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Find video protocols related to scientific articles indexed in Pubmed.
Magnetically recyclable polymer single crystal supported silver nanocatalyst.
Langmuir
PUBLISHED: 10-31-2014
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We report the immobilization of unprotected silver nanoparticle on the carboxylic acid abundant polymer single crystal surface with controllable size through photogenerated chemical reduction reaction. The resulting silver nanoparticle decorated polymer single crystal not only shows higher catalytic activity as compared to its counterpart bearing surface ligand but also exhibits size-dependent catalytic activity with the smallest size (?1.5 nm) being the most active. By further introducing iron oxide nanoparticles onto its surface, the resulting catalyst system can be magnetically recycled for up to five times with little loss in catalytic activity. These, together with the high loading originated from the high surface area to volume ratio for a polymer single crystal, make current catalyst system attractive for many industrial important catalytic applications.
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[Effect of pretreatment with qishen yiqi dropping pills on right cardiac function of patients undergoing valve replacement].
Zhongguo Zhong Yao Za Zhi
PUBLISHED: 09-11-2014
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In this study, 120 patients with rheumatic heart disease undergoing valve replacement were randomly divided into the control group and the Qishen group, with 60 cases in each group. Before the operation, the control group was given routine heart and diuretic treatments and placebo of Qishen Yiqi dropping pills for seven days (0.5 g each time, three times a day); While the Qishen group was given Qishen Yiqi dropping pills for seven days (0.5 g each time after meal, three times a day) on the basis of the routine treatments. The right ventricular end-diastolic volume (RVEDV), end-systolic volume (RVESV), stroke volume (SV) and right ventricular ejection fraction (RVEF) were detected after the operation. The results showed that patients in the two groups showed significantly lower right ventricular end diastolic volume (RVEDV), right ventricular end systolic volume (RVESV) and stroke volume (SV) decreased than that before the operation, but with significantly higher Ejection fraction (RVEF) significantly than that before the operation. However, the Qishen group showed a significantly lower right heart function reduction than the control group, with the statistical significance in the differences (P < 0.05). This indicated that the pretreatment with Qishenyiqi Drop Pills showed a remarkable efficacy in the improvement of right ventricular function after valve replacement.
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The Regenerating Spinal Cord of Gecko Maintains Unaltered Expression of ?-Catenin Following Tail Amputation.
J. Mol. Neurosci.
PUBLISHED: 09-02-2014
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The Wingless/Integrated (Wnt) signaling pathway plays important roles in central nervous system (CNS) development and regeneration, and ?-catenin, the central component, has been considered in association with adult neurogenesis. To decipher its roles on spontaneous spinal cord regeneration, we cloned ?-catenin from Gekko japonicus and examined its function in regenerating spinal cord. The protein was localized in the neurons and oligodendrocytes and maintained a stable expression levels during the spinal cord regeneration. The temporal pattern of expression has been found to be completely distinct with those of glycogen synthase kinase 3? (GSK3?). Experiments of gain-of-function by overexpression of full length ?-catenin or stabilized ?N90-?-catenin revealed that the accumulated protein attenuates the elongation of neurites and oligodendrocyte process. Knockdown of endogenous ?-catenin, however, decreased proliferation of oligodendrocytes by affecting expression of downstream lef1 and c-jun. The upregulated extracellular matrix fibronectin in injured cord was found to be inefficient in regulation of ?-catenin expression. Our results suggest that a tightly regulated stable expression of ?-catenin is required for the spontaneous spinal cord regeneration.
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Functional expression of chloride channels and their roles in the cell cycle and cell proliferation in highly differentiated nasopharyngeal carcinoma cells.
Physiol Rep
PUBLISHED: 09-01-2014
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We previously demonstrated that the growth of the poorly differentiated nasopharyngeal carcinoma cells (CNE-2Z) was more dependent on the activities of volume-activated chloride channels than that of the normal nasopharyngeal epithelial cells (NP69-SV40T). However, the activities and roles of such volume-activated chloride channels in highly differentiated nasopharyngeal carcinoma cells (CNE-1) are not clarified. In this study, it was found that a volume-activated chloride current and a regulatory volume decrease (RVD) were induced by 47% hypotonic challenges. The current density and the capacity of RVD in the highly differentiated CNE-1 cells were lower than those in the poorly differentiated CNE-2Z cells, and higher than those in the normal cells (NP69-SV40T). The chloride channel blockers, 5-nitro-2-(3-phenylpropylamino) benzoic acid (NPPB) and tamoxifen inhibited the current and RVD. Depletion of intracellular Cl(-) abolished the RVD. The chloride channel blockers reversibly inhibited cell proliferation in a concentration- and time-dependent manner, and arrested cells at the G0/G1 phases, but did not change cell viability. The sensitivity of the three cell lines to the chloride channel blockers was different, with the highest in poorly differentiated cells (CNE-2Z) and the lowest in the normal cells (NP69-SV40T). ClC-3 proteins were expressed in the three cells and distributed inside the cells as well as on the cell membrane. In conclusion, the highly differentiated nasopharyngeal carcinoma CNE-1 cells functionally expressed the volume-activated chloride channels, which may play important roles in controlling cell proliferation through modulating the cell cycle, and may be associated with cell differentiation. Chloride channels may be a potential target of anticancer therapy.
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The cytokinesis-blocked micronucleus assay as a strong predictor of lung cancer: extension of a lung cancer risk prediction model.
Cancer Epidemiol. Biomarkers Prev.
PUBLISHED: 08-29-2014
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There is an urgent need to improve lung cancer outcome by identifying and validating markers of risk. We previously reported that the cytokinesis-blocked micronucleus assay (CBMN) is a strong predictor of lung cancer risk. Here, we validate our findings in an independent external lung cancer population and test discriminatory power improvement of the Spitz risk prediction model upon extension with this biomarker.
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Proteomic response of Rhizoctonia solani GD118 suppressed by Paenibacillus kribbensis PS04.
World J. Microbiol. Biotechnol.
PUBLISHED: 08-28-2014
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Rice sheath blight, caused by Rhizoctonia solani, is considered a worldwide destructive rice disease and leads to considerable yield losses. A bio-control agent, Paenibacillus kribbensis PS04, was screened to resist against the pathogen. The inhibitory effects were investigated (>80 %) by the growth of the hyphae. Microscopic observation of the hypha structure manifested that the morphology of the pathogenic mycelium was strongly affected by P. kribbensis PS04. To explore essentially inhibitory mechanisms, proteomic approach was adopted to identify differentially expressed proteins from R. solani GD118 in response to P. kribbensis PS04 using two-dimensional gel electrophoresis. Protein profiling was used to identify 13 differential proteins: 10 proteins were found to be down-regulated while 3 proteins were up-regulated. These proteins were involved in material and energy metabolism, antioxidant activity, protein folding and degradation, and cytoskeleton regulation. Among them, material and energy metabolism was differentially regulated by P. kribbensis PS04. Protein expression was separately inhibited by the bio-control agent in oxidation resistance, protein folding and degradation, and cytoskeleton regulation. Proteome changes of the mycelium assist in understanding how the pathogen was directly suppressed by P. kribbensis PS04.
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Fibrosis progression in interferon treatment-naive Chinese plasma donors with chronic hepatitis C for 20 years: a cohort study.
Int. J. Infect. Dis.
PUBLISHED: 08-26-2014
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To evaluate the progression of fibrosis and factors influencing this in interferon (IFN) treatment-naive Chinese plasma donors infected with hepatitis C virus (HCV) for approximately 20 years.
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Prauserella shujinwangii sp. nov., from a desert environment.
Int. J. Syst. Evol. Microbiol.
PUBLISHED: 08-26-2014
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A Gram-positive, spore-forming, rod-shaped actinomycete, designated XJ46(T), was isolated from Xinjiang Uyghur Autonomous Region, China and subjected to a polyphasic taxonomic analysis. Morphological and chemotaxonomic characteristics of XJ46(T) were identified as being similar to those of members of the genus Prauserella. The phylogenetic tree based on 16S rRNA gene sequences showed that XJ46(T) shared the highest similarity (95.9?%) with Prauserella marina MS498(T). Based on its phenotypic characteristics, chemotaxonomic analysis and 16S rRNA gene sequence analysis, strain XJ46(T) is proposed to represent a novel species of the genus Prauserella, named Prauserella shujinwangii sp. nov. The type strain is XJ46(T) (?=?CGMCC 4.7125(T)?=?JCM 19736(T)).
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Molecular cloning and expression analysis of the Synaptotagmin-1 gene in the hypothalamus and pituitary of Huoyan goose during different stages of the egg-laying cycle.
Reprod. Biol. Endocrinol.
PUBLISHED: 08-21-2014
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Synaptotagmin-1 (Syt1) is an abundant, evolutionarily conserved integral membrane protein that plays essential roles in neurotransmitter release and hormone secretion. Neurotransmitters secreted by hypothalamic neurons can alter GnRH (gonadotropin-releasing hormones) neuronal activity by binding to and activating specific membrane receptors in pituitary cells and, in turn, control the release of gonadotropin hormones from the pituitary gland. To reveal the influence of Syt1 on the process of goose egg-laying, we cloned and characterized the cDNA of goose Syt1 originating from hypothalamus and pituitary tissues of Huoyan goose and investigated the mRNA expression profiles during different stages of the egg-laying cycle.
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Circulating miR-19a and miR-205 in serum may predict the sensitivity of luminal A subtype of breast cancer patients to neoadjuvant chemotherapy with epirubicin plus paclitaxel.
PLoS ONE
PUBLISHED: 08-19-2014
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The luminal A subtype of breast cancer has a good prognosis and is sensitive to endocrine therapy but is less sensitive to chemotherapy. It is necessary to identify biomarkers to predict chemosensitivity and avoid over-treatment. We hypothesized that miRNAs in the serum might be associated with chemosensitivity.
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Liquid/Liquid interfacial fabrication of thermosensitive and catalytically active Ag nanoparticle-doped block copolymer composite foam films.
Langmuir
PUBLISHED: 08-19-2014
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An aqueous solution of AgNO3 (upper phase) and a DMF/CHCl3 solution of polystyrene-b-poly(acryl acid)-b-polystyrene (PS-b-PAA-b-PS) or PS-b-PAA-b-PS/1,6-diaminohexane (DAH) (lower phase) constituted a planar liquid/liquid interface. The lower phase gradually transformed to a water-in-oil (W/O) emulsion via spontaneous emulsification due to the "ouzo effect". Polymer molecules, DAH molecules, and Ag(+) ions assembled into microcapsules around emulsion droplets that adsorbed at the planar liquid/liquid interface, resulting in formation of a foam film. DAH acted as a cross-linker during this process. Transmission electron microscopic observations indicated that Ag nanoclusters that were generated through reduction of Ag(+) ions by DMF were homogeneously dispersed in the walls of the foam structure. X-ray photoelectron spectroscopic investigations revealed that Ag(I) and Ag(0) coexisted in the film, and Ag(I) transformed to Ag(0) after further treatment. The film formed without DAH was not stable, while the film formed with DAH was very stable due to intermolecular attraction between PAA and DAH and formation of amides, as revealed by FTIR spectra. The film formed with DAH exhibited high and durable catalytic activity for hydrogenation of nitro compounds and, very interestingly, exhibited thermoresponsive catalytic behavior.
