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Find video protocols related to scientific articles indexed in Pubmed.
[Clinicopathological features associated with EGFR gene mutation in non-small cell lung cancer patients].
Zhonghua Yi Xue Za Zhi
PUBLISHED: 11-18-2014
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To explore the clinicopathological features associated with epidermal growth factor receptor (EGFR) gene mutation in non-small cell lung cancer (NSCLC) patients.
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Therapeutic effect of Jinlongshe Granule () on quality of life of stage IV gastric cancer patients using EORTC QLQ-C30: A double-blind placebo-controlled clinical trial.
Chin J Integr Med
PUBLISHED: 11-16-2014
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To evaluate the impact of Jinlongshe Granule (, JLSG) on quality of life (QOL) of stage IV gastric cancer patients.
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Investigation of drugs responsible for perioperative anaphylactic reactions using cellular allergen stimulation test.
Chin. Med. J.
PUBLISHED: 11-11-2014
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Anaphylactic reactions during anesthesia and operation are common and life threatening. Follow-up investigation is necessary for avoiding potential re-exposure of the patients to the offending drugs. The purpose of this study was to assess cellular allergen stimulation test (CAST) as a diagnostic instrument in immunoglobulin E (IgE)- and non-IgE-mediated anaphylactic reactions.
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Phylogenomics resolves the timing and pattern of insect evolution.
Bernhard Misof, Shanlin Liu, Karen Meusemann, Ralph S Peters, Alexander Donath, Christoph Mayer, Paul B Frandsen, Jessica Ware, Tomáš Flouri, Rolf G Beutel, Oliver Niehuis, Malte Petersen, Fernando Izquierdo-Carrasco, Torsten Wappler, Jes Rust, Andre J Aberer, Ulrike Aspöck, Horst Aspöck, Daniela Bartel, Alexander Blanke, Simon Berger, Alexander Böhm, Thomas R Buckley, Brett Calcott, Junqing Chen, Frank Friedrich, Makiko Fukui, Mari Fujita, Carola Greve, Peter Grobe, Shengchang Gu, Ying Huang, Lars S Jermiin, Akito Y Kawahara, Lars Krogmann, Martin Kubiak, Robert Lanfear, Harald Letsch, Yiyuan Li, Zhenyu Li, Jiguang Li, Haorong Lu, Ryuichiro Machida, Yuta Mashimo, Pashalia Kapli, Duane D McKenna, Guanliang Meng, Yasutaka Nakagaki, José Luis Navarrete-Heredia, Michael Ott, Yanxiang Ou, Günther Pass, Lars Podsiadlowski, Hans Pohl, Björn M von Reumont, Kai Schütte, Kaoru Sekiya, Shota Shimizu, Adam Slipinski, Alexandros Stamatakis, Wenhui Song, Xu Su, Nikolaus U Szucsich, Meihua Tan, Xuemei Tan, Min Tang, Jingbo Tang, Gerald Timelthaler, Shigekazu Tomizuka, Michelle Trautwein, Xiaoli Tong, Toshiki Uchifune, Manfred G Walzl, Brian M Wiegmann, Jeanne Wilbrandt, Benjamin Wipfler, Thomas K F Wong, Qiong Wu, Gengxiong Wu, Yinlong Xie, Shenzhou Yang, Qing Yang, David K Yeates, Kazunori Yoshizawa, Qing Zhang, Rui Zhang, Wenwei Zhang, Yunhui Zhang, Jing Zhao, Chengran Zhou, Lili Zhou, Tanja Ziesmann, Shijie Zou, Yingrui Li, Xun Xu, Yong Zhang, Huanming Yang, Jian Wang, Jun Wang, Karl M Kjer, Xin Zhou.
Science
PUBLISHED: 11-06-2014
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Insects are the most speciose group of animals, but the phylogenetic relationships of many major lineages remain unresolved. We inferred the phylogeny of insects from 1478 protein-coding genes. Phylogenomic analyses of nucleotide and amino acid sequences, with site-specific nucleotide or domain-specific amino acid substitution models, produced statistically robust and congruent results resolving previously controversial phylogenetic relations hips. We dated the origin of insects to the Early Ordovician [~479 million years ago (Ma)], of insect flight to the Early Devonian (~406 Ma), of major extant lineages to the Mississippian (~345 Ma), and the major diversification of holometabolous insects to the Early Cretaceous. Our phylogenomic study provides a comprehensive reliable scaffold for future comparative analyses of evolutionary innovations among insects.
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A COPD Health Management Program in a Community-Based Primary Care Setting: A Randomized Controlled Trial.
Respir Care
PUBLISHED: 11-06-2014
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A number of effective strategies have been developed to improve the quality of life in patients with COPD. However, few have been implemented in patients with COPD at all stages in a community setting. This study evaluated the efficacy of a complex COPD health management intervention in rural communities in China.
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Investigation of plasmonic whispering-gallery mode characteristics for graphene monolayer coated dielectric nanodisks.
Opt Lett
PUBLISHED: 11-01-2014
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In this Letter, we theoretically studied high-quality (Q) factor plasmonic whispering-gallery modes (WGMs) with ultrasmall mode volumes in graphene monolayer coated semiconductor nanodisks in the mid-infrared range. The influence of the chemical potential, the relaxation time of graphene, and the radius of the nanodisk on the cavity Q factor and the mode volume was numerically investigated. The numerical simulations showed that the plasmonic WGMs excited in this cavity had a deep subwavelength mode volume of 1.4×10-5(?0/2n)3, a cavity Q factor as high as 266 at a temperature lower than 250 K, and, consequently, a large Purcell factor of ?1.2×107 when the chemical potential and relaxation time were assumed to be 0.9 eV and 1.4 ps, respectively. The results provide a possible application of plasmonic WGMs in the integration of nano-optoelectronic devices based on graphene.
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High alkalinity boosts visible light driven H2 evolution activity of g-C3N4 in aqueous methanol.
Chem. Commun. (Camb.)
PUBLISHED: 10-30-2014
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A high rate of 2.23 mmol h(-1) g(-1) (quantum efficiency of 6.67% at 400 nm) for visible light driven photocatalytic H2 evolution can be achieved with g-C3N4 by alkalization of the solution to a pH of 13.3, due to accelerated transfer of photoholes to the sacrificial donor.
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Correlation of heme binding affinity and enzyme kinetics of dehaloperoxidase.
Biochemistry
PUBLISHED: 10-30-2014
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Chemical and thermal denaturation of dehaloperoxidase-hemoglobin (DHP) was investigated to test the relative stability of isoforms DHP A and DHP B and the H55V mutant of DHP A with respect to heme loss. In thermal denaturation experiments, heme loss was observed at temperatures of 54, 46, and 61 °C in DHP A, DHP B, and H55V, respectively. Guanidinium hydrochloride (GdnHCl)- and urea-induced denaturation was observed at respective concentrations of 1.15 ± 0.01 M DHP A and 1.09 ± 0.02 M DHP B, and 5.19 ± 0.05 M DHP A and 4.12 ± 0.14 M DHP B, respectively. The binding affinity of heme appears to be significantly smaller in both isoforms of DHP than in myoglobins. This observation was corroborated by heme transfer experiments, in which heme was observed to transfer for DHP A and B to horse skeletal muscle myoglobin (HSMb). GdnHCl-induced denaturation suggests a threshold of 1 mM for stabilization by binding of the inhibitor 4-bromophenol (4-BP). Concentrations of 4-BP greater than 1 mM caused destabilization. Urea-induced denaturation showed only destabilizing effects from phenolic ligand binding. Heme transfer experiments from DHP to HSMb further support the hypothesis that the binding of halophenols to DHP facilitates the removal of the heme. Thermal denaturation assessed via UV-visible spectroscopy and that assessed by differential scanning calorimetry (DSC) are both in agreement with chemical denaturation experiments and show that the denaturing abilities of the halophenols improve with the size of the para halogen atom in 4-XP, where X = iodo, bromo, chloro, or fluoro (4-IP > 4-BP > 4-CP > 4-FP), and the number of halo substituents as in 2,4,6-tribromophenol (2,4,6-TBP > 4-BP). DHP B, which differs in five amino acids, is less stable than DHP A with ?Hcal and Tm values of 165.1 kJ/mol and 47.5 °C compared to values of 183.3 kJ/mol and 50.4 °C for DHP B and DHP A, respectively. Kinetic studies verified that DHP B has a catalytic efficiency (kcat/Km) ?5-6 times greater than that of DHP A but showed an increased level of substrate inhibition in DHP B for both 2,4,6-TCP and 2,4,6-TBP. An inverse correlation between protein stability with respect to heme loss and catalytic efficiency is suggested on the basis of the fact that the heme in DHP B has a stability lower than that of DHP A but a catalytic efficiency higher than that of DHP A.
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Effect modification by gender and smoking status on the association between obesity and atopic sensitization in Chinese adults: a hospital-based case-control study.
BMC Public Health
PUBLISHED: 10-21-2014
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There is an ongoing debate on the potential association between obesity and atopy. However, no previous studies have investigated whether this relationship depends on sex and smoking status in Chinese adults.
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[Regulation mechanism of autophagy-related protein LC3 by c-Jun in methotrexate resistant human choriocarcinoma JEG-3 cells].
Zhonghua Fu Chan Ke Za Zhi
PUBLISHED: 10-21-2014
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To explore the regulation mechanism of autophagy- related protein, microtubule- associated protein 1 light chain 3 (LC3), via c-Jun in methotrexate resistant human choriocarcinoma JEG-3 cell lines.
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Glycan Imaging in Intact Rat Hearts and Glycoproteomic Analysis Reveal the Upregulation of Sialylation during Cardiac Hypertrophy.
