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Find video protocols related to scientific articles indexed in Pubmed.
Investigating the molecular genetic basis of heterosis for internode expansion in maize by microRNA transcriptomic deep sequencing.
Funct. Integr. Genomics
PUBLISHED: 11-03-2014
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Heterosis has been used widely in the breeding of maize and other crops and plays an important role in increasing yield, improving quality, and enhancing stress resistance, but its molecular mechanism is far from clear. To determine whether microRNA (miRNA)-dependent gene regulation is responsible for heterosis of elongating internodes below the ear and ear height in maize, a deep-sequencing strategy was applied to the elite hybrid Xundan20, which is currently cultivated widely in China, and its two parents. RNA was extracted from the eighth internode because it shows clear internode length heterosis. A total of 99 conserved maize miRNAs were detected in both the hybrid and parental lines. Most of these miRNAs were expressed nonadditively in the hybrid compared with its parental lines. These results indicated that miRNAs might participate in heterosis during internode expansion in maize and exert an influence on ear and plant height via the repression of their target genes. In total, eight novel miRNAs belonging to four miRNA families were predicted in the expanding internode. Global repression of miRNAs in the hybrid, which might result in enhanced gene expression, might be one reason why the hybrid shows longer internodes and taller seedlings compared with its parental lines.
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[Isolation, identification of TJ430 strain and characterization of its antifungal metabolite].
Wei Sheng Wu Xue Bao
PUBLISHED: 10-03-2014
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To screen new agro-antibiotics, rare actinomycetes were isolated by improved separation methods from soil samples and the chemical structure of the antifungal active product was elucidated.
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[Research on increasing X-ray protection capability based on photonic crystal technology].
Sheng Wu Yi Xue Gong Cheng Xue Za Zhi
PUBLISHED: 09-16-2014
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Light cannot be propagated within the range of photonic crystal band gaps. Based on this unique property, we proposed a method to improve anti-radiation capability through one-dimensional photonic crystal coating. Using transmission matrix method, we determined the appropriate dielectric materials, thickness and periodic numbers of photonic crystals through Matlab programming simulation. Then, compound one-dimensional photonic crystal coating was designed which was of high anti-radiation rate within the range of X-ray. As is shown through simulation experiments, the reflection rate against X-ray was higher than 90 percent, and the desired anti-radiation effect was achieved. Thus, this method is able to help solve the technical problems facing the inorganic lead glass such as thickness, weightiness, costliness, high lead equivalent, low transparency and high cost. This method has won China's national invention patent approval, and the patent number is 201220228549.2.
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Depolarized inactivation overcomes impaired activation to produce DRG neuron hyperexcitability in a Nav1.7 mutation in a patient with distal limb pain.
J. Neurosci.
PUBLISHED: 09-12-2014
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Sodium channel Nav1.7, encoded by SCN9A, is expressed in DRG neurons and regulates their excitability. Genetic and functional studies have established a critical contribution of Nav1.7 to human pain disorders. We have now characterized a novel Nav1.7 mutation (R1279P) from a female human subject with distal limb pain, in which depolarized fast inactivation overrides impaired activation to produce hyperexcitability and spontaneous firing in DRG neurons. Whole-cell voltage-clamp recordings in human embryonic kidney (HEK) 293 cells demonstrated that R1279P significantly depolarizes steady-state fast-, slow-, and closed-state inactivation. It accelerates deactivation, decelerates inactivation, and facilitates repriming. The mutation increases ramp currents in response to slow depolarizations. Our voltage-clamp analysis showed that R1279P depolarizes channel activation, a change that was supported by our multistate structural modeling. Because this mutation confers both gain-of-function and loss-of-function attributes on the Nav1.7 channel, we tested the impact of R1279P expression on DRG neuron excitability. Current-clamp studies reveal that R1279P depolarizes resting membrane potential, decreases current threshold, and increases firing frequency of evoked action potentials within small DRG neurons. The populations of spontaneously firing and repetitively firing neurons were increased by expressing R1279P. These observations indicate that the dominant proexcitatory gating changes associated with this mutation, including depolarized steady-state fast-, slow-, and closed-state inactivation, faster repriming, and larger ramp currents, override the depolarizing shift of activation, to produce hyperexcitability and spontaneous firing of nociceptive neurons that underlie pain.
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Sphingobacterium gobiense sp. nov., isolated from soil of the Gobi Desert.
Int. J. Syst. Evol. Microbiol.
PUBLISHED: 09-10-2014
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A Gram-stain-negative, short rod-shaped, non-motile, non-spore-forming bacterial strain, designated H7T, was isolated from the Gobi desert, Xinjiang Province of China. The temperature, NaCl and pH ranges for growth were 8-40 °C (optimum, 30 °C), NaCl 0-5% and pH 6-10 (optimum, pH 7). Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain H7T belonged to the genus Sphingobacterium and showed highest sequence similarity (91%) to Sphingobacterium composti DSM 18850.The DNA G+C content (44.3 mol%), MK-7 as the predominant respiratory quinone, and C16?:?1?7c and/or C16?:?1?6c, iso-C15:?0 and iso-C17:?03-OH as major fatty acids supported the affiliation of strain H7T to the genus Sphingobacterium while phenotypic data indicated it was a representative of a novel species for which the name Sphingobacterium gobiense sp. nov is proposed. The type strain is H7 T (?=ACCC 05757T=KCTC 32293T).
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Neuroprotective effects of viral overexpression of microRNA-22 in rat and cell models of cerebral ischemia-reperfusion injury.
J. Cell. Biochem.
PUBLISHED: 09-04-2014
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Several studies have reported that microRNA (MIR) is involved in the pathogenesis and progression of ischemic diseases, including cerebral ischemia, and that MIR-22 may inhibit the inflammatory response and cell apoptosis, which contribute to ischemia/reperfusion (I/R) injury. However, the specific function of MIR-22 in cerebral I/R injury remains far from clear. This study aimed to examine the potential protective effect of MIR-22 against cerebral I/R injury and its mechanism. As predicted, adenovirus-mediated MIR-22 overexpression markedly reduced the neurological score and infarct size (P<0.05). We demonstrated that MIR-22 overexpression resulted in a reduction in inflammatory cytokines TNF-á, IL-6, COX-2 and iNOS£¬whereas the level of IL-10 was enhanced. MIR-22 overexpression significantly inhibited NF-êB activity by decreasing NF-êB coactivator NCOA1 expres sion. Furthermore, we found that MIR-22 could reduce the apoptotic rate of cortical neurons. Caspase-3 activity was inhibited by MIR-22, and the expression of the anti-apoptosis gene Bcl-2 in neurons was increased and that of the pro-apoptosis gene Bax decreased following MIR-22 overexpression. Our results suggest that MIR-22 could be used to treat cerebral I/R injury and that its neuroprotective effect may be attributed to a reduction in inflammation and apoptosis. J. Cell. Biochem. © 2014 Wiley Periodicals, Inc.
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Addressing Criticisms of Existing Predictive Bias Research: Cognitive Ability Test Scores Still Overpredict African Americans' Job Performance.
J Appl Psychol
PUBLISHED: 08-25-2014
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Predictive bias studies have generally suggested that cognitive ability test scores overpredict job performance of African Americans, meaning these tests are not predictively biased against African Americans. However, at least 2 issues call into question existing over-/underprediction evidence: (a) a bias identified by Aguinis, Culpepper, and Pierce (2010) in the intercept test typically used to assess over-/underprediction and (b) a focus on the level of observed validity instead of operational validity. The present study developed and utilized a method of assessing over-/underprediction that draws on the math of subgroup regression intercept differences, does not rely on the biased intercept test, allows for analysis at the level of operational validity, and can use meta-analytic estimates as input values. Therefore, existing meta-analytic estimates of key parameters, corrected for relevant statistical artifacts, were used to determine whether African American job performance remains overpredicted at the level of operational validity. African American job performance was typically overpredicted by cognitive ability tests across levels of job complexity and across conditions wherein African American and White regression slopes did and did not differ. Because the present study does not rely on the biased intercept test and because appropriate statistical artifact corrections were carried out, the present study's results are not affected by the 2 issues mentioned above. The present study represents strong evidence that cognitive ability tests generally overpredict job performance of African Americans. (PsycINFO Database Record (c) 2014 APA, all rights reserved).
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Prognostic factors for patients with atypical or malignant meningiomas treated at a single center.
Neurosurg Rev
PUBLISHED: 08-21-2014
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The purpose of this study is to summarize our experience in managing patients with an atypical or malignant meningioma at our institution, with a specific focus on determining the prognostic factors for treatment outcome. We reviewed the records of 126 patients with atypical or malignant meningiomas from January 2001 to August 2011. Data collected included gender, age, Karnofsky Performance Scale (KPS) score, pathology results, cleavability, and bone invasion. The symptoms and signs were recorded for further outcome analysis. There were 37 malignant meningiomas and 89 atypical meningiomas. Total resection (Simpson grade I-II) was achieved in 80.9 % of atypical patients (n?=?72) and 67.6 % of malignant patients (n?=?25). Forty patients (44.9 %) in the atypical group underwent radiotherapy after surgery, while 26 (70.2 %) patients underwent radiotherapy in the malignant group. The median follow-up duration was 25 months. Patients with a secondary tumor had a much shorter progression-free survival (PFS) than those with a primary tumor in the malignant group. The malignant meningioma group had lower overall survival. Progression-free survival for patients in the malignant group who received postoperative radiotherapy was longer than that for those who did not receive radiotherapy. In conclusion, total resection of the tumor was important because patients with a secondary tumor were much more likely to have recurrence than patients with a primary tumor in the atypical and malignant meningioma groups. Also, radiotherapy should be performed after surgery for a malignant meningioma.
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Survival Benefit of Neoadjuvant Chemotherapy for Resectable Cancer of the Gastric and Gastroesophageal Junction: A Meta-Analysis.
J. Clin. Gastroenterol.
PUBLISHED: 08-20-2014
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The objective of the present meta-analysis was to estimate the magnitude of survival benefits of neoadjuvant chemotherapy (NAT) in resectable cancer of the gastric and gastroesophageal junction.
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Apoptotic effect of genistein on human colon cancer cells via inhibiting the nuclear factor-kappa B (NF-?B) pathway.