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Variation in Sp1 binding sites correlates with expression of survivin in breast cancer.
Mol Med Rep
PUBLISHED: 07-10-2014
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Survivin is the smallest member of the inhibitor of apoptosis (IAP) family and is deregulated in breast cancer, where it is associated with a poor overall prognosis. It is well established that survivin overexpression predominately occurs at the transcriptional level. Numerous transcription factors bind to specific sequences in the promoter regions of genes and are involved in transcriptional regulation. Specificity protein (Sp) 1 binding sites have been found in the promoter region of the survivin gene. The present study aimed to investigate whether variations in Sp1 binding sites affect survivin expression. Nested polymerase chain reaction followed by DNA sequencing were performed to analyze the survivin gene promoter region in 42 breast cancer tissue samples. Furthermore, survivin expression was assessed using immunohistochemistry. High survivin protein expression was found in 66.7% (28/42) of breast cancer tissue samples. In addition, 15 variations in seven Sp1 binding sites were detected in 12 samples and Sp1 binding site variation was found to be associated with low survivin expression in the 42 samples. These findings suggested that variations in Sp1 binding sites may be associated with survivin expression.
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A micromotor based on polymer single crystals and nanoparticles: toward functional versatility.
Nanoscale
PUBLISHED: 07-01-2014
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We report a multifunctional micromotor fabricated by the self-assembly technique using multifunctional materials, i.e. polymer single crystals and nanoparticles, as basic building blocks. Not only can this micromotor achieve autonomous and directed movement, it also possesses unprecedented functions, including enzymatic degradation-induced micromotor disassembly, sustained release and molecular detection.
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Membrane-bound complement regulatory proteins are prognostic factors of operable breast cancer treated with adjuvant trastuzumab: A retrospective study.
Oncol. Rep.
PUBLISHED: 06-25-2014
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Complement-dependent cytotoxicity (CDC) is an important antitumor mechanism of monoclonal antibodies (mAbs). However, trastuzumab, an anti-HER2 mAb, exerts only minor CDC. Overexpression of membrane-bound complement regulatory proteins (mCRPs), which suppress CDC, have been implicated in various malignant tumors. Here, we explored the predictive role of the expression levels of three mCRPs (CD55, CD59 and CD46) in the prognosis of breast cancer cases that underwent adjuvant trastuzumab treatment. We also studied the effect of mCRP downregulation on trastuzumab-induced CDC in vitro. Sixty-five HER2-positive breast cancer patients who received adjuvant therapy containing trastuzumab, were retrospectively analyzed. Levels of CD55, CD59 and CD46 expression were detected by immunohistochemistry. Chi-square test, Kaplan?Meier survival analysis and a Cox proportional hazards model were used to analyze the association between CD55, CD59 and CD46 expression and prognosis. HER2-positive SK-Br3 and BT-474 breast cancer cells were pretreated with various drugs to reduce mCRP expression. Afterwards, trastuzumab?mediated cytolytic effects were measured. Among the 65 patients, 46.2% had high expression of CD55, 44.6% had high expression of CD59 and 44.6% had high expression of CD46. The median follow-up duration was 47 months (range from 24 to 75 months). Patients with CD55 or CD59 overexpression had a higher relapse rate than those with low expression of CD55 (33.3 vs. 8.6%; P=0.013) or CD59 (31.0 vs. 11.1%; P=0.046). Similarly, mean disease-free survival of patients with CD55 or CD59 overexpression was significantly shorter than those with a low expression of CD55 (56 vs. 70 months; log-rank test, P=0.008) or CD59 (56 vs. 69 months; log-rank test, P=0.033). Multivariate analysis confirmed that CD55, but not CD59, was an independent risk factor of recurrence (HR=4.757; 95% CI, 0.985-22.974; P=0.05). In vitro, we found that tamoxifen inhibited both the protein and mRNA expression levels of CD55, but not CD59 or CD46 in SK-Br3 and BT-474 cells. After pretreatment of tamoxifen, trastuzumab-induced cytolysis was enhanced through CD55 downregulation. In conclusion, CD55 overexpression is an independent risk factor for recurrence in breast cancer patients receiving postoperative adjuvant therapy containing trastuzumab. Combined use of tamoxifen and trastuzumab for HER2-positive breast cancer treatment may enhance the antitumor effects of trastuzumab by elevated CDC, which warrants further study.
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Chitosan Degradation Products Promote Nerve Regeneration by Stimulating Schwann Cell Proliferation via miR-27a/FOXO1 Axis.
Mol. Neurobiol.
PUBLISHED: 06-16-2014
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Natural polysaccharides are biomaterials widely used for constructing scaffolds in tissue engineering. While natural polysaccharides have been shown to robustly promote tissue regeneration, the underlying molecular mechanism remains largely unknown. Here, we show that chitooligosaccharides (COS), the intermediate products of chitosan degradation, stimulate peripheral nerve regeneration in rats. Our experiment also shows that COS stimulate the proliferation of Schwann cells (SCs) during nerve regeneration. By analyzing the transcriptome and gene regulatory network, we identified the miR-27a/FOXO1 axis as the main signaling pathway for mediating the proliferative effects of COS on SCs. COS increase the expression level of miR-27a and cause a reduction of FOXO1, which subsequently accelerates the cell cycle and stimulates SC proliferation to stimulate nerve regeneration. These findings define a basic pathway for oligosaccharides-mediated cell proliferation and reveal a novel aspect of polysaccharide biomaterials in tissue engineering.
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Development of synthetic peptide-modified liposomes with LDL receptor targeting capacity and improved anticancer activity.
Mol. Pharm.
PUBLISHED: 06-03-2014
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In this study, we report an active targeting liposomal formulation directed by a novel peptide (AA13) that specifically binds to the low density lipoprotein receptor (LDLR) overexpressed on acute myeloid leukemia (AML) cells. The objectives of this study were to evaluate the in vitro and in vivo tumor drug targeting delivery of AA13-anchored liposomes on AML cells. AA13 conjugated to the distal end of DSPE-PEG2000-maleimide was incorporated into the liposomes via a postinsertion method. To study the effect of the peptide decoration and density on tumor cell targeting and internalization by AML cells (THP-1 and NB4), stealth liposomes bearing 3% (peptide/S100PC, molar ratio, LL) and 7% (peptide/S100PC, molar ratio, HL) AA13 were prepared, respectively. Higher uptake of LL (1.9- and 2.6-fold) and HL (2.3- and 3.6-fold) targeted liposomes occurred in THP-1 and NB4 cells, respectively, compared to untargeted liposomes. An LDLR inhibitor was used to confirm inhibition of the receptor-mediated cellular association of AA13 modified liposome in both cells. Daunorubicin (DNR) demonstrated a 2.2- and 3.5-fold higher cytotoxicity with the HL formulation and a 1.2- and 2.0-fold higher cytotoxicity with the LL formulation compared to the unmodified liposomal formulation in THP-1 and NB4 cells, respectively. Tumor drug accumulation of DNR-loaded HL was greater than that of the untargeted liposome in the biodistribution assay. The in vivo efficacy study in BALB/c nude mice bearing NB4 xenografts treated with DNR loaded HL also showed more tumor volume inhibition and a longer survival time compared to the untargeted formulation. In conclusion, the AA13-anchored liposomes demonstrated desirable potential as a promising vector for enhanced AML tumor drug targeting.
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Kinesin-12 influences axonal growth during zebrafish neural development.
Cytoskeleton (Hoboken)
PUBLISHED: 05-17-2014
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Kinesin-12 (also called KIF15) is a microtubule-based motor protein best known for its role in cell division. We previously reported that kinesin-12 is robustly expressed in developing terminally post-mitotic neurons, with levels diminishing as neurons reach maturity. We found that axons of cultured rodent neurons grow faster and longer if kinesin-12 is experimentally depleted, leading us to conclude that kinesin-12 plays a role in modulating axonal growth. Here we used zebrafish to explore whether these results apply to an in vivo system and whether they apply across different kinds of vertebrates. In whole mount in situ hybridization, kinesin-12 mRNA was detectable at 2-cell and 1K-cell stages. At 5.3 and 8 hours post-fertilization (hpf), hybridization signal for kinesin-12 mRNA was observed in the ectoderm. From 14 to 36 hpf, the signal had expanded to the central nervous system. At 60 hpf, the hybridization signal was concentrated in the brain. After 5 days post-fertilization, kinesin-12 expression was reduced. Kinesin-12 knockdown resulted in notably longer fast-growing axons with fewer branches by injection of a splice-blocking morpholino into Tg(huC:egfp) or Tg(hb9:gfp) zebrafish embryos. Kinesin-12 overexpression resulted in shorter axons than controls. These results are consistent with our previous observations on rodents using primary cultures for the experimental manipulations, and suggest a key role of kinesin-12 as a modulator of axonal development. © 2014 Wiley Periodicals, Inc.
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Role of text mining in early identification of potential drug safety issues.
Methods Mol. Biol.
PUBLISHED: 05-03-2014
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Drugs are an important part of today's medicine, designed to treat, control, and prevent diseases; however, besides their therapeutic effects, drugs may also cause adverse effects that range from cosmetic to severe morbidity and mortality. To identify these potential drug safety issues early, surveillance must be conducted for each drug throughout its life cycle, from drug development to different phases of clinical trials, and continued after market approval. A major aim of pharmacovigilance is to identify the potential drug-event associations that may be novel in nature, severity, and/or frequency. Currently, the state-of-the-art approach for signal detection is through automated procedures by analyzing vast quantities of data for clinical knowledge. There exists a variety of resources for the task, and many of them are textual data that require text analytics and natural language processing to derive high-quality information. This chapter focuses on the utilization of text mining techniques in identifying potential safety issues of drugs from textual sources such as biomedical literature, consumer posts in social media, and narrative electronic medical records.
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Breast carcinoma with choriocarcinomatous features: a case report and review of the literature.