J. Am. Chem. Soc.
PUBLISHED: 10-15-2014
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In the heart, glycosylation is involved in a variety of physiological and pathological processes. Cardiac glycosylation is dynamically regulated, which remains challenging to monitor in vivo. Here we describe a chemical approach for analyzing the dynamic cardiac glycome by metabolically labeling the cardiac glycans with azidosugars in living rats. The azides, serving as a chemical reporter, are chemoselectively conjugated with fluorophores using copper-free click chemistry for glycan imaging; derivatizing azides with affinity tags allows enrichment and proteomic identification of glycosylated cardiac proteins. We demonstrated this methodology by visualization of the cardiac sialylated glycans in intact hearts and identification of more than 200 cardiac proteins modified with sialic acids. We further applied this methodology to investigate the sialylation in hypertrophic hearts. The imaging results revealed an increase of sialic acid biosynthesis upon the induction of cardiac hypertrophy. Quantitative proteomic analysis identified multiple sialylated proteins including neural cell adhesion molecule 1, T-kininogens, and ?2-macroglobulin that were upregulated during hypertrophy. The methodology may be further extended to other types of glycosylation, as exemplified by the mucin-type O-linked glycosylation. Our results highlight the applications of metabolic glycan labeling coupled with bioorthogonal chemistry in probing the biosynthesis and function of cardiac glycome during pathophysiological responses.
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CD8(+)IL-17(+) T Cells Mediate Neutrophilic Airway Obliteration in T-bet Deficient Mouse Lung Allograft Recipients.
Am. J. Respir. Cell Mol. Biol.
PUBLISHED: 10-07-2014
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Acute cellular rejection is a known risk factor for the development of obliterative bronchiolitis which limits the long term survival of lung transplant recipients. However, the T cell effector mechanisms in both of these processes remain incompletely understood. Using the mouse orthotopic lung transplant model, we investigated whether C57BL/6 T-bet-/- recipients of MHC-mismatched BALB/c lung grafts develop rejection pathology and allospecific cytokine responses that differ from WT mice. T-bet-/- recipients demonstrated vigorous allograft rejection at 10 days, characterized by neutrophilic inflammation and predominantly CD8+ T cells producing allospecific IL-17 and/or IFN-?, in contrast to IFN-? dominant responses in WT mice. CD4+ T cells produced IL-17 but not IFN-? responses in T-bet-/- recipients in contrast to WT controls. Costimulation blockade using anti-CD154 Ab significantly reduced allospecific CD8+IFN-?+ responses in both T-bet-/- and WT mice, but had no attenuating effect on lung rejection pathology in Tbet-/- recipients, or the development of obliterative airway inflammation that occurred only in T-bet-/- recipients. However, neutralization of IL-17A significantly attenuated costimulation blockade-resistant rejection pathology and airway inflammation in T-bet-/- recipients. Additionally, CXCL1 (neutrophil chemokine) was increased in T-bet-/- allografts, and IL-17 induced CXCL1 from mouse lung epithelial cells in vitro. Taken together, our data show that T-bet-deficient recipients of complete MHC-mismatched lung allografts develop costimulation blockade-resistant rejection characterized by neutrophilia and obliterative airway inflammation that is predominantly mediated by CD8+IL-17+ T cells. Our data support T-bet-deficient mouse recipients of lung allografts as a viable animal model to study the immunopathogenesis of small airway injury in lung transplantion.
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Cell block from malignant pleural effusion might be a valid alternative sample for ALK detection in patients with advanced non small cell lung cancer.
Histopathology
PUBLISHED: 09-27-2014
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To evaluate the clinical value of cell block from malignant pleural effusion (MPE) as an alternative sample versus tumor tissue for anaplastic lymphoma kinase (ALK) detection in patients with advanced non small cell lung cancer (NSCLC).
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Inhibition of autophagy promoted sphingosylphosphorylcholine induced cell death in non-small cell lung cancer cells.
Biochem. Biophys. Res. Commun.
PUBLISHED: 09-25-2014
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Sphingosylphosphorylcholine (SPC) is a bioactive lipid mediated popular cell apoptosis in cancer cells. As a cell-specific sphingolipid, its function in lung cancer cells is unknown. Here we showed that SPC treatment triggered necrosis and autophagy but inhibited apoptosis in two non-small cell lung cancer cell lines: A549 cell line and H157 cell line. Then 3-methyladenine (3-MA), an autophagy inhibitor, was introduced to clarify the relationships between autophagy and necrosis or apoptosis. 3MA suppressed the survival furtherly by promoting apoptosis while had no influence on necrosis. Subsequent studies revealed that activity of AKT and mammalian target of rapamycin (mTOR) complex 1 (mTORC1) were downregulated during autophagy. Furthermore, SPC failed to promote autophagy in p53 deleted cells. Thus SPC induced autophagy in non-small cell lung cancer cells was through AKT/mTORC1 and P53 signal pathway. Besides, SPC reduced both the mitochondria membrane potential and ROS level in A549 cells. These findings provided a molecular basis of SPC-stimulated A549 cell death and support the notion that inhibition of autophagy is likely a novel anticancer mechanism.
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[Treatment of obstructive sleep apnea-hypopnea syndrome for children refractory asthma].
Zhonghua Er Bi Yan Hou Tou Jing Wai Ke Za Zhi
PUBLISHED: 09-23-2014
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The aim of this study was to understand the effect of different treatment of obstructive sleep apnea-hypopnea syndrome (OSAHS) for refractory asthma in children.
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The association between angiotensin-converting enzyme 2 polymorphisms and essential hypertension risk: A meta-analysis involving 14,122 patients.
J Renin Angiotensin Aldosterone Syst
PUBLISHED: 09-20-2014
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Some studies have evaluated the associations between the angiotensin-converting enzyme 2 (ACE2) gene polymorphisms and essential hypertension (EH) risk. However, the results remain uncertain. We carried out a meta-analysis to derive a more comprehensive estimation of these associations.
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Complete mitochondrial genome of the pink bollworm Pectinophora gossypiella (Lepidoptera: Gelechiidae).
Mitochondrial DNA
PUBLISHED: 09-19-2014
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Abstract Pectinophora gossypiella (Lepidoptera: Gelechiidae) is a key pest in many cotton-growing countries of the world. In this study, the complete mitochondrial (mt) genome of the pink bollworm P. gossypiella was determined, which is 15,202?bp in length (GenBank accession number: KM225795) containing 37 typical animal mitochondrial gene and an A?+?T-rich region. The gene order of P. gossypiella mtDNA was different from the insect ancestral gene order in the translocation of trnM, as shared by previously sequenced lepidopteran mtDNAs. The protein-coding genes (PCGs) have typical mitochondrial start codons ATN, with the exception of COI, Nad5, which uses the start codons CGA, GTT. Eight PCGs stop with complete termination codons (TAA), whereas five PCGs use incomplete stop codon T. All of the tRNA genes had typical cloverleaf secondary structures except for trnS1(AGN), in which the dihydrouridine (DHU) arm did not form a stable stem-loop structure. Like other insects, the control region is located between rrnS and trnM with a length of 309?bp and an A?+?T content of 94.8%, which is the most AT-rich region and comparatively simple, with little evidence of long tandem repeats, but harbors a conserved structure combining the motif ATAGA and a 18-bp poly-T stretch.
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[Effectiveness of inhaled hypertonic saline in children with bronchiolitis].
Zhonghua Er Ke Za Zhi
PUBLISHED: 09-17-2014
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To assess the efficacy and safety of inhaled nebulized hypertonic saline (HS) solution in infants with acute bronchiolitis.
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TSC1-mTORC1 signaling determines brown-to-white adipocyte phenotypic switch.
Diabetes
PUBLISHED: 09-13-2014
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Interconversion of white and brown adipocytes occurs between anabolic and catabolic states. The molecular mechanism regulating this phenotypic switch remains largely unknown. This study explores the role of tuberous sclerosis complex 1 (TSC1)-mechanistic target of rapamycin (mTOR) signaling in the conversion of brown to white adipose tissue. A colony of Fabp4-Tsc1-/- mice, in which the Tsc1 gene was specifically deleted by the fatty acid binding protein 4 (FABP4)-Cre, was established. Western blotting and immunostaining demonstrated the absence of TSC1 and activation of ribosomal protein S6 kinase 1, the downstream target of mTOR complex 1 (mTORC1) signaling, in the brown adipose tissues (BAT) of Fabp4-Tsc1-/- mice. Accumulation of lipid droplets in BAT was significantly increased. Levels of brown adipocyte markers were markedly down-regulated, while white adipocyte markers up-regulated. Rapamycin reversed the conversion from BAT to WAT in Fabp4-Tsc1-/- mice. Deletion of Tsc1 gene in cultured brown preadipocytes significantly increased the conversion to white adipocytes. FoxC2 mRNA, the transcriptional factor for brown adipocyte determination, was significantly decreased, while mRNAs for Rb, p107 and RIP140, the transcriptional factors for white adipocyte determination, increased in the BAT of Fabp4-Tsc1-/- mice. Our study demonstrates that TSC1-mTORC1 signaling contributes to the brown-to-white adipocyte phenotypic switch.
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[Noninvasive detection of hematocrit and the mean corpuscular hemoglobin concentration levels by Vis-NIR spectroscopy].
Guang Pu Xue Yu Guang Pu Fen Xi
PUBLISHED: 09-12-2014
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Hematocrit (HCT) and mean hemoglobin concentration(MCHC) play a very important role in preventing cardiovascular disease and anemia. A method was developed on the basis of spectroscopy to detect HCT and MCHC non-invasively and accurately. The anatomical study showed that the blood rheology abnormalities and blood viscosity's changes can cause the changes of tongue, so there is a certain correlation between tongue and blood components. Reflectance spectrums from the tongue tips of 240 volunteers were collected, then the tongue pictures were captured and the biochemical analysis results were recorded at the same time. The 240 samples were separated into two parts: calibration sample and test sample. Spectra were then subjected to a partial least squares regression (PLSR) analysis to develop mathematics models for predicting HCT levels. The correlation between the data and prediction of HCT and MCHC yielded calibration samples value of 0.998 and 0.938. HCT and MCHC levels of test samples predicted by this model from Visible-Near infrared spectra provided a coefficient of determination in prediction of 0.979 and 0.883 with an average relative error of prediction of 1.65% and 1.88%, a root mean square error of prediction of 4.066 and 4.139. From the experiment results we can see that the model which was built before can better predict the HCT and MCHC, and the results also showed that spectrometry method may provide a promising approach to the noninvasive measurement of human HCT and MCHC with a combination of PLSR analysis.
