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Find video protocols related to scientific articles indexed in Pubmed.
Synthesis of 2-Pyridyl-benzimidazole Iridium(III), Ruthenium(II), and Platinum(II) Complexes. Study of the Activity as Inhibitors of Amyloid-? Aggregation and Neurotoxicity Evaluation.
Inorg Chem
PUBLISHED: 11-20-2014
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The design of small molecules that can target the aggregation of A? as potential therapeutic agents for Alzheimer's disease is an area of study that has attracted a lot of attention recently. The novel ligand methyl 1-butyl-2-pyridyl-benzimidazole carboxylate was prepared for the synthesis of a series of new iridium(III), ruthenium(II), and platinum(II) 2-pyridyl-benzimidazole complexes. The crystal structure of the half-sandwich iridium(III) complex was established by X-ray diffraction. An arrangement of two cationic complexes in the unit cell is observed, and it seems to be organized by weak ?···? interactions that are taking place between two symmetry-related benzimidazole ring systems. All new compounds inhibited aggregation of A?1-42 in vitro as shown by both thioflavin T fluorescence assay and transmission electron microscopy. Among them the Ir compound rescued the toxicity of A?1-42 in primary cortical neurons effectively.
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A Chemical Tuned Strategy to Develop Novel Irreversible EGFR-TK Inhibitors with Improved Safety and Pharmacokinetic Profiles.
J. Med. Chem.
PUBLISHED: 11-20-2014
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Gatekeeper T790M mutation in EGFR is the most prevalent factor underlying acquired resistance. Acrylamide-bearing quinazoline derivatives are powerful irreversible inhibitors for overcoming resistance. Nevertheless, concerns about the risk of non-specific covalent modification have motivated the development of novel cysteine-targeting inhibitors. In this paper, we demonstrate that fluoro-substituted olefins can be tuned to alter Michael addition reactivity. Incorporation of these olefins into the quinazoline templates produced potent EGFR inhibitors with improved safety and pharmacokinetic properties. A lead compound 5a was validated against EGFRWT, EGFR T790M as well as A431 and H1975 cancer cell lines. Additionally, compound 5a displayed a weaker inhibition against the EGFR-independent cancer cell line SW620 when compared withafatinib. Oral administration of 5a at a dose of 30mg/kg induced tumor regression in a murine-EGFRL858R/T790M driven H1975 xenograft model. Also, 5a exhibited improved oral bioavailability and safety, as well as favorable tissue distribution properties and enhanced brain uptake. These findings provide the basis of a promising strategy toward the treatment of NSCLC patients with drug resistance.
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Using the multi-parameter variability of photoplethysmographic signals to evaluate short-term cardiovascular regulation.
J Clin Monit Comput
PUBLISHED: 11-12-2014
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Transient changes in cardiovascular regulatory activities are valuable for clinical monitoring and pathophysiological research. However, there is a lack of effective methods to evaluate short-term cardiovascular regulation. This study explores a photoplethysmography (PPG)-based multi-parameter analytical approach using the period, amplitude and baseline of PPG signals to quantitatively assess cardiovascular regulation over a 30 s period. PPG data were recorded from 31 young healthy subjects during conditions of spontaneous respiration, paced respiration with 15 breaths/min and breath holding, with each condition lasting 30 s. The three indices (SD1, SD2, SD1/SD2) in Poincaré plot of the normalized parameters and the correlations between the parameters or the first differences of the parameters were analyzed. The results showed that compared with spontaneous respiration, SD1 of PPG amplitude increased significantly but the correlations between any two parameters or between the first differences of any two parameters decreased significantly during paced respiration; SD1 and SD1/SD2 of all PPG parameters as well as the correlations of the first differences between any two PPG parameters reduced significantly during breath holding. The results indicate the respiratory induced alterations in cardiovascular autonomic function could be identified by the variability of various PPG parameters or their correlations over 30 s periods. Moreover, the indicators used to quantify the variability of the PPG parameters in this study may provide a feasible and effective way to evaluate short-term cardiovascular regulation.
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Effect of repetitive end-inspiration breath holding on very short-term heart rate variability in healthy humans.
Physiol Meas
PUBLISHED: 11-12-2014
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Very short-term heart rate variability (HRV) is thought to reflect dynamic changes in autonomic nervous activity, which is helpful in understanding the role of autonomic nervous function (ANF) in the mechanisms underlying apnea-induced cardiac arrhythmias. The goal of this study was to investigate the effect of repetitive end-inspiration breath holding on very short-term HRV. A total of 32 young healthy participants took part in the experiments. Three trials were performed, each involving seven repetitive end-inspiration breath holding and a 30?s recovery period between breath holding. Durations of breath holding in the three trials were 1:2:3. The study first evaluated the effect of analyzed data lengths on the stability of HRV indices and determined three HRV indices suitable for very short-term analysis. The results showed that in most cases, during breath holding, the square root of the mean squared differences of successive normal RR intervals (rMSSD) was significantly lower, but normalized units of the power in the low frequency band ranging from 0.04 to 0.15?Hz (nLF) and LF/high frequency (HF) were significantly higher than those during corresponding durations under the normal breathing conditions. On the contrary, during recovery after breath holding, rMSSD was significantly higher but nLF and LF/HF were lower than normal. Moreover, the durations of breath holding had no significant influence on the variations of LF/HF. In addition, as participants repeated the breath holding, HRV indices varied non-linearly. HRV changes may indicate sympathetic activation during breath holding and parasympathetic activation during recovery after breath holding. In conjunction with the existing physiological interpretation based on changes in heart rate, the results may imply that breath holding leads to both cardiac sympathetic and parasympathetic activation simultaneously, which may be a possible pathogenic factor of apnea-induced arrhythmias.
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IKK? is an IRF5 kinase that instigates inflammation.
Proc. Natl. Acad. Sci. U.S.A.
PUBLISHED: 10-19-2014
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The transcription factor interferon regulatory factor 5 (IRF5) is essential for the induction of inflammatory cytokines, but the mechanism by which IRF5 is activated is not well understood. Here we present evidence that the kinase IKK? phosphorylates and activates IRF5 in response to stimulation in several inflammatory pathways, including those emanated from Toll-like receptors and retinoic acid-inducible gene I-like receptors. IKK? phosphorylates mouse IRF5 at specific residues, including serine 445 (S446 in human IRF5 isoform 1), as evidenced by mass spectrometry analysis and detection with a phosphospecific antibody. Recombinant IKK? phosphorylated IRF5 at Ser-445 in vitro, and a point mutation of this serine abolished IRF5 activation and cytokine production. Depletion or pharmacologic inhibition of IKK? prevented IRF5 phosphorylation. These results indicate that IKK? is an IRF5 kinase that instigates inflammation.
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Spindle-like abdominal wall-lifting device in gasless laparoscopic surgery in animal experiments.
Minim Invasive Ther Allied Technol
PUBLISHED: 10-13-2014
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Abstract Introduction: CO2 pneumoperitoneum has been used to establish an operation space in laparoscopic surgery. It may have some bad consequences, such as respiratory and circulatory system changes, hypotension and hypoxia in infants and the release of free tumor cells into the abdominal cavity. Gasless laparoscopic technique can avoid these adverse effects. But present gasless laparoscopic techniques have their own disadvantages. The main shortcoming of gasless laparoscopic techniques is inadequate operative space.
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Diffuse Hepatic Amebiasis Detected by FDG PET/CT.
Clin Nucl Med
PUBLISHED: 10-03-2014
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A 39-year-old man presented with sudden decreased visual acuity in his left eye. Orbital CT and MRI revealed a soft tissue lesion in his left orbital apex. FDG PET/CT showed increased FDG uptake by the left orbital lesion, abnormal focal FDG uptake in the soft tissues of the external ears, and abnormal heterogeneous FDG activity throughout the liver. Percutaneous liver biopsy, external auditory canal discharge, and stool specimens revealed amebiasis. The patient responded to antiamebic therapy, and his lesions improved. The case demonstrates that during its early stage, hepatic amebiasis may be associated with a relatively heterogeneous pattern of FDG uptake.
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Highly enantioselective phosphination and hydrophosphonylation of azomethine imines: using chiral squaramide as a hydrogen bonding organocatalyst.
Org. Biomol. Chem.
PUBLISHED: 09-26-2014
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Enantioselective phosphination and hydrophosphonylation reactions between azomethine imines and diarylphosphine oxides or dialkyl phosphites were respectively developed by the use of a chiral squaramide as the hydrogen bonding organocatalyst, which afforded two types of phosphorus containing product in high yields with good to excellent enantioselectivities.
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Proteomic analysis and prediction of human phosphorylation sites in subcellular level reveal subcellular specificity.
Bioinformatics
PUBLISHED: 09-17-2014
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Protein phosphorylation is the most common post-translational modification (PTM) regulating major cellular processes through highly dynamic and complex signaling pathways. Large-scale comparative phosphoproteomic studies have frequently been done on whole cells or organs by conventional bottom-up mass spectrometry approaches, i.e at the phosphopeptide level. Using this approach, there is no way to know from where the phosphopeptide signal originated. Also, as a consequence of the scale of these studies, important information on the localization of phosphorylation sites in subcellular compartments (SCs) is not surveyed.
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Genomic Landscape of Ewing Sarcoma Defines an Aggressive Subtype with Co-Association of STAG2 and TP53 Mutations.