World J Surg Oncol
PUBLISHED: 04-30-2014
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Breast carcinoma with choriocarcinomatous features (BCCF) is a rare variant of breast cancer, characterized by high expression of human chorionic gonadotropin (HCG) in cancer cells such as multinucleated syncytiotrophoblast-like giant cells. The first case of BCCF was reported in 1981 by Saigo and Rosen. Only one case of BCCF was reported to show no component of breast ductal carcinoma, and only partially cancer cells, such as multinucleated syncytiotrophoblast-like giant cells, expressed HCG in all previous BCCF cases. Here, we report the first BCCF case without any component of breast ductal carcinoma in which HCG was found to express in all cancer cells.
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TolC promotes ExPEC biofilm formation and curli production in response to medium osmolarity.
Biomed Res Int
PUBLISHED: 04-12-2014
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While a high osmolarity medium activates Cpx signaling and causes CpxR to repress csgD expression, and efflux protein TolC protein plays an important role in biofilm formation in Escherichia coli, whether TolC also responds to an osmolarity change to regulate biofilm formation in extraintestinal pathogenic E. coli (ExPEC) remains unknown. In this study, we constructed ?tolC mutant and complement ExPEC strains to investigate the role of TolC in the retention of biofilm formation and curli production capability under different osmotic conditions. The ?tolC mutant showed significantly decreased biofilm formation and lost the ability to produce curli fimbriae compared to its parent ExPEC strain PPECC42 when cultured in M9 medium or 1/2 M9 medium of increased osmolarity with NaCl or sucrose at 28°C. However, biofilm formation and curli production levels were restored to wild-type levels in the ?tolC mutant in 1/2 M9 medium. We propose for the first time that TolC protein is able to form biofilm even under high osmotic stress. Our findings reveal an interplay between the role of TolC in ExPEC biofilm formation and the osmolarity of the surrounding environment, thus providing guidance for the development of a treatment for ExPEC biofilm formation.
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Experimental infection of duck origin virulent Newcastle disease virus strain in ducks.
BMC Vet. Res.
PUBLISHED: 04-09-2014
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Newcastle disease (ND) caused by virulent Newcastle disease virus (NDV) is an acute, highly contagious and fatal viral disease affecting most species of birds. Ducks are generally considered to be natural reservoirs or carriers of NDV while being resistant to NDV strains, even those most virulent for chickens; however, natural ND cases in ducks have been gradually increasing in recent years. In the present study, ducks of different breeds and ages were experimentally infected with duck origin virulent NDV strain duck/Jiangsu/JSD0812/2008 (JSD0812) by various routes to investigate the pathogenicity of NDV in ducks.
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NRAGE promotes cell proliferation by stabilizing PCNA in a ubiquitin-proteasome pathway in esophageal carcinomas.
Carcinogenesis
PUBLISHED: 04-07-2014
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Neurotrophin receptor-interacting melanoma antigen-encoding gene homolog (NRAGE) is generally recognized as a tumor suppressor as it induces cell apoptosis and suppresses cell metastasis. However, it has recently been reported that NRAGE is overexpressed in lung cancer, melanoma and colon cancer, implicating a complicated role of NRAGE as we have expected. In the study, we aim to elucidate the functional roles and molecular mechanisms of NRAGE in esophageal carcinoma. We found that both NRAGE mRNA and protein were significantly overexpressed in esophageal tumor tissues. Consistently, both in vivo and in vitro analyses demonstrated that knockdown of NRAGE apparently inhibited cell growth, and cell cycle analysis further demonstrated that NRAGE knockdown cells were mainly arrested in G2M cell phase, accompanied with an apparent reduction of S phase. In the process of exploring molecular mechanisms, we found that either knockdown in vitro or knockout in vivo of NRAGE reduced proliferating cell nuclear antigen (PCNA) protein, expression of which could completely rescue the inhibited proliferation in NRAGE defective cells. Furthermore, NRAGE physically interacted with PCNA in esophageal cancer cells through DNA polymerase III subunit, and knockdown of NRAGE facilitated PCNA K48-linked polyubiquitination, leading PCNA to the proteasome-dependent degradation and a ubiquitin-specific protease USP10 was identified to be a key regulator in the process of K48 polyubiquitination in NRAGE-deleted cells. In conclusion, our study highlights a unique role of NRAGE and implies that NRAGE is likely to be an attractive oncotarget in developing novel genetic anticancer therapeutic strategies for esophageal squamous cell carcinomas.
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Upregulation of KLF4 by methylseleninic acid in human esophageal squamous cell carcinoma cells: Modification of histone H3 acetylation through HAT/HDAC interplay.
Mol. Carcinog.
PUBLISHED: 04-04-2014
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Esophageal squamous cell carcinoma (ESCC) occurs at a very high frequency in certain areas of China. Supplementation with selenium-containing compounds was associated with a significantly lower cancer mortality rate in a study conducted in Linxia, China. Thus, selenium could be a potential anti-esophageal cancer agent. In this study, methylseleninic acid (MSA) could inhibit cell growth of ESCC cells in vitro and in vivo. Upon treated with MSA, the activity of histone deacetylases (HDACs) was decreased and general control nonrepressed protein 5 (GCN5) was upregulated in ESCC cells. Meanwhile, a significant increase of H3K9 acetylation (H3K9ac) was detected. Upregulation of Krüppel-like factor 4 (KLF4) was also observed after MSA treatment. Additionally, the acetylated histone H3 located more at KLF4 promoter region after MSA treatment, shown by chromatin immunoprecipitation (ChIP) assay. Moreover, knockdown of GCN5 decreased the protein level of both H3K9ac and KLF4, along with less cell growth inhibition. Taken all, our results indicated that MSA could inhibit ESCC cell growth, at least in part, by MSA-HDAC/GCN5-H3K9ac-KLF4 axis. To our best knowledge, this is the first report that MSA induced acetylation of histone H3 at Lys9, which might depend on the activities and the balance between HDACs and HATs. © 2014 Wiley Periodicals, Inc.
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Bilobalide induces neuronal differentiation of P19 embryonic carcinoma cells via activating Wnt/?-catenin pathway.
Cell. Mol. Neurobiol.
PUBLISHED: 03-31-2014
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Bilobalide, a natural product extracted from Ginkgo biloba leaf, is known to exhibit a number of pharmacological activities. So far, whether it could affect embryonic stem cell differentiation is still unknown. The main aim of this study was to investigate the effect of bilobalide on P19 embryonic carcinoma cells differentiation and the underlying mechanisms. Our results showed that bilobalide induced P19 cells differentiation into neurons in a concentration- and time-dependent manner. We also found that bilobalide promoted neuronal differentiation through activation of Wnt/?-catenin signaling pathway. Exposure to bilobalide increased inactive GSK-3? phosphorylation, further induced the nuclear accumulation of ?-catenin, and also up-regulated the expression of Wnt ligands Wnt1 and Wnt7a. Neuronal differentiation induced by bilobalide was totally abolished by XAV939, an inhibitor of Wnt/?-catenin pathway. These results revealed a novel role of bilobalide in neuronal differentiation from P19 embryonic cells acting through Wnt/?-catenin signaling pathway, which would provide a better insight into the beneficial effects of bilobalide in brain diseases.
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Variations in the contents of gingerols and chromatographic fingerprints of ginger root extracts prepared by different preparation methods.
J AOAC Int
PUBLISHED: 03-29-2014
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In the present study, an HPLC-DAD method was optimized for the quantitative determination of 6-gingerol, 6-shogaol, 8-gingerol, and 10-gingerol in ginger extracts. A chromatographic fingerprinting method was also established to differentiate and evaluate the ginger extracts for bioactivity. Twenty-one extracts were prepared by methods differing in ginger type (fresh versus dried), solvent, and extraction methods. The ANOVA analysis showed the methods' influence on the mean extraction yields of gingerols increased in the order of: high pressure-high temperature (HP)>blender (BD)>low pressure (LP). The optimal solvent to extract gingerols was found to be 95% ethanol. The type of ginger used had significant effects on the content of gingerols, but its overall influence depended on the solvent used. In order to maximize the extraction efficiency of gingerols, a combination of dry ginger, 95% ethanol, and the HP extraction method should be employed. The chromatographic fingerprints were obtained to differentiate the unknown components from all ginger extracts. The similarity of the chromatographic fingerprints was used to evaluate the differences among all extracts. It can be concluded that the chromatographic fingerprints are able to ensure the stability of each extract and have some correlation with the observed bioactivity.
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Direct growth of graphene on quartz substrates for label-free detection of adenosine triphosphate.
Nanotechnology
PUBLISHED: 03-26-2014
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We demonstrate that continuous, uniform graphene films can be directly synthesized on quartz substrates using a two-temperature-zone chemical vapor deposition system and that their layers can be controlled by adjusting the precursor partial pressure. Raman spectroscopy and transmission electron microscopy confirm the formation of monolayer graphene with a grain size of ?100 nm. Hall measurements show a room-temperature carrier mobility above 1500 cm2 V(-1) s(-1). The optical transmittance and conductance of the graphene films are comparable to those of transferred metal-catalyzed graphene. The method avoids the complicated and skilled post-growth transfer process and allows the graphene to be directly incorporated into a fully functional biosensor for label-free detection of adenosine triphosphate (ATP). This device shows a fast response time of a few milliseconds and achieves a high sensitivity to ATP molecules over a very wide range from 0.002 to 5 mM.
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Biotherapy for autoimmune liver diseases.
Curr Pharm Biotechnol
PUBLISHED: 03-20-2014
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Autoimmune liver diseases are chronic inflammatory conditions leading to an etiologically undefined immunemediated attack aimed at hepatocytes and the biliary epithelium. Drugs used in autoimmune liver disease such as ursodeoxycholic acid, prednisolone and azathioprine are not effective in all patients, therefore, new therapeutic approaches are needed for autoimmune liver diseases that are refractory to standard therapy. Biotherapy is a thriving area of research and development, and is used in the treatment of chronic autoimmune liver diseases. However, to date, there is no clinically validated standard biotherapy for autoimmune liver diseases. Thus, future clinical trials are required to evaluate the effectiveness and safety of biotherapy before this approach can be used in routine clinical practice for the therapy of autoimmune liver diseases. This article provides an overview of emerging biotherapy for autoimmune liver diseases.
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Prospective identification and culture of rat enteric neural stem cells (ENSCs).