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2,3,5,4'-tetrahydroxystilbene-2-O-?-d-glucoside protects human umbilical vein endothelial cells against lysophosphatidylcholine-induced apoptosis by upregulating superoxide dismutase and glutathione peroxidase.
IUBMB Life
PUBLISHED: 09-11-2014
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2,3,5,4'-Tetrahydroxystilbene-2-O-?-d-glucoside (TSG) has been shown to protect human umbilical vein endothelial cells (HUVECs) from lysophosphatidylcholine (LPC)-induced injury; however, the underlying molecular mechanism remains to be determined. The aim of this study was to investigate the protective mechanism of TSG against LPC-induced injury in HUVECs. We established a stable LPC-induced cell model by treating HUVECs with various concentrations of LPC and found 10.0 µg/mL of LPC to be optimal for inducing HUVECs injury. The effects of TSG on LPC-induced cell injury were assessed by cell counting kit-8, apoptosis assay, transmission electron microscope, and measurement of malondialdehyde (MDA), the antioxidant enzymes superoxide dismutase (SOD), reactive oxygen species (ROS), glutathione peroxidase, and mitochondrial membrane potential. The mRNA and protein levels of caspase-3, Bax, Bcl-2, PARP-1, and cytochrome C were assayed by real-time reverse transcriptase-polymerase chain reaction and immunoblotting, respectively. TSG pretreatment was able to prevent LPC-induced HUVECs injury and restore cell viability in a concentration-dependent manner. LPC treated cells showed typical apoptotic morphological changes including cytoplasmic vacuolation, swollen mitochondria, and characteristic biochemical hallmarks of apoptosis including loss of mitochondrial membrane potential, activation of caspase-3, decrease of Bcl-2, increase of PARP-1, upregulation of Bax, and release of cytochrome C, all of which were apparently inhibited by TSG pretreatment. Treatment of HUVECs with LPC led to decrease of SOD and glutathione peroxidase in addition to rapid increase of MDA and ROS levels. Pretreatment with TSG restored SOD and glutathione peroxidase levels to that of normal levels, and significantly decreased ROS and MDA levels. Our data indicate that TSG inhibits apoptosis of HUVECs mediated by LPC through blocking the mitochondrial apoptotic pathway and suggest that the mechanisms underlying the protective effects of TSG are related to the activation of SOD and glutathione peroxidase, the clearance of intracellular ROS, and reduction of lipid peroxidation. © 2014 IUBMB Life, 66(10):711-722, 2014.
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A comparative study on immunomodulatory activity of polysaccharides from two official species of ganoderma (lingzhi).
Nutr Cancer
PUBLISHED: 09-10-2014
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Two Ganoderma species, G. lucidum and G. sinense, are listed as Lingzhi in Chinese Pharmacopoeia and they are considered to have the same therapeutic effects. Polysaccharides were the main immunomodulatory and anticancer components in Ganoderma. In this study, the chemical characters and the effects of polysaccharides from G. lucidum (GLPS) and G. sinense (GSPS) on macrophage functions were investigated and compared. Chemical studies showed that GLPS and GSPS were different, displaying various molecular weight distribution and ratio of monosaccharide components. In vitro pharmacological studies showed that both GLPS and GSPS had potent effects on macrophage functions, such as promoting macrophage phagocytosis, increasing their release of nitric oxide and cytokines interleukin (IL)-1?, IL-6, IL-10, and tumor necrosis factor-?. Generally, GLPS was more powerful than GSPS. This study is helpful to elucidate the active components and pharmacological variation between the 2 Ganoderma species. The structure-activity relationship of polysaccharides from Ganoderma needs further study.
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Synthesis and Antibacterial Activity of Cinnamaldehyde Acylhydrazone with a 1,4-Benzodioxan Fragment as a Novel Class of Potent ?-Ketoacyl-Acyl Carrier Protein Synthase III (FabH) Inhibitor.
Chem. Pharm. Bull.
PUBLISHED: 09-05-2014
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Fatty acid biosynthesis is essential for bacterial survival. ?-Ketoacyl-acyl carrier protein (ACP) synthase III (FabH), is a particularly attractive antibacterial target, since it is central to the initiation of fatty acid biosynthesis. Three series of 21 cinnamaldehyde acylhydrazone derivatives, A3-9, B3-9, and C3-9, were synthesized and evaluated for FabH-inhibitory activity. Compound B6 showed the most potent biological activity against Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Bacillus subtilis (minimum inhibitory concentrations (MICs) values: 1.56-3.13?µg/mL) and was comparable with the positive control. Docking simulation by positioning compound B6 in the FabH structure active site was performed to explore the possible binding model.
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Effects of hypoxic exercise training on microRNA expression and lipid metabolism in obese rat livers.
J Zhejiang Univ Sci B
PUBLISHED: 09-04-2014
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To investigate the effects of hypoxic exercise training on microRNA (miRNA) expression and the role of miRNA expression in regulating lipid metabolism, 20 dietary-induced obese SD rats were divided into a normoxic sedentary group (N, n=10) and a hypoxic exercise training group (H, n=10). After four weeks, measurements were taken of body weight, body length, fat mass, serum lipid concentration, miRNAs differentially expressed in rat liver, and gene and protein expression levels of peroxisome proliferator activated receptor ? (PPAR?), fatty acid synthetase (FAS), and carnitine palmitoyl transferase 1A (CPT1A) in rat liver. Body weight, Lee's index, fat mass, fat/weight ratio, and serum levels of total cholesterol (TC) and high density lipoprotein cholesterol (HDL-C) were all significantly lower in the H group than in the N group (P<0.01). Six miRNAs expressed significantly differently in the liver (P<0.05). Specifically, expression levels of miR-378b were significantly lower in the H group than in the N group (P<0.05). Compared with the normoxic sedentary group, hypoxic exercise training resulted in a lower ratio of FAS mRNA to CPT1A mRNA (P<0.05), as well as lower CPT1A protein levels (P<0.01), while a higher ratio of FAS to CPT1A protein levels (P<0.01) was observed. In conclusion, hypoxic training may elevate the resistance of high fat diet induced obesity in rats by reducing the expression of miR-378b, and decrease the fatty acid mitochondrial oxidation in obese rat livers by decreasing the protein expression of CPT1A and increasing the protein expression ratio of FAS/CPT1A.
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Preparation and characterization of zwitterionic phospholipid polymer-coated poly(lactic acid) nanoparticles.
J Biomater Sci Polym Ed
PUBLISHED: 09-03-2014
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Poly(lactic acid) (PLA) nanoparticles (NPs) are the most promising polymer NPs for drug delivery and targeting. However, they are easily recognized as a foreign body and rapidly cleared from the body by the mononuclear phagocyte system. Cell membrane mimetic random copolymers, bearing both zwitterionic phosphorylcholine groups and hydrophobic butyl side chains (PMB) and additional cross-linkable trimethoxysilylpropyl side chains (PMBT), were synthesized and coated on PLA NPs. Effects of the zwitterionic copolymer coatings on the NP size distribution, dispersion stability, and drug release behavior were investigated. Furthermore, the effect of the coatings on phagocytosis was also investigated. Compared with conventional polyvinyl alcohol coating, the cell membrane mimetic copolymer coatings decreased the size and increased the stability of the PLA NPs aqueous dispersions. More importantly, doxorubicin (DOX) release was well controlled and NPs phagocytosis by mouse peritoneal macrophage was decreased to one-third when the nanoparticles were coated with PMBT. This simple and effective zwitterionic polymer coating strategy may serve as a new route to design and optimize long-circulating intravenously injectable nanoparticle drug carriers.
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Rhodium(III)-catalyzed C-H activation/[4+3] annulation of N-phenoxyacetamides and ?,?-unsaturated aldehydes: an efficient route to 1,2-oxazepines at room temperature.
Chem. Commun. (Camb.)
PUBLISHED: 09-02-2014
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An efficient Rh(III)-catalyzed coupling reaction of N-phenoxyacetamides with ?,?-unsaturated aldehydes to give 1,2-oxazepines via C-H activation/[4+3] annulation has been developed. This transformation does not require oxidants and features C-C/C-N bond formation to yield seven-membered oxazepine rings at room temperature. Further derivation of 1,2-oxazepines leads to important chroman derivatives.
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Whether sperm deoxyribonucleic acid fragmentation has an effect on pregnancy and miscarriage after in vitro fertilization/intracytoplasmic sperm injection: a systematic review and meta-analysis.
Fertil. Steril.
PUBLISHED: 09-01-2014
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To examine whether sperm DNA fragmentation has an effect on pregnancy and miscarriage after IVF and/or intracytoplasmic sperm injection (ICSI).
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Association between GSTM1 null genotype and coronary artery disease risk: a meta-analysis.
Med. Sci. Monit.
PUBLISHED: 09-01-2014
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We conducted a meta-analysis to assess the association between polymorphisms of GSTM1 null genotype and coronary artery disease (CAD) risk.
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A new cycloartane-type triterpenoid saponin xanthine oxidase inhibitor from Homonoia riparia Lour.
Molecules
PUBLISHED: 08-29-2014
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A new cycloartane-type triterpenoid saponin named riparsaponin (1) was isolated from the stem of Homonoia riparia Lour together with six known compounds. The structure of riparsaponin was determined by using NMR and mass spectroscopy and X-ray crystallography techniques. Additionally, riparsaponin has a significant inhibitory effect on xanthine oxidase in vitro, and the IC50 was 11.16 nmol/mL.
