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Metabolic shift in liver: Correlation between perfusion temperature and hypoxia inducible factor-1α

World Journal of Gastroenterology 2015年第4期21卷 1108-1116页

作者： Andrea Ferrigno Laura Giuseppina Di Pasqua Alberto Bianchi Plinio Richelmi Mariapia VairettiAndrea Ferrigno Unit of cellular and Molecular Pharmacology and Toxicology Department of Interna Medicine and TherapeuticsUniversity of PaviaPavia 27100 Italy

AIM: To study at what temperature the oxygen carried by the perfusate meets liver requirements in a model of organ perfusion. METHODS: in this study, we correlated hypoxia induciblefactor(Hi F)-1α expression to the perfusion temperature and the hepatic oxygen uptake in a model of isolated perfused rat liver. Livers from Wistar rats were perfused for 6 h with an oxygenated medium at 10, 20, 30 and 37 ℃. Oxygen uptake was measured by an oxygen probe; lactate dehydrogenase activity, lactate release and glycogen were measured spectrophotometrically; bile flow was gravitationally determined; p H of the perfusate was also evaluated; Hi F-1α m RNA and protein expression were analyzed by real time-polymerase chain reaction and ELi SA, respectively. RESULTS: Livers perfused at 10 and 20 ℃ showed no difference in lactate dehydrogenase release after 6 h of perfusion(0.96 ± 0.23 vs 0.93 ± 0.09 m U/min per g) and had lower hepatic damage as compared to 30 and 37 ℃(5.63 ± 0.76 vs 527.69 ± 45.27 m U/min per g, respectively, P s < 0.01). After 6 h, tissue ATP was significantly higher in livers perfused at 10 and 20 ℃than in livers perfused at 30 and 37 ℃(0.89 ± 0.06 and 1.16 ± 0.05 vs 0.57 ± 0.09 and 0.33 ± 0.08 nmol/mg, respectively, P s < 0.01). No sign of hypoxia was observed at 10 and 20 ℃, as highlighted by low lactate release respect to livers perfused at 30 and 37 ℃(121.4 ± 12.6 and 146.3 ± 7.3 vs 281.8 ± 45.3 and 1094.5 ± 71.7 nmol/m L, respectively, P s < 0.02), and low relative Hi F-1α m RNA(0.40 ± 0.08 and 0.20 ± 0.03 vs 0.60 ± 0.20 and 1.47 ± 0.30, respectively, P s < 0.05) and protein(3.72 ± 0.16 and 3.65 ± 0.06 vs 4.43 ± 0.41 and 6.44 ± 0.82, respectively, P s < 0.05) ***: Livers perfused at 10 and 20 ℃ show no sign of liver injury or anaerobiosis, in contrast to livers perfused at 30 and 37 ℃.

关键词： Anaerobiosis Hypoxia inducible factor-1 α ischemia

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Prognostic significance of BMP and activin membrane-bound inhibitor in colorectal cancer

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World Journal of Gastroenterology 2008年第31期14卷 4880-4888页

作者： Nozomi Togo Susumu Ohwada Shinji Sakurai Hiroyuki Toya Ichiro Sakamoto Tatsuya Yamada Tetsuhiro Nakano Ken Muroya Izumi Takeyoshi Takashi Nakajima Takashi Sekiya Yusuke Yamazumi Tsutomu Nakamura Tetsu Akiyama departments of Surgery Graduate School of Medicine Gunma University Gunma 371-8511 Japan departments of Tumor Pathology Graduate School of Medicine Gunma University Gunma 371-8511 Japan Laboratory of Molecular and genetic Information Institute for Molecular and Cellular Biosciences University of Tokyo Tokyo 113-0032 Japan

AIM： To investigate the clinical significance of BMP and activin membrane-bound inhibitor （BAMBI） which is a pseudoreceptor of transforming growth factorbeta （TGF-β） type 1 receptors and acts as a negative regulator of TGF-β signaling and expression aberrantly elevated in colorectal cancers （CRCs）. We studied BAMBI expression in CRCs. METHODS： We studied BAMBI expression in 183 surgically resected CRCs by immunochemical and immunoblotting analyses using a generated monoclonal anti-BAMBI antibody. Commercially available anti-β- catenin and anti-p53 antibodies were also applied for immunochemical analyses as a comparison ***： Immunohistochemical analysis revealed that BAMBI expression was observed in 148 （80.8%）, and strong BAMBI expression was observed in 46% of the CRCs. Strong BAMBI expression was positively correlated with histological type, depth of invasion, lymph node metastases, and tumor node metastasis （TNM） stage （P 〈 0.05）. Clear associations were found between BAMBI and β-catenin （P = 0.035） and p53 （P =0.049） expression. In curatively resected CRC, 5-year recurrence-free survival was 51.9% （P = 0.037） for strong BAMBI expression compared to 79.8% for weak BAMBI expression. In the Cox＇s multivariate analysis, lymph node metastases （relative risk 6.685; P 〈 0.001） and depth of invasion （RR 14.0; P = 0.013） were significant indicators for recurrence, and strong BAMBI expression （RR 2.26; P = 0.057） tended to be significant. CONCLUSION： BAMBI was linked to a potentially aggressive tumor phenotype and predicted tumor recurrence and cancer-related death in CRC. BAMBI expression might be applicable in the routine clinical setting of CRC.