Cytotechnology
PUBLISHED: 03-11-2014
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Hirschprung's disease (HD), a very common congenital abnormality in children, occurs mainly due to the congenital developmental defect of the enteric nervous system. The absence of enteric ganglia from the distal gut due to deletion in gut colonization by neural crest progenitor cells may lead to HD. The capacity to identify and isolate the enteric neuronal precursor cells from developing and mature tissues would enable the development of cell replacement therapies for HD. However, a mature method to culture these cells is a challenge. The present study aimed to propose a method to culture enteric neural stem cells (ENSCs) from the DsRed transgenic fetal rat gut. The culture medium used contained 15 % chicken embryo extract, basic fibroblast growth factor, and epidermal growth factor. ENSCs were cultured from embryonic day 18 in DsRed transgenic rat. Under inverted microscope and fluorescence staining, ENSCs proliferated to form small cell clusters on the second day of culture. The neurospheres-like structure were suspended in the medium, and there were some filaments between the adherent cells from day 3 to day 6 of the culture. The neurospheres were formed by ENSCs on day 8 of the culture. Network-like connections were formed between the adherent cells and differentiated cells after adding 10 % FBS. The differentiated cells were positive for neurofilament and glial fibrillary acidic protein antibodies. The present study established a method to isolate and culture ENSCs from E18 DsRed transgenic rats in the terminal stage of embryonic development. This study would offer a way to obtain plenty of cells for the future research on the transplantation of HD.
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Support vector machine-an alternative to artificial neuron network for water quality forecasting in an agricultural nonpoint source polluted river?
Environ Sci Pollut Res Int
PUBLISHED: 03-03-2014
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Water quality forecasting in agricultural drainage river basins is difficult because of the complicated nonpoint source (NPS) pollution transport processes and river self-purification processes involved in highly nonlinear problems. Artificial neural network (ANN) and support vector model (SVM) were developed to predict total nitrogen (TN) and total phosphorus (TP) concentrations for any location of the river polluted by agricultural NPS pollution in eastern China. River flow, water temperature, flow travel time, rainfall, dissolved oxygen, and upstream TN or TP concentrations were selected as initial inputs of the two models. Monthly, bimonthly, and trimonthly datasets were selected to train the two models, respectively, and the same monthly dataset which had not been used for training was chosen to test the models in order to compare their generalization performance. Trial and error analysis and genetic algorisms (GA) were employed to optimize the parameters of ANN and SVM models, respectively. The results indicated that the proposed SVM models performed better generalization ability due to avoiding the occurrence of overtraining and optimizing fewer parameters based on structural risk minimization (SRM) principle. Furthermore, both TN and TP SVM models trained by trimonthly datasets achieved greater forecasting accuracy than corresponding ANN models. Thus, SVM models will be a powerful alternative method because it is an efficient and economic tool to accurately predict water quality with low risk. The sensitivity analyses of two models indicated that decreasing upstream input concentrations during the dry season and NPS emission along the reach during average or flood season should be an effective way to improve Changle River water quality. If the necessary water quality and hydrology data and even trimonthly data are available, the SVM methodology developed here can easily be applied to other NPS-polluted rivers.
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Ginsenoside Rd ameliorates experimental autoimmune encephalomyelitis in C57BL/6 mice.
J. Neurosci. Res.
PUBLISHED: 02-26-2014
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Multiple sclerosis (MS) is a common disabling autoimmune disease without an effective treatment in young adults. Ginsenoside Rd, extracted from Panax notoginseng, has multiple pharmacological effects and potential therapeutic applications in diseases of the central nervous system. In this study, we explore the efficacy of ginsenoside Rd in experimental autoimmune encephalomyelitis (EAE), an established model of MS. EAE was induced by myelin oligodendrocyte glycoprotein 35-55-amino-acid peptide. Ginsenoside Rd (10-80 mg/kg/day) or vehicle was intraperitoneally administered on the disease onset day, and the therapy persisted throughout the experiments. The dose of 40 mg/kg/day of ginsenoside Rd was selected as optimal. Ginsenoside Rd effectively ameliorated the clinical severity in EAE mice, reduced the permeability of the blood-brain barrier, regulated the secretion of interferon-gamma and interleukin-4, promoted the Th2 shift in vivo (cerebral cortex) and in vitro (splenocytes culture supernatants), and prevented the reduction in expression of brain-derived neurotrophic factor and nerve growth factor in both cerebral cortex and lumbar spinal cord of EAE mice. This study establishes the potency of ginsenoside Rd in inhibiting the clinical course of EAE. These findings suggest that ginsenoside Rd could be a promising agent for amelioration of neuroimmune dysfunction diseases such as MS.
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The transcriptomic response to copper exposure by the gill tissue of Japanese scallops (Mizuhopecten yessoensis) using deep-sequencing technology.
Fish Shellfish Immunol.
PUBLISHED: 02-26-2014
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The bivalve Mizuhopecten yessoensis has been greatly impacted by marine pollutants in northern China. To elucidate the toxicological mechanism of copper exposure on the immune system, we investigated differentially expressed genes (DEGs) and transcript abundance in M. yessoensis gill tissue using the deep-sequencing platform Illumina HiSeq™ 2000. In total, 1312 and 2237 genes were identified as significantly up- or down-regulated, respectively. In addition, significant enrichment analysis identified 9 GO terms and 38 pathways involved in the response to copper exposure. The analysis of immune-related transcripts revealed a complex repertoire of innate recognition receptors, including toll-like receptors, NOD-like receptors and RIG-like receptors. Downstream pathway effectors, such as apoptotic, lysosomal and C-type lectin transcripts, were also analyzed. These results will provide a resource for subsequent gene expression studies regarding heavy metal exposure and the identification of copper-sensitive biomarkers to monitor the aquaculture of M. yessoensis.
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Neuroprotective effect of a formula, moschus combined with borneolum synthcticum, from traditional chinese medicine on ischemia stroke in rats.
Evid Based Complement Alternat Med
PUBLISHED: 02-22-2014
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Moschus compatible with borneolum synthcticum is a well-known herb pair in Traditional Chinese Medicine and the present study aims to assess the neuroprotective effect of a formula composed of this herb pair on ischemia stroke in rats. The middle cerebral artery occlusion model of focal cerebral ischemia in rat was performed by using intraluminal suture method. The behavioral scores, infarct volume, and neuron ultrastructure of model and formula-treated rats were investigated after the 2?h of ischemia and 24?h of reperfusion. Meanwhile the expression levels of caspase-3, caspase-9, Bcl-2, and Bax were measured by western blot analysis. The formula treatment showed obvious neuroprotective effect according to significant decrease of the neurological scores (P < 0.01) and the infarct volumes (P < 0.05) when compared to the MCAO group. We also observed that this formula had antiapoptosis activity on neuron cell under electron microscope. Furthermore, our result supported the idea that pro- and postadministration of this formula had an antiapoptosis effect by decreasing remarkably the expression of caspase-3 and caspase-9 (P < 0.05) as well as increasing significantly the ratio of Bcl-2 to Bax (P < 0.01). All evidences demonstrated the neuroprotective effect of this formula on ischemia stroke due to decrease of brain infract volume and modulation of the expression of apoptosis-related proteins.
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2-(4-Methoxyphenyl)ethyl-2-acetamido-2-deoxy-?-d-pyranoside confers neuroprotection in cell and animal models of ischemic stroke through calpain1/PKA/CREB-mediated induction of neuronal glucose transporter 3.
Toxicol. Appl. Pharmacol.
PUBLISHED: 02-20-2014
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Salidroside is proven to be a neuroprotective agent of natural origin, and its analog, 2-(4-Methoxyphenyl)ethyl-2-acetamido-2-deoxy-?-d-pyranoside (named SalA-4g), has been synthesized in our lab. In this study, we showed that SalA-4g promoted neuronal survival and inhibited neuronal apoptosis in primary hippocampal neurons exposed to oxygen and glucose deprivation (OGD) and in rats subjected to ischemia by transient middle cerebral artery occlusion (MCAO), respectively, and that SalA-4g was more neuroprotective than salidroside. We further found that SalA-4g elevated glucose uptake in OGD-injured primary hippocampal neurons and increased the expression and recruitment of glucose transporter 3 (GLUT3) in ischemic brain. Signaling analysis revealed that SalA-4g triggered the phosphorylation of CREB, and increased the expression of PKA RII in primary hippocampal neurons exposed to OGD injury, while inhibition of PKA/CREB by H-89 alleviated the elevation in glucose uptake and GLUT3 expression, and blocked the protective effects of SalA-4g. Moreover, SalA-4g was noted to inhibit intracellular Ca(2+) influx and calpain1 activation in OGD-injured primary hippocampal neurons. Our results suggest that SalA-4g neuroprotection might be mediated by increased glucose uptake and elevated GLUT3 expression through calpain1/PKA/CREB pathway.
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Using immunomic approach to enhance tumor-associated autoantibody detection in diagnosis of hepatocellular carcinoma.
Clin. Immunol.
PUBLISHED: 02-19-2014
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To explore the possibility of using a mini-array of multiple tumor-associated antigens (TAAs) as an approach to the diagnosis of hepatocellular carcinoma (HCC), 14 TAAs were selected to examine autoantibodies in sera from patients with chronic hepatitis, liver cirrhosis and HCC by immunoassays. Antibody frequency to any individual TAA in HCC varied from 6.6% to 21.1%. With the successive addition of TAAs to the panel of TAAs, there was a stepwise increase of positive antibody reactions. The sensitivity and specificity of 14 TAAs for immunodiagnosis of HCC was 69.7% and 83.0%, respectively. This TAA mini-array also identified 43.8% of HCC patients who had normal alpha-fetoprotein (AFP) levels in serum. In summary, this study further supports the hypothesis that a customized TAA array used for detecting anti-TAA autoantibodies can constitute a promising and powerful tool for immunodiagnosis of HCC and may be especially useful in patients with normal AFP levels.
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Physicochemical, mechanical and thermal properties of chitosan films with and without sorbitol.
Int. J. Biol. Macromol.
PUBLISHED: 02-10-2014
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The effect of sorbitol on the physicochemical, mechanical and thermal properties of chitosan films with different degrees of deacetylation (DD; i.e., DD85% and DD95%) was investigated. The thickness, moisture content (MC), water solubility (WS) and water-vapor permeability (WVP) of the films were evaluated. Sorbitol addition reduced MC, increased WS and significantly (p<0.01) reduced WVP of both film types. DD95% films had lower MC and WVP, and higher WS than DD85% films. Static (thermomechanical analysis) and dynamic (dynamic mechanical analysis) tests indicated that sorbitol increased the strain and decreased stress for both DD films, but DD95% could sustain higher strain and DD85% could sustain higher stress. Thermogravimetrics analysis and differential scanning calorimetry showed that sorbitol elicited a lower degradation temperature for both films, and that DD95% films exhibited higher thermal stability than DD85% films.
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Autoantibody response to murine double minute 2 protein in immunodiagnosis of hepatocellular carcinoma.
