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Find video protocols related to scientific articles indexed in Pubmed.
Multiband localized spoof plasmons in closed textured cavities.
Appl Opt
PUBLISHED: 11-18-2014
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In this work, we explore the existence of multiband localized spoof plasmons (LSPs) in closed textured cavities with multiple groove depths. It is interesting to note that the spoof LSPs in each band resemble those generated by the textured 2D cavities of the same periodicity with the corresponding single groove depth, and the field distributions and confinement characteristics of the plasmon-like modes in such a corrugated cavity are different from the conventional cavity. Hence, these multiple resonance band structures can find potential applications in the microwave and terahertz frequencies.
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[Analysis of cervical cancer incidence and mortality in cancer registries of Zhejiang province, 2000 to 2009].
Zhonghua Yu Fang Yi Xue Za Zhi
PUBLISHED: 11-13-2014
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To investigate the cervical cancer incidence and mortality in cancer registries of Zhejiang province during 2000 to 2009.
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Stitching graphene oxide sheets into a membrane at a liquid/liquid interface.
Chem. Commun. (Camb.)
PUBLISHED: 11-10-2014
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Layer-by-layer patterned GO membrane assembling at a liquid/liquid interface was realized using polyetheramine "stitching" molecules with an adjustable hydrophobicity and its thickness can be easily tuned from nanometer to micrometer by controlling reaction kinetics including reactant concentration, reaction time and reaction temperature.
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[Early diagnostic values of CD64 and CD11b indices of peripheral white blood cells for acute exacerbation of chronic obstructive pulmonary disease in older adults].
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi
PUBLISHED: 11-07-2014
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Objective To investigate the values of CD64 and CD11b indices of peripheral white blood cells in the early diagnosis of acute exacerbation of chronic obstructive pulmonary disease (AE-COPD) in older adults. Methods The study enrolled 86 aged AE-COPD patients, 82 stable-COPD patients admitted in the affiliated hospital of Jiujiang University from March 2011 to December 2013, and simultaneously 84 healthy aged volunteers as a control group. All the subjects were examined in white blood cells, hypersensitive C-reactive protein (hs-CRP), mean fluorescence intensity (MFI) of CD64 and CD11b from peripheral white blood cells within 24 hours after admission. CD64 and CD11b MFI were converted into CD64 and CD11b indices by conversion formula. The significant indicators for the diagnosis of AE-COPD were screened and the receiver operating characteristic (ROC) curve was drawn for calculating the area under the curve, critical value, sensitivity and specificity. Results Compare with stable-COPD group, the CD11b index decreased and CD64 index increased. There existed statistical difference in CD11b and CD64 indices between the AE-COPD group and the stable-COPD group (P<0.01), but not between the stable-COPD group and the healthy control group (P>0.05). Critical values of CD11b and CD64 indices were respectively less than 0.94 and more than 1.83. Their sensitively and specificity for the diagnosis of AE-COPD were 62.65% and 77.11% for CD11b and 79.52% and 98.80% for CD64. Conclusion Increased CD64 index and decreased CD11b index are the credible laboratory markers in the early diagnosis of AE-COPD, and the dynamic monitoring of them may facilitate the evaluation of therapeutic outcomes of AE-COPD.
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Formation of a G-Quartet-Fe Complex and Modulation of Electronic and Magnetic Properties of the Fe Center.
ACS Nano
PUBLISHED: 10-28-2014
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Although the G-quartet structure has been extensively investigated due to its biological importance, the formation mechanism, in particular, the necessity of metal centers, of an isolated G-quartet on solid surfaces remains ambiguous. Here, by using scanning tunneling microscopy under well-controlled ultra-high-vacuum conditions and density functional theory calculations we have been able to clarify that besides the intraquartet hydrogen bonding a metal center is mandatory for the formation of an isolated G-quartet. Furthermore, by subtly perturbing the local coordination bonding schemes within the formed G-quartet complex via local nanoscale scanning tunneling microscopy manipulations, we succeed in modulating the d orbitals and the accompanying magnetic properties of the metal center. Our results demonstrate the feasibility of forming an isolated G-quartet complex on a solid surface and that the strategy of modulating electronic and magnetic properties of the metal center can be extended to other related systems such as molecular spintronics.
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Complete mitochondrial genome of an Amynthas earthworm, Amynthas aspergillus (Oligochaeta: Megascolecidae).
Mitochondrial DNA
PUBLISHED: 10-21-2014
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Abstract We have determined the mitochondrial genome of the first Amynthas earthworm, Amynthas aspergillus (Perrier, 1872), which is a natural medical resource in Chinese traditional medicine. Its mitogenome is 15,115?bp in length containing 37 genes with the same contents and order as other sequenced earthworms. All genes are encoded by the same strand, all 13 PCGs use ATG as start codon. The content of A + T is 63.04% for A. aspergillus (33.41% A, 29.63% T, 14.56% G and 22.41% C). The complete mitochondrial genomes of A. aspergillus would be useful for the reconstruction of Oligochaeta polygenetic relationships.
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[Screening for prodromes of chemotherapy-induced vomiting and correlation between prodromes and chemotherapy-induced vomiting in lung cancer patients].
Zhonghua Zhong Liu Za Zhi
PUBLISHED: 10-21-2014
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To explore prodromes of chemotherapy-induced vomiting (CIV) and their association with CIV in lung cancer patients.
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Two nanocage anionic metal-organic frameworks with rht topology and {[M(H2O)6]6}(12+) charge aggregation for rapid and selective adsorption of cationic dyes.
Chem. Commun. (Camb.)
PUBLISHED: 10-16-2014
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Two rht anionic metal-organic frameworks were synthesized. There are six [M(H2O)6](2+) ions held together by a super-strong H-bond and arranged in a regular octahedron in each medium cage. Dye adsorption studies revealed a rapid and selective adsorption of cationic dyes and the adsorbed dyes can be released in saturated NaCl aqueous solution.
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Hemin-block copolymer micelle as an artificial peroxidase and its applications in chromogenic detection and biocatalysis.
ACS Appl Mater Interfaces
PUBLISHED: 10-16-2014
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Following an inspiration from the fine structure of natural peroxidases, such as horseradish peroxidase (HRP), an artificial peroxidase was constructed through the self-assembly of diblock copolymers and hemin, which formed a functional micelle with peroxidase-like activity. The pyridine moiety in block copolymer poly(ethylene glycol)-block-poly(4-vinylpyridine) (PEG-b-P4VP) can coordinate with hemin, and thus hemin is present in a five-coordinate complex with an open site for binding substrates, which mimics the microenvironment of heme in natural peroxidases. The amphiphilic core-shell structure of the micelle and the coordination interaction of the polymer to the hemin inhibit the formation of hemin ?-oxo dimers, and thereby enhance the stability of hemin in the water phase. Hemin-micelles exhibited excellent catalytic performance in the oxidation of phenolic and azo compounds by H2O2. In comparison with natural peroxidases, hemin-micelles have higher catalytic activity and better stability over wide temperature and pH ranges. Hemin-micelles can be used as a detection system for H2O2 with chromogenic substrates, and they anticipate the possibility of constructing new biocatalysts tailored to specific functions.
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A convenient label free colorimetric assay for pyrophosphatase activity based on a pyrophosphate-inhibited Cu(2+)-ABTS-H2O2 reaction.
Analyst
PUBLISHED: 10-15-2014
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The evaluation of pyrophosphatase (PPase) activity plays an important role in diagnosing diseases and understanding the function of PPase in the related biological events. In this work, an inhibition effect of pyrophosphate (PPi) on Cu(2+)-catalyzed H2O2-mediated oxidation of 2,2-azinobis(3-ethylbenzothiazoline)-6-sulfonic acid (ABTS) was observed. We utilize this inhibition effect to develop a convenient label free visual method for PPase activity detection. A hydroxyl radical could be generated from a Cu(2+)-based Fenton-like reaction, and then reacted with ABTS to produce colored ABTS?(+). The strong complexation between PPi and Cu(2+) disturbed this Cu(2+)-catalyzed ABTS-H2O2 reaction probably due to changing redox potentials of Cu(2+) towards H2O2. The PPase-catalyzed hydrolysis of PPi into Pi prohibited the complexation, resulting in the recovery of catalytic capability of Cu(2+). As a result, the solution color changed from colorless to green with a remarkable increase of absorbance. Compared with the traditional PPase assays, the developed visual assay is cost-effective and easy to implement. And a high sensitivity for PPase with a detection limit of 0.027 U mL(-1) was achieved. Moreover, the proposed colorimetric strategy was also applied to evaluate PPase inhibition and exhibited a good assay performance in complex biological samples.
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Synthesis of two triarylboron-functionalized metal-organic frameworks: in situ decarboxylic reaction, structure, photoluminescence, and gas adsorption properties.
Inorg Chem
PUBLISHED: 10-10-2014
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Two 3D noninterpenetrating porous metal-organic frameworks (PMOFs) [Cd3(L(1))2(DMA)2]·DMA [1, H3L(1) = tris(p-carboxylic acid)tridurylborane] and [Zn3(L(2))3(H2O)2]·5H2O·2EtOH [2, H2L(2) = 4,4'-((2,3,5,6-tetramethylphenyl)boranediyl)bis(2,3,5,6-tetramethylbenzoic acid)] were synthesized by employment of a C3-symmetric ligand (H3L(1)) to assembly with Cd(NO3)2 or Zn(NO3)2. Complex 1 exhibits a (3, 6)-connected topological network based on a Cd3 cluster and Y-shaped trinodal organic linker. Complex 2 shows a 6-connected topology, since in situ decarboxylic reaction of the initial H3L(1) occurred to generate a new ligand, H2L(2), which can be considered as a linear linker. Both 1 and 2 exhibit blue fluorescence. Significantly, complex 1 with larger channels is unstable upon the removal of guest molecules. In contrast, activated 2 exhibits higher stability and permanent porosity.