Tumour Biol.
PUBLISHED: 08-16-2014
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Genistein possesses a wide variety of biological activities, and it is best known for its ability to inhibit cancer progression. Its cancer-preventive effect has been attributed to various mechanisms, including the induction of cell cycle arrest and apoptosis as well as the antioxidant functions. Nuclear factor kappa-B (NF-?B) is a signaling pathway that controls transcriptional activation of genes important for the tight regulation of many cellular processes and is aberrantly expressed in many types of cancer. Inhibitors of NF-?B pathway have shown potential anti-tumor activities. However, it is not fully elucidated in colon cancer. In the present study, we demonstrated that genistein could induce apoptosis in human colon cancer LoVo and HT-29 cells through inhibiting NF-?B pathway, as well as downregulation of Bcl-2 and upregulation of Bax, thus providing basis for clinical application of genistein in colon cancer cases.
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A DNA-templated synthesis of encoded small molecules by DNA self-assembly.
Chem. Commun. (Camb.)
PUBLISHED: 08-06-2014
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We report a novel method for the synthesis of DNA-encoded libraries without the need for discrete DNA template. Reactant DNAs self-assemble to enable chemical reactions and photo-cleavage transfers the product to the DNA terminus, making it suitable for the subsequent affinity-based selection and hit deconvolution.
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Developmental heterogeneity of cardiac fibroblasts does not predict pathological proliferation and activation.
Circ. Res.
PUBLISHED: 07-18-2014
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Fibrosis is mediated partly by extracellular matrix-depositing fibroblasts in the heart. Although these mesenchymal cells are reported to have multiple embryonic origins, the functional consequence of this heterogeneity is unknown.
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Clostridium difficile carriage in hospitalized cancer patients: a prospective investigation in eastern China.
BMC Infect. Dis.
PUBLISHED: 07-16-2014
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Clostridium difficile carriage has been considered as a potential source for the deadly infection, but its role in cancer patients is still unclear. We aimed to identify the clinical and immunological factors that are related to C. difficile carriage in Chinese cancer patients.
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Isolation, identification, and gp85 characterization of a subgroup A avian leukosis virus from a contaminated live Newcastle Disease virus vaccine, first report in China.
Poult. Sci.
PUBLISHED: 07-07-2014
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To identify if any exogenous avian leukosis virus (ALV) exists in a live vaccine of poultry according to the directives of the Ministry of Agriculture of the People's Republic of China, a live vaccine strain of the Newcastle disease virus (NDV) was neutralized using an anti-NDV antibody, and was subsequently used to inoculate DF-1 cells to investigate the presence of exogenous ALV. The DF-1 cells were cultured for 21 d and subsequently screened using an ELISA for the p27 antigen of the ALV. An exogenous ALV, designated ALV-NDVP4, was identified. The nucleotide sequence of the gp85 gene of the ALV-NDVP4 was compared with those of the various subgroups of the ALV. The amino acid sequence identities for the predicted gp85 of the ALV-NDVP4 and those of the ALV reference strains ranged from 88.2 to 99.5% for the 12 of the subgroup A strains of ALV (ALV-A) and from 82.7 to 87.4% for the B, C, D, and E subgroup strains. The amino acid sequence identities for the gp85 of the ALV-NDVP4 and those of the subgroup J reference strains ranged from 48.7 to 49.9%. The ALV-NDVP4 shared the highest level of homology with the SDAU09C3 strain of ALV-A, which was isolated in China, suggesting a common origin. This is the first report of ALV-A contamination in a live vaccine for poultry in China. Our findings highlight the need for improved monitoring methods for poultry vaccine production.
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Comprehensive analysis of cystatin family genes suggests their putative functions in sexual reproduction, embryogenesis, and seed formation.
J. Exp. Bot.
PUBLISHED: 07-04-2014
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Cystatins are tightly bound and reversible inhibitors of cysteine proteases in C1A and C13 peptidase families, which have been identified in several species and shown to function in vegetative development and response to biotic/abiotic stresses in plants. Recent work revealed their critical role in regulating programmed cell death during embryogenesis in tobacco and suggested their more comprehensive roles in the process of sexual plant reproduction, although little is known about cystatin family genes in the processes. Here, 10 cystatin family genes in Nicotiana tabacum were identified using an expressed sequence tag (EST)-based gene clone strategy. Analysis of their biochemical properties showed that nine of them have the potency to inhibit the activities of both commercial cathepsin L-like proteases and extracted cysteine proteases from seeds, but with different K i values depending on the types of proteases and the developmental stages of the seed tested. This suggests that cystatin-dependent cathepsin L-like proteolytic pathways are probably important for early seed development. Comprehensive expression profile analysis revealed that cystatin family genes showed manifold variations in their transcription levels in different plant cell types, including the sperm, egg, and zygote, especially in the embryo and seed at different developmental stages. More interestingly, intracellular localization analysis of each cystatin revealed that most members of cystatin families are recognized as secretory proteins with signal peptides that direct them to the endoplasmic reticulum. These results suggest their widespread roles in cell fate determination and cell-cell communication in the process of sexual reproduction, especially in gamete and embryo development, as well as in seed formation.
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iTEP nanoparticle-delivered salinomycin displays an enhanced toxicity to cancer stem cells in orthotopic breast tumors.
Mol. Pharm.
PUBLISHED: 07-01-2014
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Salinomycin (Sali) has selective toxicity to cancer stem cells (CSCs), a subpopulation of cancer cells that have been recently linked with tumor multidrug resistance (MDR). To utilize its selective toxicity for cancer therapy, we sought to devise a nanoparticle (NP) carrier to deliver Sali to solid tumors through the enhanced permeability and retention effect and, hence, to increase its exposure to CSCs. First, hydrophobic Sali was conjugated to a hydrophilic, immune-tolerant, elastin-like polypeptide (iTEP); the amphiphilic iTEP-Sali conjugates self-assemble into NPs. Next, free Sali was encapsulated into the NPs alone or with two additives, N,N-dimethylhexylamine (DMHA) and ?-tocopherol. The coencapsulation significantly improved the loading efficiency and release profile of Sali. The resulting NPs of the coencapsulation, termed as iTEP-Sali NP3s, have an in vitro release half-life of 4.1 h, four times longer than iTEP-Sali NP2s, the NPs that have encapsulated Sali only. Further, the NP3 formulation increases the plasma area under curve and the tumor accumulation of Sali by 10 and 2.4 times, respectively. Lastly, these improved pharmacokinetic and tumor accumulation profiles are consistent with a boost of CSC-elimination effect of Sali in vivo. In NP3-treated 4T1 orthotopic tumors, the mean CSC frequency is 55.62%, a significant reduction from the mean frequencies of untreated tumors, 75.00%, or free Sali-treated tumors, 64.32%. The CSC-elimination effect of the NP3 can further translate to a delay of tumor growth. Given the role of CSCs in driving tumor MDR and recurrence, it could be a promising strategy to add the NP3 to conventional cancer chemotherapies to prevent or reverse the MDR.
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Enantioselective and regioselective pyrone Diels-Alder reactions of vinyl sulfones: total synthesis of (+)-cavicularin.
Angew. Chem. Int. Ed. Engl.
PUBLISHED: 06-26-2014
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The total synthesis of (+)-cavicularin is described. The synthesis features an enantio- and regioselective pyrone Diels-Alder reaction of a vinyl sulfone to construct the cyclophane architecture of the natural product. The Diels-Alder substrate was prepared by a regioselective one-pot three-component Suzuki reaction of a non-symmetric dibromoarene.
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Increased incidence of head and neck cancer in liver transplant recipients: a meta-analysis.
BMC Cancer
PUBLISHED: 06-12-2014
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It is unclear whether liver transplantation is associated with an increased incidence of post-transplant head and neck cancer. This comprehensive meta-analysis evaluated the association between liver transplantation and the risk of head and neck cancer using data from all available studies.
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Site-specific glycan microheterogeneity of inter-alpha-trypsin inhibitor heavy chain H4.
J. Proteome Res.
PUBLISHED: 06-12-2014
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Inter-alpha-trypsin inhibitor heavy chain H4 (ITIH4) is a 120 kDa acute-phase glycoprotein produced primarily in the liver, secreted into the blood, and identified in serum. ITIH4 is involved in liver development and stabilization of the extracellular matrix (ECM), and its expression is altered in liver disease. In this study, we aimed to characterize glycosylation of recombinant and serum-derived ITIH4 using analytical mass spectrometry. Recombinant ITIH4 was analyzed to optimize glycopeptide analyses, followed by serum-derived ITIH4. First, we confirmed that the four ITIH4 N-X-S/T sequons (N81, N207, N517, and N577) were glycosylated by treating ITIH4 tryptic/GluC glycopeptides with PNGaseF in the presence of (18)O water. Next, we performed glycosidase-assisted LC-MS/MS analysis of ITIH4 trypsin-GluC glycopeptides enriched via hydrophilic interaction liquid chromatography to characterize ITIH4 N-glycoforms. While microheterogeneity of N-glycoforms differed between ITIH4 protein expressed in HEK293 cells and protein isolated from serum, occupancy of N-glycosylation sites did not differ. A fifth N-glycosylation site was discovered at N274 with the rare nonconsensus NVV motif. Site N274 contained high-mannose N-linked glycans in both serum and recombinant ITIH4. We also identified isoform-specific ITIH4 O-glycoforms and documented that utilization of O-glycosylation sites on ITIH4 differed between the cell line and serum.
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Synergetic effects of subgroup J avian leukosis virus and reticuloendotheliosis virus co-infection on growth retardation and immunosuppression in SPF chickens.
Vet. Microbiol.
PUBLISHED: 06-10-2014
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To further understand the effect of co-infection of subgroup J avian leukosis virus (ALV-J) and reticuloendotheliosis virus (REV) in specific-pathogen-free (SPF) white leghorn chickens, the experiment was made to study the pathogenicity, the weight of body and immune organs, response to newcastle disease virus (NDV) and avian influenza virus subtype H9 (AIV-H9) vaccination. Chickens were randomly divided into four groups, which includes injection groups (REV, ALV-J, REV plus ALV-J), and negative control group. The pathogenesis experiments indicated that chickens co-infected with REV and ALV-J had significantly higher mortality rate than those of the chickens infected with REV or ALV-J alone (P<0.05). Chickens inoculated with REV and ALV-J had significantly lower weights than chickens in all other groups (P<0.05). There were no significant differences between the two single infection groups and co-infection group (P>0.05) on bursa and thymus over body wt ratios, however, chickens co-infected with REV and ALV-J had significantly lower titers than REV-infected chickens and ALV-J-infected chickens on HI antibody titers to ND and AIV-H9 after vaccination (P<0.05). These findings suggested that the co-infection of REV and ALV-J caused more serious growth retardation and immunosuppression in SPF chickens.