J Immunol Res
PUBLISHED: 01-20-2014
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Hepatocellular carcinoma (HCC) is the fifth most common malignancy worldwide. Although new therapeutic strategies have been continuously developed and applied to clinical treatment for HCC, the prognosis is still very poor. Thus, early detection of HCC may enhance effective and curative management. In this study, autoantibody responses to MDM2 protein in HCC patient's serum were evaluated by enzyme-linked immunosorbent assay (ELISA) and part sera were evaluated by Western blotting and indirect immunofluorescence assay. Immunohistochemistry (IHC) over tissue array slides was also performed to analyze protein expression of MDM2 in HCC and control tissues. The prevalence of autoantibodies against MDM2 was significantly higher than that in liver cirrhosis (LC), chronic hepatitis (CH), and normal human sera (NHS). The average titer of autoantibodies against MDM2 in HCC serum was higher compared to that in LC, CH, and NHS. A high titer of autoantibodies against MDM2 in ELISA could be observed in the serum in 6 to 9 months before the clinical diagnosis of HCC in the serum of several HCC patients with serial bleeding samples. Our preliminary data indicate that MDM2 and anti-MDM2 system may be a potential biomarker for early stage HCC screening and immunodiagnosis.
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Intramuscular Administration of a Synthetic CpG-Oligodeoxynucleotide Modulates Functional Responses of Neutrophils of Neonatal Foals.
PLoS ONE
PUBLISHED: 01-01-2014
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Neutrophils play an important role in protecting against infection. Foals have age-dependent deficiencies in neutrophil function that may contribute to their predisposition to infection. Thus, we investigated the ability of a CpG-ODN formulated with Emulsigen to modulate functional responses of neutrophils in neonatal foals. Eighteen foals were randomly assigned to receive either a CpG-ODN with Emulsigen (N?=?9) or saline intramuscularly at ages 1 and 7 days. At ages 1, 3, 9, 14, and 28, blood was collected and neutrophils were isolated from each foal. Neutrophils were assessed for basal and Rhodococcus equi-stimulated mRNA expression of the cytokines interferon-? (IFN-?), interleukin (IL)-4, IL-6, and IL-8 using real-time PCR, degranulation by quantifying the amount of ?-D glucuronidase activity, and reactive oxygen species (ROS) generation using flow cytometry. In vivo administration of the CpG-ODN formulation on days 1 and 7 resulted in significantly (P<0.05) increased IFN-? mRNA expression by foal neutrophils on days 3, 9, and 14. Degranulation was significantly (P<0.05) lower for foals in the CpG-ODN-treated group than the control group at days 3 and 14, but not at other days. No effect of treatment on ROS generation was detected. These results indicate that CpG-ODN administration to foals might improve innate and adaptive immune responses that could protect foals against infectious diseases and possibly improve responses to vaccination.
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Association of serum microRNA expression in hepatocellular carcinomas treated with transarterial chemoembolization and patient survival.
PLoS ONE
PUBLISHED: 01-01-2014
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Hepatocellular carcinoma (HCC) is one of the most deadly tumors. Transarterial chemoembolization (TACE) is effective for unresectable HCC. In recent years, miRNAs have been proposed as novel diagnostic and prognostic tools for HCC. This study aimed to identify whether microRNAs (miRNAs) can serve as biomarkers to reliably predict outcome before HCC patients are treated with TACE.
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Age-group-specific associations between the severity of obstructive sleep apnea and relevant risk factors in male and female patients.
PLoS ONE
PUBLISHED: 01-01-2014
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To seek accurate and credible correlation manner between gender, age, and obesity; and the severity of obstructive sleep apnea (OSA) in large-scale population.
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Suppression of Akt-mTOR pathway-a novel component of oncogene induced DNA damage response barrier in breast tumorigenesis.
PLoS ONE
PUBLISHED: 01-01-2014
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DNA damage has been thought to be directly associated with the neoplastic progression by enabling mutations in tumor suppressor genes and activating/and amplifying oncogenes ultimately resulting in genomic instability. DNA damage causes activation of the DNA damage response (DDR) that is an important cellular mechanism for maintaining genomic integrity in the face of genotoxic stress. While the cellular response to genotoxic stress has been extensively studied in cancer models, less is known about the cellular response to oncogenic stress in the premalignant context. In the present study, by using breast tissues samples from women at different risk levels for invasive breast cancer (normal, proliferative breast disease and ductal carcinoma in situ) we found that DNA damage is inversely correlated with risk of invasive breast cancer. Similarly, in MCF10A based in vitro model system where we recapitulated high DNA damage conditions as seen in patient samples by stably cloning in cyclin E, we found that high levels of oncogene induced DNA damage, by triggering inhibition of a major proliferative pathway (AKT), inhibits cell growth and causes cells to die through autophagy. These data suggest that AKT-mTOR pathway is a novel component of oncogene induced DNA damage response in immortalized 'normal-like' breast cells and its suppression may contribute to growth arrest and arrest of the breast tumorigenesis.
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Plasma sphingolipids as potential indicators of hepatic necroinflammation in patients with chronic hepatitis C and normal alanine aminotransferase level.
PLoS ONE
PUBLISHED: 01-01-2014
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Accurate estimation of hepatic necroinflammation caused by chronic hepatitis C (CHC) is crucial for prediction of prognosis and design of therapeutic strategy, which is particularly true for CHC patients with normal alanine aminotransferase (ALT) level. Recent studies have shown that sphingolipids have a close relationship with hepatitis C virus infection. The present study aimed to identify plasma sphingolipids related to hepatic necroinflammation. We included 120 treatment-naïve CHC patients and 64/120 had normal ALT levels (<40 U/L). CHC patients who underwent liver biopsies were subjected to Scheuer scoring analysis for scope of hepatic inflammation. Plasma sphingolipids were detected by high-performance liquid chromatography tandem mass spectrometry. Our results showed 44 plasma sphingolipids were detected altogether. Of all detected sphingolipids, hexosylceramide (HexCer) (d18?1/22?0) and HexCer (d18?1/24?0) showed a significant difference among G0/G1, G2, and G3/G4 (P<0.05). For identifying hepatic necroinflammation (G?2), after adjusting other factors, the odds ratio (OR) of HexCer (d18?1/22?0) reached 1.01 (95% confidence interval [CI]: 1.00-1.02). Furthermore, the area under the curve (AUC) of HexCer (d18?1/22?0) was 0.7 (P?=?0.01) and approached that of ALT (AUC?=?0.78). However, in CHC patients with normal ALT, HexCer (d18?1/22?0) was an independent factor (OR: 1.02, 95% CI: 1.01-1.03) to identify the hepatic necroinflammation (G?2). HexCer (d18?1/22?0) not only showed the largest AUC (0.78, P?=?0.001), but also exhibited the highest specificity of all indicators. These results indicate that plasma HexCer (d18?1/22?0) is a potential indicator to distinguish hepatic necroinflammation in CHC patients. For CHC with normal ALT, the ability of HexCer (d18?1/22?0) to distinguish hepatic necroinflammation might be superior to conventional serum indicators.
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Determining molecular predictors of adverse drug reactions with causality analysis based on structure learning.
J Am Med Inform Assoc
PUBLISHED: 12-11-2013
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Adverse drug reaction (ADR) can have dire consequences. However, our current understanding of the causes of drug-induced toxicity is still limited. Hence it is of paramount importance to determine molecular factors of adverse drug responses so that safer therapies can be designed.
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Adverse drug effect detection.
IEEE J Biomed Health Inform
PUBLISHED: 11-16-2013
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Large collections of electronic patient records provide abundant but under-explored information on the real-world use of medicines. Although they are maintained for patient administration, they provide a broad range of clinical information for data analysis. One growing interest is drug safety signal detection from these longitudinal observational data. In this paper, we proposed two novel algorithms-a likelihood ratio model and a Bayesian network model-for adverse drug effect discovery. Although the performance of these two algorithms is comparable to the state-of-the-art algorithm, Bayesian confidence propagation neural network, the combination of three works better due to their diversity in solutions. Since the actual adverse drug effects on a given dataset cannot be absolutely determined, we make use of the simulated observational medical outcomes partnership (OMOP) dataset constructed with the predefined adverse drug effects to evaluate our methods. Experimental results show the usefulness of the proposed pattern discovery method on the simulated OMOP dataset by improving the standard baseline algorithm-chi-square-by 23.83%.
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Basic fibroblast growth factor is a key factor that induces bone marrow mesenchymal stem cells towards cells with Schwann cell phenotype.
Neurosci. Lett.
PUBLISHED: 11-11-2013
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Bone marrow mesenchymal stem cells (MSCs) can be differentiate towards a Schwann cells (SCs) lineage when exposed to pre-inducing reagents ?-mercaptoethanol (BME) and retinoic acid (RA), followed by inducing factors: forskolin (FSK), basic fibroblast growth factor (bFGF), platelet derived growth factor (PDGF), and heregulin (HRG). However, the underlying mechanisms remain unclear. Here, we investigated the individual effects of these inducing factors on the differentiation of MSCs towards SC phenotype in rats. We show that the omission of either HRG or PDGF from the induction medium is not sufficient to change the SC-like phenotype or the expression level of the SC marker, S100?. However, the omission of bFGF from the induction medium effectively blocked neural induction of the MSCs. Moreover, only bFGF was found to inhibit MSC proliferation during differentiation. To clarify the mechanism responsible for the effect of bFGF, we also investigated the activation of the extracellular signal-regulated kinase (ERK) pathway in the induced cells. Our results suggest that morphological changes in MSCs induced by bFGF depend on the activation of ERK, and bFGF may be an indispensable factor that induces MSCs to differentiate into cells with SCs phenotype.
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The novel coding region SNPs of PPARGC1A gene and their associations with growth traits in Chinese native cattle.
Mol. Biol. Rep.
PUBLISHED: 10-26-2013
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The peroxisome proliferator-activated receptor gamma coactivator-1 alpha protein, encoded by the PPARGC1A gene, is a metabolic switch, which transcriptionally activates a complex pathway of mitochondrial biogenesis, lipid and glucose metabolism. Three SNPs (exon 3 c.396G>A, intron 9 c.1892 + 19C>T and exon 10 c.1971C>T) were found and identified in three Chinese native cattle breeds by PCR-SSCP, PCR-RFLP and DNA sequencing methods. All three populations had a low genetic diversity at SNP396 locus (PIC <0.25) while possessed a moderate genetic diversity at SNP1892 locus (0.25 < PIC < 0.5). Association study indicated that the synonymous mutation c.396G>A significantly associated with body weight and average daily gain in Nanyang cattle at the adult age (P < 0.05). Our investigation will not only extend the spectrum of genetic variation of bovine PPARGC1A gene, but also provide useful information for the marker assisted selection in beef cattle breeding program.
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Association analysis of bovine Foxa2 gene single sequence variant and haplotype combinations with growth traits in Chinese cattle.
