression in molecular subtyping of breast cancers].
Zhonghua Bing Li Xue Za Zhi
PUBLISHED: 06-18-2013
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To study the expression of carbonic anhydrase (CA) IX and its significance in molecular subtyping of breast carcinomas. METHODL MaxVision immunohistochemical staining was used to examine the expression of ER, PR, HER2, CK5/6, EGFR, and CA IX in 117 cases of breast invasive ductal carcinomas.
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Diagnostic value of the FHIT and p16 mRNA loss and the K-ras gene mutation in pleural fluids for malignant pleural effusion.
Cancer Biomark
PUBLISHED: 06-06-2013
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Inactivation of the tumor suppressor genes and activation of oncogenes are involved in the development of cancer. The aim of this study was to evaluate the diagnostic value of the fragile histidine triad (FHIT) and p16 mRNA loss and the K-ras gene mutation in distinguishing malignant from benign pleural effusion. A total of 50 patients with malignant pleural effusion and 30 patients with benign pleural effusion were enrolled in this study. All pleural fluid specimens were evaluated in parallel by cytology, reverse transcriptase-PCR for the loss of FHIT and p16 mRNA, and PCR-SSCP (single-stranded conformation polymorphism) for the mutation of K-ras gene. The detection rates of FHIT and p16 mRNA loss were significantly higher in malignant than in benign pleural effusion (P < 0.001 and P = 0.001). The K-ras mutations were more frequent in malignant than benign pleural effusion (P = 0.006). The sensitivity and specificity were 58% and 93% for FHIT loss, 48% and 90% for p16 loss, and 44% and 87% for the K-ras mutation, respectively. In combined evaluation with both FHIT and p16 loss, the sensitivity was 68%, and specificity was 90%. The combination of the three molecular markers reached 74% sensitivity, whereas the combined use of the cytology and the three markers increased the diagnostic yield of the former by 38%. More than one third of cytology negative malignant pleural effusion could be identified by at least one of the three markers. These results suggest that the detection of FHIT and p16 mRNA loss and the k-ras gene mutation in pleural fluid could be helpful adjunct to cytology in the diagnosis of malignant pleural effusion.
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Synthesis, insecticidal activities, and SAR studies of novel pyridylpyrazole acid derivatives based on amide bridge modification of anthranilic diamide insecticides.
J. Agric. Food Chem.
PUBLISHED: 05-31-2013
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Anthranilic diamides are one of the most important classes of modern agricultural insecticides. To discover new structure-modified compounds with high activity, series of novel carbonyl thioureas, carbonyl ureas, oxadiazoles, carbonyl thiophosphorylureas, oxadiazole-containing amides, and thiazoline-containing amides were designed through the modification of the amide bridge based on the structure of chlorantraniliprole and were synthesized, and bioassays were carried out. The compounds were characterized and confirmed by melting point, IR, (1)H NMR, and elemental analyses or HRMS. Preliminary bioassays indicated that some compounds exhibited significant insecticidal activities against oriental armyworm, diamondback moth, beet armyworm, corn borer, and mosquito. Among them, trifluoroethoxyl-containing carbonyl thiourea 20a showed best larvicidal activity against oriental armyworm, with LC50 and LC95 values of 0.1812 and 0.7767 mg/L, respectively. Meanwhile, 20c and 20e showed 86 and 57% death rates against diamondback moth at 0.005 mg/L, and the LC50 values of the two compounds were 0.0017 and 0.0023 mg/L, respectively, which were lower than that of the control chlorantraniliprole. The relationship between structure and insecticidal activity was discussed, and the HF calculation results indicated that the carbonyl thiourea moiety plays an important role in the insecticidal activity. The present work demonstrated that the trifluoroethoxyl-containing carbonyl thioureas can be used as lead compounds for further development of novel insecticides.
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Immune responses of orange-spotted grouper, Epinephelus coioides, against virus-like particles of betanodavirus produced in Escherichia coli.
Vet. Immunol. Immunopathol.