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Multifunctional disulfide-based cationic dextran conjugates for intravenous gene delivery targeting ovarian cancer cells.
Mol. Pharm.
PUBLISHED: 06-10-2014
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A folate-decorated, disulfide-based cationic dextran conjugate having dextran as the main chain and disulfide-linked 1,4-bis(3-aminopropyl)piperazine (BAP) residues as the grafts was designed and successfully prepared as a multifunctional gene delivery vector for targeted gene delivery to ovarian cancer SKOV-3 cells in vitro and in vivo. Initially, a new bioreducible cationic polyamide (denoted as pSSBAP) was prepared by polycondensation reaction of bis(p-nitrophenyl)-3,3'-dithiodipropanoate, a disulfide-containing monomer, and BAP. It was found that the pSSBAP was highly efficient for in vitro gene delivery against MCF-7 and SKOV-3 cell lines. Subsequently, two cationic dextran conjugates with different amounts of BAP residues (denoted as Dex-SSBAP6 and Dex-SSBAP30, respectively) were synthesized by coupling BAP to disulfide-linked carboxylated dextran or coupling pSSBAP-oligomer to p-nitrophenyl carbonated dextran. Both two conjugates were able to bind DNA to form nanosized polyplexes with an improved colloidal stability in physiological conditions. The polyplexes, however, were rapidly dissociated to liberate DNA in a reducing environment. In vitro transfection experiments revealed that the polyplexes of Dex-SSBAP30 efficiently transfected SKOV-3 cells, yielding transfection efficiency that is comparable to that of linear polyethylenimine or lipofectamine 2000. AlamarBlue assay showed that the conjugates had low cytotoxicity in vitro at a high concentration of 100 mg/L. Further, Dex-SSBAP30 has primary amine side groups and thus allows for folate (FA) conjugation, yielding FA-coupled Dex-SSBAP30 (Dex-SSBAP30-FA). It was found that Dex-SSBAP30-FA was efficient for targeted gene delivery to SKOV-3 tumor xenografted in a nude mouse model by intravenous injection, inducing a higher level of gene expression in the tumor as compared to Dex-SSBAP30 lacking FA and comparable gene expression to linear polyethylenimine as one of the most efficient polymeric vectors for intravenous gene delivery in vivo. Disulfide-based cationic dextran system thus has a high potential for intravenous gene delivery toward cancer gene therapy.
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Biweekly S-1 plus paclitaxel (SPA) as second-line chemotherapy after failure from fluoropyrimidine and platinum in advanced gastric cancer: a phase II study.
Cancer Chemother. Pharmacol.
PUBLISHED: 06-03-2014
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Second-line chemotherapy is now considered a standard therapy option in patients with advanced gastric cancer (AGC) who failed from first-line chemotherapy. Single agents, such as irinotecan, docetaxel or paclitaxel, provided an overall response rate of about 10 %. However, the efficacy was not satisfactory. The authors conducted a phase II study to investigate biweekly regimen of S-1 plus paclitaxel in Chinese AGC in second-line setting, with response rate as the primary end point.
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The mechanism for increasing the oral bioavailability of poorly water-soluble drugs using uniform mesoporous carbon spheres as a carrier.
Drug Deliv
PUBLISHED: 05-30-2014
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Abstract Uniform mesoporous carbon spheres (UMCS) were used as a carrier to improve the bioavailability of the model drug, celecoxib (CEL). Furthermore, we investigated the mechanism responsible for the improved bioavailability of CEL. The association, adhesion and uptake of UMCS by intestinal epithelial cells were studied by transmission electron microscopy (TEM), fluorescence-activated cell sorting (FACS) and laser confocal scanning microscopy (LCSM). UMCS was found to promote cellular uptake of CEL. Drug transport in Caco-2 cell monolayers proved that UMCS can significantly reduce the rate of drug efflux and improve CEL permeability. The dissolution rate of CEL from drug-loaded samples was markedly improved compared with pure crystalline CEL; moreover, oral bioavailability of CEL loaded into UMCS was also markedly improved compared with that of commercially available capsules. UMCS indicates the advantages and potential of this method to achieve improved oral absorption by increasing the dissolution rate, cellular uptake and permeability of the drug.
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A high-throughput, multiplexed assay for superfamily-wide profiling of enzyme activity.
Nat. Chem. Biol.
PUBLISHED: 05-29-2014
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The selectivity of an enzyme inhibitor is a key determinant of its usefulness as a tool compound or its safety as a drug. Yet selectivity is never assessed comprehensively in the early stages of the drug discovery process, and only rarely in the later stages, because technical limitations prohibit doing otherwise. Here, we report EnPlex, an efficient, high-throughput method for simultaneously assessing inhibitor potency and specificity, and pilot its application to 96 serine hydrolases. EnPlex analysis of widely used serine hydrolase inhibitors revealed numerous previously unrecognized off-target interactions, some of which may help to explain previously confounding adverse effects. In addition, EnPlex screening of a hydrolase-directed library of boronic acid- and nitrile-containing compounds provided structure-activity relationships in both potency and selectivity dimensions from which lead candidates could be more effectively prioritized. Follow-up of a series of dipeptidyl peptidase 4 inhibitors showed that EnPlex indeed predicted efficacy and safety in animal models. These results demonstrate the feasibility and value of high-throughput, superfamily-wide selectivity profiling and suggest that such profiling can be incorporated into the earliest stages of drug discovery.
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Association of interferon regulatory factor 4 gene polymorphisms rs12203592 and rs872071 with skin cancer and haematological malignancies susceptibility: a meta-analysis of 19 case-control studies.
BMC Cancer
PUBLISHED: 05-23-2014
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Research has indicated that the rs12203592 and rs872071 interferon regulatory factor 4 (IRF4) gene polymorphisms correlate with the risk of cancer, especially skin cancer and haematological malignancies, but the results remain controversial. To understand better the effects of these two polymorphisms on skin cancer and haematological malignancies susceptibility, a cumulative meta-analysis was performed.
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Different clinical characteristics between perimesencephalic subarachnoid hemorrhage and diffuse subarachnoid hemorrhage with negative initial angiography.
Turk Neurosurg
PUBLISHED: 05-23-2014
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The purpose of this study was to compare the different clinical features, outcome and treatment strategies in patients with perimesencephalic SAH (p-SAH) and diffuse SAH (d-SAH).
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Role of IL-10-Producing Regulatory B Cells in Chronic Hepatitis B Virus Infection.
Dig. Dis. Sci.
PUBLISHED: 05-19-2014
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A subset of interleukin (IL)-10-producing regulatory B (Breg) cells that suppress T-cell-mediated immunity was recently identified; however, their role in chronic hepatitis B (CHB) remains elusive.
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Selection of DNA-encoded small molecule libraries against unmodified and non-immobilized protein targets.
Angew. Chem. Int. Ed. Engl.
PUBLISHED: 04-30-2014
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The selection of DNA-encoded libraries against biological targets has become an important discovery method in chemical biology and drug discovery, but the requirement of modified and immobilized targets remains a significant disadvantage. With a terminal protection strategy and ligand-induced photo-crosslinking, we show that iterated selections of DNA-encoded libraries can be realized with unmodified and non-immobilized protein targets.
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Quercetin induces HepG2 cell apoptosis by inhibiting fatty acid biosynthesis.
Oncol Lett
PUBLISHED: 04-30-2014
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Quercetin can inhibit the growth of cancer cells with the ability to act as a 'chemopreventer'. Its cancer-preventive effect has been attributed to various mechanisms, including the induction of cell-cycle arrest and/or apoptosis, as well as its antioxidant functions. Quercetin can also reduce adipogenesis. Previous studies have shown that quercetin has potent inhibitory effects on animal fatty acid synthase (FASN). In the present study, activity of quercetin was evaluated in human liver cancer HepG2 cells. Intracellular FASN activity was calculated by measuring the absorption of NADPH via a spectrophotometer. MTT assay was used to test the cell viability, immunoblot analysis was performed to detect FASN expression levels and the apoptotic effect was detected by Hoechst 33258 staining. In the present study, it was found that quercetin could induce apoptosis in human liver cancer HepG2 cells with overexpression of FASN. This apoptosis was accompanied by the reduction of intracellular FASN activity and could be rescued by 25 or 50 ?M exogenous palmitic acids, the final product of FASN-catalyzed synthesis. These results suggested that the apoptosis induced by quercetin was via the inhibition of FASN. These findings suggested that quercetin may be useful for preventing human liver cancer.
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Comparative transcriptional activity of five promoters in BAC-cloned MDV for the expression of the hemagglutinin gene of H9N2 avian influenza virus.
J. Virol. Methods
PUBLISHED: 04-22-2014
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On the basis of recent studies, much attention has been given to recombinant MDV (rMDV)-based vaccines. During the construction of rMDV, the activity of promoters to transcribe foreign genes is one of the major factors that can affect protective efficacy. To investigate the transcription activity and efficacy of five different promoters, the advantage of an existing rMDV BAC infectious clone that had been previously constructed was used to construct rMDVs. The expression cassette of the hemagglutinin gene (HA) from a low pathogenic avian influenza virus (LPAIV) H9N2 strain was inserted into the US2 region under five selected promoters. These five promoters included three MDV endogenous promoters (the promoter for the gB gene and a bi-directional promoter in both directions for pp38 (ppp38) and 1.8 kb RNA transcripts (p1.8 kb)), and two exogenous promoters (CMV and SV40). Among these five promoters, the CMV promoter demonstrated the highest activity, followed by p1.8 kb and SV40, which had a similar transcriptional activity level. Two of the MDV endogenous promoters showed much lower transcriptional activities, particularly the promoter ppp38, which had the lowest activity. The results of the in vivo experiment proved that none of the three recombinant viruses of rGX-CMV-HA, rGX-SV40-HA and rGX-p1.8kb-HA provided protection in SPF chickens. Chickens vaccinated with rGX-pPP38-HA induced 50% and rGX-gB-HA induced 25% protection against the challenge with H9N2, respectively.