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The cytokines (IFN-gamma, IL-2, IL-4, IL-10, IL-17) and Treg cytokine (TGF-beta1) levels in adults with immune thrombocytopenia.
Pharmazie
PUBLISHED: 10-03-2014
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Previous studies have indicated that autoimmune diseases might be caused by an imbalance of T helper cells (Th), cytokines, and regulatory T cells (Treg) cytokines. We measured the plasma concentrations of Th1-associated cytokines (IFN-gamma, IL-2), Th2 -associated cytokines (IL-4, IL-10), Th17-associated cytokine (IL-17) and Treg -associated cytokine (TGF-beta1) in adult patients with immune thrombocytopenia (ITP) and evaluated their clinical relevance. Plasma IFN-gamma, IL-2, IL-4, IL-10, IL-17 and TGF-beta1 concentrations of 52 ITP patients and 30 age- and sex-matched healthy controls were measured by enzyme-linked immunosorbent assay method (ELISA). Concentration of Th2 cytokines (IL-4 and IL-10) were significantly higher in ITP patients compared to controls (P < 0.05). However, concentrations of Th1 cytokines (IFN-gamma, IL-2), Th17 cytokine (IL-17) and Treg cytokine (TGF-beta1) were lower in ITP patients (P < 0.05). Concentration of IL-17 was significantly higher in chronic ITP patients compared to severe ITP patients (P < 0.05), and no significant difference of cytokine concentration among the other subgroups in ITP patients was found. Among the ITP patients, concentration of IFN-gamma correlated positively and significantly with PAIgG (r = 0.48, P = 0.02). A significant correlation was neither found between other cytokine levels and platelet count, nor between cytokine levels and megakaryocytes number, nor between cytokines levels and PAIgG or GPIIb/IIIa and/or GPIb/IX autoantibodies. The present study demonstrates that an imbalance of Th and Treg cytokines may mediate the pathogenesis of ITP.
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Aggregation-induced chiral symmetry breaking of a naphthalimide-cyanostilbene dyad.
Phys Chem Chem Phys
PUBLISHED: 10-02-2014
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Spontaneously emerged supramolecular chirality and chiral symmetry breaking from achiral/racemic constituents remain poorly understood. We here report that supramolecular chirality may emerge from the structural flexibility of achiral aryl nitrogen centres which provide instantaneous chirality. Employing a naphthalimide-cyanostilbene dyad as a model, we explored the underlying mechanism of aggregation-induced chiral symmetry breaking and found that the conformations of the N-naphthylpiperazine and the N,N-dimethylaniline units facilitate the formation of ordered supramolecular structures and offer opposite handedness. Furthermore, chiral symmetry breaking of the monomers was amplified by the formation of dimers. The microscopic and the macroscopic observations from the theoretical simulations and experimental measurements are thus rationalized by connecting the population of the dihedral angles of the aryl nitrogen centres, the morphology of the self-assemblies, and the observed circular dichroism spectra.
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Improving the porosity and catalytic capacity of a zinc paddlewheel metal-organic framework (MOF) through metal-ion metathesis in a single-crystal-to-single-crystal fashion.
Inorg Chem
PUBLISHED: 09-25-2014
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Zinc paddlewheel metal-organic frameworks (MOFs) frequently exhibit low stability or complete collapse upon the removal of axial ligands. Hence, there are very few reports on gas adsorption of zinc paddlewheel MOFs. In this work, the N2 and H2 adsorption measurements were carried out for a zinc MOF (namely, SDU-1) based on two types of paddlewheel secondary building units (SBUs): [Zn2(COO)3] and [Zn2(COO)4]. Because of the existence of inherent surface instability upon removal of solvates in zinc paddlewheel SBU, SDU-1 possesses a very low surface area. Through metal-ion metathesis in a single-crystal-to-single-crystal fashion, the Zn(2+) ions in SDU-1 were exchanged by Cu(2+) ions to generate Cu-SDU-1. Through the measurements of gas adsorption and catalytic test, the porosity and catalytic capacity of Cu-SDU-1 have been improved significantly, compared to SDU-1.
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Ligustri Lucidi Fructus as a traditional Chinese medicine: a review of its phytochemistry and pharmacology.
Nat. Prod. Res.
PUBLISHED: 09-24-2014
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Ligustri Lucidi Fructus (LLF) is the fruits of Ligustrum lucidum Ait. (Oleaceae). This review based on nearly 80 literary sources discusses the knowledge of chemistry and biological effects of this species. Several types of chemical constituents considered as the characteristic and active constituents from LLF were isolated including 40 triterpenoids, 48 iridoids, 10 flavones, 10 phenylethanoid glycosides and others. Various extracts and individual compounds derived from this species have been found to possess a variety of pharmacological effects, e.g. anti-tumour, hepatoprotective, immune regulating, antioxidative and anti-ageing effects, anti-inflammation and reducing hypercholesterolaemia effects and so on. The results of data analysis on the chemical, pharmacological characteristics of LLF support the view that this species has many therapeutic properties and indicate its potential as an effective herbal remedy. Finally, some suggestions for further research on chemical and pharmacological properties are given in this review. Theoretical basis was given for further exploiting and utilising LLF.
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Mammalian cell display for rapid screening scFv antibody therapy.
Acta Biochim. Biophys. Sin. (Shanghai)
PUBLISHED: 09-22-2014
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Human antibodies are beginning to draw attention for use in immune gene therapy. The efficient generation of effective therapeutic monoclonal antibodies suitable for the treatment of cancers and infectious diseases would be enormously valuable. Antibody display methods are increasingly used to screen human monoclonal antibodies. Here we report the construction of a mammalian cell display method derived from a naive antibody repertoire, for which human single-chain variable fragments (scFv) have been transiently displayed on 293T cell surfaces based on a pDisplay vector. The sizes of the current pDisplay-scFv antibody repertoires have been estimated to be 0.74 × 10(7). An immunoblot assay confirmed the expression of the scFv antibody library. The subcellular distribution of ErbB3-scFv expression plasmid facilitated the display of ErbB3 scFv on the cell membrane surface and the efficiency of the display was evaluated by fluorescence-activated cell sorting. This method of mammalian cell display was verified by successfully screening ErbB3 scFv candidates. A published scFv control was used to confirm the feasibility of the ErbB3 scFv screening process. Three ErbB3 scFv candidates were produced and they were found to have affinity similar to the published scFv candidate. Thus, the present screening system provided an optimal alternative for rapid acquisition of a novel candidate scFv sequence to target genes with high affinity in vitro.
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[The impact of preoperative weight loss for gastric cancer patients after gastrectomy].
Zhonghua Wai Ke Za Zhi
PUBLISHED: 09-16-2014
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To elucidate the prognostic influence of preoperative weight loss for gastric cancer.
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Over 10?000 Peptide Identifications from the HeLa Proteome by Using Single-Shot Capillary Zone Electrophoresis Combined with Tandem Mass Spectrometry.
Angew. Chem. Int. Ed. Engl.
PUBLISHED: 09-13-2014
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Capillary zone electrophoresis (CZE)-tandem mass spectrometry (MS/MS) has recently attracted attention as a tool for shotgun proteomics. However, its performance for this analysis has so far fallen far below that of reversed-phase liquid chromatography (RPLC)-MS/MS. The use of a CZE method with a wide separation window (up to 90 min) and high peak capacity (ca. 300) is reported. This method was coupled to an Orbitrap Fusion mass spectrometer through an electrokinetically pumped sheath-flow interface for the analysis of complex proteome digests. Single-shot CZE-MS/MS lead to the identification of over 10?000 peptides and 2100 proteins from a HeLa cell proteome digest in approximately 100 min. This performance is nearly an order of magnitude better than earlier CZE studies and is within a factor of two to four of the state-of-the-art nano ultrahigh-pressure LC system.
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Engineering topochemical polymerizations using block copolymer templates.
J. Am. Chem. Soc.
PUBLISHED: 09-11-2014
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With the aim to achieve rapid and efficient topochemical polymerizations in the solid state, via solution-based processing of thin films, we report the integration of a diphenyldiacetylene monomer and a poly(styrene-b-acrylic acid) block copolymer template for the generation of supramolecular architectural photopolymerizable materials. This strategy takes advantage of non-covalent interactions to template a topochemical photopolymerization that yields a polydiphenyldiacetylene (PDPDA) derivative. In thin films, it was found that hierarchical self-assembly of the diacetylene monomers by microphase segregation of the block copolymer template enhances the topochemical photopolymerization, which is complete within a 20 s exposure to UV light. Moreover, UV-active cross-linkable groups were incorporated within the block copolymer template to create micropatterns of PDPDA by photolithography, in the same step as the polymerization reaction. The materials design and processing may find potential uses in the microfabrication of sensors and other important areas that benefit from solution-based processing of flexible conjugated materials.
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Species-associated differences in the inhibition of propofol glucuronidation by magnolol.