PUBLISHED: 05-28-2013
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Betanodaviruses are the causative agents of viral nervous necrosis (VNN), a serious disease of cultured marine fish worldwide. Virus-like particles (VLPs) are one of the good novel vaccine candidates to control this disease. Until now, betanodavirus vaccine studies mainly focused on the humoral immune response and mortality after virus challenge. However, little is known about the activation of genes responsible for cellular and innate immunity by vaccines. In the present study, VLPs of orange-spotted grouper nervous necrosis virus (OGNNV) were produced in prokaryotes and their ability to enter Asian sea bass cells was the same as native virus, suggesting that they possess a similar structure to OGNNV. VLPs immunogenicity was then determined by intramuscularly vaccinating Epinephelus coioides at different concentrations (1.5 or 15?gg(-1) fish body weight, FBW) and immunizing frequencies (administration once, twice and thrice). A single vaccination with the dosage of 1.5?gg(-1) FBW is enough to provoke high titer antibodies (average 3 fold higher than that of negative control) with strong neutralizing antibody titer as early as 1 week post immunization. Furthermore, quantitative PCR analysis revealed that eleven genes associated with humoral, cellular and innate immunities were up-regulated in the liver, spleen and head kidney at 12h post immunization, correlating with the early antibody response. In conclusion, we demonstrated that VLP vaccination induced humoral immune responses and activated genes associated with cellular and innate immunity against betanodavirus infection in orange-spotted grouper.
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Evolutionary increases in defense during a biological invasion.
Oecologia
PUBLISHED: 05-07-2013
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Invasive plants generally escape from specialist herbivores of their native ranges but may experience serious damage from generalists. As a result, invasive plants may evolve increased resistance to generalists and tolerance to damage. To test these hypotheses, we carried out a common garden experiment comparing 15 invasive populations with 13 native populations of Chromolaena odorata, including putative source populations identified with molecular methods and binary choice feeding experiments using three generalist herbivores. Plants from invasive populations of C. odorata had both higher resistance to three generalists and higher tolerance to simulated herbivory (shoot removal) than plants from native populations. The higher resistance of plants from invasive populations was associated with higher leaf C content and densities of leaf trichomes and glandular scales, and lower leaf N and water contents. Growth costs were detected for tolerance but not for resistance, and plants from invasive populations of C. odorata showed lower growth costs of tolerance. Our results suggest that invasive plants may evolve to increase both resistance to generalists and tolerance to damage in introduced ranges, especially when the defense traits have low or no fitness costs. Greater defenses in invasive populations may facilitate invasion by C. odorata by reducing generalist impacts and increasing compensatory growth after damage has occurred.
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The effect of the number of transferred embryos, the interval between nuclear transfer and embryo transfer, and the transfer pattern on pig cloning efficiency.
Anim. Reprod. Sci.
PUBLISHED: 05-05-2013
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To improve the efficiency of producing cloned pigs, we investigated the influence of the number of transferred embryos, the culturing interval between nuclear transfer (NT) and embryo transfer, and the transfer pattern (single oviduct or double oviduct) on cloning efficiency. The results demonstrated that transfer of either 150-200 or more than 200NT embryos compared to transfer of 100-150 embryos resulted in a significantly higher pregnancy rate (48±16, 50±16 vs. 29±5%, p<0.05) and average litter size (4.1±2.3, 7±3.6 vs. 2.5±0.5). In vitro culture of reconstructed embryos for a longer time (40h vs. 20h) resulted in higher (p<0.05) pregnancy rate (44±9 vs. 31±3%) and delivery rate (44±9 vs. 25±9%). Furthermore, double oviductal transfer dramatically increased pregnancy rate (83±6 vs. 27+8%, p<0.05), delivery rate (75±2 vs. 27+8%, p<0.05) and average litter size (6.5±2.8 vs. 2.6±1.2) compared to single oviductal transfer. Our study demonstrated that an improvement in pig cloning efficiency is achieved by adjusting the number and in vitro culture time of reconstructed embryos as well as the embryo transfer pattern.
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Effects of combined treatment with ambroxol and ciprofloxacin on catheter-associated Pseudomonas aeruginosa biofilms in a rat model.
Chemotherapy
PUBLISHED: 03-26-2013
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Pseudomonas aeruginosa is an opportunistic pathogen that causes potentially devastating infections in immunocompromised patients. These infections are particularly difficult to treat if a biofilm forms, which is likely if foreign bodies are present.
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The involvement of the JAK-STAT signaling pathway in chronic inflammatory skin disease atopic dermatitis.
JAKSTAT
PUBLISHED: 02-26-2013
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Atopic dermatitis (AD), a common chronic inflammatory skin disease, is characterized by inflammatory cell skin infiltration. The JAK-STAT pathway has been shown to play an essential role in the dysregulation of immune responses in AD, including the exaggeration of Th2 cell response, the activation of eosinophils, the maturation of B cells, and the suppression of regulatory T cells (Tregs). In addition, the JAK-STAT pathway, activated by IL-4, also plays a critical role in the pathogenesis of AD by upregulating epidermal chemokines, pro-inflammatroy cytokines, and pro-angiogenic factors as well as by downregulating antimicrobial peptides (AMPs) and factors responsible for skin barrier function. In this review, we will highlight the recent advances in our understanding of the JAK-STAT pathway in the pathogenesis of AD.