Cancer Discov
PUBLISHED: 09-15-2014
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Ewing sarcoma is a primary bone tumor initiated by EWSR1-ETS gene fusions. To identify secondary genetic lesions that contribute to tumor progression, we performed whole-genome sequencing of 112 Ewing sarcoma samples and matched germline DNA. Overall, Ewing sarcoma tumors had relatively few single-nucleotide variants, indels, structural variants, and copy-number alterations. Apart from whole chromosome arm copy-number changes, the most common somatic mutations were detected in STAG2 (17%), CDKN2A (12%), TP53 (7%), EZH2, BCOR, and ZMYM3 (2.7% each). Strikingly, STAG2 mutations and CDKN2A deletions were mutually exclusive, as confirmed in Ewing sarcoma cell lines. In an expanded cohort of 299 patients with clinical data, we discovered that STAG2 and TP53 mutations are often concurrent and are associated with poor outcome. Finally, we detected subclonal STAG2 mutations in diagnostic tumors and expansion of STAG2-immunonegative cells in relapsed tumors as compared with matched diagnostic samples.
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Targetable kinase-activating lesions in Ph-like acute lymphoblastic leukemia.
N. Engl. J. Med.
PUBLISHED: 09-11-2014
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Philadelphia chromosome-like acute lymphoblastic leukemia (Ph-like ALL) is characterized by a gene-expression profile similar to that of BCR-ABL1-positive ALL, alterations of lymphoid transcription factor genes, and a poor outcome. The frequency and spectrum of genetic alterations in Ph-like ALL and its responsiveness to tyrosine kinase inhibition are undefined, especially in adolescents and adults.
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Regulation of metabolic flux in Lactobacillus casei for lactic acid production by overexpressed ldhL gene with two-stage oxygen supply strategy.
J. Microbiol. Biotechnol.
PUBLISHED: 09-01-2014
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This study describes a novel strategy to regulate metabolic flux for lactic acid production in Lactobacillus casei. The ldhL gene encoding L-lactate dehydrogenase (L-LDH) was overexpressed in the L. casei, and a two-stage oxygen supply strategy (TOS) that maintained medium oxygen supply level (MOS) during early fermentation phase, and low oxygen supply level (LOS) in the later phase was carried out. As a consequence, maximum L-LDH activity of 95.6 U/mL was obtained in the recombinant strain, which was over 4-fold higher than that of the initial strain. Under the TOS for L. casei (pMG-ldhL), maximum lactic acid concentrations of 159.6 g/L was obtained in 36 h, corresponding to a 62.8% increase. The results presented here provide a novel way to regulate the metabolic flux of L. casei for lactic acid production in different fermentation stages, which is available to enhance organic acid production with other strains.
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Decreased calcium-activated potassium channels by hypoxia causes abnormal firing in the spontaneous firing medial vestibular nuclei neurons.
Eur Arch Otorhinolaryngol
PUBLISHED: 08-31-2014
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Vertebrobasilar insufficiency (VBI) presents complex varied clinical symptoms, including vertigo and hearing loss. Little is known, however, about how Ca(2+)-activated K(+) channel attributes to the medial vestibular nucleus (MVN) neural activity in VBI. To address this issue, we performed whole-cell patch clamp and quantitative polymerase chain reaction (qPCR) to examine the effects of hypoxia on neural activity and the changes of the large conductance Ca(2+) activated K(+) channels (BKCa channels) in the MVN neurons in brain slices of male C57BL/6 mice. Brief hypoxic stimuli of the brain slices containing MVN were administrated by switching the normoxic artificial cerebrospinal fluid (ACSF) equilibrated with 21 % O2/5 % CO2 to hypoxic ACSF equilibrated with 5 % O2/5 % CO2 (balance N2). 3-min hypoxia caused a depolarization in the resting membrane potential (RM) in 8/11 non-spontaneous firing MVN neurons. 60/72 spontaneous firing MVN neurons showed a dramatic increase in firing frequency and a depolarization in the RM following brief hypoxia. The amplitude of the afterhyperpolarization (AHPA) was significantly decreased in both type A and type B spontaneous firing MVN neurons. Hypoxia-induced firing response was alleviated by pretreatment with NS1619, a selective BKCa activator. Furthermore, brief hypoxia caused a decrease in the amplitude of iberiotoxin-sensitive outward currents and mRNA level of BKCa in MVN neurons. These results suggest that BKCa channels protect against abnormal MVN neuronal activity induced by hypoxia, and might be a key target for treatment of vertigo and hearing loss in VBI.
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Formulation, characterization and clinical evaluation of propranolol hydrochloride gel for transdermal treatment of superficial infantile hemangioma.
Drug Dev Ind Pharm
PUBLISHED: 08-25-2014
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Abstract The objective of the present study is to formulate and characterize propranolol hydrochloride (PPL?·?HCl) gel, and to evaluate the efficacy of this formulation in transdermal treatment for superficial infantile hemangioma (IH). The transdermal PPL?·?HCl gel was prepared by a direct swelling method, which chose hydroxypropyl methylcellulose (HPMC) as the matrix and used terpenes plus alcohols as permeation enhancer. Permeation studies of PPL?·?HCl were carried out with modified Franz diffusion cells through piglet skin. Our results pointed to that among all studied permeation enhancers, farnesol plus isopropanol was the most effective combination (Q24, 6027.4?±?563.1??g/cm(2), ER, 6.8), which was significantly higher than that of control gel (p?
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FOS-like antigen 1 is highly expressed in human psoriasis tissues and promotes the growth of HaCaT cells in vitro.
Mol Med Rep
PUBLISHED: 08-21-2014
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Psoriasis is a multifactorial disease and the mechanisms involved in its pathogenesis remain to be elucidated. FOS?like antigen 1 (Fra?1) is a proto?oncogene. It is a negative inhibitor of activator protein?1 activity and possesses transforming activity. The effect of and possible mechanisms underlying Fra?1 in psoriasis remain to be elucidated. In the present study, western blot analysis and reverse transcription?quantitative polymerase chain reaction (RT?qPCR) techniques were used to identify differentially expressed Fra?1 in psoriatic and in normal control tissues. Compared with the control samples, the expression of normalized Fra?1 genes in psoriasis was 12.6 times higher. Western blot analysis was used to assess the protein levels of Fra?1. The results demonstrated that the protein expression of Fra-1 was high in tissues affected by psoriasis. This also corresponded with the results of RT?qPCR. Fra?1?stable expressing HaCaT/Fra?1 or control HaCaT/vector cell lines were then generated to elucidate the function of Fra?1 in the growth of HaCaT cells. The results demonstrated that Fra?1 promoted the growth of HaCaT cells in vitro by arresting the cell cycle and inhibiting cell apoptosis. These results suggested that Fra?1 may be important in psoriasis.
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Treadmill exercise promotes angiogenesis in the ischemic penumbra of rat brains through caveolin-1/VEGF signaling pathways.
Brain Res.
PUBLISHED: 08-19-2014
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The purpose of this study was to investigate the role of caveolin-1 in treadmill-exercise-induced angiogenesis in the ischemic penumbra of rat brains, and whether caveolin-1 changes correlated with reduced brain injury induced by treadmill exercise, in rats after cerebral ischemia. Rats were randomized into five groups: sham-operated (S, n=7), model (M, n=36), exercise and model (EM, n=36), inhibitor and model (IM, n=36), and inhibitor, exercise, and model (IEM, n=36). Rats in the model groups underwent middle cerebral artery occlusion (MCAO). Rats in the inhibitor groups received an IP injection of the caveolin-1 inhibitor, daidzein (0.4mg/kg), every 24h following reperfusion. Rats were killed at 7 or 28 days after the operation. The exercise group showed better neurological recovery and smaller infarction volumes compared with the non-exercise group. Correspondingly, significant increases of caveolin-1 and vascular endothelial growth factor (VEGF) protein expression were observed compared with the non-exercise group. Additionally, the number of Flk-1/CD34 double-positive cells towards the ischemic penumbra was increased in the exercise group. Furthermore, the induction of VEGF protein, microvessel density, decrease of infarct volumes and neurological recovery was significantly inhibited by daidzein. This study indicates that treadmill exercise reduces brain injury in stroke. Our findings suggest that the caveolin-1 pathway is involved in the regulation of VEGF in association with promoted angiogenesis in the ischemic penumbra of rat brains after treadmill exercise. The caveolin-1/VEGF signaling pathway may be a potential target for therapeutic intervention in rats following MCAO.
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Remodeling of the intestinal brush border underlies adhesion and virulence of an enteric pathogen.
MBio
PUBLISHED: 08-19-2014
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Intestinal colonization by Vibrio parahaemolyticus-the most common cause of seafood-borne bacterial enteritis worldwide-induces extensive disruption of intestinal microvilli. In orogastrically infected infant rabbits, reorganization of the apical brush border membrane includes effacement of some microvilli and marked elongation of others. All diarrhea, inflammation, and intestinal pathology associated with V. parahaemolyticus infection are dependent upon one of its type 3 secretion systems (T3SS2); however, translocated effectors that directly mediate brush border restructuring and bacterial adhesion are not known. Here, we demonstrate that the effector VopV is essential for V. parahaemolyticus intestinal colonization and therefore its pathogenicity, that it induces effacement of brush border microvilli, and that this effacement is required for adhesion of V. parahaemolyticus to enterocytes. VopV contains multiple functionally independent and mechanistically distinct domains through which it disrupts microvilli. We show that interaction between VopV and filamin, as well as VopV's previously noted interaction with actin, mediates enterocyte cytoskeletal reorganization. VopV's multipronged approach to epithelial restructuring, coupled with its impact on colonization, suggests that remodeling of the epithelial brush border is a critical step in pathogenesis.