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miR-135a inhibition protects A549 cells from LPS-induced apoptosis by targeting Bcl-2.
Biochem. Biophys. Res. Commun.
PUBLISHED: 08-28-2014
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Acute lung injury (ALI) is a severe clinical condition with high morbidity and mortality. Apoptosis is a key pathologic feature of ALI, and Bcl-2 plays an important role during the pathogenesis of ALI via the regulation of apoptosis. However, the regulation of Bcl-2 during ALI, particularly through microRNAs, remains unclear. We hypothesize that certain miRNAs may play deleterious or protective roles in ALI via the regulation of Bcl-2. The LPS stimulation of A549 cells was used to mimic ALI in vitro. First, we confirmed that Bcl-2 is involved in LPS-induced apoptosis in A549 cells. Then, bioinformatic analyses and quantitative real-time polymerase chain reaction assays were performed to screen for miRNAs targeting Bcl-2. We observed that miR-135a was markedly increased in LPS-challenged A549 cells. miR-135a inhibition markedly restored Bcl-2 expression and protected A549 cells from LPS-induced apoptosis. Furthermore, bioinformatic analysis and luciferase activity assays were conducted to confirm that miR-135a binds directly to the 3'-untranslated region of Bcl-2 and suppresses its expression. Interestingly, the inhibition of miR-135a did not attenuate apoptosis under LPS-treated conditions when Bcl-2 was knocked down. Therefore, we suggest that miR-135a regulation of LPS-induced apoptosis in A549 cells may depend in part on the regulation of Bcl-2. The miR-135a/Bcl-2 signaling pathway may be a novel therapeutic target for the prevention of ALI.
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A novel signal transduction pathway that modulates rhl quorum sensing and bacterial virulence in Pseudomonas aeruginosa.
PLoS Pathog.
PUBLISHED: 08-28-2014
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The rhl quorum-sensing (QS) system plays critical roles in the pathogenesis of P. aeruginosa. However, the regulatory effects that occur directly upstream of the rhl QS system are poorly understood. Here, we show that deletion of gene encoding for the two-component sensor BfmS leads to the activation of its cognate response regulator BfmR, which in turn directly binds to the promoter and decreases the expression of the rhlR gene that encodes the QS regulator RhlR, causing the inhibition of the rhl QS system. In the absence of bfmS, the Acka-Pta pathway can modulate the regulatory activity of BfmR. In addition, BfmS tunes the expression of 202 genes that comprise 3.6% of the P. aeruginosa genome. We further demonstrate that deletion of bfmS causes substantially reduced virulence in lettuce leaf, reduced cytotoxicity, enhanced invasion, and reduced bacterial survival during acute mouse lung infection. Intriguingly, specific missense mutations, which occur naturally in the bfmS gene in P. aeruginosa cystic fibrosis (CF) isolates such as DK2 strains and RP73 strain, can produce BfmS variants (BfmSL181P, BfmSL181P/E376Q, and BfmSR393H) that no longer repress, but instead activate BfmR. As a result, BfmS variants, but not the wild-type BfmS, inhibit the rhl QS system. This study thus uncovers a previously unexplored signal transduction pathway, BfmS/BfmR/RhlR, for the regulation of rhl QS in P. aeruginosa. We propose that BfmRS TCS may have an important role in the regulation and evolution of P. aeruginosa virulence during chronic infection in CF lungs.
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Serum starvation regulates E-cadherin upregulation via activation of c-Src in non-small-cell lung cancer A549 cells.
Am. J. Physiol., Cell Physiol.
PUBLISHED: 08-27-2014
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E-cadherin is essential for the integrity of adherens junctions between lung epithelial cells, and the loss of E-cadherin allows cell motility and is thought to promote lung cancer metastasis. While the downregulation of E-cadherin expression has been well characterized and is seen with transforming growth factor-?1 (TGF-?1) exposure, few studies have focused on E-cadherin upregulation. Here, we show that serum starvation causes increased E-cadherin expression via the activation of c-Src kinase in non-small-cell lung cancer A549 cells. Serum starvation increased E-cadherin protein levels in a time- and dose-dependent manner. E-cadherin mRNA transcripts were unchanged with starvation, while protein translation inhibition with cycloheximide attenuated E-cadherin protein induction by starvation, suggesting that E-cadherin is regulated at the translational level by serum starvation. c-Src is a nonreceptor tyrosine kinase known to regulate protein translation machinery; serum starvation caused early and sustained activation of c-Src in A549 cells followed by E-cadherin upregulation. Furthermore, overexpression of a dominant negative c-Src attenuated the induction of E-cadherin by serum deprivation. Finally, we observed that TGF-?1 treatment attenuated the serum activation of c-Src as well as E-cadherin expression when cells were deprived of serum. In conclusion, our data demonstrate that the c-Src kinase is activated by serum starvation to increase E-cadherin expression in A549 cells, and these phenomena are antagonized by TGF-?1. These novel observations implicate the c-Src kinase as an upstream inducer of E-cadherin protein translation with serum starvation and TGF-?1 diametrically regulating c-Src kinase activity and thus E-cadherin abundance in A549 cells.
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Effect of diclazuril on intestinal morphology and SIgA expression in chicken infected with Eimeria tenella.
Parasitol. Res.
PUBLISHED: 08-26-2014
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Secretory immunoglobulin A (SIgA), as a vital actor involving in the mucosal immunity, plays a key role in defending a variety of pathogenic infections, such as bacteria, viruses and parasites. Eimeria tenella is an obligate intracellular apicomplexan parasite contacting with the digestive tract mucosa and specially parasitizes chicken caecum, causing a severe form of coccidiosis. Coccidiosis is currently mainly controlled using chemotherapeutic agents. Diclazuril, a classic coccidiostat, was used widely in the poultry industry. Because of the rising problem of drug resistance, it is therefore crucial to understand the pattern of the SIgA expression in the action of diclazuril against E. tenella. In this study, the intestinal morphology in the caecum was analyzed by haematoxylin-eosin (HE) staining, and the SIgA expression was examined by immunohistochemical technique. At the same time, the duodenum, jejunum and ileum tissues have also been evaluated. HE staining results showed that E. tenella infection caused severe damage characterized by structural disorder, haemorrhage, inflammatory cell infiltration, serous and fibrinous exudation in chicken caecum and invisible damage in the duodenum, jejunum and ileum. With the treatment of diclazuril, the damage in the caecum was alleviated obviously. Immunohistochemical analysis demonstrated that the SIgA level in the infected group was increased in the duodenum (p?
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Ghrelin promotes hepatic lipogenesis by activation of mTOR-PPAR? signaling pathway.
Proc. Natl. Acad. Sci. U.S.A.
PUBLISHED: 08-25-2014
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Although ghrelin has been demonstrated to stimulate energy intake and storage through a central mechanism, its effect on hepatic lipid metabolism remains largely uncharacterized. Ghrelin receptor antagonism or gene deletion significantly decreased obesity-associated hepatic steatosis by suppression of de novo lipogenesis, whereas exogenous ghrelin stimulated lipogenesis, leading to hepatic lipid accumulation in mice. The effects of ghrelin were mediated by direct activation of its receptor on hepatocytes. Cultured hepatocytes responded to ghrelin with increased lipid content and expression of lipogenesis-related genes. Ghrelin increased phosphorylation of S6, the downstream target of mammalian target of rapamycin (mTOR) signaling in cultured hepatocytes, whereas ghrelin receptor antagonism reduced hepatic phosphorylation of S6 in db/db mice. Inhibition of mTOR signaling by rapamycin markedly attenuated ghrelin-induced up-regulation of lipogenesis in hepatocytes, whereas activation of hepatic mTOR signaling by deletion of TSC1 increased hepatic lipogenesis. By interacting with peroxisome proliferator-activated receptor-? (PPAR?), mTOR mediates the ghrelin-induced up-regulation of lipogenesis in hepatocytes. The stimulatory effect of ghrelin on hepatic lipogenesis was significantly attenuated by PPAR? antagonism in cultured hepatocytes and in PPAR? gene-deficient mice. Our study indicates that ghrelin activates its receptor on hepatocytes to promote lipogenesis via a mechanism involving the mTOR-PPAR? signaling pathway.
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Unique effects of the chain lengths and anions of tetra-alkylammonium salts on quenching pyrene excimer.
ACS Appl Mater Interfaces
PUBLISHED: 08-25-2014
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Pyrene (Py) excimer, through its unique fluorescence quenching, exhibits high sensitivity and high selectivity in detecting specific electron-deficient molecules, providing a potential platform for sensing technology, optical switch, and probing hydrophobicity of molecular environment. In solution state, its quenching mechanism has been well-studied. However, there remain many unknown properties regarding the quenching mechanism of the solid-state Py excimer. In this paper, the effects of a series of tetra-alkylammonium salts (with a variety of chain lengths and anions) on Py excimer quenching are investigated to identify the controlling parameters of the fluorescence quenching in the binary system. Several experimental approaches including steady-state fluorescence spectroscopy, UV absorption, (13)C-nuclear magnetic resonance (NMR) spectra, X-ray diffraction, scanning electron microscopy, and time-dependent fluorescence decay are employed to seek for the fundamental understanding of the quenching mechanism. The result indicates a unique quenching effect of tetrabutylammonium cation on the pyrene excimer, and which is not observed in the other cations with different chain lengths (the same associated hexafluorophosphate anions). Meanwhile, hexafluorophosphate anion (in the presence of tetrabutylammonium) is able to effectively retain Py excimer fluorescence when the system is prepared by evaporating solvent at high temperature. It is also confirmed that dynamic quenching is involved in the process. Hydrophobic environment around Py molecules shows strong correlation with the formation of Py excimer. The knowledge obtained in this study provides the insights to how the interaction between salt and Py molecule affects the excimer fluorescence.