关键词： BMP and activin membrane-bound inhibitor Colorectal cancer Transforming growth factor-beta signal Prognosis Wnt signal

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Clinical features and MUT gene mutation spectrum in Chinese patients with isolated methylmalonic acidemia:identification of ten novel allelic variants

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World Journal of Pediatrics 2015年第4期11卷 358-365页

作者： Lian-Shu Han Zhuo Huang Feng Han Jun Ye Wen-Juan Qiu Hui-Wen Zhang Yu Wang Zhu-Wen Gong Xue-Fan Gu department of Pediatric Endocrinology and genetic Metabolism Xinhua HospitalShanghai Institute for Pediatric ResearchShanghai Jiaotong University School of MedicineShanghaiChina

Background: This study aims to study MUT gene mutation spectrum in Chinese patients with isolated methylmalonic academia (MMA) and their clinical features for the potential genotype-phenotype correlation. Methods: Forty-three patients were diagnosed with isolated MMA by elevated blood propionylcarnitine, propionylcarnitine to acetylcarnitine ratio, and urine methylmalonate without hyperhomocysteinemia. The MUT gene was amplifi ed by polymerase chain reaction and directly sequenced. Those patients with at least one variant allele were included. The novel missense mutations were assessed by bioinformatic analysis and screened against alleles sequenced from 50 control participants. Results: Among the 43 patients, 38 had typical clinical presentations, and the majority (30/38) experienced early-onset MMA. Eight patients died and seven were lost to follow-up. Twenty patients had poor outcomes and eight showed normal development. The 43 identified MUT gene mutations had at least one variant allele, whereas 35 had two mutant alleles. Of the 33 mutations reported before, eight recurrent mutations were identified in 32 patients, and c.729_730insTT (p.D244Lfs*39) was the most common (12/78) in the mutant alleles. Of the 10 novel mutations, six were missense mutations and four were premature termination codon mutations. The six novel missense mutations seemed to be pathogenic. Conclusions: A total of 10 novelMUT mutations were detected in the Chinese population. c.729_730insTT (p.D244Lfs*39) was the most frequent mutation. A genotype-phenotype correlation could not be found, but the genotypic characterization indicated the need of genetic counseling for MMA patients and early prenatal diagnoses for high-risk families.

关键词： methylmalonic acidemia missense mutation MUT gene

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Blockade of Na^+/H^+ exchanger type 3 causes intracellular acidification and hyperexcitability via inhibition of pH-sensitive K^+ channels in chemosensitive respiratory neurons of the dorsal vagal nucleus in rats

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Neuroscience Bulletin 2014年第1期30卷 43-52页

作者： Jing Zhang Hui Peng Sigrid C.Veasey Jing Ma Guang-Fa Wang Ke-Wei Wang department of Respiratory and Critical Care medicine Peking University First Hospital departments of Neurobiology and Molecular and cellular Pharmacology Peking University Health Science Center Center for Sleep and Circadian Neurobiology University of Pennsylvania School of Medicine department of medicine University of Pennsylvania School of Medicine

Extracellular pH （pHe） and intracellular pH （pHi） are important factors for the excitability of chemosensitive central respiratory neurons that play an important role in respiration and obstructive sleep apnea. It has been proposed that inhibition of central Na^＋/ H^＋ exchanger 3 （NHE-3）, a key pHi regulator in the brainstem, decreases the pH, leading to membrane depolarization for the maintenance of respiration. However, how intracellular pH affects the neuronal excitability of respiratory neurons remains largely unknown. In this study, we showed that NHE-3 mRNA is widely distributed in respiration-related neurons of the rat brainstem, including the dorsal vagal nucleus （DVN）. Whole-cell patch clamp recordings from DVN neurons in brain slices revealed that the standing outward current （Iso） through pH-sensitive K^＋ channels was inhibited in the presence of the specific NHE-3 inhibitor AVE0657 that decreased the pHi. Exposure of DVN neurons to an acidified PIle and AVE0657 （5 μmol/L） resulted in a stronger effect on firing rate and Iso than acidified pHe alone. Taken together, our results showed that intracellular acidification by blocking NHE-3 results in inhibition of a pH- sensitive K^＋ current, leading to synergistic excitation of chemosensitive DVN neurons for the regulation of respiration.