J. Am. Assoc. Lab. Anim. Sci.
PUBLISHED: 09-09-2014
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Magnolol, a major active constituent in herbal medicine, potently inhibits propofol glucuronidation in human liver microsomes, with inhibition constants in the nanomolar range. This study was conducted to investigate magnolol-induced inhibition of propofol glucuronidation in liver microsomes from Swiss-Hauschka mice, Sprague-Dawley rats, Chinese Bama pigs, and cynomolgus macaques. Results indicated that magnolol (10 ?M) inhibited propofol glucuronidation in liver microsomes from Bama pigs and cynomolgus macaques but not in those from mice or rats. Data from liver microsomes from Bama pigs indicated a competitive inhibition mechanism, with a Ki of 1.7 ?M. In contrast to that of pig liver microsomes, the inhibition of microsomes from cynomolgus macaques followed a noncompetitive mechanism, with a Ki of 3.4 ?M. In summary, this study indicates that magnolol-induced inhibition of propofol glucuronidation varies substantially among species, and the Ki values determined by using liver microsomes from various experimental animal species far exceed that for human liver microsomes. The inhibition of propofol glucuronidation by magnolol in liver microsomes from all animal species tested was significantly lower than the inhibition previously demonstrated in human liver microsomes. Hepatic microsomes from Swiss-Hauschka mice, Sprague-Dawley rats, Chinese Bama pigs, and cynomolgus macaques are not effective models of the inhibition of glucuronidation induced by magnolol in humans.
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Oxygen-induced self-assembly of quaterphenyl molecules on metal surfaces.
Chem. Commun. (Camb.)
PUBLISHED: 08-30-2014
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From high-resolution UHV-STM imaging and DFT calculations we have demonstrated a novel method to construct well-ordered molecular nanostructures of an unfunctionalized aromatic molecule (4Ph) on both Ag(110) and Cu(110) by introducing oxygen molecules.
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Superiority of the Ratio Between Negative and Positive Lymph Nodes for Predicting the Prognosis for Patients With Gastric Cancer.
Ann. Surg. Oncol.
PUBLISHED: 08-28-2014
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This study aimed to elucidate the prognostic prediction superiority of the ratio between negative and positive lymph nodes (RNP) in gastric cancer (GC).
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The expression of glucocorticoid receptor is negatively regulated by active androgen receptor signaling in prostate tumors.
Int. J. Cancer
PUBLISHED: 08-20-2014
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The glucocorticoid and androgen receptors (GR and AR) can commonly regulate up to 50% of their target genes in prostate cancer (PCa) cells. GR expression is stimulated by castration therapy, which has been proposed to be one mechanism that compensates for AR signaling blockade and promotes castration-resistant PCa (CRPC) progression. However, whether GR functions as a driver for CRPC or a marker reflecting AR activity remains unclear. Here, we applied PCa tissue microarrays to show that GR protein levels were elevated by castration therapy, but reduced to pre-castration levels when tumors were at the CRPC stage. Using subrenal capsule xenograft models, we showed that GR expression was inversely correlated with AR and PSA expressions. GR expression levels are not associated with tumor invasion and metastasis phenotypes. In castration-resistant C4-2 xenografts expressing AR shRNA, regressing tumors induced by AR knockdown expressed higher levels of GR and lower levels of PSA than non-regressing tumors. Immunoblotting and real-time PCR assays further showed that AR knockdown or AR antagonists increased GR expression at both mRNA and protein levels. ChIP combined with DNA sequencing techniques identified a negative androgen responsive element (nARE) 160K base pairs upstream of the GR gene. Gel shift assays confirmed that AR directly interacted with the nARE and luciferase assays demonstrated that the nARE could mediate transcription repression by ligand-activated AR. In conclusion, GR expression is negatively regulated by AR signaling and may serve as a marker for AR signaling in prostate tumors.
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Molecular analysis of proanthocyanidins related to pigmentation in brown cotton fibre (Gossypium hirsutum L.).
J. Exp. Bot.
PUBLISHED: 08-02-2014
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The structural characteristics and component differences of proanthocyanidins in brown and white cotton fibres were identified by nuclear magnetic resonance (NMR) and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) analyses. Proanthocyanidins in brown and white cotton fibres were found to contain mainly procyanidin (PC) and prodelphidin (PD) units with 2, 3-cis form (epigallocatechin and epicatechin). However, part of the proanthocyanidins in the white cotton fibres were modified by acylation and were constitutively different from the proanthocyanidins in brown cotton fibres. The relative amount of PD was similar to that of PC in white cotton fibres, while proanthocyanidins in brown cotton fibres consisted mainly of PD units with a relative ratio of 9:1. In brown cotton fibres, the proanthocyanidin monomeric composition was consistent with the expression profiles of proanthocyanidin synthase genes, suggesting that anthocyanidin reductase represented the major flow of the proanthocyanidin biosynthesis pathway. In addition, the structural characteristics and component differences of proanthocanidins in brown and white cotton fibres suggested that quinones, the oxidation products of proanthocyanidins, were the direct contributors to colour development in brown cotton fibre. This was demonstrated by vanillin-HCl staining and Borntrager's test. Collectively, these data demonstrated that the biosynthesis of proanthocyanidins is a crucial pigmentation process in brown cotton fibre, and that quinones may represent the main pigments contributing to formation of the the brown colour. This study revealed the molecular basis of pigmentation in brown cotton fibres, and provided important insights for genetic manipulation of pigment production in cotton fibres.
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[Clinical observation of abdominal regional fluorouracil implants in advanced gastric cancer patients during operation].
Zhonghua Wei Chang Wai Ke Za Zhi
PUBLISHED: 07-30-2014
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To investigate the postoperative adverse events and survival of patients with sustained-released fluorouracil implanted during operation.
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Blocking the function of inflammatory cytokines and mediators by using IL-10 and TGF-?: a potential biological immunotherapy for intervertebral disc degeneration in a beagle model.
Int J Mol Sci
PUBLISHED: 07-29-2014
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The debilitating effects of lower back pain are a major health issue worldwide. A variety of factors contribute to this, and oftentimes intervertebral disk degeneration (IDD) is an underlying cause of this disorder. Inflammation contributes to IDD, and inflammatory cytokines such as tumor necrosis factor (TNF)-? and interleukin (IL)-1?, play key roles in the pathology of IDD. Therefore, the development of treatments that inhibit the expression and/or effects of TNF-? and IL-1? in IDD patients should be a promising therapeutic approach to consider. This study characterized the potential to suppress inflammatory cytokine production in degenerative intervertebral disc (NP) cells by treatment with IL-10 and TGF-? in a canine model of IDD. IDD was induced surgically in six male beagles, and degenerative NP cells were isolated and cultured for in vitro studies on cytokine production. Cultured degenerative NP cells were divided into four experimental treatment groups: untreated control, IL-10-treated, TGF-?-treated, and IL-10- plus TGF-?-treated cells. Cultured normal NP cells served as a control group. TNF-? expression was evaluated by fluorescence activated cell sorting (FACS) analysis and enzyme-linked immunosorbent assay (ELISA); moreover, ELISA and real-time PCR were also performed to evaluate the effect of IL-10 and TGF-? on NP cell cytokine expression in vitro. Our results demonstrated that IL-10 and TGF-? treatment suppressed the expression of IL-1? and TNF-? and inhibited the development of inflammatory responses. These data suggest that IL-10 and TGF-? should be evaluated as therapeutic approaches for the treatment of lower back pain mediated by IDD.
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Simultaneous quantification of 5-hydroxyindoleacetic acid and 5-hydroxytryptamine by capillary electrophoresis with quantum dot and horseradish peroxidase enhanced chemiluminescence detection.
J. Chromatogr. B Analyt. Technol. Biomed. Life Sci.
PUBLISHED: 07-26-2014
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A capillary electrophoresis (CE) with chemiluminescence (CL) detection method was developed for the simultaneous quantification of 5-hydroxyindoleacetic acid (5-HIAA) and 5-hydroxytryptamine (5-HT). In this method, CdTe quantum dot (QD) and horseradish peroxidase (HRP) were used as enhancing reagents to co-catalyze the post-column CL reaction between luminol and hydrogen peroxide, achieving highly efficient CL emission. 5-HIAA and 5-HT inhibit the CL emission resulting to the formation of negative peaks in electropherogram. The degree of CL suppression is proportional to the concentration of 5-HT and 5-HIAA. The linear ranges for the determination of 5-HIAA and 5-HT were 2.5×10(-8)-2.5×10(-6) M and 2.5×10(-8)-5.0×10(-6) M with detection limits (signal/noise=3) of 7.0×10(-9) M and 6.0×10(-9) M, respectively. Intraday precision do not exceed 5.0%. The accuracy was confirmed by the recoveries ranged from 98% to 104%. The present method was successfully applied for the quantification of 5-HIAA and 5-HT in human urine. The concentrations of 5-HT and 5-HIAA in human urine were found to be in the range of 0.78-1.2 ?M and 3.2-5.1 ?M, respectively.
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[Hemocompatibility of polyoxymethylene used for bileaflet heart valve].
Sheng Wu Yi Xue Gong Cheng Xue Za Zhi
PUBLISHED: 07-22-2014
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The possibility of polyoxymethylene (POM) as heart valve leaflet material was investigated by comparing the hemocompatibility with that of 316L stainless steel and low-temperature isotropic pyrolytic carbon (LTIC). Surface hydrophobicity was characterized by water contact angle measurement. Platelet adhesion, APTT/PT/TT and hemolysis rate tests were applied for evaluating hemocompatibility. The results showed that POM was hydrophobic and had a low hemolytic rate, adhesion amount and activation degree of platelets on POM surface were less than 316L stainless steel, and was similar to LTIC. This research pointed out potential application of POM as heart valve leaflets.