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Quercetin but not quercitrin ameliorates tumor necrosis factor-alpha-induced insulin resistance in C2C12 skeletal muscle cells.
Biol. Pharm. Bull.
PUBLISHED: 02-23-2013
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Skeletal muscle is a major site for glucose metabolism and its injury by cytokines can induce insulin resistance leading to type 2 diabetes. It has been suggested that quercetin may act as an anti-diabetic agent, however, the effects of quercetin on insulin resistance in skeletal muscle remain unknown. We aimed to investigate the role of quercetin and its glycoside, quercitrin in tumor necrosis factor-alpha (TNF-?) induced C2C12 skeletal muscle cell impairment. Quercetin, but not quercitrin moderately attenuated the effects of TNF-? and enhanced the basal and insulin stimulated uptake of glucose in a dose-dependent manner via the activation of the protein kinase B (Akt) and AMP-activated protein kinase (AMPK) pathways. Furthermore, the underlying mechanism also involved the suppression of nuclear factor-?B (NF-?B) signaling and the nitric oxide (NO)/inducible nitric oxide synthase (iNOS) system, downstream of AMPK transduction. In summary, quercetin exhibited its effect of improving glucose uptake and insulin sensitivity in skeletal muscle cells via the two independent signaling pathways of Akt and AMPK, and can be developed as a potential anti-diabetic agent.
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A Pan-BCL2 inhibitor renders bone-marrow-resident human leukemia stem cells sensitive to tyrosine kinase inhibition.
Cell Stem Cell
PUBLISHED: 01-17-2013
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Leukemia stem cells (LSCs) play a pivotal role in the resistance of chronic myeloid leukemia (CML) to tyrosine kinase inhibitors (TKIs) and its progression to blast crisis (BC), in part, through the alternative splicing of self-renewal and survival genes. To elucidate splice-isoform regulators of human BC LSC maintenance, we performed whole-transcriptome RNA sequencing, splice-isoform-specific quantitative RT-PCR (qRT-PCR), nanoproteomics, stromal coculture, and BC LSC xenotransplantation analyses. Cumulatively, these studies show that the alternative splicing of multiple prosurvival BCL2 family genes promotes malignant transformation of myeloid progenitors into BC LSCS that are quiescent in the marrow niche and that contribute to therapeutic resistance. Notably, sabutoclax, a pan-BCL2 inhibitor, renders marrow-niche-resident BC LSCs sensitive to TKIs at doses that spare normal progenitors. These findings underscore the importance of alternative BCL2 family splice-isoform expression in BC LSC maintenance and suggest that the combinatorial inhibition of prosurvival BCL2 family proteins and BCR-ABL may eliminate dormant LSCs and obviate resistance.
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Grape seed proanthocyanidins ameliorate pancreatic beta-cell dysfunction and death in low-dose streptozotocin- and high-carbohydrate/high-fat diet-induced diabetic rats partially by regulating endoplasmic reticulum stress.
Nutr Metab (Lond)
PUBLISHED: 01-01-2013
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It is increasingly being realized that failure of pancreatic beta cells to secrete enough insulin to adequately compensate for obesity and insulin resistance is the primary defects of type 2 diabetes mellitus (T2DM). Pancreatic beta cells possess a highly developed and active endoplasmic reticulum (ER), reflecting their role in folding, export and processing of newly synthesized insulin. ER stress-induced pancreatic beta-cell failure is a novel event in the pathogenesis of T2DM. Some studies with antioxidants indicated a beneficial impact on ER stress. Our previous study found that strong antioxidants, grape seed proanthocyanidins (GSPs), ameliorated ER stress to protect skeletal muscle from cell death in type 2 diabetic rats. The present study continued to investigate the effect of GSPs on beta-cell failure and ER stress in diabetic pancreas.
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Molecular characterization and functional analysis of Cashmere goat mammalian target of rapamycin.
DNA Cell Biol.
PUBLISHED: 12-16-2011
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The mammalian target of rapamycin (mTOR) is an evolutionarily conserved protein kinase that belongs to the phosphatidylinositol kinase-related kinase family. We describe our molecular characterization of mTOR and its function (GenBank accession HM114224) in Cashmere goat (Capra hircus). The goat mTOR complementary DNA is 8617 bp, comprising an open reading frame of 7650 bp--corresponding to a polypeptide of 2549 amino acids--and a 909 bp 3 untranslated region with a polyA tract and a polyadenylation signal at nucleotides 8575-8580. In a bioinformatics analysis, goat mTOR has typical sites of activity and domains. mTOR mRNA was measured in brain, heart, testis, liver, spleen, kidney, and lung by real-time polymerase chain reaction, and the expression of mTOR in fetal fibroblasts was detected by western blot. The viability of fetal fibroblasts was inhibited on treatment with CCI-779, a specific inhibitor of mTOR. Our data supplied evidence that the transcription of mTOR was detected in the seven tissues in Cashmere goat, and mTOR protein was translated in fetal fibroblasts. The proliferation of fetal fibroblasts decreases on inhibition of mTOR.