关键词： Na^+/H^+ exchange potassium channel dorsal vagal nucleus in situ hybridization respiration

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Characterization of Binding Sites of Eukaryotic Transcription Factors

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Genomics, Proteomics & Bioinformatics 2006年第2期4卷 67-79页

作者： Jiang Qian Jimmy Lin Donald J. Zack The Wilmer Institute department of Molecular Biology and genetics department of Neuroscience McKusick-Nathans Institute of genetic medicine Johns Hopkins University School of Medicine Baltimore MD 21287 USA.

To explore the nature of eukaryotic transcription factor （TF） binding sites and determine how they differ from surrounding DNA sequences, we examined four features associated with DNA binding sites： G＋C content, pattern complexity, palindromic structure, and Markov sequence ordering. Our analysis of the regulatory motifs obtained from the TRANSFAC databases using yeast intergenic sequences as background, revealed that these four features show variable enrichment in motif sequences. For example, motif sequences were more likely to have palindromic structure than were background sequences. In addition, these features were tightly localized to the regulatory motifs, indicating that they are a property of the motif sequences themselves and are not shared by the general promoter ＂environment＂ in which the regulatory motifs reside. By breaking down the motif sequences according to the TF classes to which they bind, more specific associations were identified. Finally, we found that some correlations, such as G＋C content enrichment, were species-specific, while others, such as complexity enrichment, were universal across the species examined. The quantitative analysis provided here should increase our understanding of protein-DNA interactions and also help facilitate the discovery of regulatory motifs through bioinformatics.

关键词： transcription factor promoter gene regulation bioinformatics

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Chikungunya infection:A potential re-emerging global threat

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Asian Pacific Journal of Tropical medicine 2016年第10期9卷 910-915页

作者： Shanmugaraj Bala Murugan Ramalingam Sathishkumar Plant genetic Engineering Laboratory Department of BiotechnologyBharathiar University

Infectious diseases are indeed a lifelong threat to everyone irrespective of age, sex, lifestyle and socio-economic status. The infectious diseases have persisted among the prominent causes of death globally. Recently, re-emergence of Chikungunya viral infection harmed many in Asian and African countries. Chikungunya was considered as a major threat in developing and underdeveloped countries; the recent epidemiological outbreak of Chikungunya in La Reunion urges the global researchers to develop effective vaccine against this viral disease. In this review, Chikungunya, pathogenesis and epidemiology were briefly described.

关键词： Aedes mosquitoes Alphavirus Arthropod-borne disease Chikungunya Non-Structural Proteins Structural Proteins

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Spliceosomal genes in the *** genome:a comparison with those in ***,***,*** and ***

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Protein & Cell 2011年第5期2卷 395-409页

作者： Bing Yu Petra Fey Karen E.Kestin-Pilcher Alexei Fedorov Ashwin Prakash Rex L.Chisholm Jane Y.Wu department of Molecular and Clinical genetics Royal Prince Alfred Hospital and Sydney Medical School(Central)the University of SydneyNSW 2006Australia dictyBase Center for Genetic MedicineNorthwestern UniversityChicagoIL 60611USA department of Neurology and Lurie Comprehensive Cancer Center Center for Genetic MedicineNorthwestern University Feinberg Medical SchoolChicagoIL 60611USA department of medicine and Program in Bioinformatics and Proteomics/Genomics The University of ToledoToledoOH 43614USA

Little is known about pre-mRNA splicing in Dictyostelium discoideum although its genome has been completely *** analysis suggests that pre-mRNA splicing plays an important role in *** gene expression as two thirds of its genes contain at least one *** curation of the genome to date has revealed 40 genes in *** with clear evidence of alternative splicing,supporting the existence of alternative splicing in this unicellular *** identified 160 candidate U2-type spliceosomal proteins and related factors in *** based on 264 known human genes involved in *** small ribonucleoproteins(snRNPs),PRP19 complex proteins and late-acting proteins are highly conserved in *** and throughout the *** non-snRNP and hnRNP families,*** orthologs are closer to those in ***,*** and *** than to their counterparts in *** splicing regulators,including SR proteins and CUGbinding proteins,were found in ***,but not in *** comprehensive catalog of spliceosomal proteins provides useful information for future studies of splicing in *** where the efficient genetic and biochemical manipulation will also further our general understanding of pre-mRNA splicing.