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Metabolite changes during the life history of Porphyra haitanensis.
Plant Biol (Stuttg)
PUBLISHED: 04-17-2014
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Plant metabolomics is essentially the comprehensive analysis of complex metabolites of plant extracts. Metabolic fingerprinting is an important part of plant metabolomics research. In this study, metabolic fingerprinting of different stages of the life history of the red alga Porphyra haitanensis was performed. The stages included conchocelis filaments, sporangial branchlets, conchosporangia, discharged conchospores and conchosporangial branchlets after conchospore discharge. Metabolite extracts were analysed with ultra-performance liquid chromatography coupled with electrospray ionisation quadrupole-time of flight mass spectrometry. Analyses profiles were subjected to principal components analysis and orthogonal projection to latent structures discriminant analysis using the SIMCA-P software for biomarker selection and identification. Based on the MS/MS spectra and data from the literature, potential biomarkers, mainly of phosphatidylcholine and lysophosphatidylcholine, were identified. Identification of these biomarkers suggested that plasma membrane phospholipids underwent major changes during the life history of P. haitanensis. The levels of phosphatidylcholine and lysophosphatidylcholine increased in sporangial branchlets and decreased in discharged conchospores. Moreover, levels of sphingaine (d18:0) decreased in sporangial branchlets and increased in discharged conchospores, which indicates that membrane lipids were increasingly synthesised as energy storage in sporangial branchlets, while energy was consumed in sporangial branchlets to discharged conchospores. A metabolomic study of different growth phases of P. haitanensis will enhance our understanding of its physiology and ecology. This article is protected by copyright. All rights reserved.
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Terpenoids from Tripterygium hypoglaucum and their inhibition of LPS-induced NO production.
Biosci. Biotechnol. Biochem.
PUBLISHED: 04-16-2014
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One new (1) and three known (2-4) sesquiterpenes and four known diterpenes (5-8) were isolated from the root bark of Tripterygium hypoglaucum. Their structures were elucidated on the basis of extensive spectroscopic analyses (IR, ESI-MS, HR-ESI-MS, 1D-NMR, and 2D-NMR). The inhibitory activity toward LPS-induced NO production of these terpenoids was evaluated, all the compounds showing inhibitory effects.
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Sequential treatment of icotinib after first-line pemetrexed in advanced lung adenocarcinoma with unknown EGFR gene status.
J Thorac Dis
PUBLISHED: 04-12-2014
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In non-small cell lung cancer (NSCLC), the well-developed epidermal growth factor receptor (EGFR) is an important therapeutic target. EGFR activating gene mutations have been proved strongly predictive of response to EGFR-tyrosine kinase inhibitors (TKI) in NSCLC. However, both in daily clinical practice and clinical trials, patients with unknown EGFR gene status (UN-EGFR-GS) are very common. In this study, we assessed efficacy and tolerability of sequential treatment of first-line pemetrexed followed by icotinib in Chinese advanced lung adenocarcinoma with UN-EGFR-GS.
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Transthyretin as a potential biomarker for the differential diagnosis between lung cancer and lung infection.
Biomed Rep
PUBLISHED: 04-10-2014
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Satisfactory biomarkers for screening and early diagnosis of lung cancer remain scarce and require further investigation. The aim of the present study was to examine the changes of the biochemical and protein composition in the serum and pleural effusion from lung cancer and lung infection (bacterial pneumonia) patients. A total of 92 patients with lung cancer, 38 with bacterial pneumonia and 42 healthy controls were enrolled in the study. The serum levels of cholesterol, apolipoprotein A and transthyretin (TTR) in the lung cancer patients were higher than that of the lung infection patients (P<0.05). The levels of TTR were higher, whereas the activity of adenosine deaminase (ADA) was lower in the pleural effusion from the lung cancer patients compared to the lung infection patients (P<0.05). Furthermore, the pleural effusion/serum TTR ratios in the lung cancer patients were higher, whereas the ratios of ADA were lower (P<0.05). By matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis, four major peaks corresponding to native TTR, Sul-TTR, Cys-TTR and Cysgly-TTR were observed in the serum of the lung cancer and lung infection patients. A significant increase was found in the proportion of Cysgly-TTR in the pleural effusion from the patients with lung cancer. The data indicated that a combination of pleural effusion/serum TTR ratios and modified TTR may be beneficial for the differential diagnosis between lung cancer and lung infection.
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The human embryonic stem cell proteome revealed by multidimensional fractionation followed by tandem mass spectrometry.
Proteomics
PUBLISHED: 04-09-2014
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Human embryonic stem cells (hESCs) have received considerable attention due to their therapeutic potential and their usefulness in understanding early development and cell fate commitment. In order to appreciate the unique properties of these pluripotent, self-renewing cells, we have performed an in-depth multidimensional fractionation followed by LC-MS/MS analysis of the hESCs harvested from defined media to elucidate expressed, phosphorylated, O-GlcNAc modified, and secreted proteins. From the triplicate analysis, we were able to assign more than 3000 proteins with less than 1% false-discovery rate. This analysis also allowed us to identify nearly 500 phosphorylation sites and 68 sites of O-GlcNAc modification with the same high confidence. Investigation of the phosphorylation sites allowed us to deduce the set of kinases that are likely active in these cells. We also identified more than 100 secreted proteins of hESCs that likely play a role in extracellular matrix formation and remodeling, as well as autocrine signaling for self-renewal and maintenance of the undifferentiated state. Finally, by performing in-depth analysis in triplicate, spectral counts were obtained for these proteins and post-translationally modified peptides, which will allow us to perform relative quantitative analysis between these cells and any derived cell type in the future. This article is protected by copyright. All rights reserved.
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MiR-451 is decreased in hypertrophic cardiomyopathy and regulates autophagy by targeting TSC1.
J. Cell. Mol. Med.
PUBLISHED: 04-09-2014
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The molecular mechanisms that drive the development of cardiac hypertrophy in hypertrophic cardiomyopathy (HCM) remain elusive. Accumulated evidence suggests that microRNAs are essential regulators of cardiac remodelling. We have been suggested that microRNAs could play a role in the process of HCM. To uncover which microRNAs were changed in their expression, microRNA microarrays were performed on heart tissue from HCM patients (n = 7) and from healthy donors (n = 5). Among the 13 microRNAs that were differentially expressed in HCM, miR-451 was the most down-regulated. Ectopic overexpression of miR-451 in neonatal rat cardiomyocytes (NRCM) decreased the cell size, whereas knockdown of endogenous miR-451 increased the cell surface area. Luciferase reporter assay analyses demonstrated that tuberous sclerosis complex 1 (TSC1) was a direct target of miR-451. Overexpression of miR-451 in both HeLa cells and NRCM suppressed the expression of TSC1. Furthermore, TSC1 was significantly up-regulated in HCM myocardia, which correlated with the decreased levels of miR-451. As TSC1 is a known positive regulator of autophagy, we examined the role of miR-451 in the regulation of autophagy. Overexpression of miR-451 in vitro inhibited the formation of the autophagosome. Conversely, miR-451 knockdown accelerated autophagosome formation. Consistently, an increased number of autophagosomes was observed in HCM myocardia, accompanied by up-regulated autophagy markers, and the lipidated form of LC3 and Beclin-1. Taken together, our findings indicate that miR-451 regulates cardiac hypertrophy and cardiac autophagy by targeting TSC1. The down-regulation of miR-451 may contribute to the development of HCM and may be a potential therapeutic target for this disease.
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Platelet-derived growth factor-? expression in rabbit models of cerebral vasospasm following subarachnoid hemorrhage.
Mol Med Rep
PUBLISHED: 04-01-2014
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Subarachnoid hemorrhage (SAH), one of the serious types of stroke incurred by bleeding into the space surrounding the brain, occurs when brains are deprived of oxygen by various factors, particularly an interruption to the blood supply or a ruptured aneurysm. Cerebral vasospasm (CVS) is one of the most common complications of SAH. It has been proposed that platelet?derived growth factor (PDGF) is involved in CVS. The aim of the present study was to analyze expression of PDGF in rabbit models of CVS. Post-SAH CVS rabbit models were created using endovascular puncture and employed to analyze the expression patterns of PDGF by enzyme-linked immunosorbent assay and immunohistochemistry. The results indicated that the creation of the rabbit model of CVS induced using endovascular puncture was successful and demonstrated the double phase changes observed in human CVS. The acute stage started at 12 h post-SAH with narrowing of the vascular lumen diameter. This narrowing appeared again on the seventh day in delayed CVS alongside increased thickness of vessel walls. PDGF-? expression was observed in vascular smooth muscle cells of the rabbit models. PDGF-? was expressed as early as 3 h post-SAH, it was evident after 1 day and reached a peak in 7 days, suggesting that PDGF-? is involved in the early stages of CVS. In the current study, it was confirmed that PDGF-? expression was present in the rabbit models of CVS, which may aid the elucidation of the pathogenesis of CVS, and also provide useful information for diagnosis and treatment of CVS.
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Development of spiropyran-based electrochemical sensor via simultaneous photochemical and target-activatable electron transfer.
Biosens Bioelectron
PUBLISHED: 03-20-2014
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In traditional electrochemical sensors, the electrochemical signal transduction of the redox-active material is usually controlled by the analytical target. Due to non-specific interaction between the redox mediator and the target, false signal by single stimulus may not be avoided. To address this issue, we have developed a new electrochemical sensor that uses a functional spiropyran, an important class of photo and thermochromic compounds, as both recognition receptor and latent redox mediator, to realize simultaneous photochemical and target-modulated electron transfer. As a proof of principle, ?-galactosidase was chosen as a model target. The new synthesized spiropyran probe, SP-?-gal, undergoes reversibly structural isomerization to form merocyanine under UV light irradiation. After the glycosidic bond being cleaved by ?-galactosidase, the opened merocyanine of SP-?-gal forms redox-active 2-(2.5-dihydroxystyryl)-1.3.3-trimethyl-3H-indolium, and thus produces a pair of reversible redox current peaks under the electrochemical scanning. To amplify the detection signal, SP-?-gal was self-assembled with single-walled carbon nanotubes (SWCNTs) on the surface of glass carbon electrode. Kinetics experiments confirm that the probe is an ideal candidate for the determination of different concentrations of ?-galactosidase digestion kinetics. Further, the SP-?-gal/SWCNTs-modified electrode is chemically stable in complex biological fluids. It was successfully applied to monitor ?-galactosidase activity in the 10% calf thymus. This work represents not only a significant step forward in the further development of low-dimensional carbon nanomaterials/small organic molecular probes-based electrochemical biosensors, but also a new platform which may be extended to the assay of other enzyme such as ?-D-glycosidase and so on by translating the biorecognition into electrochemical signal responses.