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Mechanical compression insults induce nanoscale changes of membrane-skeleton arrangement which could cause apoptosis and necrosis in dorsal root ganglion neurons.
Biosci. Biotechnol. Biochem.
PUBLISHED: 07-14-2014
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In a primary spinal cord injury, the amount of mechanical compression insult that the neurons experience is one of the most critical factors in determining the extent of the injury. The ultrastructural changes that neurons undergo when subjected to mechanical compression are largely unknown. In the present study, using a compression-driven instrument that can simulate mechanical compression insult, we applied mechanical compression stimulation at 0.3, 0.5, and 0.7 MPa to dorsal root ganglion (DRG) neurons for 10 min. Combined with atomic force microscopy, we investigated nanoscale changes in the membrane-skeleton, cytoskeleton alterations, and apoptosis induced by mechanical compression injury. The results indicated that mechanical compression injury leads to rearrangement of the membrane-skeleton compared with the control group. In addition, mechanical compression stimulation induced apoptosis and necrosis and also changed the distribution of the cytoskeleton in DRG neurons. Thus, the membrane-skeleton may play an important role in the response to mechanical insults in DRG neurons. Moreover, sudden insults caused by high mechanical compression, which is most likely conducted by the membrane-skeleton, may induce necrosis, apoptosis, and cytoskeletal alterations.
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Porous zirconium metal-organic framework constructed from 2D ? 3D interpenetration based on a 3,6-connected kgd net.
Inorg Chem
PUBLISHED: 07-07-2014
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A new porous zirconium metal-organic framework (Zr MOF), Zr6(?3-O)4(?3-OH)4(OH)6(H2O)6(BTB)2·6DMF·H2O (1; H3BTB = 5'-(4-carboxyphenyl)[1,1':3',1?-terphenyl]-4,4?-dicarboxylic acid), based on Zr6 clusters and tricarboxylate ligands has been constructed and characterized. The Zr6 clusters were linked by BTB ligands to generate a 2D network of kgd topology. The interpenetrations among the 2D networks gave rise to a 3D porous framework, which represents the first Zr MOF constructed from 2D ? 3D interpenetration. The gas uptake and catalytic properties for 1 have also been studied.
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Thermally activated delayed fluorescence of fluorescein derivative for time-resolved and confocal fluorescence imaging.
J. Am. Chem. Soc.
PUBLISHED: 06-26-2014
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Compared with fluorescence imaging utilizing fluorophores whose lifetimes are in the order of nanoseconds, time-resolved fluorescence microscopy has more advantages in monitoring target fluorescence. In this work, compound DCF-MPYM, which is based on a fluorescein derivative, showed long-lived luminescence (22.11 ?s in deaerated ethanol) and was used in time-resolved fluorescence imaging in living cells. Both nanosecond time-resolved transient difference absorption spectra and time-correlated single-photon counting (TCSPC) were employed to explain the long lifetime of the compound, which is rare in pure organic fluorophores without rare earth metals and heavy atoms. A mechanism of thermally activated delayed fluorescence (TADF) that considers the long wavelength fluorescence, large Stokes shift, and long-lived triplet state of DCF-MPYM was proposed. The energy gap (?EST) of DCF-MPYM between the singlet and triplet state was determined to be 28.36 meV by the decay rate of DF as a function of temperature. The ?E(ST) was small enough to allow efficient intersystem crossing (ISC) and reverse ISC, leading to efficient TADF at room temperature. The straightforward synthesis of DCF-MPYM and wide availability of its starting materials contribute to the excellent potential of the compound to replace luminescent lanthanide complexes in future time-resolved imaging technologies.
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Acid hydrolysis of crude tannins from infructescence of Platycarya strobilacea Sieb. et Zucc to produce ellagic acid.
Nat. Prod. Res.
PUBLISHED: 06-09-2014
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The infructescence of Platycarya strobilacea Sieb. et Zucc is a well-known traditional medicine in China, Japan and Korea. The infructescence of P. strobilacea Sieb. et Zucc is a rich source of ellagitannins that are composed of ellagic acid (EA) and gallic acid, linked to a sugar moiety. The aim of this study was to prepare EA by acid hydrolysis of crude tannins from the infructescence of P. strobilacea Sieb. et Zucc, and establish a new technological processing method for EA. The natural antioxidant EA was prepared by using the water extraction of infructescence of P. strobilacea Sieb. et Zucc, evaporation, condensation, acid hydrolysis and prepared by the process of crystallisation. The yield percentage of EA from crude EA was more than 20% and the purity of the product was more than 98%, as identified by using HPLC. The structure was identified on the basis of spectroscopic analysis and comparison with authentic compound.
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Bottom-up proteomics of Escherichia coli using dynamic pH junction preconcentration and capillary zone electrophoresis-electrospray ionization-tandem mass spectrometry.
Anal. Chem.
PUBLISHED: 06-09-2014
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We report the use of the dynamic pH junction based capillary zone electrophoresis-electrospray ionization-tandem mass spectrometry (CZE-ESI-MS/MS) for bottom-up proteomics with an electrokinetically pumped sheath-flow nanospray capillary electrophoresis-mass spectrometry (CE-MS) interface and both LTQ-XL and LTQ-Orbitrap-Velos mass spectrometers. Conventional injection of 20 nL of a 1 mg/mL BSA digest identified 37 peptides and produced 66% sequence coverage. In contrast, pH junction injection of 130 nL (or larger) of a 0.05 mg/mL BSA digest identified 40 peptides and produced 70% coverage using a pH 6.5 sample buffer and the LTQ. A 20 nL conventional injection of a 1 mg/mL Escherichia coli digest identified 508 peptides and 199 proteins with the LTQ. A 400 nL pH junction injection of a 0.1 mg/mL E. coli digest identified 527 peptides and 179 proteins with the LTQ. Triplicate technical replicates of a 0.01 mg/mL sample with 400-nL injection volume using a pH junction identified 288 ± 9 peptides and 121 ± 5 proteins with the LTQ. There was outstanding concordance in migration time between the pH junction and normal injection. The pH junction produced narrower peaks and significant concentration for all but the most acidic components in the sample. Compared with the conventional stacking method, the pH junction method can generate comparable performance for small injection volume (20 nL) and significantly better concentration performance for a large injection volume (200 nL). We also applied the pH junction to three intact standard proteins and observed a >10× increase in peak intensity compared to conventional injection.
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U0126 promotes osteogenesis of rat bone-marrow-derived mesenchymal stem cells by activating BMP/Smad signaling pathway.
Cell Tissue Res.
PUBLISHED: 05-30-2014
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U0126 has been reported as a specific inhibitor of the ERK1/2 signaling pathway, which plays a vital role during the osteogenic differentiation of mesenchymal stem cells (MSCs). We report the positive effect of U0126 on the osteogenesis of rat MSCs. We find that U0126 promotes the osteogenic differentiation of rat MSCs as demonstrated by the quantitative real-time polymerase chain reaction for osteogenic markers, alkaline phosphatase activity and calcium nodule formation. Our data indicate that U0126 enhances the BMP/Smad signaling pathway in rat MSCs, while inhibiting the ERK1/2 signaling pathway. Furthermore, Western blot results demonstrate that U0126 increases Smad1/5/8 phosphorylation synergistically with ?-glycerophosphate. In addition, U0126 significantly increases the expression of BMP2 during the process of osteogenesis in rat MSCs and the level of phosphorylated Smad1/5/8 is significantly reduced by BMP2 antibody, suggesting that U0126 also promotes the expression of BMP2 to enhance Smad proteins phosphorylation. Thus, we demonstrate a novel function for U0126 in promoting osteogenic differentiation of rat MSCs by the activation of the BMP/Smad signaling pathway.
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Nicking enzyme and graphene oxide-based dual signal amplification for ultrasensitive aptamer-based fluorescence polarization assays.
Biosens Bioelectron
PUBLISHED: 05-21-2014
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In this work, two different configurations for novel amplified fluorescence polarization (FP) aptasensors based on nicking enzyme signal amplification (NESA) and graphene oxide (GO) enhancement have been developed for ultrasensitive and selective detection of biomolecules in homogeneous solution. One approach involves the aptamer-target binding induced the stable hybridization between an aptamer probe and a fluorophore-labeled DNA probe linked to GO, and forms a nicking site-containing duplex DNA region due to the enhancement of base stacking. The second analytical method involves the target induced the assembly of two aptamer subunits into an aptamer-target complex, and then hybridizes with a fluorophore-labeled DNA probe linked to GO, forming a nicking site-containing duplex DNA region. The formation of the duplex DNA region in both methods triggers the NESA process, resulting in the release of many short DNA fragments carrying the fluorophore from GO, generating a significant decrease of the FP value that provides the readout signal for the amplified sensing process. By using the NESA coupled GO enhancement path, the sensitivity of the developed aptasensors can be significantly improved by four orders of magnitude over traditional aptamer-based homogeneous assays. Moreover, these aptasensors also exhibit high specificity for target molecules, which are capable of detecting target molecule in biological samples. Considering these qualities, the proposed FP aptasensors based NESA and GO enhancement can be expected to provide an ultrasensitive platform for amplified analysis of target molecules.
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The positive impact of surgeon specialization on survival for gastric cancer patients after surgery with curative intent.
Gastric Cancer
PUBLISHED: 05-17-2014
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Many studies have affirmed the survival benefit for cancer patients treated by specialized surgeons.