关键词： pre-mRNA splicing spliceosomal genes Dictyostelium discoideum comparative genomics splicing regulators

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Eight novel MUT loss-of-function missense mutations in Chinese patients with isolated methylmalonic academia

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World Journal of Pediatrics 2017年第4期13卷 381-386页

作者： Lian-Shu Han Zhuo Huang Feng Han Yu Wang Zhu-Wen Gong Xue-Fan Gu department of Pediatric Endocrinology and genetic Metabolism Xinhua HospitalShanghai Institute for Pediatric ResearchShanghai Jiaotong University School of MedicineShanghaiChina

Background:Isolated methylmalonic acidemia is a rare autosomal recessive metabolic disorder mostly caused by mutations in the methylmalonyl coenzyme A mutase (MCM) gene (MUT).This study aimed to verify whether missense mutations in MUT in Chinese patients affect the stability and enzymatic activity of ***:Eight Chinese patients were identified with novel *** carrying the wild-type and mutated MUT cDNA were constructed and transfected into HEK293T cells for functional *** expression and activity of MCM were determined by western blot and ultra-performance liquid chromatography,***:All patients had high levels of blood propionylcarnitine and urinary methylmalonyl *** the end of the study,two patients were lost to follow-up,three died,and three survived with mental *** to the wild-type protein,the expression levels of all missense mutations of in vitro MCM protein were decreased (P<0.05) except those for I597R,and the MCM activity of the mutations was reduced in a permissive ***:The missense mutations L140P,A141T,G161V,W309G,I505T,Q514K,I597R and G723D affected the stability and enzymatic activity of MCM,indicating that they had a disease-causing capacity.

关键词： methylmalonic acid methylmalonic acidemia methylmalonyl-CoA mutation missense mutation

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First Complete Genome Sequence of a Probiotic Enterococcus faecium Strain T-110 and Its Comparative Genome Analysis with Pathogenic and Non-pathogenic Enterococcus faecium Genomes

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Journal of genetics and Genomics 2015年第1期42卷 43-46页

作者： Purushothaman Natarajan Madasamy Parani Genomics Laboratory Department of Genetic EngineeringSRM University

Enterococci bacteria are important in environmental, food and clinical microbiology. Enterococcus faecium is a nosocomial pathogen that causes bacteremia, endocarditis and other infections. It is among the most prevalent organisms encountered in hospital-associated infections accounting for approximately 12% of nosocomial infections in the USA （Linden and Miller, 1999）. However, certain strains of E. faecium are not only non-pathogenic but also have beneficial effects on human health with probiotic potential. For example, E. faecium T-110 is a consortium member in several probiotic products including BIO-THREE~ which is widely prescribed for human, animal and aqua-cultural use. This strain was originally developed by TOA Pharmaceuticals in Japan, and later used in the probiotic products of several other companies.

关键词： 110 First Complete Genome Sequence of a Probiotic Enterococcus faecium Strain T-110 and Its Comparative Genome Analysis with Pathogenic and Non-pathogenic Enterococcus faecium Genomes ORFs

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Phosphorylation of human Sgo1 by NEK2A is essential for chromosome congression in mitosis

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Cell Research 2007年第7期17卷 608-618页

作者： Guosheng Fu1 Xia Ding2,3 Kai Yuan1,2 Felix Aikhionbare2 Jianhui Yao1 Xin Cai1,2 Kai Jiang1,2 Xuebiao Yao1,21 Laboratory of cellular Dynamics, Hefei National Laboratory, University of Science & Technology of China, Hefei 230027, China2 department of Physiology and Cancer Biology Program, Morehouse School of medicine, Atlanta, GA 30310, USA3 department of medicine, Beijing University of Chinese medicine, Beijing 100029, China Laboratory of cellular Dynamics Hefei National Laboratory University of Science & Technology of China Hefei China department of Physiology and Cancer Biology Program Morehouse School of Medicine Atlanta USA department of medicine Beijing University of Chinese Medicine Beijing China

Chromosome segregation in mitosis is orchestrated by the interaction of the kinetochore with spindle microtubules. Ourrecent study shows that NEK2A interacts with MAD1 at the kinetochore and possibly functions as a novel integrator ofspindle checkpoint signaling. However, it is unclear how NEK2 A regulates kinetochore-microtubule attachment in *** we show that NEK2A phosphorylates human Sgol and such phosphorylation is essential for faithful chromosomecongression in mitosis. NEK2A binds directly to HsSgol in vitro and co-distributes with HsSgol to the kinetochore ofmitotic cells. Our in vitro phosphorylation experiment demonstrated that HsSgol is a substrate of NEK2A and the phos-phorylation sites were mapped to Ser14and Ser507as judged by the incorporation of32P. Although such phosphorylation isnot required for assembly of HsSgol to the kinetochore, expression of non-phosphorylatable mutant HsSgol perturbedchromosome congression and resulted in a dramatic increase in microtubule attachment errors, including syntelic andmonotelic attachments. These findings reveal a key role for the NEK2A-mediated phosphorylation of HsSgol in orches-trating dynamic kinetochore-microtubule interaction. We propose that NEK2 A-mediated phosphorylation of human Sgolprovides a link between centromeric cohesion and spindle microtubule attachment at the kinetochores.

关键词： Sgo1 NEK2A phosphorylation microtubule kinetochore

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