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Use of a multifunctional column for the determination of deoxynivalenol in grains, grain products, and processed foods.
J AOAC Int
PUBLISHED: 12-15-2011
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Deoxynivalenol (DON), also known as vomitoxin, belongs to a class of naturally occurring mycotoxins produced by Fusarium spp. DON, 12, 13-epoxy-3,7 trihydroxytrichothec-9-en-8-one, is one of the most frequently detected mycotoxins in agricultural commodities worldwide. A method consisting of extraction, filtration, column cleanup, and RP-HPLC-UV separation and quantitation was validated for the determination of DON in grains (rice and barley), grain products (whole wheat flour, white flour, wheat germ, and wheat bran), and processed foods (bread, breakfast cereals, and pretzels). A 25 g test portion was extracted with 100 mL acetonitrile-water (84 + 16, v/v). After blending for 3 min, the supernatant was applied to a multifunctional column (MycoSep 225). The purified filtrate (2 mL) was evaporated to dryness and redissolved in the mobile phase. The toxins were then subjected to RP-HPLC-UV analysis. The accuracy and repeatability characteristics of the method were determined. Recoveries of DON added at levels ranging from 0.5 to 1.5 microg/g for all test matrixes were from 75 to 98%. SD and RSD(r) ranged from 0.7 to 11.6% and 0.9 to 12.7%, respectively. Within-laboratory HorRat values were from 0.1 to 0.7 for all matrixes analyzed. The method was found to meet AOAC method performance criteria for grains, grain products, and processed foods. The identity of DON in naturally contaminated test sample extracts was confirmed by HPLC/MS/MS analysis.
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Estimation of sample size and testing power (Part 3).
Zhong Xi Yi Jie He Xue Bao
PUBLISHED: 12-14-2011
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This article introduces the definition and sample size estimation of three special tests (namely, non-inferiority test, equivalence test and superiority test) for qualitative data with the design of one factor with two levels having a binary response variable. Non-inferiority test refers to the research design of which the objective is to verify that the efficacy of the experimental drug is not clinically inferior to that of the positive control drug. Equivalence test refers to the research design of which the objective is to verify that the experimental drug and the control drug have clinically equivalent efficacy. Superiority test refers to the research design of which the objective is to verify that the efficacy of the experimental drug is clinically superior to that of the control drug. By specific examples, this article introduces formulas of sample size estimation for the three special tests, and their SAS realization in detail.
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Estimation of sample size and testing power (part 2).
Zhong Xi Yi Jie He Xue Bao
PUBLISHED: 11-18-2011
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This article introduces definitions of three special tests, namely, non-inferiority test (to verify that the efficacy of the experimental drug is clinically not inferior to that of the positive control drug), equivalence test (to verify that the efficacy of the experimental drug is equivalent to that of the control drug) and superiority test (to verify that the efficacy of the experimental drug is superior to that of the control drug), and methods of sample size estimation under the three different conditions. By specific examples, the article introduces formulas of sample size estimation for the three special tests, and their SAS realization in detail.
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Estimation of sample size and testing power (Part 1).
Zhong Xi Yi Jie He Xue Bao
PUBLISHED: 10-22-2011
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This article introduces the general concepts and methods of sample size estimation and testing power analysis. It focuses on parametric methods of sample size estimation, including sample size estimation of estimating the population mean and the population probability. It also provides estimation formulas and introduces how to realize sample size estimation manually and by SAS software.
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Aptamer biosensor based on fluorescence resonance energy transfer from upconverting phosphors to carbon nanoparticles for thrombin detection in human plasma.
Anal. Chem.