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Inhibition of Rac1 activity induces G1/S phase arrest through the GSK3/cyclin D1 pathway in human cancer cells.
Oncol. Rep.
PUBLISHED: 08-07-2014
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Rac1 has been shown to regulate the cell cycle in cancer cells. Yet, the related mechanism remains unclear. Thus, the present study aimed to investigate the mechanism involved in the regulation of G1/S phase transition by Rac1 in cancer cells. Inhibition of Rac1 by inhibitor NSC23766 induced G1/S phase arrest and inhibited the proliferation of A431, SW480 and U2-OS cells. Suppression of GSK3 by shRNA partially rescued G1/S phase arrest and inhibition of proliferation. Incubation of cells with NSC23766 reduced p-AKT and inactivated p-GSK3? and p-GSK3?, increased p-cyclin D1 expression and decreased the level of cyclin D1 protein. Consequently, cyclin D1 targeting transcriptional factor E2F1 expression, which promotes G1 to S phase transition, was also reduced. In contrast, constitutive active Rac1 resulted in increased p-AKT and inactivated p-GSK3? and p-GSK3?, decreased p-cyclin D1 expression and enhanced levels of cyclin D1 and E2F1 expression. Moreover, suppression of GSK3 did not alter p-AKT or Rac1 activity, but decreased p-cyclin D1 and increased total cyclin D1 protein. However, neither Rac1 nor GSK3 inhibition altered cyclin D1 at the RNA level. Moreover, after inhibition of Rac1 or GSK3 following proteasome inhibitor MG132 treatment, cyclin D1 expression at the protein level remained constant, indicating that Rac1 and GSK3 may regulate cyclin D1 turnover through phosphorylation and degradation. Therefore, our findings suggest that inhibition of Rac1 induces cell cycle G1/S arrest in cancer cells by regulation of the GSK3/cyclin D1 pathway.
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Overexpression of HMGB1 in melanoma predicts patient survival and suppression of HMGB1 induces cell cycle arrest and senescence in association with p21 (Waf1/Cip1) up-regulation via a p53-independent, Sp1-dependent pathway.
Oncotarget
PUBLISHED: 07-23-2014
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Although laboratory studies have implicated the high mobility group box 1 (HMGB1) in melanoma, its clinical relevance remains unclear. We analyzed nearly 100 cases of human melanoma and found that HMGB1 was highly overexpressed in melanoma samples relative to normal skin and nevi tissues. Significantly, higher levels of HMGB1 correlated with more advanced disease stages and with poorer survival in melanoma patients. Unlike the well-documented pro-inflammatory role of the extracellular HMGB1, we found that its intracellular activity is necessary for melanoma cell proliferation. An absolute dependency of melanoma cell proliferation on HMGB1 was underscored by the marked response of cell cycle arrest and senescence to HMGB1 knockdown. We demonstrated that HMGB1 deficiency-induced inhibition of cell proliferation was mediated by p21, which was induced via a Sp1-dependent mechanism. Taken together, our data demonstrate a novel oncogenic role of HMGB1 in promoting human melanoma cell proliferation and have important implications in melanoma patient care.
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One-pot synthesis of macro-mesoporous bioactive glasses/polylactic acid for bone tissue engineering.
Mater Sci Eng C Mater Biol Appl
PUBLISHED: 07-19-2014
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The macro-mesoporous bioactive glasses/polylactic acid nanofibers were synthesized via electrospun method followed by acid treatment processing. It was identified to be an effective and simple synthetic strategy to form the uniform nanofibers about 350 nm in size. The non-ionic triblock copolymer, poly(ethylene glycol)-block-poly(propylene glycol)-block-poly(ethylene glycol) (P123), was used as the template for mesoporous structure (5 nm) and the macroporous structure about 10 ?m in size derived from the overlapping of the nanofibers. Furthermore, the surface hydrophilic-hydrophobic property can be adjusted by varying the amount of mesoporous bioglass precursor (MBG-p). With the outstanding structure characters and the suitable hydrophilic property, these nanofiber composites show controlled drug release and the fast hydroxyapatite (HAP) mineralization performance. Herein, the novel materials are expected to have potential application for bone tissue engineering.
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MiR-20a inhibits cutaneous squamous cell carcinoma metastasis and proliferation by directly targeting LIMK1.
Cancer Biol. Ther.
PUBLISHED: 07-14-2014
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MicroRNA-20a (miR-20a) plays a key role in tumorigenesis and progression. But its function is reverse in different kinds of malignant tumor, and its role and mechanism in cutaneous squamous cell carcinoma (CSCC) remains unclear.
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Identification of FLOT2 as a novel target for microRNA-34a in melanoma.
J. Cancer Res. Clin. Oncol.
PUBLISHED: 07-11-2014
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To confirm whether flotillin 2 (FLOT2) is a direct target of miR-34a and miR-34a/FLOT2 pathway plays a key role in melanoma proliferation and metastasis.
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[Prokaryotic expression and immunological characteristics of Mycobacterium tuberculosis Rv1886c].
Wei Sheng Wu Xue Bao
PUBLISHED: 07-03-2014
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Ag85B (Rv1886c) is secreted during the early stages of infection by Mycobacterium tuberculosis. The purpose of this study was probed into the immune response against Ag85B in vivo.
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Escherichia coli isolates from sick chickens in China: Changes in antimicrobial resistance between 1993 and 2013.
Vet. J.
PUBLISHED: 06-24-2014
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The use of antimicrobials for the control of infectious disease has increased in recent decades. Understanding trends in antimicrobial resistance provides clues about the relationship between antimicrobial use and the emergence of resistance. We examined the resistance of 540 Escherichia coli isolates to 19 antimicrobials that represent 11 classes of antimicrobial agents. The isolates were collected from chickens between 1993 and 2013 in China. Overall, >96.7% of the isolates were resistant to at least one of the tested compounds, and 87.2% of them displayed multidrug resistance (MDR) representing five to six antimicrobial classes. A high proportion of E. coli isolates were resistant to tetracycline (90.6%), nalidixic acid (80.6%), ampicillin (77.2%), trimethoprim-sulfamethoxazole (76.9%), and streptomycin (72.8%). Only 3.0% of the isolates were resistant to nitrofurantoin, and none was resistant to meropenem. Resistance to amikacin, ampicillin, aztreonam, ceftazidime, cefotaxime, cephalothin, chloramphenicol, ciprofloxacin, fosfomycin, levofloxacin, norfloxacin, nalidixic acid, piperacillin, and trimethoprim-sulfamethoxazole significantly increased from 1993 to 2013 (P?<0.01). There was an increasing trend in MDR over the 20?year period.
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[Development of skin moisture and body fat measurement system for mobile application].
Zhongguo Yi Liao Qi Xie Za Zhi
PUBLISHED: 06-20-2014
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Integrating physiological parameters measurement into mobile devices is a development tendency of mobile healthcare. Measurement methods for skin moisture and body fat content are studied in this paper. Electrodes are designed for easy integration into mobile devices, and can be embedded in the cover of the mobile phone. Experiments were conducted to obtain a fast and easy measurement method. The results of evaluation show that the measurement system can achieve the same accuracy as commercial products (with correlation above 0.9 and root mean squared error below 4%) in skin moisture and body fat content measurement. Measurement of local-area body fat content showed a nearly linear positive correlation between local-area body fat content and local-area body impedance.
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[Immunological characteristics of Mycobacterium tuberculosis antigen Rv2628].
Sheng Wu Gong Cheng Xue Bao
PUBLISHED: 06-20-2014
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Antigen Rv2628 of Mycobacterium tuberculosis is associated with latent tuberculosis infection. In this study, Rv2628 was prokaryotic expressed and purified, its immunological characteristics was evaluated with macrophage cell line RAW264.7 and BALB/c mice. The results show that Rv2628 was mainly expressed in form of inclusion body confirmed by SDS-PAGE, and could react with rabbit anti-H37Rv polyclonal antibody detected by Western blotting assay, indicating that the protein had an effective immunoreactivity. The interactions between Rv2628 and macrophage cell line RAW264.7 confirmed that it could effectively induce cells to produce pro-inflammatory cytokines, the relative expression level of IL-6 mRNA was higher than the control group in 1-12 h. BALB/c mice were subcutaneously immunized with Rv2628 protein, the production of IFN-gamma and IL-4 in the spleen cells was determined by Sandwich ELISA, in the Rv2628 immunized group, the level of IFN-gamma was significantly higher than that of IL-4 (P < 0.000 1). It indicated the protein induced Th1-tendency immune responses. At the same time, Rv2628(11-30) peptide used as coating antigen, the murine serum antibody titer detected by indirect-ELISA was 1:1 600, which demonstrated that Rv2628 could also induce humoral immune responses. In summary, Rv2628 could induce specific pro-inflammatory cytokines, affectively induce strongly Th1-tendency immune response and humoral response, it could be a potential target for developing subunit vaccine against TB. In addition, it laid foundation for probing the cross-talk between M. tb and host.
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microRNA-194 suppresses osteosarcoma cell proliferation and metastasis in vitro and in vivo by targeting CDH2 and IGF1R.
Int. J. Oncol.