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A DArT marker-based linkage map for wild potato Solanum bulbocastanum facilitates structural comparisons between Solanum A and B genomes.
BMC Genet.
PUBLISHED: 05-16-2014
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BackgroundWild potato Solanum bulbocastanum is a rich source of genetic resistance against a variety of pathogens. It belongs to a taxonomic group of wild potato species sexually isolated from cultivated potato. Consistent with genetic isolation, previous studies suggested that the genome of S. bulbocastanum (B genome) is structurally distinct from that of cultivated potato (A genome). However, the genome architecture of the species remains largely uncharacterized. The current study employed Diversity Arrays Technology (DArT) to generate a linkage map for S. bulbocastanum and compare its genome architecture with those of potato and tomato.ResultsTwo S. bulbocastanum parental linkage maps comprising 458 and 138 DArT markers were constructed. The integrated map comprises 401 non-redundant markers distributed across 12 linkage groups for a total length of 645 cM. Sequencing and alignment of DArT clones to reference physical maps from tomato and cultivated potato allowed direct comparison of marker orders between species. A total of nine genomic segments informative in comparative genomic studies were identified. Seven genome rearrangements correspond to previously-reported structural changes that have occurred since the speciation of tomato and potato. We also identified two S. bulbocastanum genomic regions that differ from cultivated potato, suggesting possible chromosome divergence between Solanum A and B genomes.ConclusionsThe linkage map developed here is the first medium density map of S. bulbocastanum and will assist mapping of agronomical genes and QTLs. The structural comparison with potato and tomato physical maps is the first genome wide comparison between Solanum A and B genomes and establishes a foundation for further investigation of B genome-specific structural chromosome rearrangements.
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Capillary zone electrophoresis-electrospray ionization-tandem mass spectrometry for top-down characterization of the Mycobacterium marinum secretome.
Anal. Chem.
PUBLISHED: 04-28-2014
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Capillary zone electrophoresis (CZE) with an electrokinetically pumped sheath-flow nanospray interface was coupled with a high-resolution Q-Exactive mass spectrometer for the analysis of culture filtrates from Mycobacterium marinum. We confidently identified 22 gene products from the wildtype M. marinum secretome in a single CZE-tandem mass spectrometry (MS/MS) run. A total of 58 proteoforms were observed with post-translational modifications including signal peptide removal, N-terminal methionine excision, and acetylation. The conductivities of aqueous acetic acid and formic acid solutions were measured from 0.1% to 100% concentration (v/v). Acetic acid (70%) provided lower conductivity than 0.25% formic acid and was evaluated as low ionic-strength and a CZE-MS compatible sample buffer with good protein solubility.
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Capillary zone electrophoresis-electrospray ionization-tandem mass spectrometry for quantitative parallel reaction monitoring of peptide abundance and single-shot proteomic analysis of a human cell line.
J Chromatogr A
PUBLISHED: 04-09-2014
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We coupled capillary zone electrophoresis (CZE) with an ultrasensitive electrokinetically pumped nanospray ionization source for tandem mass spectrometry (MS/MS) analysis of complex proteomes. We first used the system for the parallel reaction monitoring (PRM) analysis of angiotensin II spiked in 0.45mg/mL of bovine serum albumin (BSA) digest. A calibration curve was generated between the loading amount of angiotensin II and intensity of angiotensin II fragment ions. CZE-PRM generated a linear calibration curve across over 4.5 orders of magnitude dynamic range corresponding to angiotensin II loading amount from 2amole to 150fmole. The relative standard deviations (RSDs) of migration time were <4% and the RSDs of fragment ion intensity were ?20% or less except 150fmole angiotensin II loading amount data (?36% RSD). We further applied the system for the first bottom up proteomic analysis of a human cell line using CZE-MS/MS. We generated 283 protein identifications from a 1h long, single-shot CZE MS/MS analysis of the MCF7 breast cancer cell line digest, corresponding to ?80ng loading amount. The MCF7 digest was fractionated using a C18 solid phase extraction column; single-shot analysis of a single fraction resulted in 468 protein identifications, which is by far the largest number of protein identifications reported for a mammalian proteomic sample using CZE.
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Investigation of the effect of pore size on gas uptake in two metal-organic frameworks.
Chem. Commun. (Camb.)
PUBLISHED: 04-04-2014
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Two porous metal–organic frameworks (1 and 2) with a fsc topology based on mixed ligands have been assembled and characterized. The different pillared ligands (pyrazine for 1 and 4,4?-bipyridine for 2) significantly influence the pore size of the frameworks. Gas uptake measurements reveal that complex 1 possesses higher H2, CO2, and CH4 uptake capacities than 2, although the surface area of 1 is lower than that of complex 2. These results further experimentally prove that the pore size plays an important role in gas uptake in porous MOFs, and the slit pore with a size of ~6 Å exhibits stronger interactions with gas molecules.
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Investigation on interaction between Ligupurpuroside A and pepsin by spectroscopic and docking methods.
Spectrochim Acta A Mol Biomol Spectrosc
PUBLISHED: 04-01-2014
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Ligupurpuroside A is one of the major glycoside in Ku-Din-Cha, a type of Chinese functional tea. In order to better understand its digestion and metabolism in humans, the interaction between Ligupurpuroside A and pepsin has been investigated by fluorescence spectra, UV-vis absorption spectra and synchronous fluorescence spectra along with molecular docking method. The fluorescence experiments indicate that Ligupurpuroside A can effectively quench the intrinsic fluorescence of pepsin through a combined quenching way at the low concentration of Ligupurpuroside A, and a static quenching procedure at the high concentration. The binding constant, binding sites of Ligupurpuroside A with pepsin have been calculated. The thermodynamic analysis suggests that non-covalent reactions, including electrostatic force, hydrophobic interaction and hydrogen bond are the main forces stabilizing the complex. According to the Förster's non-radiation energy transfer theory, the binding distance between pepsin and Ligupurpuroside A was calculated to be 3.15 nm, which implies that energy transfer occurs between pepsin and Ligupurpuroside A. Conformation change of pepsin was observed from UV-vis absorption spectra and synchronous fluorescence spectra under experimental conditions. In addition, all these experimental results have been validated by the protein-ligand docking studies which show that Ligupurpuroside A is located in the cleft between the domains of pepsin.
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A high throughput screening assay for identifying glycation inhibitors on MALDI-TOF target.
Food Chem
PUBLISHED: 03-26-2014
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The Maillard reaction plays an important role in the food industry, however, the deleterious effects generated by the advanced glycation end-products (AGEs) have been well recognized. Many efforts have been made to seek new AGE inhibitors, in particular those natural ones without adverse effect. We have developed a rapid, mass spectrometry based, on-plate screening assay for novel AGE inhibitors. The glycation reaction, inhibition feedback as well as the subsequent MALDI mass spectrometric analysis occurred on one single MALDI plate. At 1:10M ratio of peptide to sugar, as little as 4h incubation time allowed the screening test to be ready for analysis. DSP, inhibition and IC50 were calculated to evaluate selected inhibitors and resulting inhibition efficiencies were consistent with available references. We demonstrated that this method provide a potential high throughput screening assay to analyze and identify the anti-glycation agents.
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Alteronol Induces Differentiation of Melanoma B16F0 Cells.
Recent Pat Anticancer Drug Discov
PUBLISHED: 03-25-2014
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Alteronol, isolated from a microbial mutation strains, have been applied for Chinese and international patents for tumor treatment. The aim of this project study is to investigate the characteristics of proliferation and redifferentiation induced by alteronol in melanoma B16F0 cells. Cell proliferation is determined by tetrazolium salt colorimetric method (MTT assay). Morphological changes were analyzed by using Giemsa staining. The levels of Melanin and tyrosinase were measured by spectrophotometry. The mRNA expressions of tyrosinase-related protein Trp1 and Trp2 were evaluated by reverse transcription-polymerase chain reaction (RT-PCR). The anchorage-independent proliferation of B16F0 was monitored by the colony formation assay. Tumorigenicity was characterized by an animal model in vivo. The results showed that the proliferation of B16F0 cells was inhibited by alteronol in a concentration and time dependent manner. All well-known markers of melanoma cell differentiation, including morphological changes, the extended dendrites, and the change of tyrosinase activity, were enhanced greatly with the increase of alteronol concentrations. Taken together, the expression of tyrosinase related gene, decreased cell colony formation rate and the tumorigenicity in vivo, all of these revealed that alteronol play a key role in inducing differentiation and suppressing the proliferation of B16F0 tumor cells in vitro and in vivo.
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Adoptive immunotherapy for non-small cell lung cancer by NK and cytotoxic T lymphocytes mixed effector cells: retrospective clinical observation.
Int. Immunopharmacol.