PUBLISHED: 09-30-2011
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We presented a new aptamer biosensor for thrombin in this work, which was based on fluorescence resonance energy transfer (FRET) from upconverting phosphors (UCPs) to carbon nanoparticles (CNPs). The poly(acrylic acid) (PAA) functionalized UCPs were covalently tagged with a thrombin aptamer (5-NH(2)- GGTTGGTGTGGTTGG-3), which bound to the surface of CNPs through ?-? stacking interaction. As a result, the energy donor and acceptor were taken into close proximity, leading to the quenching of fluorescence of UCPs. A maximum fluorescence quenching rate of 89% was acquired under optimized conditions. In the presence of thrombin, which induced the aptamer to form quadruplex structure, the ?-? interaction was weakened, and thus, the acceptor was separated from the donor blocking the FRET process. The fluorescence of UCPs was therefore restored in a thrombin concentration-dependent manner, which built the foundation of thrombin quantification. The sensor provided a linear range from 0.5 to 20 nM for thrombin with a detection limit of 0.18 nM in an aqueous buffer. The same linear range was obtained in spiked human serum samples with a slightly higher detection limit (0.25 nM), demonstrating high robustness of the sensor in a complex biological sample matrix. As a practical application, the sensor was used to monitor thrombin level in human plasma with satisfactory results obtained. This is the first time that UCPs and CNPs were employed as a donor-acceptor pair to construct FRET-based biosensors, which utilized both the photophysical merits of UCPs and the superquenching ability of CNPs and thus afforded favorable analytical performances. This work also opened the opportunity to develop biosensors for other targets using this UCPs-CNPs system.
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The repetition principle in scientific research.
Zhong Xi Yi Jie He Xue Bao
PUBLISHED: 09-13-2011
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The repetition principle is important in scientific research, because the observational indexes are random variables, which require a certain amount of samples to reveal their changing regularity. The repetition principle stabilizes the mean and the standard variation, so that statistics of the sample can well represent the parameters of the population. Thus, the statistical inference will be reliable. This article discussed the repetition principle from the perspective of common sense and specialty with examples.
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Detection of low prevalence somatic mutations in solid tumors with ultra-deep targeted sequencing.
Genome Biol.
PUBLISHED: 08-31-2011
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Ultra-deep targeted sequencing (UDT-Seq) can identify subclonal somatic mutations in tumor samples. Early assays limited breadth and depth restrict their clinical utility. Here, we target 71 kb of mutational hotspots in 42 cancer genes. We present novel methods enhancing both laboratory workflow and mutation detection. We evaluate UDT-Seq true sensitivity and specificity (> 94% and > 99%, respectively) for low prevalence mutations in a mixing experiment and demonstrate its utility using six tumor samples. With an improved performance when run on the Illumina Miseq, the UDT-Seq assay is well suited for clinical applications to guide therapy and study clonal selection in heterogeneous samples.
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The control principle in scientific research.
Zhong Xi Yi Jie He Xue Bao
PUBLISHED: 08-19-2011
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The control principle is one of the four basic principles of research design. Without a control group, the conclusion of research will be unconvincing; furthermore, if the control group is not set properly, the conclusion will be unreliable. Generally, there is more than one control group in a multi-factor design. Problems like incomplete control and excessive control should be avoided. This article introduces the meaning and function of the control principle, common forms of control, common errors that researchers tend to make as well as analysis and differentiation of these errors.
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The structure-activity relationship in herbicidal monosubstituted sulfonylureas.
Pest Manag. Sci.
PUBLISHED: 08-09-2011
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The herbicide sulfonylurea (SU) belongs to one of the most important class of herbicides worldwide. It is well known for its ecofriendly, extreme low toxicity towards mammals and ultralow dosage application. The original inventor, G Levitt, set out structure-activity relationship (SAR) guidelines for SU structural design to attain superhigh bioactivity. A new approach to SU molecular design has been developed.
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Overexpression of tyrosine kinase receptor B promotes metastasis of ovarian serous adenocarcinoma by lymphangiogenesis.
Tumori
PUBLISHED: 07-29-2011
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The aim of this study was to detect the expression features of tyrosine kinase receptor B (TrkB) and analyze the possible correlation between TrkB expression and lymph vessel density (LVD) in metastasis of ovarian serous adenocarcinoma.
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Electrochemical tuning of luminescent carbon nanodots: from preparation to luminescence mechanism.
Adv. Mater. Weinheim
PUBLISHED: 07-26-2011
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The size of C-nanodots can be electrochemically tuned by changing the applied potential during their preparation. The higher the applied potential, the smaller the resulting C-nanodots. Moreover, the surface oxidation degree of the C-nanodots can also be electrochemically tuned. The red-shift of emission independent of the size provides an insight into the luminescence mechanism of C-nanodots.
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Methods and analysis of realizing randomized grouping.
Zhong Xi Yi Jie He Xue Bao
PUBLISHED: 07-14-2011
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Randomization is one of the four basic principles of research design. The meaning of randomization includes two aspects: one is to randomly select samples from the population, which is known as random sampling; the other is to randomly group all the samples, which is called randomized grouping. Randomized grouping can be subdivided into three categories: completely, stratified and dynamically randomized grouping. This article mainly introduces the steps of complete randomization, the definition of dynamic randomization and the realization of random sampling and grouping by SAS software.