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'Druggable' alterations detected by Ion Torrent in metastatic colorectal cancer patients.
Oncol Lett
PUBLISHED: 03-20-2014
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The frequency and poor prognosis of patients with metastatic colorectal cancer (mCRC) emphasizes the requirement for improved biomarkers for use in the treatment and prognosis of mCRC. In the present study, somatic variants in exonic regions of key cancer genes were identified in mCRC patients. Formalin-fixed, paraffin-embedded tissues obtained by biopsy of the metastases of mCRC patients were collected, and the DNA was extracted and sequenced using the Ion Torrent Personal Genome Machine. For the targeted amplification of known cancer genes, the Ion AmpliSeq™ Cancer Panel, which is designed to detect 739 Catalogue of Somatic Mutations in Cancer (COSMIC) mutations in 604 loci from 46 oncogenes and tumor suppressor genes using as little as 10 ng of input DNA, was used. The sequencing results were then analyzed using the Ampliseq™ Variant Caller plug-in within the Ion Torrent Suite software. In addition, Ingenuity Pathway software was used to perform a pathway analysis. The Cox regression analysis was also conducted to investigate the potential correlation between alteration numbers and clinical factors, including response rate, disease-free survival and overall survival. Among 10 specimens, 65 genetic alterations were identified in 24 genes following the exclusion of germline mutations using the SNP database, whereby 41% of the alterations were also present in the COSMIC database. No clinical factors were found to significantly correlate with the alteration numbers in the patients by statistical analysis. However, pathway analysis identified 'colorectal cancer metastasis signaling' as the most commonly mutated canonical pathway. This analysis further revealed mutated genes in the Wnt, phosphoinositide 3-kinase (PI3K)/AKT and transforming growth factor (TGF)-?/SMAD signaling pathways. Notably, 11 genes, including the expected APC, BRAF, KRAS, PIK3CA and TP53 genes, were mutated in at least two samples. Notably, 90% (9/10) of mCRC patients harbored at least one 'druggable' alteration (range, 1-6 alterations) that has been linked to a clinical treatment option or is currently being investigated in clinical trials of novel targeted therapies. These results indicated that DNA sequencing of key oncogenes and tumor suppressors enables the identification of 'druggable' alterations for individual colorectal cancer patients.
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Transcriptional activity comparison of different sites in recombinant Marek's disease virus for the expression of the H9N2 avian influenza virus hemagglutinin gene.
J. Virol. Methods
PUBLISHED: 03-03-2014
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Over the last two decades, much attention has been paid to MDV-vectored recombinant vaccines. Many factors have influenced their protective efficacy, and insertion site has been among the main influential factors for the expression of foreign genes in recombinant Marek's disease virus (rMDV). To compare the transcriptional activity of different sites of rMDV, an H9N2 avian influenza virus hemagglutinin gene (AIV-H9N2-HA) expression cassette that used the bi-directional promoter of serotype 1 MDV (MDV1) in the 1.8kb RNA transcript direction (p1.8kb) as a promoter was inserted into 4 different regions of MDV using the bacterial artificial chromosome (BAC) vector and FLP/FRT recombination technique. The insertion regions included 3 of its own sites (US2, US10 and one of Meq genes) in the MDV genome and a foreign site (gpt gene) in the BAC vector. Quantitative PCR and enzyme-linked immunosorbent assay (ELISA) were used to analyze and compare the H9N2-HA expression levels of these different rMDVs both at the mRNA level and at the protein level. The results indicated that among the four tested insertion regions, the HA expression cassette in the US2 region demonstrated the highest activity, followed by that in the Meq region, which was almost equal to that of US10. Further, the expression cassette had the lowest activity in the foreign region gpt gene. The above data could be useful for choosing proper recombinant insertion regions in the construction of rMDV to express different foreign genes, and it is a prerequisite for developing effective MDV-vectored recombinant vaccines.
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S phase cell percentage normalized BrdU incorporation rate, a new parameter for determining S arrest.
Biomed. Environ. Sci.
PUBLISHED: 03-03-2014
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In this study, a new parameter, S phase cell percentage (S fraction) normalized BrdU (SFN-BrdU) incorporation rate, was introduced to detect S arrest. The results showed a positive linear correlation between the BrdU incorporation rate and the S fraction in unperturbed 16HBE cells. Theoretical analysis indicated that only S arrest could result in a decrease in the SFN-BrdU incorporation rate. Additionally, the decrease in SFN-BrdU incorporation rate and the activation of DNA damage checkpoints further demonstrated that S arrest was induced by diethyl sulfate treatment of 16HBE cells. In conclusion, SFN-BrdU incorporation rate can be used to detecting S arrest.
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A pan inhibitor of DASH family enzymes induces immunogenic modulation and sensitizes murine and human carcinoma cells to antigen-specific cytotoxic T lymphocyte killing: implications for combination therapy with cancer vaccines.
Vaccine
PUBLISHED: 02-26-2014
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Recent studies have suggested that pan inhibitors of dipeptidyl peptidase-4 activity and/or structure homologs (DASH), including ARI-4175, can mediate tumor regression by immune-mediated mechanisms. This study assessed the potential of combining ARI-4175 with cancer vaccines. We evaluated ARI-4175's effect on immunogenic modulation, ability to sensitize tumor cells to antigen-specific CTL killing, effect on immune-cell subsets and function, and antitumor activity in 2 tumor models, both as a monotherapy and in combination with a recombinant viral or dendritic cell (DC)-based tumor-cell vaccine. ARI-4175's effects on the growth, surface phenotype, and antigen-specific CTL-mediated lysis of murine and human carcinoma cell lines were assessed in vitro. In vivo, C57BL-6 mice were treated orally with ARI-4175, after which splenocytes were assessed by flow cytometry and functional assays. Antitumor studies were performed in murine models of colon carcinoma (MC38-CEA(+) in CEA-transgenic C57BL-6 mice) and rhabdomyosarcoma (M3-9-M in C57BL-6 mice). Mice received oral ARI-4175 alone or in combination with a vaccine consisting of recombinant vaccinia/fowlpox CEA-TRICOM (colon model) or a DC-based tumor-cell vaccine (rhabdomyosarcoma model). Exposure to ARI-4175 had no effect on the proliferation or viability of carcinoma cells in vitro; however, it did alter tumor phenotype, making murine and human tumor cells more sensitive to antigen-specific CTL killing. Assessment of immune-cell subsets and function indicated that ARI-4175 increased levels of natural killer cells and DCs. Detrimental immune effects, including reduced T effector cells and increased immunosuppressive cells (Tregs, MDSCs), were normalized when treatment stopped, suggesting that scheduling is critical when combining this agent with vaccine. As a monotherapy, ARI-4175 had potent antitumor activity in both tumor models, and had even greater effects when combined with a vaccine (either DC-based or poxviral vector based). These findings provide the rationale for the combined use of cancer immunotherapy with DASH enzyme inhibitors such as ARI-4175.
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SOX2 oncogenes amplified and operate to activate AKT signaling in gastric cancer and predict immunotherapy responsiveness.
J. Cancer Res. Clin. Oncol.
PUBLISHED: 02-22-2014
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Gastric cancer is the second leading cause of cancer mortality in the world. Whether the oncogene, amplified on chromosome 3q26, SOX2, a master transcriptional regulator of stemness, operate to drive strong growth phenotype in gastric cancer were unknown.
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The preparation, characterization and evaluation of regenerated cellulose/collagen composite hydrogel films.
Carbohydr Polym
PUBLISHED: 02-05-2014
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Porous structured regenerated cellulose films were oxidized by periodate oxidation to obtain 2,3-dialdehyde cellulose (DARC) films, which were then reacted with collagen to obtain DARC/Col composite films. The subsequent FT-IR spectra indicated that collagen was immobilized on the DARC matrix via the Schiff base reaction between NH2 in collagen and CHO in DARC backbone. Scanning electron microscopy revealed that DARC/Col exhibited a refined 3D network structure and its porosity and pore size decreased with increasing of collagen concentration. The composite films demonstrated a good equilibrium-swelling ratio, air permeability and water retention properties. The composite films also showed excellent mechanical properties, which was vital for practical application. Finally, the cytotoxicity of the composite film was evaluated using NIH3T3 mice fibroblast cells, the results revealed that DARC/Col composite films have good biocompatibility for use as scaffold material in tissue engineering.
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A pH and redox dual responsive 4-arm poly(ethylene glycol)-block-poly(disulfide histamine) copolymer for non-viral gene transfection in vitro and in vivo.
Int J Mol Sci
PUBLISHED: 01-28-2014
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A novel 4-arm poly(ethylene glycol)-b-poly(disulfide histamine) copolymer was synthesized by Michael addition reaction of poly(ethylene glycol) (PEG) vinyl sulfone and amine-capped poly(disulfide histamine) oligomer, being denoted as 4-arm PEG-SSPHIS. This copolymer was able to condense DNA into nanoscale polyplexes (<200 nm in average diameter) with almost neutral surface charge (+(5-10) mV). Besides, these polyplexes were colloidal stable within 4 h in HEPES buffer saline at pH 7.4 (physiological environment), but rapidly dissociated to liberate DNA in the presence of 10 mM glutathione (intracellular reducing environment). The polyplexes also revealed pH-responsive surface charges which markedly increased with reducing pH values from 7.4-6.3 (tumor microenvironment). In vitro transfection experiments showed that polyplexes of 4-arm PEG-SSPHIS were capable of exerting enhanced transfection efficacy in MCF-7 and HepG2 cancer cells under acidic conditions (pH 6.3-7.0). Moreover, intravenous administration of the polyplexes to nude mice bearing HepG2-tumor yielded high transgene expression largely in tumor rather other normal organs. Importantly, this copolymer and its polyplexes had low cytotoxicity against the cells in vitro and caused no death of the mice. The results of this study indicate that 4-arm PEG-SSPHIS has high potential as a dual responsive gene delivery vector for cancer gene therapy.