PUBLISHED: 03-17-2014
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The objective of the current study was to retrospectively investigate the efficacy of adaptive transfer of natural killer (NK) and cytotoxic T lymphocytes mixed effector (NKTm) cells in non-small cell lung cancer (NSCLC) patients in comparison to a control group of NSCLC patients. NKTm cells were obtained by ex vivo expansion of peripheral blood mononuclear cells (PBMCs) of patients followed with phenotype determination. Primary end point was overall survival (OS). Ex vivo expansion caused significant enrichment of CD3(+), CD3(+)CD8(+), CD45RO(+), CD25(+), CD29(+) and CD3(+)CD16(+)/CD56(+) cells (P<0.05). The OS of the immunotherapy group was significantly longer than that of the control group and the risk of death decreased by 43.8% (31.1 months vs 18.1 months, P=0.008, HR=0.562, 95% CI 0.367-0.860). Two-year survival rate of patients in the immunotherapy group was better than in the control group (62.95% vs 35.44%, P<0.05). Gender, clinical stage, application of TKI, number of chemotherapy cycle, and application of NKTm immunotherapy were independent prognostic factors for NSCLC patients. The OS in subgroups of males, <60 years age, clinical stage IIIb+IV, no brain metastases, without radiotherapy, chemotherapy of >6 cycles, no application of TKI and TKI invalid was prolonged after NKTm cellular immunotherapy (P<0.05). Ex vivo expansion of NKTm cells was effective and had no adverse safety concerns, thus highlight that adaptive transfer of NKTm cells may prolong the OS of NSCLC patients and increase 2 year survival rate.
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Complete mitochondrial genome of a Pheretimoid earthworm Metaphire vulgaris (Oligochaeta: Megascolecidae).
Mitochondrial DNA
PUBLISHED: 03-13-2014
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Abstract We have determined the mitochondrial genome of the first Pheretimoid earthworm, Metaphire vulgaris (Chen, 1930). This mitogenome is 15,061?bp in length containing 37 genes typical of other annelid. All genes are encoded by the same strand, ATP8 is not adjacent to ATP6, all 13 PCGs use ATG as a start codon. These features are consistent with first determined earthworm Lumbricus terrestris, but unusual among animal mtDNAs.
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Comparison of the staging of regional lymph nodes using the sixth and seventh editions of the tumor-node-metastasis (TNM) classification system for the evaluation of overall survival in gastric cancer patients: findings of a case-control analysis involving a single inst
Surgery
PUBLISHED: 03-11-2014
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It has recently been reported that the sixth edition of the tumor-node-metastasis (TNM) classification system for gastric cancer involving the staging of regional lymph nodes (N) has inappropriate cut-offs with regard to counts of metastatic lymph nodes. It remains controversial, however, as to whether the seventh edition of this classification system is completely accurate in staging N for the prediction of the prognosis of gastric cancer. Our aim was to determine which of these two editions of the TNM classification system was superior with regard to the prediction of the prognosis of Chinese patients with gastric cancer.
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Salvianolic acid B promotes osteogenesis of human mesenchymal stem cells through activating ERK signaling pathway.
Int. J. Biochem. Cell Biol.
PUBLISHED: 02-18-2014
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Salvianolic acid B, a major bioactive component of Chinese medicine herb, Salvia miltiorrhiza, is widely used for treatment of cardiovascular diseases. Our recent studies have shown that Salvianolic acid B can prevent development of osteoporosis. However, the underlying mechanisms are still not clarified clearly. In the present study, we aim to investigate the effects of Salvianolic acid B on viability and osteogenic differentiation of human mesenchymal stem cells (hMSCs). The results showed Salvianolic acid B (Sal B) had no obvious toxic effects on hMSCs, whereas Sal B supplementation (5?M) increased the alkaline phosphatase activity, osteopontin, Runx2 and osterix expression in hMSCs. Under osteogenic induction condition, Sal B (5?M) significantly promoted mineralization; and when the extracellular-signal-regulated kinases signaling (ERK) pathway was blocked, the anabolic effects of Sal B were diminished, indicating that Sal B promoted osteogenesis of hMSCs through activating ERK signaling pathway. The current study confirms that Sal B promotes osteogenesis of hMSCs with no cytotoxicity, and it may be used as a potential therapeutic agent for the management of osteoporosis.
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Absolute quantitation of host cell proteins in recombinant human monoclonal antibodies with an automated CZE-ESI-MS/MS system.
Electrophoresis
PUBLISHED: 02-13-2014
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We report the first use of CZE for absolute characterization of host cell proteins (HCPs) in recombinant human monoclonal antibodies. An electrokinetically pumped nanoelectrospray interface was used to couple CZE with a tandem mass spectrometer. Three isotopic-labeled peptides (LSFDKDAMVAR, VDIVENQAMDTR, and LVSDEMVVELIEK) were synthesized by direct incorporation of an isotope-labeled lysine or arginine. The heavy-labeled peptides were spiked in the HCP digests at known concentrations. After CZE-ESI-MS/MS analysis, the peaks of native and isotopic-labeled peptides were extracted with mass tolerance ? 5 ppm from the electropherograms, and the ratios of peak area between native and isotopic-labeled peptides pairs were calculated. Calibration curves (the ratios of peak area versus spiked peptide amount) with R(2) values of 0.999, 0.997, and 0.999 were obtained for the three HCP peptides, and the absolute amounts of the three proteins present were determined to be at the picomole level in a 20 ?g sample of digested HCPs. The target proteins were present at the 7-30 ppt level in the purified HCP samples.
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Uncovering immobilized trypsin digestion features from large-scale proteome data generated by high-resolution mass spectrometry.
J Chromatogr A
PUBLISHED: 02-01-2014
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Immobilized trypsin produces very fast protein digestion, which is attractive for application to high throughput bottom-up proteomics. While there is a rich literature on the preparation of immobilized trypsin, there are very few studies that investigate its application to complex proteomic samples. In this work, we compared solution-phase trypsin with trypsin immobilized on magnetic microspheres for digestion of two complex proteomes, Escherichia coli and the MCF7 cell line. The digests were separated by HPLC, and detected with a Q-Exactive mass spectrometer, which generated high resolution and high quality parent- and fragment-ion mass spectra. The data were analyzed using MaxQuant. We make several conclusions about the features of immobilized trypsin digestion of complex proteomes. First, both immobilized and solution-phase trypsin generate peptides that sample the same protein pool. Second, immobilized trypsin can digest complex proteomes two orders of magnitude faster than solution-phase trypsin while retaining similar numbers of protein identifications and proteome depth. Digestion using immobilized trypsin for 5-min produces a similar number of missed cleavages as solution-based trypsin digestion for 4-h; digestion using immobilized trypsin for 20-min produces a similar number of missed cleavages as solution-based trypsin digestion for 12-h. Third, immobilized trypsin produces quantitatively reproducible digestion of complex proteomes. Finally, there is small but measurable loss of peptide due to non-specific adsorption to the immobilization matrix. This adsorption generates a bias against detection of basic peptides.
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Taurine promotes human mesenchymal stem cells to differentiate into osteoblast through the ERK pathway.
Amino Acids
PUBLISHED: 01-28-2014
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Taurine has been reported to influence bone metabolism, but the role of taurine on osteogenic differentiation of human mesenchymal stem cells (hMSCs) remains unclear. In the present study, we investigated the effect of taurine on osteogenic differentiation of hMSCs. The results showed that taurine increased the alkaline phosphatase (ALP) activity and mineralized nodules in hMSCs induced by osteogenic induced medium. Meanwhile, RT-PCR analysis showed that taurine up-regulated the mRNA expression of ALP, osteopontin, Runt-related transcription factor 2 (Runx2) and Osterix in a dose-dependent manner. Furthermore, taurine induced activation of extracellular signal regulated kinase (ERK) and pretreatment with the ERK inhibitor U0126 abolished the taurine-induced osteogenesis of hMSCs. Taken together, our study reveals that taurine promotes the osteogenesis of hMSCs by activating the ERK pathway.
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Maintenance of amyloid ? peptide homeostasis by artificial chaperones based on mixed-shell polymeric micelles.
Angew. Chem. Int. Ed. Engl.
PUBLISHED: 01-23-2014
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The disruption of A? homeostasis, which results in the accumulation of neurotoxic amyloids, is the fundamental cause of Alzheimer's disease (AD). Molecular chaperones play a critical role in controlling undesired protein misfolding and maintaining intricate proteostasis in?vivo. Inspired by a natural molecular chaperone, an artificial chaperone consisting of mixed-shell polymeric micelles (MSPMs) has been devised with tunable surface properties, serving as a suppressor of AD. Taking advantage of biocompatibility, selectivity toward aberrant proteins, and long blood circulation, these MSPM-based chaperones can maintain A? homeostasis by a combination of inhibiting A? fibrillation and facilitating A? aggregate clearance and simultaneously reducing A?-mediated neurotoxicity. The balance of hydrophilic/hydrophobic moieties on the surface of MSPMs is important for their enhanced therapeutic effect.
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Stable, reproducible, and automated capillary zone electrophoresis-tandem mass spectrometry system with an electrokinetically pumped sheath-flow nanospray interface.
Anal. Chim. Acta
PUBLISHED: 01-21-2014
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A PrinCE autosampler was coupled to a Q-Exactive mass spectrometer by an electrokinetically pumped sheath-flow nanospray interface to perform automated capillary zone electrophoresis-electrospray ionization-tandem mass spectrometry (CZE-ESI-MS/MS). 20ng aliquots of an Escherichia coli digest were injected to evaluate the system. Eight sequential injections over an 8-h period identified 1115±70 (relative standard deviation, RSD=6%) peptides and 270±8 (RSD=3%) proteins per run. The average RSDs of migration time, peak intensity, and peak area were 3%, 24% and 19%, respectively, for 340 peptides with high intensity. This is the first report of an automated CZE-ESI-MS/MS system using the electrokinetically pumped sheath-flow nanospray interface. The results demonstrate that this system is capable of reproducibly identifying over 1000 peptides from an E. coli tryptic digest in a 1-h analysis time.