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Hydrogen peroxide modulates the dynamic microtubule cytoskeleton during the defence responses to Verticillium dahliae toxins in Arabidopsis.
Plant Cell Environ.
PUBLISHED: 06-28-2011
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The molecular mechanisms of signal transduction of plants in response to infection by Verticillium dahliae (VD) are not well understood. We previously showed that NO may act as an upstream signalling molecule to trigger the depolymerization of cortical microtubules in Arabidopsis. In the present study, we used the wild-type, and atrbohD and atrbohF mutants of Arabidopsis to explore the mechanisms of action of H(2)O(2) signals and the dynamic microtubule cytoskeleton in defence responses. We demonstrated that H(2)O(2) may also act as an upstream signalling molecule to regulate cortical microtubule depolymerization. The depolymerization of the cortical microtubules played a functional role in the signalling pathway to mediate the expression of defence genes. The results indicate that H(2)O(2) modulates the dynamic microtubule cytoskeleton to trigger the expression of defence genes against V. dahliae toxins (VD-toxins) in Arabidopsis.
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The principle of randomization in scientific research.
Zhong Xi Yi Jie He Xue Bao
PUBLISHED: 06-15-2011
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Scientific research design includes specialty design and statistics design which can be subdivided into experimental design, clinical trial design and survey design. Usually, statistics textbooks introduce the core aspects of experimental design as the three key elements, the four principles and the design types, which run through the whole scientific research design and determine the overall success of the research. This article discusses the principle of randomization, which is one of the four principles, and focuses on the following two issues--the definition and function of randomization and the real life examples which go against the randomization principle, thereby demonstrating that strict adherence to the randomization principle leads to meaningful and valuable scientific research.
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Psychometric properties of the Chinese version of Sense of Coherence Scale in women with cervical cancer.
Psychooncology
PUBLISHED: 06-08-2011
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The purpose of this study was to test the psychometric properties of a Chinese version of the Sense of Coherence Scale (C-SOC-13) in women with cervical cancer in Mainland China.
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Invasive Eupatorium adenophorum has ecophysiological advantages over native congeners but similar responses to CO(2) enrichment.
Physiol Plant
PUBLISHED: 06-08-2011
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Both global change and biological invasions threaten biodiversity worldwide. However, their interactions and related mechanisms are still not well elucidated. To elucidate potential traits contributing to invasiveness and whether ongoing increase in CO(2) aggravates invasions, noxious invasive Eupatorium adenophorum and native E. japonicum and E. chinensis were compared under ambient and doubled atmospheric CO(2) concentrations in terms of growth, biomass allocation, morphology, and physiology. The invader had consistently higher leaf mass fraction and specific leaf area than the natives, contributing to a higher leaf area ratio, and therefore to faster growth and invasiveness. The higher leaf mass fraction of the invader was associated with lower total root mass fraction. The invader allocated a higher fraction of leaf nitrogen (N) to photosynthesis, contributing to higher area-based N content in photosynthetic apparatus, photosynthetic rate, nitrogen- and water-use efficiencies, and invasiveness. CO(2) enrichment increased growth of all studied plants by increasing actual photosynthesis, although it decreased photosynthetic capacities due to decreased area-based leaf and photosynthetic N contents. Responses of the invasive and native plants to elevated CO(2) were not significantly different, indicating that the ongoing increase in CO(2) may not exacerbate biological invasions, inconsistent with the prevailing results in references. The difference may be associated with the fact that almost all previous studies compared phylogenetically unrelated invasive and native plants. More comparative studies of sympatric, related invasive and native plants are needed to elucidate whether CO(2) enrichment facilitates invasions.
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Synthesis, structure and biological activity of novel 1,2,4-triazole mannich bases containing a substituted benzylpiperazine moiety.
Chem Biol Drug Des
PUBLISHED: 05-31-2011
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A series of novel Mannich bases with trifluoromethyl-1,2,4-triazole and substituted benzylpiperazine moieties were synthesized. Their structures were confirmed by IR, (1) H NMR and elemental analysis. The single crystal structure of compound 4r was also determined. The preliminary bioassays showed that most of the lead compounds had low herbicidal activity against Brassica campestris, Echinochloa crusgalli, and KARI enzyme. However, most of them exhibited significant fungicidal activity at the dosage of 50 ?g/mL toward five test fungi. Among the 18 novel compounds, several showed superiority over the commercial fungicide Triadimefon against Cercospora arachidicola and Fusarium oxysporum f. sp. cucumerinum during this study. Meanwhile, some compounds displayed plant growth regulatory activity at the dosage of 10 ?g/mL.
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[Study on the integrated monitoring program regarding mouse and main mouse-borne disease in Zhejiang province].