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Effects of oxysophoridine on amino acids after cerebral ischemic injury in mice.
Ann Indian Acad Neurol
PUBLISHED: 01-26-2014
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Our previous studies demonstrated that oxysophoridine (OSR) had neuroprotective effects on mice through antioxidant and anti-apoptotic mechanisms. In this study, we investigated whether OSR could influence the release of amino acids in ischemic mice brains.
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Serum adropin levels are decreased in patients with acute myocardial infarction.
Regul. Pept.
PUBLISHED: 01-22-2014
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Adropin is a recently identified bioactive protein that is important for energy homeostasis and maintaining insulin sensitivity. We sought to detect serum adropin levels in acute myocardial infarction (AMI) patients.
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Development of colloidal gold-based immunochromatographic assay for rapid detection of Mycoplasma suis in porcine plasma.
Biosens Bioelectron
PUBLISHED: 01-18-2014
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A one-step immunochromatographic assay using gold nanoparticles coated with polyclonal antibody (pAb) against Mycoplasma suis (M. suis) was developed in this study for the detection of M. suis in porcine plasma. The colloidal gold was prepared by the reduction of gold salt with sodium citrate coupled with pAb against M. suis. The pAb was produced by immunizing the BALB/c mice with recombinant MSG1 (rMSG1) protein from M. suis expressed in Escherichia coli. The optimal concentrations of the capture antibody and the coating antibody were 12 ?g/ml and 1.5 mg/ml, respectively, and that of the blocking buffer was 1% bovine serum albumin. The lower detection limit of the immunochromatographic assay test was 100 ng/ml with visual detection under optimal conditions of analysis. Classical swine fever virus, porcine reproductive and respiratory syndrome virus, swine pneumonia mycoplasma, swine toxoplasma, and porcine parvovirus were used to evaluate the specificity of the immunochromatographic strips. No cross-reaction of the antibodies with other related swine pathogens was observed. This qualitative test based on the visual evaluation of the results did not require any equipment. The assay time for M. suis detection was less than 10 min, suitable for rapid detection at the grassroots level. The one-step colloidal gold immunochromatographic strips that we developed had high specificity and sensitivity. Therefore, this method would be feasible, convenient, rapid, and effective for detecting M. suis in porcine plasma.
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Construction of recombinant Marek's disease virus (MDV) lacking the meq oncogene and co-expressing AIV-H9N2 HA and NA genes under control of exogenous promoters.
J. Biotechnol.
PUBLISHED: 01-14-2014
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To develop a recombinant Marek's disease virus (rMDV1) co-expressing the hemagglutinin gene (HA) and neuramidinase gene (NA) from a low pathogenic avian influenza virus (LPAIV) H9N2 strain and lacking the meq oncogene that shares homology with the Jun/Fos family of transcriptional factors, a wild strain of MDV GX0101 was used as parental virus, the HA and NA genes co-expression cassette under control of the CMV and SV40 early promoters was inserted at two meq sites of GX0101 to form a new meq knock-out mutant MDV (MZC12HA/NA) through homologous recombination. MZC12HA/NA was reconstituted by transfection of recombinant BAC-MDV DNA into the secondary chicken embryo fibroblast (CEF) cells. Highly purified MZC12HA/NA was obtained after four rounds of plaque purification and proliferation. In vitro growth properties of recombinant virus were also inspected and concluded that the MZC12HA/NA had the same growth kinetics in CEF cultures as its parental wild type virus GX0101. Southern blot indicated that co-expression cassette was successfully inserted at two copies sites of meq gene, so two meq genes were knocked-out completely. RT-qPCR showed transcription and expression levels of the HA and NA genes were both significantly higher than that of GX0101 own pp38 gene. Indirect fluorescence antibody (IFA) test, and Western blot analyses indicated that HA and NA genes were co-expressed simultaneously under control of the different promoters but meq genes were not. These results herald a new and effective recombinant meq-deleted MDV-based AIV-H9N2 vaccine may be useful in protecting chickens from very virulent MDV and H9N2 challenges.
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C-reactive protein as a prognostic marker in chronic obstructive pulmonary disease.
Exp Ther Med
PUBLISHED: 01-08-2014
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The present study aimed to evaluate whether circulating C-reactive protein (CRP) levels are a biomarker of systemic inflammation and a significant predictor of future chronic obstructive pulmonary disease (COPD) outcome. During the study, 116 patients with stable COPD and 35 age- and gender-matched healthy subjects with normal pulmonary function were observed. Patient follow-up was also performed to evaluate the strength of the associations between CRP levels and future outcomes. The observations from the present study showed that serum CRP levels were significantly higher in stable COPD patients than in control subjects (4.48±0.83 vs. 1.01±0.27 mg/l, respectively; P<0.05). In addition, it was identified that a serum CRP concentration of >3 mg/l is a poor prognostic variable of COPD compared with a CRP concentration of ?3 mg/l [hazard ratio (HR), 2.71; 95% confidence interval (CI), 1.05-6.99; P<0.05]. A quantitative synthesis of four studies including 1,750 COPD patients was performed and statistically similar results were obtained (HR, 1.54; 95% CI, 1.14-2.07; P<0.01). The present study showed that circulating CRP levels are higher in stable COPD patients and, therefore, may be used as a long-term predictor of future outcomes. These observations highlight the importance of high sensitivity CRP assays in patients with stable COPD.
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Increased expression of cathepsin L: a novel independent prognostic marker of worse outcome in hepatocellular carcinoma patients.
PLoS ONE
PUBLISHED: 01-01-2014
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To investigate the expression and role of Cathepsin L (CTSL) in Hepatocellular carcinoma (HCC) tissue and cell line (MHCC-97H), and to evaluate the clinical and prognostic significance of CTSL protein in patients with HCC.
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Optimized spatial priorities for biodiversity conservation in China: a systematic conservation planning perspective.
PLoS ONE
PUBLISHED: 01-01-2014
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By addressing several key features overlooked in previous studies, i.e. human disturbance, integration of ecosystem- and species-level conservation features, and principles of complementarity and representativeness, we present the first national-scale systematic conservation planning for China to determine the optimized spatial priorities for biodiversity conservation. We compiled a spatial database on the distributions of ecosystem- and species-level conservation features, and modeled a human disturbance index (HDI) by aggregating information using several socioeconomic proxies. We ran Marxan with two scenarios (HDI-ignored and HDI-considered) to investigate the effects of human disturbance, and explored the geographic patterns of the optimized spatial conservation priorities. Compared to when HDI was ignored, the HDI-considered scenario resulted in (1) a marked reduction (?9%) in the total HDI score and a slight increase (?7%) in the total area of the portfolio of priority units, (2) a significant increase (?43%) in the total irreplaceable area and (3) more irreplaceable units being identified in almost all environmental zones and highly-disturbed provinces. Thus the inclusion of human disturbance is essential for cost-effective priority-setting. Attention should be targeted to the areas that are characterized as moderately-disturbed, <2,000 m in altitude, and/or intermediately- to extremely-rugged in terrain to identify potentially important regions for implementing cost-effective conservation. We delineated 23 primary large-scale priority areas that are significant for conserving China's biodiversity, but those isolated priority units in disturbed regions are in more urgent need of conservation actions so as to prevent immediate and severe biodiversity loss. This study presents a spatially optimized national-scale portfolio of conservation priorities--effectively representing the overall biodiversity of China while minimizing conflicts with economic development. Our results offer critical insights for current conservation and strategic land-use planning in China. The approach is transferable and easy to implement by end-users, and applicable for national- and local-scale systematic conservation prioritization practices.
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Theranostic nanoparticles based on bioreducible polyethylenimine-coated iron oxide for reduction-responsive gene delivery and magnetic resonance imaging.
Int J Nanomedicine
PUBLISHED: 01-01-2014
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Theranostic nanoparticles based on superparamagnetic iron oxide (SPIO) have a great promise for tumor diagnosis and gene therapy. However, the availability of theranostic nanoparticles with efficient gene transfection and minimal toxicity remains a big challenge. In this study, we construct an intelligent SPIO-based nanoparticle comprising a SPIO inner core and a disulfide-containing polyethylenimine (SSPEI) outer layer, which is referred to as a SSPEI-SPIO nanoparticle, for redox-triggered gene release in response to an intracellular reducing environment. We reveal that SSPEI-SPIO nanoparticles are capable of binding genes to form nano-complexes and mediating a facilitated gene release in the presence of dithiothreitol (5-20 mM), thereby leading to high transfection efficiency against different cancer cells. The SSPEI-SPIO nanoparticles are also able to deliver small interfering RNA (siRNA) for the silencing of human telomerase reverse transcriptase genes in HepG2 cells, causing their apoptosis and growth inhibition. Further, the nanoparticles are applicable as T2-negative contrast agents for magnetic resonance (MR) imaging of a tumor xenografted in a nude mouse. Importantly, SSPEI-SPIO nanoparticles have relatively low cytotoxicity in vitro at a high concentration of 100 ?g/mL. The results of this study demonstrate the utility of a disulfide-containing cationic polymer-decorated SPIO nanoparticle as highly potent and low-toxic theranostic nano-system for specific nucleic acid delivery inside cancer cells.
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Autologous tumor lysate-pulsed dendritic cell immunotherapy with cytokine-induced killer cells improves survival in gastric and colorectal cancer patients.