PUBLISHED: 06-18-2014
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Studies have shown that miR-194 functions as a tumor suppressor and is associated with tumor growth and metastasis. We studied the effects of miR-194 in osteosarcoma and the possible mechanism by which miR-194 affected the survival, apoptosis and metastasis of osteosarcoma. Both human osteosarcoma cell lines SOSP-9607 and U2-OS were transfected with recombinant lentiviruses to regulate miR-194 expression. Overexpression of miR-194 partially inhibited the proliferation, migration, and invasion of osteosarcoma cells in vitro, as well as tumor growth and pulmonary metastasis of osteosarcoma cells in vivo. Potential miR-194 target genes were predicted using bioinformatics. Luciferase reporter assay, real-time quantitative PCR and western blotting confirmed that CDH2 (N-cadherin) and IGF1R were targets of miR-194. Using real-time quantitative PCR, we evaluated the expression of miR-194 and two miR-194 target genes, CDH2 and IGF1R in osteosarcoma samples from 107 patients and 99 formalin- or paraformalin-fixed paraffin-embedded tissues. The expressions of the target genes were also examined in osteosarcoma samples using immunohistochemistry. Overexpression of miR-194 inhibited tumor growth and metastasis of osteosarcoma probably by downregulating CDH2 and IGF1R. miR-194 may prove to be a promising therapeutic agent for osteosarcoma.
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Fumanjian, a Classic Chinese Herbal Formula, Can Ameliorate the Impairment of Spatial Learning and Memory through Apoptotic Signaling Pathway in the Hippocampus of Rats with A? 1-40 -Induced Alzheimer's Disease.
Evid Based Complement Alternat Med
PUBLISHED: 05-28-2014
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Alzheimer's disease (AD) is the most common form of dementia and lacks disease-altering treatments. Fumanjian (FMJ), a famous classic Chinese herbal prescription for dementia, was first recorded in the Complete Works of Jingyue during the Ming Dynasty. This study aimed to investigate whether FMJ could prevent cognitive deficit and take neuroprotective effects in A? 1-40-induced rat model through apoptotic signaling pathway. AD model was established by bilateral injection of A? 1-40 into hippocampus in rat. All rats were tested for their capabilities of spatial navigation and memorization by Morris water maze. Apoptosis was tested using TUNEL staining in hippocampus neuronal cells; RT-PCR tested expression of Bcl-2 and Bax mRNA; western blotting tested protein level of cleaved caspase-3. After 14 days of treatment, FMJ significantly improved the escape latency and enhanced platform-cross number compared with the A? 1-40-injected group (P < 0.05 or P < 0.01). FMJ also significantly decreased number of TUNEL-positive neuronal apoptosis and the expressions of Bax and cleaved Caspase-3 and increased the expression of Bcl-2 (P < 0.01) compared with AD model group. In conclusion, FMJ exerts a protective effect against A? 1-40-induced learning and memory deficits and neuronal apoptosis, suggesting that FMJ could be used as a potential therapeutic formula for AD.
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Expression of Nfic during root formation in first mandibular molar of rat.
J. Mol. Histol.
PUBLISHED: 05-22-2014
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The transcription factor Nfic is a key regulator during tooth development. Nfic deficient mice exhibit short and abnormal molar roots and severely deformed incisors. Dental epithelial cells, known as Hertwig's epithelial root sheath (HERS), participate in root formation. However, whether Nfic is involved in HERS-mesenchyme interaction remains unclear. In this study, the detail temporal and spatial expression pattern of Nfic during rat molar development was examined using immunohistochemistry and in situ hybridization. Nfic was detected in ameloblasts, dental follicle cells (DFCs) and dental papilla cells (DPCs), especially the DPCs close to dentin, from postnatal day 5 to day 16. Nfic expression in DPCs, DFCs and HERS cells was also examined by western blot and RT-PCR. Nfic was detected in DPCs and DFCs, but not in HERS cells. Co-culture experiment further indicated that Nfic mRNA expression in DPCs was elevated by the presence of HERS cells. Our results revealed that Nfic could be a marker gene for root odontoblasts differentiation initiation and its expression might be regulated through epithelial-mesenchymal interaction.
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[Portable instrument for arteriosclerosis assessment].
Zhongguo Yi Liao Qi Xie Za Zhi
PUBLISHED: 05-21-2014
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A portable instrument for arteriosclerosis assessment containing sensor module, acquisition board and embedded module was developed for home care in this paper. The sensor module consists of one ECG module and three pulse wave extraction modules, synchronously acquiring human ECG and pulse wave signal of carotid, radial, and dorsal, respectively. The acquisition board converts the sensor module's analog output signals into digital signals and transmits them to the embedded module. The embedded module realizes the functions including signal display, storage and the calculation and output of pulse wave velocity. The structure of the proposed portable instrument is simple, easy to use, and easy to expand. Small size, low cost, and low power consumption are also the advantages of this device. Experimental results demonstrated that the proposed portable instrument for arteriosclerosis assessment has high accuracy, good repeatability and can assess the degree of atherosclerosis appropriately.
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Detection of circulating methylated opioid binding protein/cell adhesion molecule-like gene as a biomarker for ovarian carcinoma.
Clin. Lab.
PUBLISHED: 05-21-2014
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Hypermethylation of the opioid binding protein/cell adhesion molecule-like (OPCML) gene is frequently observed in ovarian carcinoma. We evaluated the detection of circulating hypermethylated OPCML for detecting ovarian carcinoma and assessing its prognosis.
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Laparoendoscopic single-site ureteroureterostomy with intraoperative retrograde ureteroscopy-assisted technique for benign proximal and middle ureteral strictures: a single-center experience.
J Laparoendosc Adv Surg Tech A
PUBLISHED: 05-20-2014
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To report a minimally invasive and reproducible technique that greatly facilitates the identification of the stricture during laparoendoscopic single-site ureteroureterostomy (LESS-UU) for benign proximal and middle ureteral strictures, using the intraoperative retrograde ureteroscopy-assisted technique.
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[Antigenic determinants analysis and detection of virulence factors in F18 fimbriae Escherichia coli strains isolated from pigs].
Wei Sheng Wu Xue Bao
PUBLISHED: 05-14-2014
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We determined the present distribution of serogroups and virulence factors among F18 Escherichia coli isolates from pigs with diarrhea and/or edema disease during 1998 to 2006. Epitope of F18 fimbriae was also analyzed.
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As a novel p53 direct target, bidirectional gene HspB2/?B-crystallin regulates the ROS level and Warburg effect.
Biochim. Biophys. Acta
PUBLISHED: 05-13-2014
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Many mammalian genes are composed of bidirectional gene pairs with the two genes separated by less than 1.0kb. The transcriptional regulation and function of these bidirectional genes remain largely unclear. Here, we report that bidirectional gene pair HspB2/?B-crystallin, both of which are members of the small heat shock protein gene family, is a novel direct target gene of p53. Two potential binding sites of p53 are present in the intergenic region of HspB2/?B-crystallin. p53 up-regulated the bidirectional promoter activities of HspB2/?B-crystallin. Actinomycin D (ActD), an activator of p53, induces the promoter and protein activities of HspB2/?B-crystallin. p53 binds to two p53 binding sites in the intergenic region of HspB2/?B-crystallin in vitro and in vivo. Moreover, the products of bidirectional gene pair HspB2/?B-crystallin regulate glucose metabolism, intracellular reactive oxygen species (ROS) level and the Warburg effect by affecting metabolic genes, including the synthesis of cytochrome c oxidase 2 (SCO2), hexokinase II (HK2), and TP53-induced glycolysis and apoptosis regulator (TIGAR). The ROS level and the Warburg effect are affected after the depletion of p53, HspB2 and ?B-crystallin respectively. Finally, we show that both HspB2 and ?B-crystallin are linked with human renal carcinogenesis. These findings provide novel insights into the role of p53 as a regulator of bidirectional gene pair HspB2/?B-crystallin-mediated ROS and the Warburg effect.
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Electrochemical impedance immunosensor for sub-picogram level detection of bovine interferon gamma based on cylinder-shaped TiO? nanorods.
Biosens Bioelectron
PUBLISHED: 05-08-2014
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Bovine interferon gamma (BoIFN-?) released by T cells plays very important roles in early diagnosis of Mycobacterium tuberculosis (MTB) infections and control of bovine tuberculosis. In this work, a label-free electrochemical impedance immunosensor is for the first time developed for highly sensitive determination of BoIFN-?. Cylinder-shaped TiO2 nanorods are synthesized by a facile hydrothermal method, and show high surface area and good hydrophicility. The immunosensor is fabricated by the immobilization of BoIFN-? monoclonal antibody on the TiO2 nanorods modified glassy carbon electrode. The prepared TiO2 and immunosensor are characterized using transmission electron microscopy, scanning electron microscopy, X-ray diffraction, contact angle measurement, cyclic voltammetry, and electrochemical impedance spectra. The BoIFN-? concentration is measured through the relative increase of impedance values in corresponding specific binding of BoIFN-? antigen and BoIFN-? antibody. The relative increased impedance values are proportional to the logarithmic value of BoIFN-? concentrations in a wide range of 0.0001 to 0.1 ng/mL with a low detection limit of 0.1 pg/mL. The developed BoIFN-? immunosensor shows a 249-fold decrease in detection limit in comparison with current enzyme-linked immunosorbent assay. This study provides a new, simple, and highly sensitive approach for very potential application in early diagnosis of MTB infections and control of bovine tuberculosis.
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Synthesis of N-alkyl isatins via oxidative cyclization of N-alkyl 2-bromo(chloro)acetanilides.
Org. Biomol. Chem.