Zhonghua Liu Xing Bing Xue Za Zhi
PUBLISHED: 05-17-2011
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To study the integrated monitoring program regarding mouse and plague, hemorrhagic fever of renal syndrome (HFRS) and leptospirosis.
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How to appropriately choose observed indexes.
Zhong Xi Yi Jie He Xue Bao
PUBLISHED: 05-14-2011
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Observed index is a very important element in a research design, because it is a specific reflection of the effects of research factors on the research subjects and is indispensable in any research. Generally, there are two types of observed indexes: the indexes that reflect natural attributes, habits or states of the research subjects and the indexes that reflect the effects of different drugs or treatments on research subjects. This article mainly introduces the definition, characteristics, selection and observation of research indexes and the major and minor indexes.
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MAGE-A1, MAGE-A3, and NY-ESO-1 can be upregulated on neuroblastoma cells to facilitate cytotoxic T lymphocyte-mediated tumor cell killing.
Cancer Immunol. Immunother.
PUBLISHED: 05-12-2011
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Approximately half of patients with stage IV neuroblastoma are expected to relapse despite current therapy, and when this occurs, there is little likelihood of achieving a cure. Very few clinical trials have been conducted to determine whether cellular immune responses could be harnessed to fight this tumor, largely because potential tumor antigens for cytotoxic T lymphocytes (CTL) are limited. MAGE-A1, MAGE-A3, and NY-ESO-1 are cancer-testis (CT) antigens expressed on a number of malignant solid tumors, including neuroblastoma, but many tumor cell lines down-regulate the expression of CT antigens as well as major histocompatibility (MHC) antigens, precluding recognition by antigen-specific T cells. If expression of cancer antigens on neuroblastoma could be enhanced pharmacologically, CT antigen-specific immunotherapy could be considered for this tumor. We have demonstrated that the expression of MAGE-A1, MAGE-A3, and NY-ESO-1 can be upregulated on neuroblastoma cells following exposure to pharmacologic levels of the demethylating agent 5-aza-2-deoxycytidine (decitabine, DAC). Expression of NY-ESO-1, MAGE-A1, or MAGE-A3 was induced in 10/10 neuroblastoma cell lines after 5 days of exposure to DAC. Culture of neuroblastoma cell lines with IFN-? was also associated with an increased expression of either MHC Class I or II by cytofluorometry, as reported by other groups. MAGE-A1, MAGE-A3, and NY-ESO-1-specific CTL were cultured from volunteer donors by stimulating peripheral blood mononuclear cells with dendritic cells pulsed with overlapping peptide mixes derived from full-length proteins, and these CTL preferentially lysed HLA partially matched, DAC-treated neuroblastoma and glioblastoma cell lines. These studies show that demethylating chemotherapy can be combined with IFN-? to increase the expression of CT antigens and MHC molecules on neuroblastoma cells, and pre-treatment with these agents makes tumor cell lines more susceptible to CTL-mediated killing. These data provide a basis to consider the use of demethylating chemotherapy in neuroblastoma patients, in conjunction with immune therapies that facilitate the expansion of CT antigen-specific CTL.
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Cloning and characterization of two ?-glucosidase/xylosidase enzymes from yak rumen metagenome.
Appl. Biochem. Biotechnol.
PUBLISHED: 04-20-2011
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Two ?-glucosidase/xylosidase genes, Rubg3A and Rubg3B, were cloned from yak rumen uncultured microorganisms by metagenome method and function-based screening. Recombinant RuBG3A and RuBG3B purified from Escherichia coli were characterized for enzymatic properties, and they exhibited activity against 4-nitrophenyl-?-D: -glucopyranoside and 4-nitrophenyl-?-D: -xylopyranoside, suggesting bifunctional ?-glucosidase/xylosidase activity. Chromatography analysis showed that they could effectively hydrolyze cellooligosaccharide substrates, indicating the facilitation in saccharification of cellulose. RuBG3A and RuBG3B can also increase the reducing sugar released in xylan hydrolysis to 218% and 169%, respectively, through synergism with xylanase, suggesting their application in hemicellulose saccharification. Molecular modeling and substrate docking showed that there should be one active center responsible for the bifunctional activity in each enzyme, since the active site pocket is substantially wide to allow the entry of both ?-glucosidic or ?-xylosidic substrates, which elucidated the structure-function relationship in substrate specificities. Therefore, the enzymatic properties, the participation in hydrolysis of cellooligosaccharides, and the synergism with xylanase make RuBG3A and RuBG3B very interesting candidates for saccharification of both cellulose and hemicellulose.
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How to appropriately choose and arrange research factors.