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Sulfiredoxin-1 Attenuates Oxidative Stress via Nrf2/ARE Pathway and 2-Cys Prdxs After Oxygen-Glucose Deprivation in Astrocytes.
J. Mol. Neurosci.
PUBLISHED: 08-22-2014
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Sulfiredoxin-1 (Srxn1), an endogenous antioxidant protein, is involved in keeping the balance of the cell's oxidation/reduction and can resist oxidative stress. However, the exact antioxidant effects of Srxn1 remain fully unclear. The study aims to examine the effects of Srxn1 on oxidative stress and explore the potential mechanisms in astrocytes with 6 h/oxygen-glucose deprivation (OGD), 24 h/respiration. In the study, silencing Srxn1 was performed before exposure to 6 h/OGD, 24 h/respiration in primary astrocytes. Decreased cell viability and increased cellular damage measured by CellTiter 96H AQueous Non-Radioactive Cell Proliferation Assay (MTS) and lactate dehydrogenase (LDH) were observed in Srxn1 silencing astrocytes. In addition, Srxn1 silencing resulted in a decrease in both intracellular superoxide dismutase (SOD) and glutathione (GSH). NF-E2-related factor 2 (Nrf2), a transcription factor known to influence susceptibility to oxidative stress, upregulated Srxn1 expression during oxidative stress caused by OGD in the astrocytes. Electromobility shift assay (EMSA) demonstrated a decreased binding of Nrf2 to oligomers containing Srxn1 ter-specific antioxidant response element (ARE)-binding site in Nrf2 silencing astrocytes. We also found that a reduction of peroxiredoxin (Prdx)-SO3 was closely dependent on Srxn1. In addition, 2-Cys Prdxs protein levels were increased in the astrocytes exposed to OGD, as evaluated by immunoblot analysis. All taken together, the study suggested that silencing Srxn1 would result into increasing sensitivity to OGD-induced oxidative stress injury in astrocytes. Furthermore, Nrf2/ARE pathway was involved into Srxn1, playing its antioxidant protection against oxidative stress, all of which would provide a novel therapeutic theory for treating acute ischemic brain injury.
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Aryl hydrocarbon receptor negatively regulates NLRP3 inflammasome activity by inhibiting NLRP3 transcription.
Nat Commun
PUBLISHED: 08-20-2014
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NLRP3 inflammasome is a multi-protein complex, which plays crucial roles in host defense against pathogens. The NLRP3 protein level is considered rate limiting for the activation of the inflammasome, thus its expression must be tightly controlled to maintain immune homeostasis. However, the molecular mechanisms that modulate NLRP3 expression, especially at the transcriptional level, remain largely unknown. In the present study, we show that aryl hydrocarbon receptor (AhR) activation inhibits NLRP3 expression, caspase-1 activation and subsequent IL-1? secretion in peritoneal macrophages, whereas siRNA knockdown of AhR has opposite effects. AhR could bind to the xenobiotic response element (XRE) in the NLRP3 promoter and inhibit NLRP3 transcription. Furthermore, AhR activation suppresses Alum-induced peritonitis in vivo. Therefore, we identified AhR as a negative regulator of NLRP3 inflammasome activity by inhibiting the transcription of NLRP3 and suggested AhR as a potential target for the intervention of diseases with uncontrolled inflammasome activation.
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Functionalized carbon nanotube via distillation precipitation polymerization and its application in nafion-based composite membranes.
ACS Appl Mater Interfaces
PUBLISHED: 08-19-2014
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The objective of this study is to develop a novel approach to in situ functionalizing multiwalled carbon nanotubes (MWCNTs) and exploring their application in Nafion-based composite membranes for efficient proton conduction. Covalent grafting of acrylate-modified MWCNTs with poly(methacrylic acid-co-ethylene glycol dimethacrylate), poly(vinylphosphonic acid-co-ethylene glycol dimethacrylate), and sulfonated poly(styrene-co-divinylbenzene) was achieved via surface-initiated distillation precipitation polymerization. The formation of core-shell structure was verified by TEM images, and polymer layers with thickness around 30 nm were uniformly covered on the MWCNTs. The graft yield reached up to 93.3 wt % after 80 min of polymerization. The functionalized CNTs (FCNTs) were incorporated into the Nafion matrix to prepare composite membranes. The influence of various functional groups (-COOH, -PO3H2, and -SO3H) in FCNTs on proton transport of the composite membranes was studied. The incorporation of FCNTs afforded the composite membranes significantly enhanced proton conductivities under reduced relative humidity. The composite membrane containing 5 wt % phosphorylated MWCNTs (PCNTs) showed the highest proton conductivity, which was attributed to the construction of lower-energy-barrier proton transport pathways by PCNTs, and excellent water-retention and proton-conduction properties of the cross-linked polymer in PCNTs. Moreover, the composite membranes exhibited an enhanced mechanical stability.
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Changes in corneal deformation parameters after lenticule creation and extraction during small incision lenticule extraction (SMILE) procedure.
PLoS ONE
PUBLISHED: 08-14-2014
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To investigate the effects of lenticule creation and subsequent corneal lenticule extraction on corneal deformation parameters during small incision lenticule extraction (SMILE) procedure.
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A multitasking functional group leads to structural diversity using designer C-H activation reaction cascades.
Nat Commun
PUBLISHED: 08-14-2014
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The C-H activation strategy has become one of the preferred methods to introduce chemical functionality to a chemically inert carbon atom. Intensive efforts have been devoted to developing either versatile bond formations (product structural diversity) or effective directing groups (substrate site selectivity). From the views of medicinal and synthetic practitioners, the C-H activation approach remains inadequate due to its limitation to point-to-point derivatization. Direct assembly of 3D molecular complexity in a single step remains elusive for this strategy. Towards this goal, a multitasking functional group is required to accomplish several missions in one pot: site selecitivity, cleavability and redox versatility. We demonstrate that an oxyacetamide group is such a multifunctional warhead that enables a series of C-H functionalization cascades and allows direct access to structurally diverse polycyclic heterocyles in one pot. The progress of these reaction cascades were fully controlled by oxidants and temperature. The proliferation of the reaction chain can be extended to a four-step cascade.
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AKR1C3 overexpression mediates methotrexate resistance in choriocarcinoma cells.
Int J Med Sci
PUBLISHED: 08-13-2014
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Chemotherapy is typically used to treat choriocarcinoma, but a small proportion of tumors develop resistance to chemotherapy. Similarly, methotrexate (MTX) is a first-line chemotherapy used to treat choriocarcinoma; although ~30% of patients are drug-resistant for MTX mono-therapy. Thus, we sought to elucidate the mechanism of chemotherapeutic-resistance of MTX.
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Heterogeneous nuclear ribonucleoprotein E1 regulates protein disulfide isomerase translation in oxidized low-density lipoprotein -activated endothelial cells.
Acta Physiol (Oxf)
PUBLISHED: 08-08-2014
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Endothelium-derived protein disulfide isomerase (PDI) is required for thrombus formation in vivo. But, how to control PDI overproduction in oxidized low-density lipoprotein (oxLDL)-activated vascular endothelial cells (VECs) is not well understood. In this study, we try to answer this question by using our newly identified activator of mTOC1 3-benzyl-5-((2-nitrophenoxy) methyl)-dihydrofuran-2 (3H)-one (3BDO) that has been shown to protect VECs.
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Distinct subfunctionalization and neofunctionalization of the B-class MADS-box genes in Physalis floridana.
Planta
PUBLISHED: 08-08-2014
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This work suggested that in Physalis PFGLO1-PFDEF primarily determined corolla and androecium identity, and acquired a novel role in gynoecia functionality, while PFGLO2-PFTM6 functioned in pollen maturation only. The B-class MADS-box genes play a crucial role in determining the organ identity of the corolla and androecium. Two GLOBOSA-like (GLO-like) PFGLO1 and PFGLO2 and two DEFICIENS-like (DEF-like) PFDEF and PFTM6 genes were present in Physalis floridana. However, the double-layered-lantern1 (doll1) mutant is the result of a single recessive mutation in PFGLO1, hinting a distinct divergent pattern of B-class genes. In this work, we utilized the tobacco rattle virus (TRV)-mediated gene silencing approach to further verify this assumption in P. floridana. Silencing of PFGLO1 or/and PFDEF demonstrated their primary role in determining corolla and androecium identity. However, specific PFGLO2 or/and PFTM6 silencing did not affect any organ identity but showed a reduction in mature pollen. These results suggested that both PFGLO2 and PFTM6 had lost their role in organ identity determination but functioned in pollen maturation. Evaluation of fruit setting in reciprocal crosses suggested that both PFGLO1 and PFDEF might have acquired an essential and novel role in the functionality of gynoecia. Such a divergence of the duplicated GLO-DEF heterodimer genes in floral development is different from the existing observations within Solanaceae. Therefore, our research sheds new light on the functional evolution of the duplicated B-class MADS-box genes in angiosperms.
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Metallothioneins attenuate paraquat-induced acute lung injury in mice through the mechanisms of anti-oxidation and anti-apoptosis.
Food Chem. Toxicol.
PUBLISHED: 08-08-2014
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Paraquat (PQ) is a widely used herbicide, and lung is the primary target of PQ poisoning. Metallothionein (MT) is a potent antioxidant and free radical scavenger, and has been shown to play a protective role in lung injury induced by different stressors. This study was undertaken to evaluate the protective potential of MT against PQ-induced acute lung injury using MT-I/II null (MT(-/-)) mice. Wild-type (MT(+/+)) mice and MT(-/-) mice were given one intragastric administration of 50mg/kg PQ for 24h, and it was revealed that MT(-/-) mice were more susceptible to PQ-induced acute lung injury than MT(+/+) mice evidenced by the following findings. As compared with MT(+/+) mice, MT(-/-) mice presented more severe histopathological lesions in the lung, higher pulmonary malondialdehyde content, and more reduced pulmonary antioxidative enzymes activities. PQ also induced more apoptosis in pneumocytes from MT(-/-) mice, and the expressions of apoptosis-related proteins Bax, Bcl-2, cleaved-caspase-3, and the ratio of Bax/Bcl-2 were all more significantly increased in PQ-treated MT(-/-) mice. Our results clearly demonstrate that endogenous MT can attenuate PQ-induced acute lung injury, possibly through the mechanisms of anti-oxidation and anti-apoptosis.