PUBLISHED: 04-17-2014
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A highly efficient method for the synthesis of N-alkyl isatins starting from N-alkyl 2-bromo or 2-chloro acetanilides is described. The starting materials are easy to prepare and the yields of isatins are generally high. Operationally the reaction is very simple to run. Even though best results were obtained with a catalytic amount of CuI, the reactions of N-alkyl 2-bromo acetanilides actually performed well even in the absence of any metal catalyst.
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Self-powered ultraviolet photodetectors based on selectively grown ZnO nanowire arrays with thermal tuning performance.
Phys Chem Chem Phys
PUBLISHED: 04-15-2014
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A self-powered Schottky-type ultraviolet photodetector with Al-Pt interdigitated electrodes has been fabricated based on selectively grown ZnO nanowire arrays. At zero bias, the fabricated photodetector exhibited high sensitivity and excellent selectivity to UV light illumination with a fast response time of 81 ms. By tuning the Schottky barrier height through the thermally induced variation of the interface chemisorbed oxygen, an ultrahigh sensitivity of 3.1 × 10(4) was achieved at 340 K without an external power source, which was 82% higher than that obtained at room temperature. According to the thermionic emission-diffusion theory and the solar cell theory, the changes in the photocurrent of the photodetector at zero bias with various system temperatures were calculated, which agreed well with the experimental data. This work demonstrates a promising approach to modulating the performance of a self-powered photodetector by heating and provides theoretical support for studying the thermal effect on the future photoelectric device.
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High Expression of SOX2 and OCT4 Indicates Radiation Resistance and an Independent Negative Prognosis in Cervical Squamous Cell Carcinoma.
J. Histochem. Cytochem.
PUBLISHED: 04-07-2014
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Radiotherapy (RT) as a preoperative or postoperative adjuvant or primary treatment is the most common management modality for locally advanced cervical cancer. Radioresistance of tumor cells remains a major therapeutic problem. Consequently, we aimed to explore if the stem cell biomarkers SOX2 and OCT4 protein could be used to predict radioresistance in patients with locally advanced cervical squamous cell carcinoma (LACSCC). These 132 patients were divided into two groups (radiation-resistant and radiation-sensitive groups) according to progress-free survival (PFS). Using pretreatment paraffin-embedded tissues, we evaluated SOX2 and OCT4 expression using immunohistochemical staining. The percentage of overexpression of SOX2 and OCT4 in the radiation-resistant group was much higher than that in the radiation-sensitive group (p<0.001 and p <0.001, respectively). The patients with high expression of SOX2 and OCT4 showed a shorter PFS than those with low expression. Our study suggests that the expression of SOX2 and OCT4 in tumor cells indicates resistance to radiotherapy and that these two factors were important predictors of poor survival in patients with LACSCC (hazard ratio [95% CI], 2.294 [1.013, 5.195] and 2.300 [1.050, 5.037], respectively; p=0.046 and p=0.037, respectively).
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The landscape of somatic mutations in epigenetic regulators across 1,000 paediatric cancer genomes.
Nat Commun
PUBLISHED: 03-12-2014
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Studies of paediatric cancers have shown a high frequency of mutation across epigenetic regulators. Here we sequence 633 genes, encoding the majority of known epigenetic regulatory proteins, in over 1,000 paediatric tumours to define the landscape of somatic mutations in epigenetic regulators in paediatric cancer. Our results demonstrate a marked variation in the frequency of gene mutations across 21 different paediatric cancer subtypes, with the highest frequency of mutations detected in high-grade gliomas, T-lineage acute lymphoblastic leukaemia and medulloblastoma, and a paucity of mutations in low-grade glioma and retinoblastoma. The most frequently mutated genes are H3F3A, PHF6, ATRX, KDM6A, SMARCA4, ASXL2, CREBBP, EZH2, MLL2, USP7, ASXL1, NSD2, SETD2, SMC1A and ZMYM3. We identify novel loss-of-function mutations in the ubiquitin-specific processing protease 7 (USP7) in paediatric leukaemia, which result in decreased deubiquitination activity. Collectively, our results help to define the landscape of mutations in epigenetic regulatory genes in paediatric cancer and yield a valuable new database for investigating the role of epigenetic dysregulations in cancer.
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3D porous chitosan scaffolds suit survival and neural differentiation of dental pulp stem cells.
Cell. Mol. Neurobiol.
PUBLISHED: 02-26-2014
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A key aspect of cell replacement therapy in brain injury treatment is construction of a suitable biomaterial scaffold that can effectively carry and transport the therapeutic cells to the target area. In the present study, we created small 3D porous chitosan scaffolds through freeze-drying, and showed that these can support and enhance the differentiation of dental pulp stem cells (DPSCs) to nerve cells in vitro. The DPSCs were collected from the dental pulp of adult human third molars. At a swelling rate of ~84.33 ± 10.92 %, the scaffold displayed high porosity and interconnectivity of pores, as revealed by SEM. Cell counting kit-8 assay established the biocompatibility of the chitosan scaffold, supporting the growth and survival of DPSCs. The successful neural differentiation of DPSCs was assayed by RT-PCR, western blotting, and immunofluorescence. We found that the scaffold-attached DPSCs showed high expression of Nestin that decreased sharply following induction of differentiation. Exposure to the differentiation media also increased the expression of neural molecular markers Microtubule-associated protein 2, glial fibrillary acidic protein, and 2',3'-cyclic nucleotide phosphodiesterase. This study demonstrates that the granular 3D chitosan scaffolds are non-cytotoxic, biocompatible, and provide a conducive and favorable micro-environment for attachment, survival, and neural differentiation of DPSCs. These scaffolds have enormous potential to facilitate future advances in treatment of brain injury.
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Cyclin D2 plays a regulatory role in HBV replication.
Virology
PUBLISHED: 02-25-2014
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Hepatitis B virus (HBV) infection is the leading cause of liver diseases. However, the molecular mechanisms of HBV infection and carcinogenesis have not been fully elucidated. In this study, we found that cyclin D2 was upregualted in HBV-expressing cells and liver tissues of HBV-transgenic mice. Gene silencing of cyclin D2 inhibited HBV DNA replicative intermediates, 3.5 kb mRNA, core protein level, as well as the secretions of HBsAg and HBeAg. On the contrary, overexpression of cyclin D2 promoted HBV replication. Furthermore, cyclin D2 regulated HBV replication by enhancing the activity of HBV core and Sp1 promoters by targeting transcription factor CREB2. Silencing of CREB2 abolished enhancement of HBV replication induced by cyclin D2. Together, our study has uncovered a positive role of cyclin D2 in HBV replication. It is conceivable that therapeutic application of cyclin D2 inhibitor in HBV infection therapy.
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LIM mineralization protein-1 inhibits the malignant phenotypes of human osteosarcoma cells.
Int J Mol Sci
PUBLISHED: 01-28-2014
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Osteosarcoma (OS), also known as osteogenic sarcoma, is the most common primary malignancy of bone tumor in children and adolescents. However, its underlying molecular pathogenesis is still only vaguely understood. Recently, LIM mineralization protein-1 (LMP-1) was reported to be an essential positive regulator of osteoblast differentiation. In the present study, we found that the expression of LMP-1 is downregulated in OS tissues compared with adjacent normal tissues. Moreover, we restored the expression of LMP-1 through a recombinant adenovirus. Overexpression of LMP-1 inhibited cell proliferation and invasion, arrested cell cycle progression, and induced apoptosis in vitro. Finally, ectopic LMP-1 expression suppressed the expression of Runx2 and BMP-2 in OS cells. These data demonstrate that LMP-1 is an essential tumor suppressor in the OS pathological process, which will provide a new opportunity for discovering and identifying novel effective treatment strategies.
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Label-free detection of adenosine based on fluorescence resonance energy transfer between fluorescent silica nanoparticles and unmodified gold nanoparticles.
Anal. Chim. Acta
PUBLISHED: 01-17-2014
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A sensitive and convenient strategy was developed for label-free assay of adenosine. The strategy adapted the fluorescence resonance energy transfer property between Rhodamine B doped fluorescent silica nanoparticles (SiNPs) and gold nanoparticles (AuNPs) to generate signal. The different affinities of AuNPs toward the unfolded and folded aptamers were employed for the signal transfer in the system. In the presence of adenosine, the split aptamer fragments react with adenosine to form a structured complex. The folded aptamer cannot be adsorbed on the surface of AuNPs, which induces the aggregation of AuNPs under high ionic concentration conditions, and the aggregation of AuNPs leads to the decrease of the quenching ability. Therefore, the fluorescence intensity of Rhodamine B doped fluorescent SiNPs increased along with the concentration of adenosine. Because of the highly specific recognition ability of the aptamer toward adenosine and the strong quenching ability of AuNPs, the proposed strategy demonstrated good selectivity and high sensitivity for the detection of adenosine. Under the optimum conditions in the experiments, a linear range from 98nM to 100?M was obtained with a detection limit of 45nM. As this strategy is convenient, practical and sensitive, it will provide a promising potential for label-free aptamer-based protein detection.
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D-Aminoacylase-initiated cascade Aldol condensation/Robinson annulation for synthesis of substituted cyclohex-2-enones from simple aldehydes and acetone.
Amino Acids
PUBLISHED: 01-16-2014
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As an important building block, developing efficient and green synthesis strategy of cyclohex-2-enones is of great importance. In this present work, a general approach to the mild synthesis of substituted cyclohex-2-enones derivatives starting fro m simple aldehydes and acetone have been achieved via D-aminoacylase-initiated Aldol condensation/Robinson annulation cascade reaction using imidazole as an additive in organic media. The influences of reaction conditions including solvents, enzyme concentration, additives type, molar ratio of enzyme to additive, and substrate scopes were systematically investigated. Furthermore, some experiments were designed to explore the catalytic roles of D-aminoacylase and imidazole in the multistep cascade process, and one possible mechanism was proposed.