Gene
PUBLISHED: 10-03-2013
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Forkhead box A2 (Foxa2) has been recognized as one of the most potent transcriptional activators that is implicated in the control of feeding behavior and energy homeostasis. However, similar researches about the effects of genetic variations of Foxa2 gene on growth traits are lacking. Therefore, this study detected Foxa2 gene polymorphisms by DNA pool sequencing, PCR-RFLP and PCR-ACRS methods in 822 individuals from three Chinese cattle breeds. The results showed that four sequence variants (SVs) were screened, including two mutations (SV1, g. 7005 C>T and SV2, g. 7044 C>G) in intron 4, one mutation (SV3, g. 8449 A>G) in exon 5 and one mutation (SV4, g. 8537 T>C) in the 3UTR. Notably, association analysis of the single mutations with growth traits in total individuals (at 24months) revealed that significant statistical difference was found in four SVs, and SV4 locus was highly significantly associated with growth traits throughout all three breeds (P<0.05 or P<0.01). Meanwhile, haplotype combination CCCCAGTC also indicated remarkably associated to better chest girth and body weight in Jiaxian Red cattle (P<0.05). We herein described a comprehensive study on the variability of bovine Foxa2 gene that was predictive of molecular markers in cattle breeding for the first time.
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Solution of export coefficients of nitrogen from different land-use patterns based on Bayesian analysis.
Water Sci. Technol.
PUBLISHED: 08-09-2013
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The export coefficient model has been applied worldwide to the estimation of non-point source (NPS) pollution. Determining the export coefficients (ECs) from each pollution source and different space-time progressions is problematic because of uncertainty in the ECs of nitrogen from different land-use patterns. Bayesian theory uses the prior probability distribution and likelihood data to generate a posterior probability distribution. The total nitrogen (TN) ECs and stream loss rates K (d(-1)) for five land-use patterns were estimated by combining published results with monthly data for ChangLe River system for 2004-08. After 10(4) iterations, the results had small Markov chain Monte Carlo errors and convergence was obtained. Average TN ECs for the entire watershed were 26.1 ± 8.8, 70.3 ± 9.4, 41.7 ± 6.9, 8.9 ± 1.6 and 6.2 ± 0.5 kg ha(-1) yr(-1) for paddy field, dry land, residential land, woodland and barren land with coefficients of variation (CVs) of 16.9, 6.31, 8.91, 13.3 and 27.9% among sub-catchments respectively. The average K value was 0.33 d(-1) with a CV of 11.3%. Estimated ECs, K and the coupling water quality model were used to predict the years 2008 and 2009; the results validated the model. This Bayesian model can determine ECs using prior knowledge and monitored data, overcoming the problems of the regression model. The model facilitates explicit consideration of uncertainty in NPS management.
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Peroxiredoxin 1 is a tumor-associated antigen in esophageal squamous cell carcinoma.
Oncol. Rep.
PUBLISHED: 06-28-2013
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Peroxiredoxin 1 (Prdx1) is an antioxidant and plays an important role in H2O2-mediated cell signaling. We previously found that the expression level of Prdx1 was elevated in esophagus squamous cell carcinoma (ESCC) tissue using a proteomics approach. Since overexpressed protein can induce an autoimmune response, to further examine whether serum from ESCC patients exhibits immunoreactivity against Prdx1, autoantibody responses to Prdx1 were evaluated by ELISA, western blotting and indirect immunofluorescence assay in sera from patients with ESCC and normal individuals. Immunohistochemical study with tissue array slides and western blot analysis with cancer cell lines were also performed to analyze the protein expression profiles of Prdx1 in ESCC tissues and cancer cell lines. The results demonstrated that the positive rate of autoantibody against Prdx1 in ESCC sera was 13.2% (9/68), whereas this rate was 0% (0/89) in normal individuals. Data also showed that expression of Prdx1 was significantly increased in ESCC tissues when compared to expression in paired adjacent normal tissues (P<0.05). The data indicate that Prdx1 may contribute to malignant transformation of the esophagus, and may be used as a biomarker in the immunodiagnosis of ESCC.
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The anti-inflammatory effect of donepezil on experimental autoimmune encephalomyelitis in C57 BL/6 mice.
Neuropharmacology
PUBLISHED: 06-23-2013
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Donepezil is a potent and selective acetylcholinesterase inhibitor. It has been reported to restore cognitive performance in multiple sclerosis (MS) patients and experimental autoimmune encephalomyelitis (EAE) mice, an established model of MS. However, there are no reports about the anti-inflammatory effects of donepezil on EAE. In this study, the donepezil treatments on EAE mice were initiated at day 7 post immunization (7 p.i., subclinical periods, early donepezil treatment) and day 13 p.i. (clinical periods, late donepezil treatment) with the dosage of 1, 2 and 4 mg/kg/d respectively and the treatments persisted throughout the experiments. Blood-brain barrier (BBB) permeability was detected by Evans blue content, the expression of matrix metalloproteinase-2 (MMP-2) and MMP-9, Akt and phosphorylated Akt (p-Akt) as well as nerve growth factor (NGF) and its precursor form (proNGF) in the brains of EAE mice were detected by Western blot, and the levels of interferon-? and interleukin-4 in the splenocytes culture supernatants and brains of EAE mice were evaluated by ELISA. The results showed that the 2 mg/kg/d late donepezil treatment was the optimal dosage and could ameliorate clinical and pathological parameters, improve magnetic resonance imaging outcomes, reduce the permeability of BBB, inhibit the production of MMP-2 and MMP-9, modulate the expression of NGF and proNGF, increase Th2 bias and the phosphorylation of Akt in the brains of EAE mice. Our data suggested that the anti-inflammatory effects of donepezil may be a novel mechanism on treating EAE and provided further insights to understand the donepezils neuroprotective activities in MS.
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A tetragonal prismatic {Co32} nanocage based on thiacalixarene.
Chem. Commun. (Camb.)
PUBLISHED: 06-21-2013
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A novel coordination nanocage was obtained with in situ-generated tetrazole ligands from the cobalt-thiacalix[4]arene system. Four in situ-generated 1,3-bis(2H-tetrazol-5-yl)benzene ligands bolster eight Co4-calix shuttle-cock-like secondary building units (SBUs) into a hollow tetragonal prism with a high surface area. The unprecedented {Co32} nanocage presents the highest nuclearity example.
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Predicting drug responsiveness in human cancers using genetically engineered mice.
Clin. Cancer Res.
PUBLISHED: 06-18-2013
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To use genetically engineered mouse models (GEMM) and orthotopic syngeneic murine transplants (OST) to develop gene expression-based predictors of response to anticancer drugs in human tumors. These mouse models offer advantages including precise genetics and an intact microenvironment/immune system.
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Stream nitrogen sources apportionment and pollution control scheme development in an agricultural watershed in eastern China.
Environ Manage
PUBLISHED: 06-08-2013
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A modeling system that couples a land-usebased export coefficient model, a stream nutrient transport equation, and Bayesian statistics was developed for stream nitrogen source apportionment. It divides a watershed into several sub-catchments, and then considers the major landuse categories as stream nitrogen sources in each subcatchment. The runoff depth and stream water depth are considered as the major factors influencing delivery of nitrogen from land to downstream stream node within each sub-catchment. The nitrogen sources and delivery processes are lumped into several constant parameters that were calibrated using Bayesian statistics from commonly available stream monitoring and land-use datasets. This modeling system was successfully applied to total nitrogen (TN) pollution control scheme development for the ChangLe River watershed containing six sub-catchments and four land-use categories. The temporal (across months and years) and spatial (across sub-catchments and land-use categories) variability of nonpoint source (NPS) TN export to stream channels and delivery to the watershed outlet were assessed. After adjustment for in-stream TNretention, the time periods and watershed areas with disproportionately high-TN contributions to the stream were identified. Aimed at a target stream TN level of 2 mg L-1, a quantitative TN pollution control scheme was further developed to determine which sub-catchments, which land-use categories in a sub-catchment, which time periods, and how large of NPS TN export reduction were required. This modeling system provides a powerful tool for stream nitrogen source apportionment and pollution control scheme development at the watershed scale and has only limited data requirements.
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Molecular Cloning of Hemocyanin cDNA from Fenneropenaeus chinensis and Antimicrobial Analysis of Two C-terminal Fragments.
Mar. Biotechnol.
PUBLISHED: 06-07-2013
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Peptides derived from shrimp hemocyanin have antimicrobial properties. This is the first report of hemocyanin cDNA (FCHc) cloned from Fenneropenaeus chinensis and recombinant expression of two C-terminal fragments. Based on sequence analysis of Fenneropenaeus chinensis hemocyanin FCHc, we subcloned two FCHc fragments by designing special primers. Two antimicrobial peptides (AMPs) were derived from FCHc (FCHc-C1 and FCHc-C2). The recombinant sequence of FCHc-C1 consisted of 207 bp encoding 69 amino acids and the recombinant sequence of FCHc-C2 consisted of 120 bp encoding 40 amino acids. The results of Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting indicated that recombinant FCHc-C1 and FCHc-C2 peptides (rFCHc-C1 and rFCHc-C2) were expressed successfully. An inhibition assay showed that FCHc-C1 and FCHc-C2 were anionic AMPs with antifungal and antibacterial activities.
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Using immunoproteomics to identify tumor-associated antigens (TAAs) as biomarkers in cancer immunodiagnosis.
Autoimmun Rev
PUBLISHED: 05-26-2013
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Since intracellular proteins involved in carcinogenesis have been shown to provoke autoantibody responses, autoantibodies can be used as probes in immunoproteomics to isolate, identify, and characterize potential tumor-associated antigens (TAAs). Once a TAA is identified, several approaches will be used to comprehensively characterize and validate the identified TAA/anti-TAA systems that are potential biomarkers in certain types of cancer. Our ultimate goal is to establish rigorous criteria for designation of an autoantibody to a TAA as a cancer biomarker, examine candidate TAAs for sensitivity and specificity of anti-TAA antibody response, and further develop customized TAA arrays that can be used to enhance anti-TAA antibody detection in cancer. This review will mainly focus on the recent advances in our studies using immunoproteomic approach to identify and characterize TAAs as biomarkers in cancer.
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Expression of autophagy and UPR genes in the developing brain during ethanol-sensitive and resistant periods.