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Objective optical quality and intraocular scattering in myopic adults.
Invest. Ophthalmol. Vis. Sci.
PUBLISHED: 08-07-2014
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To evaluate objective optical quality and intraocular scattering in adults with myopia.
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Ultrasensitive electrochemical detection of microRNA with star trigon structure and endonuclease mediated signal amplification.
Biosens Bioelectron
PUBLISHED: 08-04-2014
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MicroRNAs play important roles in gene regulation. They can be used as effective biomarkers for diagnosis and prognosis of diseases like cancers. Due to their intrinsic properties of short length, low abundance and sequence homology among family members, it is difficult to realize sensitive and selective detection with economical use of time and cost. Herein, we report an ultrasensitive electrochemical method for microRNA analysis employing two oligonucleotides and one endonuclease. Generally, a glassy carbon electrode is first covered with gold nanoparticles (AuNPs) mediated by poly(diallyldimethylammonium chloride) (PDDA). Then, thiolated capture probe (CP) with methylene blue (MB) labeled at 5' end is modified on the pretreated electrode. Hybridization occurs among target microRNA, CP and auxiliary probe (AP), forming a star trigon structure on the electrode surface. Subsequently, endonuclease recognizes and cleaves CP on CP/AP duplex, releasing microRNA and AP back to the solution. The two regenerated elements can then form another star trigon with other CP molecules, initiating cycles of CP cleavage and MB departure. Significant decrease of electrochemical signals is thus observed, which can be used to reflect the concentration of microRNA. This proposed method has a linear response to microRNA in a wide range from 100 aM to 1 nM and the sensitivity of attomolar level can be achieved. Moreover, it has high selectivity against single-base mismatch sequences and can be used directly in serum samples. Therefore, this method shows great feasibility for the detection of microRNA and may have potential applications in cancer diagnosis and prognosis.
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Serum anti-osteopontin autoantibody as a novel diagnostic and prognostic biomarker in patients with hepatocellular carcinoma.
Oncol. Rep.
PUBLISHED: 07-30-2014
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Osteopontin (OPN) is a secreted phosphorylated and glycosylated protein, which plays an important role in carcinogenesis and metastasis. In hepatocellular carcinoma (HCC), OPN is being investigated either as a therapeutic target gene or as a biomarker for diagnosis. Yet, the role of the anti-OPN autoantibody in HCC remains unclear. In the present study, the level of serum anti-OPN autoantibody in HCC was analyzed by enzyme-linked immunosorbent assay. Immunohistochemistry (IHC) was also performed to analyze protein expression profiles and the prognostic significance of OPN in HCC. In this study, the prevalence and titer of anti-OPN autoantibodies in HCC were significantly higher than these values in normal human serum (NHS) (P=0.001, P=0.000, respectively). When both ?-fetoprotein and the autoantibody against OPN were used simultaneously as diagnostic biomarkers, the sensitivity was up to 65%. In IHC, 59 of the 83 (65.6%) HCC specimens expressed OPN with cytoplasmic positive staining. The overall survival (OS) of HCC patients with OPN-positive tumors was 28.81 months compared to 39.37 months for HCC patients with OPN-negative tumors (P<0.01). Furthermore, multivariate analysis showed that OPN overexpression was the strongest independent adverse prognostic factor for OS (P=0.02). Taken together, our data indicate that the anti-OPN autoantibody may be a supplementary serological biomarker for HCC, and is correlated with poor prognosis in HCC patients.
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Catechol-O-methyltransferase Val158Met polymorphism: modulation of wearing-off susceptibility in a Chinese cohort of Parkinson's disease.
Parkinsonism Relat. Disord.
PUBLISHED: 07-27-2014
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Catechol-O-methyltransferase (COMT) is one of the cardinal enzymes that metabolize dopamine and other catecholamine neurotransmitters in the central and peripheral nervous system. Recent studies have shown that the impact of COMT haplotypes on the development of wearing-off phenomenon is in dispute, while the relationship between COMT haplotypes and wearing-off phenomenon in ethnic Chinese population is lacking. The purpose of this study was to characterize the correlation between the Val158Met polymorphism in the COMT gene and the motor complication "wearing-off" in Chinese PD patients. We have sequenced the COMT gene in 259 PD patients and 257 healthy controls. Our results demonstrated that Met/Met homozygosity of the COMT Val158Met polymorphism was related to a decreased risk of developing wearing-off. This finding suggests that COMT Val158Met may affect susceptibility to wearing-off in PD.
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First Acyclic Diene Metathesis Polymerization Under Biphasic Conditions Using a Dicationic Ruthenium Alkylidene: Access to High-Molecular-Weight Polymers with Very Low Ruthenium Contamination.
Macromol Rapid Commun
PUBLISHED: 07-24-2014
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The acyclic diene metathesis (ADMET) polymerization of 6-hydroxy-1,10-undecadiene (M1) and 6-acetoxy-1,10-undecadiene (M2) by the action of two different catalysts, i.e., the second-generation Grubbs-Hoveyda system ([RuCl2 (IMesH2 )(CH-2-(2-PrO-C6 H4 )]) (1) and the dicationic ruthenium alkylidene [Ru(DMF)3 (IMesH2 )(CH-2-(2-PrO-C6 H4 )] (2, IMesH2 = 1,3-dimesitylimidazolin-2-ylidene) is reported. Biphasic conditions using 1-butyl-2,3-dimethylimidazolium tetrafluoroborate ([BDMIM(+) BF4 (-) ]) and 1,2,4-trichlorobenzene (TCB) are applied. Under the chosen conditions (T = 75 °C, 20 mbar), the use of catalyst 1 results only in the formation of low-molecular-weight polymers (M¯n ? 10 000 g mol(-1) ), while catalyst 2 allows for the high yield synthesis of high-molecular-weight polymers (M¯n ? 40 000 g mol(-1) , yields ? 99%). Irrespective of the catalyst used, all polymers display a high trans-content (>95%). Notably, Ru-contamination of the target polymers without any additional purification is as low as 1.2 ppm with catalyst 2. Together with the high yields and high molecular weights, the low Ru-contaminations clearly illustrate the advantages of the biphasic setup.
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Increased Circulating Th1/Th2 Ratios But Not Other Lymphocyte Subsets During Controlled Ovarian Stimulation are Linked to Subsequent Implantation Failure after Transfer of In Vitro Fertilized Embryos.
Am. J. Reprod. Immunol.
PUBLISHED: 07-10-2014
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To investigate alterations of circulating lymphocyte subsets in women undergoing controlled ovarian stimulation (COS) and survey their relations with pregnancy outcome.
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Synthesis, molecular docking and biological evaluation of coumarin derivatives containing piperazine skeleton as potential antibacterial agents.
Bioorg. Med. Chem.
PUBLISHED: 07-05-2014
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A series of 4-hydroxycoumarin derivatives were designed and synthesized in order to find some more potent antibacterial drugs. Their antibacterial activities against Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis and Staphylococcus aureus were tested. These compounds showed good antibacterial activities against Gram-positive strains. Compound 4g represented the most potent antibacterial activity against Bacillus subtilis and S. aureus with MIC of 0.236, 0.355?g/mL, respectively. What's more, it showed the most potent activity against SaFabI with IC50 of 0.57?M. Molecular docking of 4g into S. aureus Enoyl-ACP-reductase active site were performed to determine the probable binding mode, while the QSAR model was built to check the previous work as well as to introduce new directions.
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Comparative proteomic analysis of antagonistic Bacillus amyloliquefaciens Q-426 cultivated under different pH conditions.
Biotechnol. Appl. Biochem.
PUBLISHED: 06-28-2014
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Bacillus amyloliquefaciens Q-426 produces lipopeptide compounds with antifungal activities. Initial pH value was proved to have a significant influence on production of lipopeptide compounds. The correlation between pH and intrinsic mechanism of lipopeptide production was rarely discussed. In this research, comparative proteomics, using two dimensional gel electrophoresis and mass spectrometry, was applied to identify B. amyloliquefaciens Q-426 intracellular proteins differentially expressed under initial pH 5.0 and pH 7.3. A total of 24 differential spots (8 down-regulated and 16 up-regulated) under pH 5.0 were identified. Certain proteins involve in the regulation of bacillomycin and fengycin production by B. amyloliquefaciens Q-426. These proteins include four induced proteins related to stress response, TPP-dependent acetoin dehydrogenase, butanediol dehydrogenase, two ABC-type oligopeptide transport system proteins, two-component response regulator DegU and chorismate mutase PheB. These results indicated intrinsic differences of antagonistic B. amyloliquefaciens Q-426 under different pH conditions. This article is protected by copyright. All rights reserved.
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Coping styles and social support among depressed Chinese family caregivers of patients with esophageal cancer.
Eur J Oncol Nurs
PUBLISHED: 06-22-2014
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To determine the coping styles of family caregivers of patients with esophageal cancer and examine the relationships between depression, coping styles and social support.
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Radiographic, clinical, and patients' assessment of segmental direct vertebral body derotation versus simple rod derotation in main thoracic adolescent idiopathic scoliosis: a prospective, comparative cohort study.
Eur Spine J
PUBLISHED: 06-20-2014
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The application of vertebral body derotation (DVBD) is still controversial by now; the purpose of this prospective cohort study was to compare comprehensive outcomes between segmental DVBD and simple rod derotation (SRD) especially in main thoracic adolescent idiopathic scoliosis.