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PSD-93 deletion inhibits Fyn-mediated phosphorylation of NR2B and protects against focal cerebral ischemia.
Neurobiol. Dis.
PUBLISHED: 01-12-2014
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Modification of N-methyl-d-aspartate receptor (NMDAR)-mediated excitotoxicity appears to be a potential target in the treatment of ischemic stroke. Postsynaptic density protein-93 (PSD-93) specifically binds the C-terminal tails of the NMDAR, which is critical to couple NMDAR activity to specific intracellular signaling. This study is to investigate whether PSD-93 disruption displays neuroprotection in a focal ischemic stroke model of adult mice and, if it does, to explore possible mechanisms. It was found that, following middle cerebral artery occlusion (MCAO), PSD-93 knockout (KO) mice manifested significant reductions in infarcted volume, neurological deficits and number of degenerated neurons. PSD-93 deletion also reduced cultured cortical neuronal death caused by glucose and oxygen deprivation (OGD). Ischemic long term potentiation (i-LTP) could not be induced in the PSD-93 KO group and wild type (WT) groups pretreated with either AP-5 (NMDAR inhibitor) or PP2 (Src family inhibitor). PSD-93 KO decreased the phosphorylation of the NR2B at Tyr1472 and the interaction between NR2B and Fyn after MCAO. Together, our study demonstrated that PSD-93 KO confers profound neuroprotection against ischemic brain injury, which probably links to the inhibitory effect on Fyn-mediated phosphorylation of NR2B caused by PSD-93 deletion. These findings may provide a novel avenue for the treatment of ischemic stroke.
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LGR5 is required for the maintenance of spheroid-derived colon cancer stem cells.
Int. J. Mol. Med.
PUBLISHED: 01-10-2014
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Colon cancer stem cells (CCSCs) are involved in colon cancer and promote tumor progression and recurrence. LGR5, a marker for intestinal stem cells (ISCs), is also considered to serve as a marker for CCSCs. However, the precise function of LGR5 in CCSCs is unclear. In this study, we demonstrated that LGR5 was highly expressed in CCSCs-enriched HT29 spheroid cells. Downregulation of LGR5 with small interfering RNA (siRNA) decreased the expression of stem the cell markers CD133 and CD44 in HT29 spheroid cells. In addition, silencing of LGR5 inhibited cell proliferation, secondary tumor sphere formation and induced cell apoptosis, and G0/G1 phase arrest in vitro by modulating Bcl-2, Bcl-xL and Bax. Knockdown of LGR5 enhanced chemosensitivity and reduced the invasive ability of HT29 spheroid cells. Moreover, LGR5-siRNA suppressed tumorigenicity of HT29 spheroid cells in vivo. The findings suggested that LGR5 plays a vital role in the maintenance of CCSCs and is a potential therapeutic target for colon cancer.
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Generation and application of a 293 cell line stably expressing bovine interferon-gamma.
Protein Expr. Purif.
PUBLISHED: 01-09-2014
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A stable mammalian cell line expressing highly active bovine interferon-gamma (BoIFN-?) was generated using Flp recombinase-mediated integration. This recombinant 293 cell line (B1) efficiently secreted FLAG-tagged BoIFN-? protein into the culture supernatant, as determined by ELISA and Western blot. The recombinant BoIFN-? exhibited high anti-viral activity, suggesting that the 293 cells expressed BoIFN-? that structurally and biologically resembled the natural protein. Two monoclonal antibodies (mAbs) with high affinity for the 293 cell-expressed BoIFN-? were identified using this cell line, and these mAbs can be used for the development of diagnostic kits. Thus, this work demonstrates the successful generation of a 293 cell line that produces large quantities of highly active BoIFN-? and demonstrates its potential application in the research of bovine infectious diseases.
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Comparison of high-solids to liquid anaerobic co-digestion of food waste and green waste.
Bioresour. Technol.
PUBLISHED: 01-09-2014
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Co-digestion of food waste and green waste was conducted with six feedstock mixing ratios to evaluate biogas production. Increasing the food waste percentage in the feedstock resulted in an increased methane yield, while shorter retention time was achieved by increasing the green waste percentage. Food waste/green waste ratio of 40:60 was determined as preferred ratio for optimal biogas production. About 90% of methane yield was obtained after 24.5 days of digestion, with total methane yield of 272.1 mL/g VS. Based the preferred ratio, effect of total solids (TS) content on co-digestion of food waste and green waste was evaluated over a TS range of 5-25%. Results showed that methane yields from high-solids anaerobic digestion (15-20% TS) were higher than the output of liquid anaerobic digestion (5-10% TS), while methanogenesis was inhibited by further increasing the TS content to 25%. The inhibition may be caused by organic overloading and excess ammonia.
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miR-199a-5p regulates the expression of metastasis-associated genes in B16F10 melanoma cells.
Int J Clin Exp Pathol
PUBLISHED: 01-01-2014
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MicroRNAs are regulatory factors that play important roles in tumor development, invasion and metastasis. Previously, we showed that miR-199a is abnormally expressed in clinical melanoma specimens and expression was closely associated with clinical features of metastasis. However, the exact molecular mechanisms by which miR-199a-5p influences melanoma invasion and metastasis remains unclear. In this study, we investigated gene expression changes of metastasis-associated genes in B16F10 melanoma cells following targeted silencing or overexpression of miR-199a-5p, using mouse tumor metastasis PCR arrays. Comparison of gene expression changes in miR-199a-5p-silenced versus overexpressing cells identified a set of upregulated genes (> 2-fold) including Cd44, Cdh1, Cxcr4, Etv4, Fxyd5, Rpsa, Mmp3, Myc, Rb1, Tcf20, Hprt1, Actb1 and downregulated genes (> 2-fold) including Ctsk, Itga7 and Tnfsf10. Regulation of a subset of these genes (Myc, Tnfsf10 and Cd44) following miR-199a-5p silencing or overexpression was validated by reverse transcription-polymerase chain reaction (RT-PCR) and western blot. In conclusion, our study demonstrates that miR-199a-5p regulates melanoma metastasis-related genes, and may provide a basis for the development of novel, molecularly targeted drugs.
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A Simplified and Versatile System for the Simultaneous Expression of Multiple siRNAs in Mammalian Cells Using Gibson DNA Assembly.
PLoS ONE
PUBLISHED: 01-01-2014
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RNA interference (RNAi) denotes sequence-specific mRNA degradation induced by short interfering double-stranded RNA (siRNA) and has become a revolutionary tool for functional annotation of mammalian genes, as well as for development of novel therapeutics. The practical applications of RNAi are usually achieved by expressing short hairpin RNAs (shRNAs) or siRNAs in cells. However, a major technical challenge is to simultaneously express multiple siRNAs to silence one or more genes. We previously developed pSOS system, in which siRNA duplexes are made from oligo templates driven by opposing U6 and H1 promoters. While effective, it is not equipped to express multiple siRNAs in a single vector. Gibson DNA Assembly (GDA) is an in vitro recombination system that has the capacity to assemble multiple overlapping DNA molecules in a single isothermal step. Here, we developed a GDA-based pSOK assembly system for constructing single vectors that express multiple siRNA sites. The assembly fragments were generated by PCR amplifications from the U6-H1 template vector pB2B. GDA assembly specificity was conferred by the overlapping unique siRNA sequences of insert fragments. To prove the technical feasibility, we constructed pSOK vectors that contain four siRNA sites and three siRNA sites targeting human and mouse ?-catenin, respectively. The assembly reactions were efficient, and candidate clones were readily identified by PCR screening. Multiple ?-catenin siRNAs effectively silenced endogenous ?-catenin expression, inhibited Wnt3A-induced ?-catenin/Tcf4 reporter activity and expression of Wnt/?-catenin downstream genes. Silencing ?-catenin in mesenchymal stem cells inhibited Wnt3A-induced early osteogenic differentiation and significantly diminished synergistic osteogenic activity between BMP9 and Wnt3A in vitro and in vivo. These findings demonstrate that the GDA-based pSOK system has been proven simplistic, effective and versatile for simultaneous expression of multiple siRNAs. Thus, the reported pSOK system should be a valuable tool for gene function studies and development of novel therapeutics.
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Transcatheter pulmonary valve replacement by hybrid approach using a novel polymeric prosthetic heart valve: proof of concept in sheep.
PLoS ONE
PUBLISHED: 01-01-2014
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Since 2000, transcatheter pulmonary valve replacement has steadily advanced. However, the available prosthetic valves are restricted to bioprosthesis which have defects like poor durability. Polymeric heart valve is thought as a promising alternative to bioprosthesis. In this study, we introduced a novel polymeric transcatheter pulmonary valve and evaluated its feasibility and safety in sheep by a hybrid approach.
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The piggyBac transposon-mediated expression of SV40 T antigen efficiently immortalizes mouse embryonic fibroblasts (MEFs).