Zhong Xi Yi Jie He Xue Bao
PUBLISHED: 04-14-2011
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Research factors are a very important element in any research design. Research factors include experimental and non-experimental factors. The former is the general term used to describe the similar experimental conditions that researchers are interested in, while the latter are other factors that researchers have little interest in but may influence the result. This article mainly focuses on the following issues: the definition of research factors, the selection and arrangement of experimental factors and non-experimental factors, the interaction between research factors, the standardization of research factors and the common mistakes frequently made by researchers.
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Performance and clinical evaluation of the 92-gene real-time PCR assay for tumor classification.
J Mol Diagn
PUBLISHED: 03-29-2011
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Accurate determination of cancer origin is necessary to guide optimal treatment but remains a diagnostic challenge. Gene expression profiling technologies have aided the classification of tumors and, therefore, could be applied in conjunction with clinicopathologic correlates to improve accuracy. We report an expanded version of the previously described 92-gene assay to classify 30 main tumor types and 54 histological subtypes, with coverage of ?95% of all solid tumors based on incidence. Increased tissue coverage was achieved through expansion of a reference tumor database containing 2,206 specimens, with a median of 62 samples per main tumor type. The 92-gene classification algorithm demonstrated sensitivities of 87% and 85% for 30 main types and 54 histological subtypes, respectively, in leave-one-out cross validation, and 83% in a test set of 187 tumors representing 28 of the 30 main cancer types. These findings provide further support that broad and diverse tumor classification can be performed using a relatively compact gene set. An additional 300 consecutive cases submitted for clinical testing were profiled to characterize clinical utility in a real-world setting: the 92-gene assay confirmed 78% of samples having a single suspected primary tumor and provided a single molecular prediction in 74% of cases with two or more differential diagnoses. Further development of the 92-gene RT-PCR assay has resulted in a significant expansion in reportable tumor types and histological features with strong performance characteristics and supports the use of molecular classification as an objective standardized adjunct to current methods.
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How to appropriately choose research subjects.
Zhong Xi Yi Jie He Xue Bao
PUBLISHED: 03-23-2011
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The research subject is the first key element of the three key elements in the research design. An appropriate selection of research subjects is crucial to the success of the research. This article summarizes the general principles for the selection of research subjects, the types and numbers of research subjects and the common mistakes that researchers tend to make in the selection of the research subjects. This article also provides the methodology suggestions for the selection of research subjects.
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Accumulation of nerve growth factor and its receptors in the uterus and dorsal root ganglia in a mouse model of adenomyosis.
Reprod. Biol. Endocrinol.
PUBLISHED: 03-08-2011
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Adenomyosis is a common gynecological disease, which is accompanied by a series of immunological and neuroendocrinological changes. Nerve growth factor (NGF) plays a critical role in producing pain, neural plasticity, immunocyte aggregation and release of inflammatory factors. This study aimed to investigate the expression of NGF and its two receptors in uteri and dorsal root ganglia (DRG) in an adenomyosis mouse model, as well as their relationship with the severity of adenomyosis.
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Competition-based transfer of carbohydrate expression information from a cell-adhered surface to a secondary surface.
Chem. Commun. (Camb.)
PUBLISHED: 03-07-2011
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An information transfer strategy was developed for the visualization of carbohydrate expression by the competition of a primary cell-adhered solid surface with a carbohydrate assembled surface as an artificial secondary surface for one species. The strategy could be effectively utilized for in situ monitoring of dynamic carbohydrate expression on an adhesive cell surface.
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Wilms tumor 1-specific cytotoxic T lymphocytes can be expanded from adult donors and cord blood.
Leuk. Res.
PUBLISHED: 03-05-2011
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The use of WT1-specific CTL is one potential strategy to treat leukemic relapse following allogeneic stem cell transplant (SCT). Previous studies have largely focused on generating WT1-CTL from adult donors by cloning. We demonstrate that WT1-CTL can be generated from healthy adult donors and from cord blood by stimulating with an overlapping pool of peptides derived from full length WT1 and selecting antigen-specific cells based on the expression of CD137. The rapid expansion with anti-CD3 and IL-2 resulted in a 100-200-fold expansion. These CTL lysed WT1 expressing targets, including leukemia lines, in a HLA restricted manner.
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JoVE Visualize is a tool created to match the last 5 years of PubMed publications to methods in JoVE's video library.

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In developing our video relationships, we compare around 5 million PubMed articles to our library of over 4,500 methods videos. In some cases the language used in the PubMed abstracts makes matching that content to a JoVE video difficult. In other cases, there happens not to be any content in our video library that is relevant to the topic of a given abstract. In these cases, our algorithms are trying their best to display videos with relevant content, which can sometimes result in matched videos with only a slight relation.