Metab Brain Dis
PUBLISHED: 05-16-2013
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Fetal alcohol spectrum disorders (FASD) results from ethanol exposure to the developing fetus and is the leading cause of mental retardation. FASD is associated with a broad range of neurobehavioral deficits which may be mediated by ethanol-induced neurodegeneration in the developing brain. An immature brain is more susceptible to ethanol neurotoxicity. We hypothesize that the enhanced sensitivity of the immature brain to ethanol is due to a limited capacity to alleviate cellular stress. Using a third trimester equivalent mouse model of ethanol exposure, we demonstrated that subcutaneous injection of ethanol induced a wide-spread neuroapoptosis in postnatal day 4 (PD4) C57BL/6 mice, but had little effect on the brain of PD12 mice. We analyzed the expression profile of genes regulating apoptosis, and the pathways of ER stress response (also known as unfolded protein response, UPR) and autophagy during these ethanol-sensitive and resistant periods (PD4 versus PD12) using PCR microarray. The expression of pro-apoptotic genes, such as caspase-3, was much higher on PD4 than PD12; in contrast, the expression of genes that regulate UPR and autophagy, such as atf6, atg4, atg9, atg10, beclin1, bnip3, cebpb, ctsb, ctsd, ctss, grp78, ire1?, lamp, lc3 perk, pik3c3, and sqstm1 was significantly higher on PD12 than PD4. These results suggest that the vulnerability of the immature brain to ethanol could result from high expression of pro-apoptotic proteins and a deficiency in the stress responsive system, such as UPR and autophagy.
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Genetic variation in the PNPLA3 gene and hepatocellular carcinoma in USA: risk and prognosis prediction.
Mol. Carcinog.
PUBLISHED: 05-08-2013
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Nonalcoholic fatty liver disease (NAFLD) is an emerging epidemic with high prevalence in Western countries. Genome-wide association studies had reported that a variation in the patatin-like phospholipase domain containing 3 (PNPLA3) gene is associated with high susceptibility to NAFLD. However, the relationship between this variation and hepatocellular carcinoma (HCC) has not been well established. We investigated the impact of PNPLA3 genetic variation (rs738409: C>G) on HCC risk and prognosis in the United States by conducting a case-control study that included 257 newly diagnosed and pathologically confirmed Caucasian patients with HCC (cases) and 494 healthy controls. Multivariate logistics and Cox regression models were used to control for the confounding effects of HCC risk and prognostic factors. We observed higher risk of HCC for subjects with a homozygous GG genotype than for those with CC or CG genotypes, the adjusted odds ratio (OR) was 3.21 (95% confidence interval [CI], 1.68-6.41). We observed risk modification among individuals with diabetes mellitus (OR = 19.11; 95% CI, 5.13-71.20). The PNPLA3 GG genotype was significantly associated with underlying cirrhosis in HCC patients (OR = 2.48; 95% CI, 1.05-5.87). Moreover, GG allele represents an independent risk factor for death. The adjusted hazard ratio of the GG genotype was 2.11 (95% CI, 1.26-3.52) compared with CC and CG genotypes. PNPLA3 genetic variation (rs738409: C>G) may determine individual susceptibility to HCC development and poor prognosis. Further experimental investigations are necessary for thorough assessment of the hepatocarcinogenic role of PNPLA3.
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Cloning and characterization of the gene cluster required for beauvericin biosynthesis in Fusarium proliferatum.
Sci China Life Sci
PUBLISHED: 04-24-2013
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Beauvericin, a cyclohexadepsipeptide-possessing natural product with synergistic antifungal, insecticidal, and cytotoxic activities. We isolated and characterized the fpBeas gene cluster, devoted to beauvericin biosynthesis, from the filamentous fungus Fusarium proliferatum LF061. Targeted inactivation of the F. proliferatum genomic copy of fpBeas abolished the production of beauvericin. Comparative sequence analysis of the FpBEAS showed 74% similarity with the BbBEAS that synthesizes the cyclic trimeric ester beauvericin in Beauveria bassiana, which assembles N-methyl-dipeptidol monomer intermediates by the programmed iterative use of the nonribosomal peptide synthetase modules. Differences between the organization of the beauvericin loci in F. proliferaturm and B. bassiana revealed the mechanism for high production of beauvericin in F. proliferatum. Our work provides new insights into beauvericin biosynthesis, and may lead to beauvericin overproduction and creation of new analogs via synthetic biology approaches.
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Anionic polythiophene derivative as peroxidase mimetics and their application for detection of hydrogen peroxide and glucose.
Talanta
PUBLISHED: 04-19-2013
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In this paper, we discovered that the anionic polythiophenes derivative, poly[2-(3-thienyl)ethyloxy-4-butylsulfonate] (PTEBS), possesses intrinsic peroxidase-like activity that can catalyze the reaction of peroxidase substrate 3,3,5,5-tetramethylbenzidine (TMB) in the presence of H2O2 to produce a blue color reaction, which provides colorimetric detection of H2O2. PTEBS exhibits several advantages such as high catalytic efficiency, good stability, and rapid response over horseradish peroxidase (HRP). By coupling the oxidation of glucose catalyzed by glucose oxidase, a simple, inexpensive, highly sensitive and selective colorimetric method for glucose detection has been developed. The absorbance was proportional to the concentration of glucose in the range from 0.01 to 0.5mM with a detection limit of 0.004 mM. This work is not only of importance for a better understanding of the unique properties of polythiophenes derivative but also of great potential for medical diagnostics and biotechnology.
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Effects of ARHI on breast cancer cell biological behavior regulated by microRNA-221.
Tumour Biol.
PUBLISHED: 04-03-2013
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The aplysia ras homolog member I (ARHI) is a tumor suppressor gene and is downregulated in various cancers. The downregulation of ARHI was regulated by miR-221 in prostate cancer cell lines. However, it has not been reported whether ARHI is regulated by miR-221 in breast cancer. Here, we reported that the ARHI protein level was downregulated in breast cancer tissues and breast cancer cell lines. The overexpression of ARHI could inhibit cell proliferation and invasion and induce cell apoptosis. To address whether ARHI is regulated by miR-221 in breast cancer cell lines, the results in this study showed that a significant inverse correlation existed between ARHI and miR-221. MiR-221 displayed an upregulation in breast cancer tissues and breast cancer cell lines. The inhibition of miR-221 induced a significant upregulation of ARHI in MCF-7 cells. To prove a direct interaction between miR-221 and ARHI mRNA, ARHI 3UTR, which includes the potential target site for miR-221, was cloned downstream of the luciferase reporter gene of the pMIR-REPORT vector to generate the pMIR-ARHI-3UTR vector. The results confirmed a direct interaction of miR-221 with a target site on the 3UTR of ARHI. In conclusion, ARHI is a tumor suppressor gene that is downregulated in breast cancer. The overexpression of ARHI could inhibit breast cancer cell proliferation and invasion and induce cell apoptosis. This study demonstrated for the first time that the downregulation of ARHI in breast cancer cells could be regulated by miR-221.
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The antitumor activity of zinc(II) and copper(II) complexes with 5,7-dihalo-substituted-8-quinolinoline.
Eur J Med Chem
PUBLISHED: 03-04-2013
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[Zn?(ClQ)?(CH?OH)?] (1), [Zn(BrQ)?(H?O)?] (2), [Zn?(ClIQ)?] (3) and [Cu(BrQ)?] (4) (H-ClQ = 5,7-dichloro-8-hydroxylquinoline, H-BrQ = 5,7-dibromo-8-hydroxylquinoline, and H-ClIQ = 5-chloro-7-iodo-8-hydroxylquinoline) were synthesized. Compounds 1-4 showed high anti-proliferative cytotoxicities against BEL-7404, SK-OV-3, NCI-H460 tumor cells, and HL-7702 normal cells in vitro, with IC?? values in the 1.4 nM to 32.13 ?M range. Compounds 2-4 exhibited significantly enhanced cytotoxicity against BEL-7404 cell line, comparing with free 5,7-dihalo-8-quinolinol. Western blotting analysis showed that 2, 3 depleted mutant p53 protein in MDA-MB-231, and compound 2 decreased the ratio of Bcl-2/Bax in NCI-H460 significantly. The binding abilities of 1-4 to DNA were stronger than that of free quinolinol ligand. Intercalation is the probable binding mode for the complexes and free quinolinol ligands with DNA.
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Molecular characterization and expression analysis of a KIFC1-like kinesin gene in the testis of Eumeces chinensis.
Mol. Biol. Rep.
PUBLISHED: 02-19-2013
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The member of the kinesin-14 subfamily, KIFC1, is a carboxyl-terminal motor protein that plays an important role in the elongation of nucleus and acrosome biogenesis during the spermiogenesis of mammals. Here, we had cloned and sequenced the cDNA of a mammalian KIFC1 homologue (termed ec-KIFC1) from the total RNA of the testis of the reptile Eumeces chinensis. The full-length sequence was 2,339 bp that contained a 216 bp 5-untranslated region (5UTR), a 194 bp 3-untranslated region (3UTR) and a 1,929 bp open reading frame that encoded a special protein of 643 amino acids (aa). The calculated molecular weight of the putative ec-KIFC1 was 71 kDa and its estimated isoelectric point was 9.47. The putative ec-KIFC1 protein owns a tail domain from 1 to 116 aa, a stalk domain from 117 to 291 aa and a conserved carboxyl motor domain from 292 to 642 aa. Protein alignment demonstrated that ec-KIFC1 had 45.6, 42.8, 44.6, 36.9, 43.7, 46.4, 45.1, 55.6 and 49.8 % identity with its homologues in Mus musculus, Salmo salar, Danio rerio, Eriocheir sinensis, Rattus norvegicus, Homo sapiens, Bos taurus, Gallus gallus and Xenopus laevis, respectively. Tissue expression analysis showed the presence of ovary, heart, liver, intestine, oviduct, testis and muscle. The phylogenetic tree revealed that ec-KIFC1 was more closely related to vertebrate KIFC1 than to invertebrate KIFC1. In situ hybridization showed that the ec-KIFC1 mRNA was localized in the periphery of the nuclear membrane and the center of the nucleus in early spermatids. In mid spermatids, the ec-KIFC1 had abundant expression in the center of nucleus, and was expressed in the tail and the anterior part of spermatids. In the late spermatid, the nucleus gradually became elongated, and the ec-KIFC1 mRNA signal was still centralized in the nucleus. In mature spermatids, the signal of the ec-KIFC1 gradually became weak, and was mainly located at the tail of spermatids. Therefore, the ec-KIFC1 probably plays a critical role in the spermatogenesis of E. chinensis.
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High antitumor activity of 5,7-dihalo-8-quinolinolato cerium complexes.
Eur J Med Chem
PUBLISHED: 02-04-2013
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Three cerium complexes: [Ce(ClQ)4] (1) (H-ClQ=5,7-dichloro-8-hydroxylquinoline), [Ce(ClIQ)4]·CH2Cl2·0.5H2O (2) (H-ClIQ=5-chloro-7-iodo-8-hydroxylquinoline) and [Ce2(BrQ)4(H-BrQ)(H2O)3Cl2]·1.5H2O (3) (H-BrQ=5,7-dibromo-8-hydroxylquinoline) were synthesized. The structures of 1 and 2 are mononuclear whereas 3 has a binuclear structure. Compared with the H-ClQ, H-ClIQ and H-BrQ, complexes 1-3 exhibited significantly higher cytotoxicity (IC50=0.09-5.23 ?M) to SK-OV-3 and BEL-7404, 1 and 2 exhibited higher cytotoxicity to NCI-H460. Most the complexes and ligands exhibited higher cytotoxicity than cisplatin. Complexes 1-3 are much more sensitive to SK-OV-3 than to human normal liver cell HL-7702. Their antitumor activities were achieved through cell apoptosis and arrest at G0/G1-phase. Studies on the binding properties of 1-3 to DNA indicate that intercalation is the most probable binding mode.