PLoS ONE
PUBLISHED: 01-01-2014
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Mouse embryonic fibroblasts (MEFs) are mesenchymal stem cell (MSC)-like multipotent progenitor cells and can undergo self-renewal and differentiate into to multiple lineages, including bone, cartilage and adipose. Primary MEFs have limited life span in culture, which thus hampers MEFs' basic research and translational applications. To overcome this challenge, we investigate if piggyBac transposon-mediated expression of SV40 T antigen can effectively immortalize mouse MEFs and that the immortalized MEFs can maintain long-term cell proliferation without compromising their multipotency. Using the piggyBac vector MPH86 which expresses SV40 T antigen flanked with flippase (FLP) recognition target (FRT) sites, we demonstrate that mouse embryonic fibroblasts (MEFs) can be efficiently immortalized. The immortalized MEFs (piMEFs) exhibit an enhanced proliferative activity and maintain long-term cell proliferation, which can be reversed by FLP recombinase. The piMEFs express most MEF markers and retain multipotency as they can differentiate into osteogenic, chondrogenic and adipogenic lineages upon BMP9 stimulation in vitro. Stem cell implantation studies indicate that piMEFs can form bone, cartilage and adipose tissues upon BMP9 stimulation, whereas FLP-mediated removal of SV40 T antigen diminishes the ability of piMEFs to differentiate into these lineages, possibly due to the reduced expansion of progenitor populations. Our results demonstrate that piggyBac transposon-mediated expression of SV40 T can effectively immortalize MEFs and that the reversibly immortalized piMEFs not only maintain long-term cell proliferation but also retain their multipotency. Thus, the high transposition efficiency and the potential footprint-free natures may render piggyBac transposition an effective and safe strategy to immortalize progenitor cells isolated from limited tissue supplies, which is essential for basic and translational studies.
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Co-expression of CD147 and GLUT-1 indicates radiation resistance and poor prognosis in cervical squamous cell carcinoma.
Int J Clin Exp Pathol
PUBLISHED: 01-01-2014
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The aim of this study was to investigate the association of CD147 and GLUT-1, which play important roles in glycolysis in response to radiotherapy and clinical outcomes in patients with locally advanced cervical squamous cell carcinoma (LACSCC). The records of 132 female patients who received primary radiation therapy to treat LACSCC at FIGO stages IB-IVA were retrospectively reviewed. Forty-seven patients with PFS (progression-free survival) of less than 36 months were regarded as radiation-resistant. Eighty-five patients with PFS longer than 36 months were regarded as radiation-sensitive. Using pretreatment paraffin-embedded tissues, we evaluated CD147 and GLUT-1 expression by immunohistochemistry. Overexpression of CD147, GLUT-1, and CD147 and GLUT-1 combined were 44.7%, 52.9% and 36.5%, respectively, in the radiation-sensitive group, and 91.5%, 89.4% and 83.0%, respectively, in the radiation-resistant group. The 5-year progress free survival (PFS) rates in the CD147-low, CD147-high, GLUT-1-low, GLUT-1-high, CD147- and/or GLUT-1-low and CD147- and GLUT-1- dual high expression groups were 66.79%, 87.10%, 52.78%, 85.82%, 55.94%, 82.90% and 50.82%, respectively. CD147 and GLUT-1 co-expression, FIGO stage and tumor diameter were independent poor prognostic factors for patients with LACSCC in multivariate Cox regression analysis. Patients with high expression of CD147 alone, GLUT-1 alone or co-expression of CD147 and GLUT-1 showed greater resistance to radiotherapy and a shorter PFS than those with low expression. In particular, co-expression of CD147 and GLUT-1 can be considered as a negative independent prognostic factor.
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Gene Therapy with Beta-Defensin 2 Induces Antitumor Immunity and Enhances Local Antitumor Effects.
Hum. Gene Ther.
PUBLISHED: 12-19-2013
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Abstract Beta-defensins, small antimicrobial peptides, are involved in host immune responses to tumors. In this study, we used beta-defensin 2 (BD2) to explore the possible role of beta-defensins in cancer gene therapy. A recombinant plasmid expressing a secretable form of BD2 was constructed. The biological activities of BD2 in immature dendritic cells (iDCs) were tested in vitro and in vivo. The antitumor effects were investigated in three established tumor models. The secreted BD2 was detected and exhibited chemotactic activity in iDCs both in vitro and in vivo. Recruitment and activation of iDCs in tumor niches resulted in significant tumor growth inhibition. Adoptive transfer of splenocytes and depletion of immune cell subsets revealed that CD8(+) T lymphocyte responses mediated the increased tumor inhibition. Furthermore, we also found that chemotactic and maturation-inducing activities in iDCs in tumor milieu contributed to enhanced local antitumor effects. Our study indicates that gene therapy with BD2 can mediate specific antitumor immunity and augment local antitumor effects. Our study also suggested that beta-defensins may merit further exploration for cancer immunotherapy as promising immunogenes.
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Tivozanib reverses multidrug resistance mediated by ABCB1 (P-glycoprotein) and ABCG2 (BCRP).
Future Oncol
PUBLISHED: 12-03-2013
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 Aim: This study aimed to investigate the mechanism of reversal of multidrug resistance mediated by ABC transporters with tivozanib (AV-951 and KRN-951). Tivozanib is a potent inhibitor of VEGF-1, -2 and -3 receptors. Materials & methods: ABCB1- and ABCG2-overexpressing cell lines were treated with respective substrate antineoplastic agents in the presence or absence of tivozanib. Results: The results indicate that tivozanib can significantly reverse ABCB1-mediated resistance to paclitaxel, vinblastine and colchicine, as well as ABCG2-mediated resistance to mitoxantrone, SN-38 and doxorubicin. Drug efflux assays showed that tivozanib increased the intracellular accumulation of substrates by inhibiting the ABCB1 and ABCG2 efflux activity. Furthermore, at a higher concentration, tivozanib inhibited the ATPase activity of both ABCB1 and ABCG2 and inhibited the photolabeling of ABCB1 or ABCG2. Conclusion: We conclude that tivozanib at noncytotoxic concentrations has the previously unknown activity of reversing multidrug resistance mediated by ABCB1 and ABCG2 transporters.
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Using proteomics to identify the HBx interactome in hepatitis B virus: how can this inform the clinic?
Expert Rev Proteomics
PUBLISHED: 11-29-2013
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Hepatitis B virus (HBV) is a small and enveloped DNA virus, of which chronic infection is the main risk factor of liver cirrhosis and hepatocellular carcinoma. Hepatitis B virus X protein (HBx) is a multifunctional protein encoded by HBV genome, which have significant effects on HBV replication and pathogenesis. Through directly interacting with cellular proteins, HBx is capable to promote HBV replication, regulate transcription of host genes, disrupt protein degradation, modulate signaling pathway, manipulate cell death and deregulate cell cycle. In this review, we briefly discuss the diversified effects of HBx-interactome and their potential clinical significances.
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Prostaglandins temporally regulate cytoplasmic actin bundle formation during Drosophila oogenesis.
Mol. Biol. Cell
PUBLISHED: 11-27-2013
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While Prostaglandins (PGs), lipid signals produced downstream of cyclooxygenase (COX) enzymes, regulate actin dynamics in cell culture and platelets, their roles during development are largely unknown. Here we define a new role for Pxt, the Drosophila COX-like enzyme, in regulating the actin cytoskeleton-temporal restriction of actin remodeling during oogenesis. PGs are required for actin filament bundle formation during stage 10B (S10B). Additionally, loss of Pxt results in early actin remodeling, including extensive actin filaments and aggregates, within the posterior nurse cells of stage 9 (S9) follicles; wild-type follicles exhibit similar structures at a low frequency. Hu li tai shao (Hts), the homolog of Adducin, and Villin (Quail), an actin bundler, localize to all early actin structures, while Enabled (Ena), an actin elongation factor, preferentially localizes to those in pxt mutants. Reduced Ena levels strongly suppress early actin remodeling in pxt mutants. Furthermore, loss of Pxt results in reduced Ena localization to the sites of bundle formation during S10B. Together these data lead to the model that PGs temporally regulate actin remodeling during Drosophila oogenesis by controlling Ena localization/activity, such that in S9, PG signaling inhibits, while at S10B, it promotes Ena-dependent actin remodeling.
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Multiscale entropy analysis of different spontaneous motor unit discharge patterns.
IEEE J Biomed Health Inform
PUBLISHED: 11-16-2013
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This study explores a novel application of multiscale entropy (MSE) analysis for characterizing different patterns of spontaneous electromyogram (EMG) signals including sporadic, tonic and repetitive spontaneous motor unit discharges, and normal surface EMG baseline. Two algorithms for MSE analysis, namely, the standard MSE and the intrinsic mode entropy (IMEn) (based on the recently developed multivariate empirical mode decomposition method), were applied to different patterns of spontaneous EMG. Significant differences were observed in multiple scales of the standard MSE and IMEn analyses (<;i>p<;/i> <; 0.001) for any two of the spontaneous EMG patterns, while such significance may not be observed from the single-scale entropy analysis. Compared to the standard MSE, the IMEn analysis facilitates usage of a relatively low scale number to discern entropy difference among various patterns of spontaneous EMG signals. The findings from this study contribute to our understanding of the nonlinear dynamic properties of different spontaneous EMG patterns, which may be related to spinal motoneuron or motor unit health.
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[Resection of parapharyngeal neoplasms via styloid diaphragm approach].
Zhonghua Er Bi Yan Hou Tou Jing Wai Ke Za Zhi
PUBLISHED: 11-08-2013
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To evaluate the surgical technique and efficacy of the resection of parapharyngeal space neoplasm via styloid diaphragm approach.
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Graphene oxide doped conducting polymer nanocomposite film for electrode-tissue interface.