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Chitosan Degradation Products Promote Nerve Regeneration by Stimulating Schwann Cell Proliferation via miR-27a/FOXO1 Axis.
Mol. Neurobiol.
PUBLISHED: 06-16-2014
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Natural polysaccharides are biomaterials widely used for constructing scaffolds in tissue engineering. While natural polysaccharides have been shown to robustly promote tissue regeneration, the underlying molecular mechanism remains largely unknown. Here, we show that chitooligosaccharides (COS), the intermediate products of chitosan degradation, stimulate peripheral nerve regeneration in rats. Our experiment also shows that COS stimulate the proliferation of Schwann cells (SCs) during nerve regeneration. By analyzing the transcriptome and gene regulatory network, we identified the miR-27a/FOXO1 axis as the main signaling pathway for mediating the proliferative effects of COS on SCs. COS increase the expression level of miR-27a and cause a reduction of FOXO1, which subsequently accelerates the cell cycle and stimulates SC proliferation to stimulate nerve regeneration. These findings define a basic pathway for oligosaccharides-mediated cell proliferation and reveal a novel aspect of polysaccharide biomaterials in tissue engineering.
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Biocompatible click chemistry enabled compartment-specific pH measurement inside E. coli.
Nat Commun
PUBLISHED: 06-01-2014
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Bioorthogonal reactions, especially the Cu(I)-catalysed azide-alkyne cycloaddition, have revolutionized our ability to label and manipulate biomolecules under living conditions. The cytotoxicity of Cu(I) ions, however, has hindered the application of this reaction in the internal space of living cells. By systematically surveying a panel of Cu(I)-stabilizing ligands in promoting protein labelling within the cytoplasm of Escherichia coli, we identify a highly efficient and biocompatible catalyst for intracellular modification of proteins by azide-alkyne cycloaddition. This reaction permits us to conjugate an environment-sensitive fluorophore site specifically onto HdeA, an acid-stress chaperone that adopts pH-dependent conformational changes, in both the periplasm and cytoplasm of E. coli. The resulting protein-fluorophore hybrid pH indicators enable compartment-specific pH measurement to determine the pH gradient across the E. coli cytoplasmic membrane. This construct also allows the measurement of E. coli transmembrane potential, and the determination of the proton motive force across its inner membrane under normal and acid-stress conditions.
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Preparation and Optimization of Resveratrol Nanosuspensions by Antisolvent Precipitation Using Box-Behnken Design.
AAPS PharmSciTech
PUBLISHED: 05-18-2014
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Resveratrol, a natural polyphenolic component, has inspired considerable interest for its extensive physiological activities. However, the poor solubility of resveratrol circumscribes its therapeutic applications. The purpose of this study was to optimize and prepare resveratrol nanosuspensions using the antisolvent precipitation method. The effects of crucial formulation and process variables (drug concentration, stabilizer, and surfactant contents) on particle size were investigated by utilizing a three-factor three-level Box-Behnken design (BBD) to perform this experiment. Different mathematical polynomial models were used to identify the impact of selected parameters and to evaluate their interrelationship for predictive formulation purposes. The optimal formulation consisted of drug 29.2 (mg/ml), polyvinylpyrrolidone (PVP) K17 0.38%, and F188 3.63%, respectively. The morphology of nanosuspensions was found to be near-spherical shaped by scanning electron microscopy (SEM) observation. The X-ray powder diffraction (XRPD) and differential scanning calorimetry (DSC) analysis confirmed that the nanoparticles were in the amorphous state. Furthermore, in comparison to raw material, resveratrol nanosuspensions showed significantly enhanced saturation solubility and accelerated dissolution rate resulting from the decrease in particle size and the amorphous status of nanoparticles. Meanwhile, resveratrol nanosuspensions exhibited the similar antioxidant potency to that of raw resveratrol. The in vivo pharmacokinetic study revealed that the C max and AUC0?? values of nanosuspension were approximately 3.35- and 1.27-fold greater than those of reference preparation, respectively. Taken together, these results suggest that this study provides a beneficial approach to address the poor solubility issue of the resveratrol and affords a rational strategy to widen the application range of this interesting substance.
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Nuclear factor of activated T cells 5 maintained by Hotair suppression of miR-568 upregulates S100 calcium binding protein A4 to promote breast cancer metastasis.
Breast Cancer Res.
PUBLISHED: 05-13-2014
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IntroductionThe onset of distal metastasis, which underlies the high mortality of breast cancers, warrants substantial studies to depict its molecular basis. Nuclear factor of activated T cells 5 (NFAT5) is upregulated in various malignancies and is critically involved in migration and invasion of neoplastic cells. Nevertheless, the metastasis-related events potentiated by this transcriptional factor and the mechanism responsible for NFAT5 elevation in carcinoma cells remain to be fully elucidated.MethodsThe correlation of NFAT5 with breast cancer invasiveness was investigated in vitro and clinically. The genes transcriptionally activated by NFAT5 were probed and their roles in breast cancer progression were dissected. The upstream regulators of NFAT5 were studied with particular attempt to explore the involvement of non-coding RNAs, and the mechanism underlying the maintenance of NFAT5 expression was deciphered.ResultsIn metastatic breast cancers, NFAT5 promotes epithelial-mesenchymal transition (EMT) and invasion of cells by switching on the expression of the calcium binding protein S100A4, and facilitates the angiogenesis of breast epithelial cells and thus the development of metastases by transcriptionally activating vascular endothelial growth factor C (VEGF-C). NFAT5 is directly targeted by miR-568, which is in turn suppressed by the long non-coding RNA, Hotair, via a documented in trans gene silencing pattern, that is recruitment of the polycomb complex (Polycomb Repressive Complex 2; PRC2) and LSD1, and consequently methylation of histone H3K27 and demethylation of H3K4 on the miR-568 loci.ConclusionThis study unravels a detailed role of NFAT5 in mediating metastatic signaling, and provides broad insights into the involvement of Hotair, in particular, by transcriptionally regulating the expression of microRNA(s), in the metastasis of breast cancers.
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Association mapping of brassinosteroid candidate genes and plant architecture in a diverse panel of Sorghum bicolor.
Theor. Appl. Genet.
PUBLISHED: 05-04-2014
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This first association analysis between plant architecture and BR candidate genes in sorghum suggests that natural allelic variation has significant and pleiotropic effects on plant architecture phenotypes. Sorghum bicolor (L) Moench is a self-pollinated species traditionally used as a staple crop for human consumption and as a forage crop for livestock feed. Recently, sorghum has received attention as a bioenergy crop due to its water use efficiency and biomass yield potential. Breeding for superior bioenergy-type lines requires knowledge of the genetic mechanisms controlling plant architecture. Brassinosteroids (BRs) are a group of hormones that determine plant growth, development, and architecture. Biochemical and genetic information on BRs are available from model species but the application of that knowledge to crop species has been very limited. A candidate gene association mapping approach and a diverse sorghum collection of 315 accessions were used to assess marker-trait associations between BR biosynthesis and signaling genes and six plant architecture traits. A total of 263 single nucleotide polymorphisms (SNPs) from 26 BR genes were tested, 73 SNPs were significantly associated with the phenotypes of interest and 18 of those were associated with more than one trait. An analysis of the phenotypic variation explained by each BR pathway revealed that the signaling pathway had a larger effect for most phenotypes (R (2) = 0.05-0.23). This study constitutes the first association analysis between plant architecture and BR genes in sorghum and the first LD mapping for leaf angle, stem circumference, panicle exsertion and panicle length. Markers on or close to BKI1 associated with all phenotypes and thus, they are the most important outcomes of this study and will be further validated for their future application in breeding programs.
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Core-shell hollow microspheres of magnetic iron oxide@amorphous calcium phosphate: synthesis using adenosine 5'-triphosphate and application in pH-responsive drug delivery.
Chem Asian J
PUBLISHED: 04-02-2014
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Drug nanocarriers with magnetic targeting and pH-responsive drug-release behavior are promising for applications in controlled drug delivery. Magnetic iron oxides show excellent magnetism, but their application in drug delivery is limited by low drug-loading capacity and poor control over drug release. Herein, core-shell hollow microspheres of magnetic iron oxide@amorphous calcium phosphate (MIO@ACP) were prepared and investigated as magnetic, pH-responsive drug nanocarriers. Hollow microspheres of magnetic iron oxide (HMIOs) were prepared by etching solid MIO microspheres in hydrochloric acid/ethanol solution. After loading a drug into the HMIOs, the drug-loaded HMIOs were coated with a protective layer of ACP by using adenosine 5'-triphosphate (ATP) disodium salt (Na2 ATP) as stabilizer, and drug-loaded core-shell hollow microspheres of MIO@ACP (HMIOs/drug/ACP) were obtained. The as-prepared HMIOs/drug/ACP drug-delivery system exhibits superparamagnetism and pH-responsive drug-release behavior. In a medium with pH?7.4, drug release was slow, but it was significantly accelerated at pH?4.5 due to dissolution of the ACP shell. Docetaxel-loaded core-shell hollow microspheres of MIO@ACP exhibited high anticancer activity.
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Prognostic significance of NPM1 mutations in acute myeloid leukemia: A meta-analysis.
Mol Clin Oncol
PUBLISHED: 03-21-2014
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Nucleophosmin 1 (NPM1) mutations have been identified in a substantial number of patients with acute myeloid leukemia (AML). Favorable outcomes in AML cases with NPM1 mutations have been previously reported. However, widely differing survival estimates have been indicated. Therefore, a meta-analysis of nine studies including a total of 4509 subjects was performed. The frequency of NPM1 mutations was found to be 6.45-56.08%. NPM1-mutation type (NPM1-mt) patients had >2-fold higher odds of achieving complete remission compared with NPM1-wild-type (NPM1-wt). The summary hazard ratio (HR) of NPM1-mt/NPM1-wt for disease-free survival (DFS) and OS was 0.67 and 0.63, respectively. In conclusion, these findings suggest that the NPM1 mutation has a favorable effect on the outcome for AML. The present meta-analysis was based on data abstracted from observational studies. However, the results obtained may justify the risk-adapted therapeutic strategies for AML according to the NPM1 status.