PLoS ONE
PUBLISHED: 01-01-2014
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Gastric and colorectal cancers (GC and CRC) have poor prognosis and are resistant to chemo- and/or radiotherapy. In the present study, the prophylactic effects of dendritic cell (DC) vaccination are evaluated on disease progression and clinical benefits in a group of 54 GC and CRC patients treated with DC immunotherapy combined with cytokine-induced killer (CIK) cells after surgery with or without chemo-radiotherapy. DCs were prepared from the mononuclear cells isolated from patients using IL-2/GM-CSF and loaded with tumor antigens; CIK cells were prepared by incubating peripheral blood lymphocytes with IL-2, IFN-?, and CD3 antibodies. The DC/CIK therapy started 3 days after low-dose chemotherapy and was repeated 3-5 times in 2 weeks as one cycle with a total of 188.3 ± 79.8 × 10(6) DCs and 58.8 ± 22.3 × 10(8) CIK cells. Cytokine levels in patients' sera before and after treatments were measured and the follow-up was conducted for 98 months to determine disease-free survival (DFS) and overall survival (OS). The results demonstrate that all cytokines tested were elevated with significantly higher levels of IFN-? and IL-12 in both GC and CRC cohorts of DC/CIK treated patients. By Cox regression analysis, DC/CIK therapy reduced the risk of post-operative disease progression (p<0.01) with an increased OS (<0.01). These results demonstrate that in addition to chemo- and/or radiotherapy, DC/CIK immunotherapy is a potential effective approach in the control of tumor growth for post-operative GC and CRC patients.
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Discrimination between Adenocarcinoma and Normal Pancreatic Ductal Fluid by Proteomic and Glycomic Analysis.
J. Proteome Res.
PUBLISHED: 12-12-2013
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Sensitive and specific biomarkers for pancreatic cancer are currently unavailable. The high mortality associated with adenocarcinoma of the pancreatic epithelium justifies the broadest possible search for new biomarkers that can facilitate early detection or monitor treatment efficacy. Protein glycosylation is altered in many cancers, leading many to propose that glycoproteomic changes may provide suitable biomarkers. In order to assess this possibility for pancreatic cancer, we have performed an in-depth LC-MS/MS analysis of the proteome and MS(n)-based characterization of the N-linked glycome of a small set of pancreatic ductal fluid obtained from normal, pancreatitis, intraductal papillary mucinous neoplasm (IPMN), and pancreatic adenocarcinoma patients. Our results identify a set of seven proteins that were consistently increased in cancer ductal fluid compared to normal (AMYP, PRSS1, GP2-1, CCDC132, REG1A, REG1B, and REG3A) and one protein that was consistently decreased (LIPR2). These proteins are all directly or indirectly associated with the secretory pathway in normal pancreatic cells. Validation of these changes in abundance by Western blotting revealed increased REG protein glycoform diversity in cancer. Characterization of the total N-linked glycome of normal, IPMN, and adenocarcinoma ductal fluid clustered samples into three discrete groups based on the prevalence of six dominant glycans. Within each group, the profiles of less prevalent glycans were able to distinguish normal from cancer on this small set of samples. Our results emphasize that individual variation in protein glycosylation must be considered when assessing the value of a glycoproteomic marker, but also indicate that glycosylation diversity across human subjects can be reduced to simpler clusters of individuals whose N-linked glycans share structural features.
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The Effects of ABCC2 G1249A Polymorphism on the Risk of Resistance to Antiepileptic Drugs: A Meta-Analysis of the Literature.
Genet Test Mol Biomarkers
PUBLISHED: 12-10-2013
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Aims: The G1249A variant of the multidrug resistance-associated protein 2 (ABCC2) gene may be associated with the development of antiepileptic drug (AED) resistance. Although numerous studies have investigated the association between the G1249A variant and the risk of drug resistance in epilepsy, the results of these studies have been inconclusive. To assess the role of G1249A polymorphism in drug resistance in epilepsy, a meta-analysis was performed. Materials and Methods: We systematically reviewed relevant studies retrieved by the PubMed and Embase. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated based on the date extracted from the studies to evaluate the strength of association. We also analyzed the heterogeneity and sensitivity of each report and the publication bias of the studies. Results: A total of 6 published studies, involving 2213 patients (1100 patients with drug-resistant epilepsy and 1113 controls with drug-responsive epilepsy) were reviewed in the present meta-analysis. The overall results indicated that the variant genotypes were associated with a significantly decreased risk of AED resistance (AA vs. GG: OR=0.372, 95% CI=0.182-0.762; recessive model: OR=0.399, 95% CI=0.200-0.795) (fixed-effects model). A stratified analysis by ethnicity showed similar findings for Caucasians in an additive model (A vs. G: OR=0.700, 95% CI=0.494-0.992). Conclusions: The meta-analysis suggests that the ABCC2 G1249A polymorphism is significantly associated with a decreased risk of AED resistance. However, further functional investigations are warranted to validate the association.
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Sodium channels contribute to degeneration of dorsal root ganglion neurites induced by mitochondrial dysfunction in an in vitro model of axonal injury.
J. Neurosci.
PUBLISHED: 12-06-2013
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Axonal degeneration occurs in multiple neurodegenerative disorders of the central and peripheral nervous system. Although the underlying molecular pathways leading to axonal degeneration are incompletely understood, accumulating evidence suggests contributions of impaired mitochondrial function, disrupted axonal transport, and/or dysfunctional intracellular Ca(2+)-homeostasis in the injurious cascade associated with axonal degeneration. Utilizing an in vitro model of axonal degeneration, we studied a subset of mouse peripheral sensory neurons in which neurites were exposed selectively to conditions associated with the pathogenesis of axonal neuropathies in vivo. Rotenone-induced mitochondrial dysfunction resulted in neurite degeneration accompanied by reduced ATP levels and increased ROS levels in neurites. Blockade of voltage-gated sodium channels with TTX and reverse (Ca(2+)-importing) mode of the sodium-calcium exchanger (NCX) with KB-R7943 partially protected rotenone-treated neurites from degeneration, suggesting a contribution of sodium channels and reverse NCX activity to the degeneration of neurites resulting from impaired mitochondrial function. Pharmacological inhibition of the Na(+)/K(+)-ATPase with ouabain induced neurite degeneration, which was attenuated by TTX and KB-R7943, supporting a contribution of sodium channels in axonal degenerative pathways accompanying impaired Na(+)/K(+)-ATPase activity. Conversely, oxidant stress (H2O2)-induced neurite degeneration was not attenuated by TTX. Our results demonstrate that both energetic and oxidative stress targeted selectively to neurites induces neurite degeneration and that blockade of sodium channels and of reverse NCX activity blockade partially protects neurites from injury due to energetic stress, but not from oxidative stress induced by H2O2.
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Multistep DNA-Templated Synthesis Using a Universal Template.
J. Am. Chem. Soc.
PUBLISHED: 11-18-2013
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We report a DNA-templated synthesis method that allows construction of the entire DNA-encoded library with a single DNA template. Taking advantage of deoxyinosines indiscriminate base-pairing property, we designed a "universal template" that is capable of directing chemical reactions with multiple reactant DNAs with different sequences. In combination with other design features including photocleavable linkers and direct encoding by the reactant DNA, we demonstrated the capabilities of the universal template in library synthesis, target selection, and hit decoding. Our method can be generally and straightforwardly applied to prepare a variety of chemically diverse DNA-encoded libraries.
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Parabiosis in mice: a detailed protocol.
J Vis Exp
PUBLISHED: 10-23-2013
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Parabiosis is a surgical union of two organisms allowing sharing of the blood circulation. Attaching the skin of two animals promotes formation of microvasculature at the site of inflammation. Parabiotic partners share their circulating antigens and thus are free of adverse immune reaction. First described by Paul Bert in 1864(1), the parabiosis surgery was refined by Bunster and Meyer in 1933 to improve animal survival(2). In the current protocol, two mice are surgically joined following a modification of the Bunster and Meyer technique. Animals are connected through the elbow and knee joints followed by attachment of the skin allowing firm support that prevents strain on the sutured skin. Herein, we describe in detail the parabiotic joining of a ubiquitous GFP expressing mouse to a wild type (WT) mouse. Two weeks after the procedure, the pair is separated and GFP positive cells can be detected by flow cytometric analysis in the blood circulation of the WT mouse. The blood chimerism allows one to examine the contribution of the circulating cells from one animal in the other.
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A General Method for Making Peptide Therapeutics Resistant to Serine Protease Degradation: Application to Dipeptidyl Peptidase IV Substrates.
J. Med. Chem.
PUBLISHED: 10-16-2013
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Bioactive peptides have evolved to optimally fulfill specific biological functions, a fact which has long attracted attention for their use as therapeutic agents. While there have been some recent commercial successes fostered in part by advances in large-scale peptide synthesis, development of peptides as therapeutic agents has been significantly impeded by their inherent susceptibility to protease degradation in the bloodstream. Here we report that incorporation of specially designed amino acid analogues at the P1 position, directly C-terminal of the enzyme cleavage site, renders peptides, including glucagon-like peptide-1 (7-36) amide (GLP-1) and six other examples, highly resistant to serine protease degradation without significant alteration of their biological activity. We demonstrate the applicability of the method to a variety of proteases, including dipeptidyl peptidase IV (DPP IV), dipeptidyl peptidase 8 (DPP8), fibroblast activation protein ? (FAP?), ?-lytic protease (?LP), trypsin, and chymotrypsin. In summary, the "P1 modification" represents a simple, general, and highly adaptable method of generating enzymatically stable peptide-based therapeutics.
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Identification of STAT target genes in adipocytes.
JAKSTAT
PUBLISHED: 09-24-2013
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Adipocytes play important roles in lipid storage, energy homeostasis and whole body insulin sensitivity. Studies in the last two decades have identified the hormones and cytokines that activate specific STATs in adipocytes in vitro and in vivo. Five of the seven STAT family members are expressed in adipocyte (STATs 1, 3, 5A, 5B and 6). Many transcription factors, including STATs, have been shown to play an important role in adipose tissue development and function. This review will summarize the importance of adipocytes, indicate the cytokines and hormones that utilize the JAK-STAT signaling pathway in fat cells and focus on the identification of STAT target genes in mature adipocytes. To date, specific target genes have been identified for STATs, 1, 5A and 5B, but not for STATs 3 and 6.
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Rediscovering the lost art of internal podalic version: two case reports.
Trop Doct
PUBLISHED: 09-20-2013
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Internal podalic version was once widely used for the management of placenta praevia. In modern obstetrics, it has been replaced by caesarean section which can lead to uterine rupture in a subsequent pregnancy. However, if the foetus is dead or very small, internal podalic version still may be carried out without major complication to avoid unnecessary caesarean section in developing countries where follow up is not satisfactory and the foetal prognosis is poor.
