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Parkinson’s disease-associated VPS35 mutant reduces mitochondrial membrane potential and impairs PINK1/Parkinmediated mitophagy

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Translational Neurodegeneration 2021年第2期10卷 240-256页

作者： Kai Yu Ma Michiel R.Fokkens Fulvio Reggiori Muriel Mari Dineke S.Verbeek department of genetics University of GroningenUniversity Medical Center GroningenGroningenThe Netherlands department of Biomedical Sciences of Cells and Systems University of GroningenUniversity Medical Center GroningenGroningenThe Netherlands

Background:Mitochondrial dysfunction plays a prominent role in the pathogenesis of Parkinson’s disease(PD),and several genes linked to familial PD,including PINK1(encoding PTEN-induced putative kinase 1[PINK1])and PARK2(encoding the E3 ubiquitin ligase Parkin),are directly involved in processes such as mitophagy that maintain mitochondrial *** dominant p.D620N variant of vacuolar protein sorting 35 ortholog(VPS35)gene is also associated with familial PD but has not been functionally connected to PINK1 and ***:To better mimic and study the patient situation,we used CRISPR-Cas9 to generate heterozygous human SH-SY5Y cells carrying the PD-associated D620N variant of *** cells were treated with a protonophore carbonyl cyanide m-chlorophenylhydrazone(CCCP)to induce the PINK1/Parkin-mediated mitophagy,which was assessed using biochemical and microscopy ***:Mitochondria in the VPS35-D620N cells exhibited reduced mitochondrial membrane potential and appeared to already be damaged at steady *** a result,the mitochondria of these cells were desensitized to the CCCPinduced collapse in mitochondrial potential,as they displayed altered fragmentation and were unable to accumulate PINK1 at their surface upon this ***,Parkin recruitment to the cell surface was inhibited and initiation of the PINK1/Parkin-dependent mitophagy was ***:Our findings extend the pool of evidence that the p.D620N mutation of VPS35 causes mitochondrial dysfunction and suggest a converging pathogenic mechanism among VPS35,PINK1 and Parkin in PD.

关键词： VPS35 PINK1 Parkin Mitophagy Mitochondrial membrane potential Parkinson’s disease

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Isolation of Cr(Ⅵ) reducing bacteria from industrial effuents and their potential use in bioremediation of chromium containing wastewater

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Journal of Environmental Sciences 2009年第6期21卷 814-820页

作者： Ahmed Zahoor Abdul Rehman department of Microbiology and Molecular genetics University of the PunjabNew Campus

The present study was aimed to assess the ability of Bacillus ***-2-1 and Staphylococcus capitis to reduce hexavalent chromium into its trivalent *** ***-2-1 could tolerate Cr（Ⅵ）（4800 μg/mL） and *** could tolerate Cr（Ⅵ）（2800 μg/mL）.Both organisms were able to resist Cd^2＋（50 μg/mL）,Cu^2＋（200 μg/mL）,Pb^2＋（800 μg/mL）,Hg^2＋（50 μg/mL） and Ni2＋（4000 μg/mL）.*** resisted Zn^2＋ at 700 μg/mL while Bacillus ***-2-1 only showed resistance up to 50 μg/*** ***-2-1 and *** showed optimum growth at pH 6 and 7,respectively,while both bacteria showed optimum growth at 37°*** ***-2-1 and *** could reduce 85% and 81% of hexavalent chromium from the medium after 96 h and were also capable of reducing hexavalent chromium 86% and 89%,respectively,from the industrial effuents after 144 *** free extracts of Bacillus ***-2-1 and *** showed reduction of 83% and 70% at concentration of 10 μg Cr（Ⅵ）/mL,*** presence of an induced protein having molecular weight around 25 kDa in the presence of chromium points out a possible role of this protein in chromium *** bacterial isolates can be exploited for the bioremediation of hexavalent chromium containing wastes,since they seem to have a potential to reduce the toxic hexavalent form to its nontoxic trivalent form.

关键词： Cr(Ⅵ) reducing bacteria bioremediation Bacillus sp. JDM-2-1 Staphylococcus capitis

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Growth phase dependent changes in the structure and protein composition of nucleoid in Escherichia coli

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Science China(Life Sciences) 2015年第9期58卷 902-911页

作者： TALUKDER AliAzam ISHIHAMA Akira department of Microbiology Jahangirnagar University Micro-Nano Technology Research Center Hosei University department of Molecular genetics National Institute of Genetics

The genomic DNA of bacteria is highly compacted in a single or a few bodies known as nucleoids. Here, we have isolated Escherichia coli nucleoid by sucrose density gradient centrifugation. The sedimentation rates, structures as well as pro- tein/DNA composition of isolated nucleoids were then compared under various growth phases. The nucleoid structures were found to undergo changes during the cell growth; i. e., the nucleoid structure in the stationary phase was more tightly com- pacted than that in the exponential phase. In addition to factor for inversion stimulation （Fis）, histone-like nucleoid structuring protein （H-NS）, heat-unstable nucleoid protein （HU） and integration host factor （IHF） here we have identified, three new can- didates of E. coli nucleoid, namely DNA-binding protein from starved cells （Dps）, host factor for phage QJ3 （Hfq） and sup- pressor of taC phenotype A （StpA）. Our results reveal that the major components of exponential phase nucleoid are Fis, HU, H-NS, StpA and Hfq, while Dps occupies more than half of the stationary phase nucleoid. It has been known for a while that Dps is the main nucleoid-associated protein at stationary phase. From these results and the prevailing information, we propose a model for growth phase dependent changes in the structure and protein composition of nucleoid in E. coli.