Biomaterials
PUBLISHED: 10-29-2013
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One of the most significant components for implantable bioelectronic devices is the interface between the microelectrodes and the tissue or cells for disease diagnosis or treatment. To make the devices work efficiently and safely in vivo, the electrode-tissue interface should not only be confined in micro scale, but also possesses excellent electrochemical characteristic, stability and biocompatibility. Considering the enhancement of many composite materials by combining graphene oxide (GO) for its multiple advantages, we dope graphene oxide into poly(3,4-ethylenedioxythiophene) (PEDOT) forming a composite film by electrochemical deposition for electrode site modification. As a consequence, not only the enlargement of efficient surface area, but also the development of impedance, charge storage capacity and charge injection limit contribute to the excellent electrochemical performance. Furthermore, the stability and biocompatibility are confirmed by numerously repeated usage test and cell proliferation and attachment examination, respectively. As electrode-tissue interface, this biomaterial opens a new gate for tissue engineering and implantable electrophysiological devices.
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Laparoendoscopic single-site retroperitoneoscopic adrenalectomy for pheochromocytoma: case selection, surgical technique, and short-term outcome.
J. Endourol.
PUBLISHED: 10-23-2013
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Abstract Purpose: To present our experience with case selection and operative skills of laparoendoscopic single-site (LESS) retroperitoneoscopic adrenalectomy for pheochromocytoma and evaluate its feasibility. Patients and Methods: Between June 2011 and December 2012, we performed LESS retroperitoneoscopic adrenalectomy for 16 patients with pheochromocytoma. In all patients, the diameter of the pheochromocytoma was less than 4.0?cm. During the operation, a single-port access was inserted through a 2.5-3.0?cm transverse incision below the tip of the 12th rib. Internally, the operative procedure duplicates the conventional retroperitoneoscopic adrenalectomy for pheochromocytoma. Results: No conversions to open surgery or standard laparoscopy with additional trocars were necessary. The mean operative duration was 68.1 minutes (range 41-125?min). The mean blood loss was negligible (<50?mL), and no patient needed blood transfusion. Intraoperative hypertension (SBP>180?mmHg) occurred in 12.5% (2/16) of the patients. No patient had sustained hypertension, and none experienced intraoperative hypotension (systolic blood pressure <80?mm Hg). The only postoperative complication was one case of pneumonia successfully treated with antibiotics. The average postoperative hospital stay was 3.1 days (range 2-5 days). All patients left the hospital with a good cosmetic appearance. Conclusions: In properly selected patients, LESS retroperitoneoscopic adrenalectomy is a feasible and safe procedure for pheochromocytoma.
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DNAJB1 stabilizes MDM2 and contributes to cancer cell proliferation in a p53-dependent manner.
Biochim. Biophys. Acta
PUBLISHED: 10-20-2013
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Both MDM2 and MDMX regulate p53, but these proteins play different roles in this process. To clarify the difference, we performed a yeast 2 hybrid (Y2H) screen using the MDM2 acidic domain as bait. DNAJB1 was found to specifically bind to MDM2, but not MDMX, in vitro and in vivo. Further investigation revealed that DNAJB1 stabilizes MDM2 at the post-translational level. The C-terminus of DNAJB1 is essential for its interaction with MDM2 and for MDM2 accumulation. MDM2 was degraded faster by a ubiquitin-mediated pathway when DNAJB1 was depleted. DNAJB1 inhibited the MDM2-mediated ubiquitination and degradation of p53 and contributed to p53 activation in cancer cells. Depletion of DNAJB1 in cancer cells inhibited activity of the p53 pathway, enhanced the activity of the Rb/E2F pathway, and promoted cancer cell growth in vitro and in vivo. This function was p53 dependent, and either human papillomavirus (HPV) E6 protein or siRNA against p53 was able to block the contribution caused by DNAJB1 depletion. In this study, we discovered a new MDM2 interacting protein, DNAJB1, and provided evidence to support its p53-dependent tumor suppressor function.
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Virulence determinants of Salmonella Gallinarum biovar Pullorum identified by PCR signature-tagged mutagenesis and the spiC mutant as a candidate live attenuated vaccine.
Vet. Microbiol.
PUBLISHED: 10-03-2013
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Salmonella Gallinarum biovar Pullorum (S. Gallinarum biovar Pullorum) is the causative agent of pullorum disease (PD) in chickens which results in considerable economic losses to the poultry industries in developing countries. PCR-Signature Tagged Mutagenesis was used to identify virulence determinants of S. Gallinarum biovar Pullorum and novel attenuated live vaccine candidates for use against this disease. A library of 1800 signature-tagged S. Gallinarum biovar Pullorum mutants was constructed and screened for virulence-associated genes in chickens. The attenuation of 10 mutants was confirmed by in vivo and in vitro competitive index (CI) studies. The transposons were found to be located in SPI-1 (2/10 mutants), SPI-2 (3/10), the virulence plasmid (1/10) and non-SPI genes (4/10). One highly attenuated spiC mutant persisted in spleen and liver for less than 10 days and induced high levels of circulating antibody and protective immunity against oral challenge in young broiler chickens. The spiC mutant is a potential new vaccine candidate for use with chickens against this disease.
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[Protective mechanism of cerebrospinal fluid containing qingxin kaiqiao recipe on PC12 cell injury induced by glutamate].
Zhongguo Zhong Yao Za Zhi
PUBLISHED: 09-27-2013
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To study the protective effect of cerebrospinal fluid containing Qingxin Kaiqiao recipe on PC12 cell injury induced by glutamate (Glu), in order to provide basis for the conical application of the recipe.
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[Effects of qingxin kaiqiao recipe volatile oil on expressions of GFAP and caspase-3 in the cortex and hippocampus of AD rats].
Zhongguo Zhong Xi Yi Jie He Za Zhi
PUBLISHED: 09-26-2013
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To study effects of Qingxin Kaiqiao Recipe (QKR) and its volatile oil on the expressions of Abeta(25-35) glial fibrillary acidic protein (GFAP), beta-amyloid (Abeta), beta-amyloid precursor protein (betaAPP), and Caspase-3 in the cortex and hippocampus of Alzheimers disease (AD) rats induced by injecting Abeta(25-35) into the bilateral amygdala.
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[Study on protective effect of cerebrospinal fluid containing Qingxin Kaiqiao Fang on sodium dithionite-induced PC12 cell injury].
Zhongguo Zhong Yao Za Zhi
PUBLISHED: 08-16-2013
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To study the protective effect of cerebrospinal fluid containing Qingxin Kaiqiao Fang on sodium dithionite (Na2S2O4)-induced PC12 cell injury, in order to provide basis for clinical application of the prescription.
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Adverse events during 2 years of daily wear of silicone hydrogels in children.
Optom Vis Sci
PUBLISHED: 08-14-2013
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Type and incidence of adverse events and rate of discontinuations for 2 years of daily wear with silicone hydrogel contact lenses in Chinese children with myopia.
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An NAC transcription factor controls ethylene-regulated cell expansion in flower petals.
Plant Physiol.
PUBLISHED: 08-09-2013
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Cell expansion is crucial for plant growth. It is well known that the phytohormone ethylene functions in plant development as a key modulator of cell expansion. However, the role of ethylene in the regulation of this process remains unclear. In this study, 2,189 ethylene-responsive transcripts were identified in rose (Rosa hybrida) petals using transcriptome sequencing and microarray analysis. Among these transcripts, an NAC (for no apical meristem [NAM], Arabidopsis transcription activation factor [ATAF], and cup-shaped cotyledon [CUC])-domain transcription factor gene, RhNAC100, was rapidly and dramatically induced by ethylene in the petals. Interestingly, accumulation of the RhNAC100 transcript was modulated by ethylene via microRNA164-dependent posttranscriptional regulation. Overexpression of RhNAC100 in Arabidopsis (Arabidopsis thaliana) substantially reduced the petal size by repressing petal cell expansion. By contrast, silencing of RhNAC100 in rose petals using virus-induced gene silencing significantly increased petal size and promoted cell expansion in the petal abaxial subepidermis (P < 0.05). Expression analysis showed that 22 out of the 29 cell expansion-related genes tested exhibited changes in expression in RhNAC100-silenced rose petals. Moreover, of those genes, one cellulose synthase and two aquaporin genes (Rosa hybrida Cellulose Synthase2 and R. hybrida Plasma Membrane Intrinsic Protein1;1/2;1) were identified as targets of RhNAC100. Our results suggest that ethylene regulates cell expansion by fine-tuning the microRNA164/RhNAC100 module and also provide new insights into the function of NAC transcription factors.
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Integrating UIMA annotators in a web-based text processing framework.
Stud Health Technol Inform
PUBLISHED: 08-08-2013
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The Unstructured Information Management Architecture (UIMA) [1] framework is a growing platform for natural language processing (NLP) applications. However, such applications may be difficult for non-technical users deploy. This project presents a web-based framework that wraps UIMA-based annotator systems into a graphical user interface for researchers and clinicians, and a web service for developers. An annotator that extracts data elements from lung cancer radiology reports is presented to illustrate the use of the system. Annotation results from the web system can be exported to multiple formats for users to utilize in other aspects of their research and workflow. This project demonstrates the benefits of a lay-user interface for complex NLP applications. Efforts such as this can lead to increased interest and support for NLP work in the clinical domain.
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What is Visualize?

JoVE Visualize is a tool created to match the last 5 years of PubMed publications to methods in JoVE's video library.

How does it work?

We use abstracts found on PubMed and match them to JoVE videos to create a list of 10 to 30 related methods videos.

Video X seems to be unrelated to Abstract Y...

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.