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Magneto-fluorescent core-shell supernanoparticles.
Nat Commun
PUBLISHED: 03-12-2014
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Magneto-fluorescent particles have been recognized as an emerging class of materials that exhibit great potential in advanced applications. However, synthesizing such magneto-fluorescent nanomaterials that simultaneously exhibit uniform and tunable sizes, high magnetic content loading, maximized fluorophore coverage at the surface and a versatile surface functionality has proven challenging. Here we report a simple approach for co-assembling magnetic nanoparticles with fluorescent quantum dots to form colloidal magneto-fluorescent supernanoparticles. Importantly, these supernanoparticles exhibit a superstructure consisting of a close-packed magnetic nanoparticle 'core', which is fully surrounded by a 'shell' of fluorescent quantum dots. A thin layer of silica coating provides high colloidal stability and biocompatibility, and a versatile surface functionality. We demonstrate that after surface pegylation, these silica-coated magneto-fluorescent supernanoparticles can be magnetically manipulated inside living cells while being optically tracked. Moreover, our silica-coated magneto-fluorescent supernanoparticles can also serve as an in vivo multi-photon and magnetic resonance dual-modal imaging probe.
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Characterization and discrimination of polysaccharides from different species of Cordyceps using saccharide mapping based on PACE and HPTLC.
Carbohydr Polym
PUBLISHED: 02-18-2014
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Polysaccharides from seven species of natural and cultured Cordyceps were firstly investigated and compared using saccharide mapping, partially acidic/enzymatic (?-amylase, ?-glucanase and pectinase) digestion followed with polysaccharide analysis by using carbohydrate gel electrophoresis (PACE) and high performance thin layer chromatography (HPTLC) analysis, respectively, to obtain the comprehensive profiles of hydrolysates of the polysaccharides and their characters. The results showed that 1,4-?-D-glucosidic, 1,4-?-D-glucosidic and 1,4-?-D-galactosidic linkages were existed in natural and cultured Cordyceps sinensis, cultured Cordyceps militaris, natural Cordyceps gracilis and Cordyceps ciecadae. The similarity of polysaccharides from cultured C. militaris to natural C. sinensis was relatively high, which might contribute to the rational use of C. militaris. Moreover, different species of natural and cultured Cordyceps can be differentiated based on the saccharide mapping, which is helpful to well understand the structural characters of polysaccharides from different species of Cordyceps and to improve the quality control of polysaccharides in natural and cultured Cordyceps.
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Homologous recombination efficiency enhanced by inhibition of MEK and GSK3?
Genesis
PUBLISHED: 02-10-2014
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Homologous recombination in embryonic stem cells (ESCs) is widely utilized in genome engineering, particularly in the generation of gene targeted mice. However, genome engineering is often plagued by the problem of low homologous recombination efficiency. In this study, we developed a novel method to increase the efficiency of homologous recombination in ESCs by changing its culture conditions. By comparing the efficiency of different ESCs in various culture conditions, we determined that chemicals that inhibit the MEK and GSK3? pathways (2i condition) enhance homologous recombination and eliminate differences in efficiencies among cell lines. Analysis of gene expression patterns in ESCs maintained in different culture conditions has identified several homologous recombination-related candidates, including the pluripotent markers Eras and Tbx3. The results of this study suggest that homologous recombination is associated with ESC pluripotency. genesis 00:1-8, 2014. © 2014 Wiley Periodicals, Inc.
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Chitosan-coated mesoporous microspheres of calcium silicate hydrate: environmentally friendly synthesis and application as a highly efficient adsorbent for heavy metal ions.
J Colloid Interface Sci
PUBLISHED: 01-28-2014
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Chitosan-coated calcium silicate hydrate (CSH/chitosan) mesoporous microspheres formed by self-assembly of nanosheets have been synthesized in aqueous solution under ambient conditions without using any toxic surfactant or organic solvent. The method reported herein has advantages of simplicity, low cost and being environmentally friendly. The BET specific surface area of CSH/chitosan mesoporous microspheres is measured to be as high as ~356 m(2) g(-1), which is considerably high among calcium silicate materials. The as-prepared CSH/chitosan mesoporous microspheres are promising adsorbent and exhibit a quick and highly efficient adsorption behavior toward heavy metal ions of Ni(2+), Zn(2+), Cr(3+), Pb(2+) Cu(2+) and Cd(2+) in aqueous solution. The adsorption kinetics can be well fitted by the pseudo second-order model. The maximum adsorption amounts of Ni(2+), Zn(2+), Pb(2+), Cu(2+) and Cd(2+) on CSH/chitosan mesoporous microspheres are extremely high, which are 406.6, 400, 796, 425 and 578 mg/g, respectively. The CSH/chitosan adsorbent exhibits the highest affinity for Pb(2+) ions among five heavy metal ions. The adsorption capacities of the CSH/chitosan adsorbent toward heavy metal ions are relatively high compared with those reported in the literature.
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Upper airway cough syndrome in children and two inflammatory factors: TRPV1 and TGF-?2.
Int. J. Pediatr. Otorhinolaryngol.
PUBLISHED: 01-22-2014
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The aim of the study was to investigate upper airway cough syndrome (UACS) in children and to determine alternative methods to explore the relationships among TRPV1, TGF-?2, and UACS.
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Giant cell tumor of the clivus: A case report and review of the literature.
Oncol Lett
PUBLISHED: 01-15-2014
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Giant cell tumors (GCTs) mainly occur in metaphyses of long bones and are generally considered histologically benign; however, GCTs may be locally aggressive with a high rate of local recurrence and exhibit the potential for distant metastasis. Primary GCT of the clivus is extremely rare and is easily misdiagnosed and, thus, treatment remains controversial. The present report describes the case of a 22-year-old male with GCT located in the skull base originating from the clivus, with the involvement of multiple cranial nerves, which was successfully treated with transnasal transsphenoidal surgery following adjuvant radiotherapy and intravenous bisphosphonate administration. The patient remains symptom free at two years of follow-up. This report contributes to the limited literature regarding GCTs of the skull.
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Bt Rice Expressing Cry2Aa Does Not Harm Cyrtorhinus lividipennis, a Main Predator of the Nontarget Herbivore Nilapavarta lugens.
PLoS ONE
PUBLISHED: 01-01-2014
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T2A-1 is a newly developed transgenic rice that expresses a synthesized cry2Aa gene driven by the maize ubiquitin promoter. T2A-1 exhibits high resistance against lepidopteran pests of rice. The brown planthopper, Nilapavarta lugens (Stål), is a main nontarget sap-sucking insect pest of rice, and Cyrtorhinus lividipennis (Reuter) is the major predator of the eggs and young nymphs of planthoppers. As C. lividipennis may expose to the Cry2Aa protein via N. lugens, it is therefore essential to assess the potential effects of transgenic cry2Aa rice on this predator. In the present study, three experiments were conducted to evaluate the ecological risk of transgenic cry2Aa rice to C. lividipennis: (1) a direct feeding experiment in which C. lividipennis was fed an artificial diet containing Cry2Aa at the dose of 10-time higher than that it may encounter in the realistic field condition; (2) a tritrophic experiment in which the Cry2Aa protein was delivered to C. lividipennis indirectly through prey eggs or nymphs; (3) a realistic field experiment in which the population dynamics of C. lividipennis were investigated using vacuum-suction. Both direct exposure to elevated doses of the Cry2Aa protein and prey-mediated exposure to realistic doses of the protein did not result in significant detrimental effects on the development, survival, female ratio and body weight of C. lividipennis. No significant differences in population density and population dynamics were observed between C. lividipennis in transgenic cry2Aa and nontransgenic rice fields. It may be concluded that transgenic cry2Aa rice had no detrimental effects on C. lividipennis. This study represents the first report of an assessment continuum for the effects of transgenic cry2Aa rice on C. lividipennis.
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Fine Epitope Mapping of the Central Immunodominant Region of Nucleoprotein from Crimean-Congo Hemorrhagic Fever Virus (CCHFV).
PLoS ONE
PUBLISHED: 01-01-2014
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Crimean-Congo hemorrhagic fever (CCHF), a severe viral disease known to have occurred in over 30 countries and distinct regions, is caused by the tick-borne CCHF virus (CCHFV). Nucleocapsid protein (NP), which is encoded by the S gene, is the primary antigen detectable in infected cells. The goal of the present study was to map the minimal motifs of B-cell epitopes (BCEs) on NP. Five precise BCEs (E1, 247FDEAKK252; E2a, 254VEAL257; E2b, 258NGYLNKH264; E3, 267EVDKA271; and E4, 274DSMITN279) identified through the use of rabbit antiserum, and one BCE (E5, 258NGYL261) recognized using a mouse monoclonal antibody, were confirmed to be within the central region of NP and were partially represented among the predicted epitopes. Notably, the five BCEs identified using the rabbit sera were able to react with positive serum mixtures from five sheep which had been infected naturally with CCHFV. The multiple sequence alignment (MSA) revealed high conservation of the identified BCEs among ten CCHFV strains from different areas. Interestingly, the identified BCEs with only one residue variation can apparently be recognized by the positive sera of sheep naturally infected with CCHFV. Computer-generated three-dimensional structural models indicated that all the antigenic motifs are located on the surface of the NP stalk domain. This report represents the first identification and mapping of the minimal BCEs of CCHFV-NP along with an analysis of their primary and structural properties. Our identification of the minimal linear BCEs of CCHFV-NP may provide fundamental data for developing rapid diagnostic reagents and illuminating the pathogenic mechanism of CCHFV.
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