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Preparative isolation and analysis of alcohol dehydrogenase inhibitors from Glycyrrhiza uralensis root using ultrafiltration combined with high-performance liquid chromatography and high-speed countercurrent chromatography.
J Sep Sci
PUBLISHED: 01-19-2014
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A simple, rapid, and effective assay based on ultrafiltration combined with high-performance liquid chromatography and high-speed countercurrent chromatography was developed for screening and purifying alcohol dehydrogenase inhibitors from Glycyrrhiza uralensis root extract. Experiments were carried out to optimize binding conditions including alcohol dehydrogenase concentration, incubation time, temperature, and pH. By comparing the chromatograms, three compounds were found possessing alcohol dehydrogenase binding activity in Glycyrrhiza uralensis root. Under the target-guidance of ultrafiltration combined with the high-performance liquid chromatography experiment, liquiritin (1), isoliquiritin (2), and liquiritigenin (3) were separated by high-speed countercurrent chromatography using ethyl acetate/methanol/water (5:1:4) as the solvent system. The alcohol dehydrogenase inhibitory activities of these three isolated compounds were assessed; compound 2 showed strongest inhibitory activity with an IC50 of 8.95 ?M. The results of the present study indicated that the combinative method using ultrafiltration, high-performance liquid chromatography and high-speed countercurrent chromatography could be widely applied for the rapid screening and isolation of enzyme inhibitors from complex mixtures.
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Antigenic mapping of the hemagglutinin of an H9N2 avian influenza virus reveals novel critical amino acid positions in antigenic sites.
J. Virol.
PUBLISHED: 01-15-2014
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H9N2 influenza virus is undergoing extensive genetic and antigenic evolution, warranting detailed antigenic mapping of its hemagglutinin (HA). Through examining antibody escape mutants of an Asian avian H9N2 virus, we identified 9 critical amino acid positions in H9 antigenic sites. Five of these positions, 164, 167, 168, 196, and 207, have not been reported previously and, thus, represent novel molecular markers for monitoring the antigenic change of H9N2 virus.
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Alteronol inhibits the invasion and metastasis of B16F10 and B16F1 melanoma cells in vitro and in vivo.
Life Sci.
PUBLISHED: 01-08-2014
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The purpose of this study is to evaluate the anti-metastatic effects of alteronol on melanoma B16F10 and B16F1 cells in vitro and in vivo.
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Self-assembly synthesis, structural features, and photophysical properties of dilanthanide complexes derived from a novel amide type ligand: energy transfer from Tb(III) to Eu(III) in a heterodinuclear derivative.
Inorg Chem
PUBLISHED: 01-08-2014
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A novel amide type ligand benzyl-N,N-bis[(2'-furfurylaminoformyl)phenoxyl)ethyl]-amine (L) has been designed and applied for the self-assembly generation of homodinuclear lanthanide coordination compounds [Ln2(?2-L)2(NO3)6(EtOH)2] [Ln = Eu (1), Tb (2), and Gd (3)] and a heterodinuclear derivative [EuTb(?2-L)2(NO3)6(EtOH)2] (4). All the complexes have been characterized by the X-ray single-crystal diffraction analyses. They are isostructural, crystallize in a monoclinic space group P21/c, and form [2 + 2] rectangular macrocycle structures. Compound 4 is the first example of a [2 + 2] rectangular macrocycle heterodinuclear EuTb complex assembled from an amide type ligand. In 4, the discrete 0D dimeric [EuTb(?2-L)2(NO3)6(EtOH)2] units are extended, via the multiple N-H···O hydrogen bonds, into a 2D supramolecular network that has been topologically classified as a uninodal 4-connected underlying net with the sql [Shubnikov tetragonal plane net] topology. The triplet state ((3)??*) of L studied by the Gd(III) complex 3 demonstrated that the ligand beautifully populates Tb(III) emission (? = 52%), whereas the corresponding Eu(III) derivative 1 shows weak luminescence efficiency (? = 0.7%) because the triplet state of L has a poor match with (5)D1 energy level of Eu(III). Furthermore, the photoluminescent properties of heterodinuclear complex 4 have been compared with those of the analogous homodinuclear compounds. The quantum yield and lifetime measurements prove that energy transfer from Tb(III) to Eu(III) is being achieved, namely, that the Tb(III) center is also acting to sensitize the Eu(III) and enhancing Eu(III) emission in 4.
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Quantitative proteomics of Xenopus laevis embryos: expression kinetics of nearly 4000 proteins during early development.
Sci Rep
PUBLISHED: 01-03-2014
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While there is a rich literature on transcription dynamics during the development of many organisms, protein data is limited. We used iTRAQ isotopic labeling and mass spectrometry to generate the largest developmental proteomic dataset for any animal. Expression dynamics of nearly 4,000 proteins of Xenopus laevis was generated from fertilized egg to neurula embryo. Expression clusters into groups. The cluster profiles accurately reflect the major events that mark changes in gene expression patterns during early Xenopus development. We observed decline in the expression of ten DNA replication factors after the midblastula transition (MBT), including a marked decline of the licensing factor XCdc6. Ectopic expression of XCdc6 leads to apoptosis; temporal changes in this protein are critical for proper development. Measurement of expression in single embryos provided no evidence for significant protein heterogeneity between embryos at the same stage of development.
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Complex Impacts of PI3K/AKT Inhibitors to Androgen Receptor Gene Expression in Prostate Cancer Cells.
PLoS ONE
PUBLISHED: 01-01-2014
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Androgen deprivation therapy (ADT) is the first-line treatment to metastatic prostate cancer (PCa). However, sustained expression and function of the androgen receptor (AR) gene contribute to the progression of castration resistant prostate cancers (CRPC). Additionally, tumors can adapt the PI3K/AKT survival pathway to escape ADT. Co-targeting AR and PI3K/AKT signaling has been proposed to be a more effective therapeutic means for CRPC patients. Many clinical trials are ongoing to test whether PI3K/AKT inhibitors are beneficial to PCa patients. However whether these inhibitors have any impacts on the expressions of full length AR (AR-FL) and its splice variant (AR-V7) remains unclear.
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Surface modification and characterization of carbon spheres by grafting polyelectrolyte brushes.
Nanoscale Res Lett
PUBLISHED: 01-01-2014
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Modified carbon spheres (CSPBs) were obtained by grafting poly(diallyl dimethyl ammonium chloride) (p-DMDAAC) on the surface of carbon spheres (CSs). It can be viewed as a kind of cation spherical polyelectrolyte brushes (CSPBs), which consist of carbon spheres as core and polyelectrolytes as shell. The method of synthesizing carbon spheres was hydrothermal reaction. Before the polyelectrolyte brushes were grafted, azo initiator [4,4'-Azobis(4-cyanovaleric acyl chloride)] was attached to the carbon spheres' surface through hydroxyl groups. CSPBs were characterized by scanning electron microscope (SEM), Fourier transform infrared spectroscopy (FTIR), gel permeation chromatography (GPC), conductivity meter, and system zeta potential. The results showed that compared with carbon spheres, the conductivity and zeta potential on CSPBs increased from 9.98 to 49.24 ?S/cm and 11.6 to 42.5 mV, respectively, after the polyelectrolyte brushes were grafted. The colloidal stability in water was enhanced, and at the same time, the average diameter of the CSPBs was found to be 173 nm, and the average molecular weight and grafted density of the grafted polyelectrolyte brushes were 780,138 g/mol and 4.026?×?10(9)/nm(2,) respectively.
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Baicalin protects the cardiomyocytes from ER stress-induced apoptosis: inhibition of CHOP through induction of endothelial nitric oxide synthase.
PLoS ONE
PUBLISHED: 01-01-2014
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Baicalin, the main active ingredient of the Scutellaria root, exerts anti-oxidant and anti-apoptotic effects in cardiovascular diseases. However, the therapeutic mechanism of baicalin remains unknown. Cultured neonatal rat cardiomyocytes were pre-treated with baicalin (0-50 µM) for 24 h, and subsequently treated with tunicamycin (100 ng/ml). Cell viability was detected by MTT assay, and cell damage was determined by LDH release and TUNEL assay. The expression of CHOP, JNK, caspase-3, eNOS was analyzed by western blot. NO was measured by DAF-FM staining. As a result, treatment with baicalin significantly reduced apoptosis induced by ER stress inducer tunicamycin in cardiomyocytes. Molecularly, baicalin ameliorated tunicamycin-induced ER stress by downregulation of CHOP. In addition, baicalin inverted tunicamycin-induced decreases of eNOS mRNA and protein levels, phospho eNOS and NO production through CHOP pathway. However, the protective effects of baicalin were significantly decreased in cardiomyocytes treated with L-NAME, which suppressed activation of nitric oxide synthase. In conclusion, our results implicate that baicalin could protect cardiomyocytes from ER stress-induced apoptosis via CHOP/eNOS/NO pathway, and suggest the therapeutic values of baicalin against ER stress-associated cardiomyocyte apoptosis.
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New Yellow-Emitting Nitride Phosphor SrAlSi4N7:Ce(3+) and Important Role of Excessive AlN in Material Synthesis.
ACS Appl Mater Interfaces
PUBLISHED: 12-13-2013
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Synthesis and luminescent properties of Ce(3+)-doped SrAlSi4N7 yellow-emitting phosphor are reported. In comparison with YAG: Ce(3+), the phosphor exhibits smaller thermal quenching and a broader emission band centering at 555 nm with a bandwidth as large as 115 nm, being suitable for fabricating high color rendering white LED. It is observed in material synthesis that intense luminescence can be achieved only in case of excessive AlN in the raw materials. The role of the excessive AlN is studied. The mechanism for existence of edge-sharing [AlN4] tetrahedral, which is unreasonable according to the aluminum avoidance principle, is discussed in detail.