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A pan-inhibitor of DASH family enzymes induces immune-mediated regression of murine sarcoma and is a potent adjuvant to dendritic cell vaccination and adoptive T-cell therapy.
J. Immunother.
PUBLISHED: 09-03-2013
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Multimodality therapy consisting of surgery, chemotherapy, and radiation will fail in approximately 40% of patients with pediatric sarcomas and result in substantial long-term morbidity in those who are cured. Immunotherapeutic regimens for the treatment of solid tumors typically generate antigen-specific responses too weak to overcome considerable tumor burden and tumor suppressive mechanisms and are in need of adjuvant assistance. Previous work suggests that inhibitors of DASH (dipeptidyl peptidase IV activity and/or structural homologs) enzymes can mediate tumor regression by immune-mediated mechanisms. Herein, we demonstrate that the DASH inhibitor, ARI-4175, can induce regression and eradication of well-established solid tumors, both as a single agent and as an adjuvant to a dendritic cell (DC) vaccine and adoptive cell therapy (ACT) in mice implanted with the M3-9-M rhabdomyosarcoma cell line. Treatment with effective doses of ARI-4175 correlated with recruitment of myeloid (CD11b) cells, particularly myeloid DCs, to secondary lymphoid tissues and with reduced frequency of intratumoral monocytic (CD11bLy6-CLy6-G) myeloid-derived suppressor cells. In immunocompetent mice, combining ARI-4175 with a DC vaccine or ACT with tumor-primed T cells produced significant improvements in tumor responses against well-established M3-9-M tumors. In M3-9-M-bearing immunodeficient (Rag1) mice, ACT combined with ARI-4175 produced greater tumor responses and significantly improved survival compared with either treatment alone. These studies warrant the clinical investigation of ARI-4175 for treatment of sarcomas and other malignancies, particularly as an adjuvant to tumor vaccines and ACT.
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A bipartite molecular module controls cell death activation in the Basal cell lineage of plant embryos.
PLoS Biol.
PUBLISHED: 09-01-2013
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Plant zygote divides asymmetrically into an apical cell that develops into the embryo proper and a basal cell that generates the suspensor, a vital organ functioning as a conduit of nutrients and growth factors to the embryo proper. After the suspensor has fulfilled its function, it is removed by programmed cell death (PCD) at the late stages of embryogenesis. The molecular trigger of this PCD is unknown. Here we use tobacco (Nicotiana tabacum) embryogenesis as a model system to demonstrate that the mechanism triggering suspensor PCD is based on the antagonistic action of two proteins: a protease inhibitor, cystatin NtCYS, and its target, cathepsin H-like protease NtCP14. NtCYS is expressed in the basal cell of the proembryo, where encoded cystatin binds to and inhibits NtCP14, thereby preventing precocious onset of PCD. The anti-cell death effect of NtCYS is transcriptionally regulated and is repressed at the 32-celled embryo stage, leading to increased NtCP14 activity and initiation of PCD. Silencing of NtCYS or overexpression of NtCP14 induces precocious cell death in the basal cell lineage causing embryonic arrest and seed abortion. Conversely, overexpression of NtCYS or silencing of NtCP14 leads to profound delay of suspensor PCD. Our results demonstrate that NtCYS-mediated inhibition of NtCP14 protease acts as a bipartite molecular module to control initiation of PCD in the basal cell lineage of plant embryos.
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Small-fiber neuropathy Nav1.8 mutation shifts activation to hyperpolarized potentials and increases excitability of dorsal root ganglion neurons.
J. Neurosci.
PUBLISHED: 08-30-2013
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Idiopathic small-fiber neuropathy (I-SFN), clinically characterized by burning pain in distal extremities and autonomic dysfunction, is a disorder of small-caliber nerve fibers of unknown etiology with limited treatment options. Functional variants of voltage-gated sodium channel Nav1.7, encoded by SCN9A, have been identified in approximately one-third of I-SFN patients. These variants render dorsal root ganglion (DRG) neurons hyperexcitable. Sodium channel Nav1.8, encoded by SCN10A, is preferentially expressed in small-diameter DRG neurons, and produces most of the current underlying the upstroke of action potentials in these neurons. We previously demonstrated two functional variants of Nav1.8 that either enhance ramp current or shift activation in a hyperpolarizing direction, and render DRG neurons hyperexcitable, in I-SFN patients with no mutations of SCN9A. We have now evaluated additional I-SFN patients with no mutations in SCN9A, and report a novel I-SFN-related Nav1.8 mutation I1706V in a patient with painful I-SFN. Whole-cell voltage-clamp recordings in small DRG neurons demonstrate that the mutation hyperpolarizes activation and the response to slow ramp depolarizations. However, it decreases fractional channels resistant to fast inactivation and reduces persistent currents. Current-clamp studies reveal that mutant channels decrease current threshold and increase the firing frequency of evoked action potentials within small DRG neurons. These observations suggest that the effects of this mutation on activation and ramp current are dominant over the reduced persistent current, and show that these pro-excitatory gating changes confer hyperexcitability on peripheral sensory neurons, which may contribute to pain in this individual with I-SFN.
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STAT1, NF-?B and ERKs play a role in the induction of lipocalin-2 expression in adipocytes.
Mol Metab
PUBLISHED: 08-01-2013
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Lipocalin-2 (LCN2) is induced in conditions of obesity and Type 2 diabetes (T2DM). IFN? and TNF? induce LCN2 expression in adipocytes in a manner that is dependent on transcription. The effects of these cytokines are additive. IFN? induced STAT1 and TNF? induced NF-?B play a role in the induction of LCN2. In the LCN2 promoter, one NF-?B binding site and four STAT1 binding sites were identified by in silico and in vitro approaches. MAPK (ERKs 1 and 2) activation was required for the IFN? and TNF? induction of LCN2 expression, but did not affect the nuclear translocation or DNA binding activity of STAT1 or NF-?B. The NF-?B binding site and the STAT1 binding sites we identified in vitro were confirmed by in vivo studies. Transfection of a LCN2 promoter/luciferase reporter construct confirmed acute activation by IFN? and TNF?. Our studies identify mechanisms involved in the actions of cytokines secreted from immune cells in adipose tissue that induce LCN2 expression in conditions of obesity and T2DM.
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Increased number of Th22 cells and correlation with Th17 cells in peripheral blood of patients with IgA nephropathy.
Hum. Immunol.
PUBLISHED: 07-22-2013
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T-helper (Th) 22 and Th17 cells are involved in the pathogenesis of autoimmune diseases. However, the role of Th22 and correlation with Th17 cells in the pathophysiology of IgA nephropathy (IgAN) remain unknown. In our study, Th22 and Th17 cells in peripheral blood of IgAN patients, non-IgA mesangial proliferative glomerulonephritis (non-IgA MsPGN) patients, and healthy controls were measured by flow cytometry. The concentration of plasma interleukin-22 (IL-22) was examined by enzyme linked immunosorbent assay (ELISA). The results showed that Th22 cells, Th17 cells, and plasma IL-22 were significantly elevated in IgAN patients compared with non-IgA MsPGN patients and healthy controls. Th22 cells showed a positive correlation with the levels of plasma IL-22 in IgAN patients. Moreover, a significantly positive correlation between Th22 cells and Th17 in IgAN patients was observed. Furthermore, IgAN patients with proteinuria showed a higher percentage of Th22 cells than IgAN patients without proteinuria. Our data demonstrated that IgAN had increased frequencies of peripheral Th22, Th17 cells and plasma IL-22, indicating that Th22 along with Th17 cells are involved in the immune responses of IgAN.
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XRCC1 Arg399Gln variation and leukemia susceptibility: evidence from 2,647 cases and 5,518 controls.
Tumour Biol.
PUBLISHED: 07-18-2013
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Previous reports implicate XRCC1 Arg399Gln polymorphism as a possible risk factor for several cancers. Increasing studies have been conducted on the association of XRCC1 Arg399Gln polymorphisms with susceptibility to leukemia. However, conflicting results have been generated. The goal of the present study was to derive a more precise estimation of the relationship. Meta-analyses assessing the association of XRCC1 Arg399Gln variation with leukemia were conducted, and subgroup analyses on ethnicity and clinical types were further performed. Eligible studies were identified for the period up to February 2013. Consequently, 16 publications including 17 case-control studies with 2,647 cases and 5,518 controls were selected for analysis. The overall data indicated a significant association of XRCC1 Arg399Gln polymorphism with leukemia risk (Gln/Gln versus Arg/Arg: OR?=?1.37, 95% confidence interval (CI)?=?1.08-1.74; dominant model: OR?=?1.23, 95%CI?=?1.03-1.46; recessive model: OR?=?1.23, 95%CI?=?1.06-1.44). In the subgroup analysis by ethnicity, Gln allele may increase leukemia susceptibility among Asians (Gln/Gln versus Arg/Arg: OR?=?1.82, 95%CI?=?1.19-2.78; dominant model: OR?=?1.53, 95%CI?=?1.00-2.33; recessive model: OR?=?1.51, 95%CI?=?1.11-2.06), but not Caucasians or mixed ethnicities. In the subgroup analysis by clinical types, increased risk was observed in acute lymphocytic leukemia (ALL) subgroup (Gln/Gln versus Arg/Arg: OR?=?1.45, 95%CI?=?1.09-1.93; recessive model: OR?=?1.30, 95%CI?=?1.00-1.69), but not in acute myeloid leukemia, chronic lymphocytic leukemia, or chronic myeloid leukemia subgroups, respectively. Collectively, the results of the present study suggest that XRCC1 Arg399Gln polymorphism might be a low-penetrant risk factor for leukemia, particularly among Asians. Homozygous Gln/Gln alleles might have a correlation with increased ALL susceptibility.
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[Clinical analyses of perinuclear antineutrophil cytoplasmic antibody associated hypertrophic pachymeningitis].
Zhonghua Yi Xue Za Zhi
PUBLISHED: 07-18-2013
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To explore the clinical characteristics, diagnosis, treatment and prognosis of perinuclear antineutrophil cytoplasmic antibody (p-ANCA) associated hypertrophic pachymeningitis.
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What is Visualize?

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

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

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