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Hypoxia/oxidative stress alters the pharmacokinetics of CPU86017-RS through mitochondrial dysfunction and NADPH oxidase activation.
Acta Pharmacol. Sin.
PUBLISHED: 01-24-2013
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Aim:Hypoxia/oxidative stress can alter the pharmacokinetics (PK) of CPU86017-RS, a novel antiarrhythmic agent. The aim of this study was to investigate the mechanisms underlying the alteration of PK of CPU86017-RS by hypoxia/oxidative stress.Methods:Male SD rats exposed to normal or intermittent hypoxia (10% O2) were administered CPU86017-RS (20, 40 or 80 mg/kg, ig) for 8 consecutive days. The PK parameters of CPU86017-RS were examined on d 8. In a separate set of experiments, female SD rats were injected with isoproterenol (ISO) for 5 consecutive days to induce a stress-related status, then CPU86017-RS (80 mg/kg, ig) was administered, and the tissue distributions were examined. The levels of Mn-SOD (manganese containing superoxide dismutase), endoplasmic reticulum (ER) stress sensor proteins (ATF-6, activating transcription factor 6 and PERK, PRK-like ER kinase) and activation of NADPH oxidase (NOX) were detected with Western blotting. Rat liver microsomes were incubated under N2 for in vitro study.Results:The Cmax, t1/2, MRT (mean residence time) and AUC (area under the curve) of CPU86017-RS were significantly increased in the hypoxic rats receiving the 3 different doses of CPU86017-RS. The hypoxia-induced alteration of PK was associated with significantly reduced Mn-SOD level, and increased ATF-6, PERK and NOX levels. In ISO-treated rats, the distributions of CPU86017-RS in plasma, heart, kidney, and liver were markedly increased, and NOX levels in heart, kidney, and liver were significantly upregulated. Co-administration of the NOX blocker apocynin eliminated the abnormalities in the PK and tissue distributions of CPU86017-RS induced by hypoxia/oxidative stress. The metabolism of CPU86017-RS in the N2-treated liver microsomes was significantly reduced, addition of N-acetylcysteine (NAC), but not vitamin C, effectively reversed this change.Conclusion:The altered PK and metabolism of CPU86017-RS induced by hypoxia/oxidative stress are produced by mitochondrial abnormalities, NOX activation and ER stress; these abnormalities are significantly alleviated by apocynin or NAC.
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Polarity in migrating neurons is related to a mechanism analogous to cytokinesis.
Curr. Biol.
PUBLISHED: 01-11-2013
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Migrating neurons are bipolar, with a leading process and a trailing process [1]. The proximal region of the leading process displays a concentration of F-actin that contributes to the advance of the soma and the centrosome [2-7]. Here, we show that kinesin-6, a microtubule-based motor protein best known for its role in cytokinesis, also concentrates in this region. Depletion of kinesin-6 results in multipolar neurons that either are stationary or continuously change their direction of movement. In such neurons, F-actin no longer concentrates in a single process. During cytokinesis, kinesin-6 forms a complex with a Rho-family GTPase-activating protein called MgcRacGAP to signal to the actin cytoskeleton so that cortical movements are concentrated in the cleavage furrow [8-13]. During neuronal migration, MgcRacGap also concentrates in the proximal region of the leading process, and inhibition of its activity results in a phenotype similar to kinesin-6 depletion. We conclude that neuronal migration utilizes a cytoskeletal pathway analogous to cytokinesis, with kinesin-6 signaling through MgcRacGap to the actin cytoskeleton to constrain process number and restrict protrusive activity to a single leading process, thus resulting in a bipolar neuron able to move in a directed fashion.
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An Antibody-based Blood Test Utilizing a Panel of Biomarkers as a New Method for Improved Breast Cancer Diagnosis.
Biomark Cancer
PUBLISHED: 01-01-2013
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In order to develop a new tool for diagnosis of breast cancer based on autoantibodies against a panel of biomarkers, a clinical trial including blood samples from 507 subjects was conducted. All subjects showed a breast abnormality on exam or breast imaging and final biopsy pathology of either breast cancer patients or healthy controls. Using an enzyme-linked immunosorbent assay, the samples were tested for autoantibodies against a predetermined number of biomarkers in various models that were used to determine a diagnosis, which was compared to the clinical status. Our new assay achieved a sensitivity of 95.2% [CI = 92.8-96.8%] at a fixed specificity of 49.5%. Receiver-operator characteristic curve analysis showed an area under the curve of 80.1% [CI = 72.6-87.6%]. These results suggest that a blood test which is based on models comprising several autoantibodies to specific biomarkers may be a new and novel tool for improving the diagnostic evaluation of breast cancer.
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?3 Integrin Promotes TGF-?1/H2O2/HOCl-Mediated Induction of Metastatic Phenotype of Hepatocellular Carcinoma Cells by Enhancing TGF-?1 Signaling.
PLoS ONE
PUBLISHED: 01-01-2013
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In addition to being an important mediator of migration and invasion of tumor cells, ?3 integrin can also enhance TGF-?1 signaling. However, it is not known whether ?3 might influence the induction of metastatic phenotype of tumor cells, especially non-metastatic tumor cells which express low level of ?3. Here we report that H2O2 and HOCl, the reactive oxygen species produced by neutrophils, could cooperate with TGF-?1 to induce metastatic phenotype of non-metastatic hepatocellular carcinoma (HCC) cells. TGF-?1/H2O2/HOCl, but not TGF-?1 or H2O2/HOCl, induced ?3 expression by triggering the enhanced activation of p38 MAPK. Intriguingly, ?3 in turn promoted TGF-?1/H2O2/HOCl-mediated induction of metastatic phenotype of HCC cells by enhancing TGF-?1 signaling. ?3 promoted TGF-?1/H2O2/HOCl-induced expression of itself via positive feed-back effect on p38 MAPK activation, and also promoted TGF-?1/H2O2/HOCl-induced expression of ?3 and SNAI2 by enhancing the activation of ERK pathway, thus resulting in higher invasive capacity of HCC cells. By enhancing MAPK activation, ?3 enabled TGF-?1 to augment the promoting effect of H2O2/HOCl on anoikis-resistance of HCC cells. TGF-?1/H2O2/HOCl-induced metastatic phenotype was sufficient for HCC cells to extravasate from circulation and form metastatic foci in an experimental metastasis model in nude mice. Inhibiting the function of ?3 could suppress or abrogate the promoting effects of TGF-?1/H2O2/HOCl on invasive capacity, anoikis-resistance, and extravasation of HCC cells. These results suggest that ?3 could function as a modulator to promote TGF-?1/H2O2/HOCl-mediated induction of metastatic phenotype of non-metastatic tumor cells, and that targeting ?3 might be a potential approach in preventing the induction of metastatic phenotype of non-metastatic tumor cells.
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Toxicity and Immunogenicity of Enterotoxigenic Escherichia coli Heat-Labile and Heat-Stable Toxoid Fusion 3xSTaA14Q-LTS63K/R192G/L211A in a Murine Model.
PLoS ONE
PUBLISHED: 01-01-2013
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Diarrhea is the second leading cause of death to young children. Enterotoxigenic Escherichia coli (ETEC) are the most common bacteria causing diarrhea. Adhesins and enterotoxins are the virulence determinants in ETEC diarrhea. Adhesins mediate bacterial attachment and colonization, and enterotoxins including heat-labile (LT) and heat-stable type Ib toxin (STa) disrupt fluid homeostasis in host cells that leads to fluid hyper-secretion and diarrhea. Thus, adhesins and enterotoxins have been primarily targeted in ETEC vaccine development. A recent study reported toxoid fusions with STa toxoid (STaP13F) fused at the N- or C-terminus, or inside the A subunit of LTR192G elicited neutralizing antitoxin antibodies, and suggested application of toxoid fusions in ETEC vaccine development (Liu et al., Infect. Immun. 79:4002-4009, 2011). In this study, we generated a different STa toxoid (STaA14Q) and a triple-mutant LT toxoid (LTS63K/R192G/L211A, tmLT), constructed a toxoid fusion (3xSTaA14Q-tmLT) that carried 3 copies of STaA14Q for further facilitation of anti-STa immunogenicity, and assessed antigen safety and immunogenicity in a murine model to explore its potential for ETEC vaccine development. Mice immunized with this fusion antigen showed no adverse effects, and developed antitoxin antibodies particularly through the IP route. Anti-LT antibodies were detected and were shown neutralizing against CT in vitro. Anti-STa antibodies were also detected in the immunized mice, and serum from the IP immunized mice neutralized STa toxin in vitro. Data from this study indicated that toxoid fusion 3xSTaA14Q-tmLT is safe and can induce neutralizing antitoxin antibodies, and provided helpful information for vaccine development against ETEC diarrhea.
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Clinical and imaging features of pulmonary artery sling in infants without significant hemodynamic changes.
Chin. Med. J.
PUBLISHED: 11-18-2011
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Pulmonary artery sling (PAS) is a rare congenital heart anomaly and may cause unexplained respiratory symptoms in infants. Since the non-specific respiratory symptoms of PAS may lead to misdiagnosis, the aim of this study was to clarify the clinical and imaging features of this disease for timely diagnosis and treatment.
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Modeling drug exposure data in electronic medical records: an application to warfarin.
AMIA Annu Symp Proc
PUBLISHED: 10-22-2011
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Identification of patients drug exposure information is critical to drug-related research that is based on electronic medical records (EMRs). Drug information is often embedded in clinical narratives and drug regimens change frequently because of various reasons like intolerance or insurance issues, making accurate modeling challenging. Here, we developed an informatics framework to determine patient drug exposure histories from EMRs by combining natural language processing (NLP) and machine learning (ML) technologies. Our framework consists of three phases: 1) drug entity recognition - identifying drug mentions; 2) drug event detection - labeling drug mentions with a status (e.g., "on" or "stop"); and 3) drug exposure modeling - predicting if a patient is taking a drug at a given time using the status and temporal information associated with the mentions. We applied the framework to determine patient warfarin exposure at hospital admissions and achieved 87% precision, 79% recall, and an area under the receiver-operator characteristic curve of 0.93.
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