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Controllable atomistic graphene oxide model and its application in hydrogen sulfide removal.
J Chem Phys
PUBLISHED: 12-11-2013
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The determination of an atomistic graphene oxide (GO) model has been challenging due to the structural dependence on different synthesis methods. In this work we combine temperature-programmed molecular dynamics simulation techniques and the ReaxFF reactive force field to generate realistic atomistic GO structures. By grafting a mixture of epoxy and hydroxyl groups to the basal graphene surface and fine-tuning their initial concentrations, we produce in a controllable manner the GO structures with different functional groups and defects. The models agree with structural experimental data and with other ab initio quantum calculations. Using the generated atomistic models, we perform reactive adsorption calculations for H2S and H2O?H2S mixtures on GO materials and compare the results with experiment. We find that H2S molecules dissociate on the carbonyl functional groups, and H2O, CO2, and CO molecules are released as reaction products from the GO surface. The calculation reveals that for the H2O?H2S mixtures, H2O molecules are preferentially adsorbed to the carbonyl sites and block the potential active sites for H2S decomposition. The calculation agrees well with the experiments. The methodology and the procedure applied in this work open a new door to the theoretical studies of GO and can be extended to the research on other amorphous materials.
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Tuning the K(+) Concentration in the Tunnel of OMS-2 Nanorods Leads to a Significant Enhancement of the Catalytic Activity for Benzene Oxidation.
Environ. Sci. Technol.
PUBLISHED: 11-14-2013
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OMS-2 nanorods with tunable K(+) concentration were prepared by a facile hydrothermal redox reaction of MnSO4, (NH4)2S2O8, and (NH4)2SO4 at 120 °C by adding KNO3 at different KNO3/MnSO4 molar ratios. The OMS-2 nanorod catalysts are characterized by X-ray diffraction, transmission electron microscopy, N2 adsorption and desorption, inductively coupled plasma, and X-ray photoelectron spectrometry. The effect of K(+) concentration on the lattice oxygen activity of OMS-2 is theoretically and experimentally studied by density functional theory calculations and CO temperature-programmed reduction. The results show that increasing the K(+) concentration leads to a considerable enhancement of the lattice oxygen activity in OMS-2 nanorods. An enormous decrease (?T50 = 89 °C; ?T90 > 160 °C) in reaction temperatures T50 and T90 (corresponding to 50 and 90% benzene conversion, respectively) for benzene oxidation has been achieved by increasing the K(+) concentration in the K(+)-doped OMS-2 nanorods due to the considerable enhancement of the lattice oxygen activity.
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Accurate determination of peptide phosphorylation stoichiometry via automated diagonal capillary electrophoresis coupled with mass spectrometry: proof of principle.
Anal. Chem.
PUBLISHED: 11-05-2013
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While reversible protein phosphorylation plays an important role in many cellular processes, simple and reliable measurement of the stoichiometry of phosphorylation can be challenging. This measurement is confounded by differences in the ionization efficiency of phosphorylated and unphosphorylated sites during analysis by mass spectrometry. Here, we demonstrate diagonal capillary electrophoresis-mass spectrometry for the accurate determination of this stoichiometry. Diagonal capillary electrophoresis is a two-dimensional separation method that incorporates an immobilized alkaline phosphatase microreactor at the distal end of the first capillary and employs identical electrophoretic separation modes in both dimensions. The first dimension is used to separate a mixture of the phosphorylated and unphosphorylated forms of a peptide. Fractions are parked in the reactor where they undergo complete dephosphorylation. The products are then periodically transferred to the second capillary and analyzed by mass spectrometry (MS). Because the phosphorylated and unphosphorylated forms differ in charge, they are well resolved in the first dimension separation. Because the unphosphorylated and dephosphorylated peptides are identical, there is no bias in ionization efficiency, and phosphorylation stoichiometry can be determined by the ratio of the signal of the two forms. A calibration curve was generated from mixtures of a phosphorylated standard peptide and its unphosphorylated form, prepared in a bovine serum albumin tryptic digest. This proof of principle experiment demonstrated a linear response across nearly 2 orders of magnitude in stoichiometry.
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Achieving a Rare Breathing Behavior in a Polycatenated 2?D to 3?D Net through a Pillar-Ligand Extension Strategy.
Chemistry
PUBLISHED: 11-04-2013
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Through a pillar-ligand extension strategy, a rare breathing behavior in polycatenated 2D?3D nets has been achieved. Three variants exhibit interesting sorption properties that range from non-breathing to breathing behaviors, which is influenced by the angles between the pillars and the single honeycomb layers. The increase in pillar length does not lead to an increase in polycatenation multiplicity, which is controlled by the length of intralayer tripodal carboxylate. It also does not induce obviously expanded interlayer separations but occupies much more the free voids, and as a consequence, a smaller pore volume is obtained. This suggests that in 2D?3D polycatenated bilayer metal-organic frameworks, the porosity is not always enhanced by increasing the length of the interlayer pillars with the intralayer linker remaining unchanged.
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A tubular europium-organic framework exhibiting selective sensing of Fe3+ and Al3+ over mixed metal ions.
Chem. Commun. (Camb.)
PUBLISHED: 11-02-2013
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A luminescent europium-organic framework with tubular channels based on the H4BTMIPA ligand (H4BTMIPA = 5,5-methylenebis(2,4,6-trimethylisophthalic acid)) was assembled and characterized. The [H2N(CH3)2](+) ions as counterions are located in the channels. The cation exchange between [H2N(CH3)2](+) and metal ions resulted in complex that can selectively sense Fe(3+) and Al(3+) ions through fluorescence quenching and enhancement, respectively.
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Aqp1 Enhances Migration of Bone Marrow Mesenchymal Stem Cells Through Regulation of FAK and ?-Catenin.
Stem Cells Dev.
PUBLISHED: 09-27-2013
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Bone marrow mesenchymal stem cells (MSCs) have the potential to migrate to the site of injury and regulate the repair process. Aquaporin 1 (Aqp1) is a water channel molecule and a regulator of endothelial cell migration. To study the role of Apq1 in MSC migration, we manipulated the expression of the Aqp1 gene in MSCs and explored its effects on MSC migration both in vitro and in vivo. Overexpression of Aqp1 promoted MSC migration, while depletion of Aqp1 impaired MSC migration in vitro. When the green fluorescent protein (GFP) labeled Aqp1 overexpressing MSCs were systemically injected into rats with a femoral fracture, there were significantly more GFP-MSCs found at the fracture gap in the Aqp1-GFP-MSC-treated group compared to the GFP-MSC group. To elucidate the underlying mechanism, we screened several migration-related regulators. The results showed that ?-catenin and focal adhesion kinase (FAK) were upregulated in the Aqp1-MSCs and downregulated in the Aqp1-depleted MSCs, while C-X-C chemokine receptor type 4 had no change. Furthermore, ?-catenin and FAK were co-immunoprecipitated with Aqp1, and depletion of FAK abolished the Aqp1 effects on MSC migration. This study demonstrates that Aqp1 enhances MSC migration ability mainly through the FAK pathway and partially through the ?-catenin pathway. Our finding suggests a novel function of Aqp1 in governing MSC migration, and this may aid MSC therapeutic applications.
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A "strongly" self-catenated metal-organic framework with the highest topological density among 3,4-coordinated nets.
Inorg Chem
PUBLISHED: 09-23-2013
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A new type of 3D "strongly" self-catenated metal-organic framework (SDU-9) has been constructed from [Cu2(COO)4] paddlewheel secondary building units and a tripodal carboxylate linker. SDU-9 ([Cu6(H2O)6L4]·24H2O, where [H3L = 4,4,4"-(hydroxysilanetriyl)tris(triphenyl-4-carboxylic acid), represents a rare example of a highly symmetrical coordination network and extremely tight self-catenation. To the best of our knowledge, SDU-9 has the highest topological density among all known 3,4-coordinated nets.
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N stages of the seventh edition of TNM Classification are the most intensive variables for predictions of the overall survival of gastric cancer patients who underwent limited lymphadenectomy.
Tumour Biol.
PUBLISHED: 09-19-2013
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The objective of this study was to explore the prognostic prediction rationality of the seventh edition N stage for gastric cancer (GC) patients who underwent the limited lymphadenectomy. Clinicopathological data of 769 GC patients who underwent the curative resection between 1997 and 2006 were analyzed for demonstration that the seventh edition N stage had the significant superiorities of prognostic prediction to the patients who underwent the limited lymphadenectomy. Although the extent of lymphadenectomy was associated with the overall survival (OS) of gastric cancer (GC) patients, the N stages of the seventh edition of the TNM Classification were identified as the most intensively independent predictors of GC prognosis. Using stratum analysis, the 5-year survival rate of patients who underwent limited lymphadenectomy was observed to be significantly different from that of patients who underwent extended lymphadenectomy, regardless of the extent of lymph node metastasis. Multinomial logistic regression analysis revealed that combining the extents of lymph node metastasis and lymphadenectomy could improve the prediction accuracy of patient survival status. Case control analysis showed that regardless of the extent of lymphadenectomy, the seventh edition N stages featured significant superiority for OS evaluation of GC patients. The seventh edition N stage had the prediction rationality for the OS of GC patients who underwent the limited lymphadenectomy.
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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.