关键词： growth phase sucrose gradient bacterial nucleoid DNA binding protein DNA compaction

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Vascular endothelial growth factor gene polymorphisms in age-related macular degeneration in a Turkish population

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International Journal of Ophthalmology(English edition) 2014年第5期7卷 773-777页

作者： Yunus Bulgu Gokhan Ozan Cetin Vildan Caner Ebru Nevin Cetin Volkan Yaylali Cem Yildirim department of Ophthalmology School of MedicinePamukkale University department of medical genetics School of MedicinePamukkale University department of medical Biology School of MedicinePamukkale University

AIM:To assess the association between age-related macular degeneration(AMD) and three single nucleotide polymorphisms(SNPS) related to the vascular endothelial growth factor(VEGF) ***:The patients who were diagnosed with AMD were included in this prospective study. Three SNPs(rs1413711, rs2146323, and rs3025033) of the VEGF gene were genotyped by real-time polymerase chain reaction in the genomic DNA isolated from peripheral blood samples of the 82 patients and 80 ***:The genotype frequencies of rs1413711 and rs2146323 were not significantly different between the study group and the control group(P =0.072 and P =0.058).However, there was a significant difference in the genotype frequencies of these SNPs between the wet type AMD and dry type AMD(P =0.005 and P =0.010,respectively). One of the SNPs(rs1413711) was also found to be associated with the severity of AMD(P =0.001)with significant genotype distribution between early,intermediate, and advanced stages of the disease. The ancestral alleles were protective for both SNPs while the polymorphic alleles increased the risk for dry ***:VEGF SNPs rs1413711 and rs2146323 polymorphisms are significantly associated with AMD subtypes in our population.

关键词： age-related macular degeneration vascular endothelial growth factor single nucleotide polymorphism

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Impact of copy number of distinct SV40PolyA segments on expression of a GFP reporter gene

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Science China(Life Sciences) 2010年第5期53卷 606-612页

作者： YIN Kang,WANG XiuFang,MA Huan,XIE Ying,FENG JingJing,YANG QinQing & Lü ZhanJun Hebei Key Lab of Laboratory Animal,department of genetics,Hebei medical University,Shijiazhuang 050017,China 1. Hebei Key Lab of Laboratory Animal Department of Genetics Hebei Medical University Shijiazhuang 050017 China

The presence of Alu repeats downregulates the expression of the green fluorescent protein(GFP) *** found that SV40PolyA(PolyA,240 bp),in either orientation,eliminated the inhibition of GFP gene expression induced by Alu repeats when it was placed between the GFP gene and the Alu *** this study,4 different segments(each 60 bp) were amplified from antisense PolyA(PolyAas) by PCR,and inserted upstream of Alu14 in pAlu14 plasmid(14 Alu repeats inserted downstream of the GFP gene in vector pEGFP-C1 in a head-tail tandem manner).Segments 1F1R(the first 60 bp segment at the 5' end of PolyAas) and 4F4R(the fourth 60 bp segment from the 5' end of PolyAas) did not activate GFP gene expression,whereas 2F2R and 3F3R(the middle two segments) did(as detected by Northern blot analysis and fluorescent microscopy).Different copy numbers of 2F2R and 3F3R segments,in a head and tail tandem manner,were inserted downstream of the GFP gene in pAlu14.p2F2R*4-Alu28,p3F3R*4-Alu18 and p3F3R*4-Alu28 were used as length controls to verify that the decrease in the expression of GFP was not due to the increased length of the inserted segment in the expression *** found that 2 and 4 copies of 2F2R or 3F3R activated the GFP gene more strongly than one copy of ***,more than 8 copies of 2F2R or 3F3R reduced the activation of the GFP *** concluded that SV40PolyAas contained at least two gene-activating elements(2F2R and 3F3R) and 2-4 copies of 2F2R or 3F3R were optimal for the expression of the GFP gene.

关键词： copy number SV40PolyA repeat sequence GFP Alu

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Chemotherapy suppresses SHH gene expression via a specific enhancer

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Journal of genetics and Genomics 2023年第1期50卷 27-37页

作者： Yafei Zhang Jianqiong Lin Kaibin Yang Zhicao Yue department of Cell Biology and medical genetics International Cancer Centerand Guangdong Key Laboratory for Genome Stability and Disease PreventionShenzhen University Medical SchoolShenzhenGuangdong 518060China

Sonic hedgehog(SHH)signaling is a key regulator of embryonic development and tissue homeostasis that is involved in gastrointestinal(GI)cancer *** of SHH gene expression is a paradigm of long-range enhancer *** the classical chemotherapy drug 5-fluorouracil(5FU)as an example,here we show that SHH gene expression is suppressed by *** is downstream of immediate early genes(IEGs),including Early growth response 1(Egr1).A specific 139 kb upstream enhancer is responsible for its *** down EGR1 expression or blocking its binding to this enhancer renders SHH unresponsive to *** further demonstrate that down-regulation of SHH expression does not depend on 5FU’s impact on nucleotide metabolism or DNA damage;rather,a sustained oxidative stress response mediates this rapid *** enhancer is present in a wide range of tumors and normal tissues,thus providing a target for cancer chemotherapy and its adverse effects on normal *** propose that SHH is a stress-responsive gene downstream of IEGs,and that traditional chemotherapy targets a specific enhancer to suppress its expression.

关键词： Sonic hedgehog Chemotherapy Immediate early genes EGR1 5-Fluorouracil

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Novel CDH1 germline mutations identified in Chinese gastric cancer patients

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World Journal of Gastroenterology 2013年第6期19卷 909-916页

作者： Qin-Hua Chen Wei Deng Xiao-Wei Li Xiu-Fang Liu Jing-Mei Wang Li-Feng Wang Nong Xiao Qiong He Ya-Ping Wang Yi-Mei Fan department of medical genetics Medical SchoolNanjing University Jiangsu Key Laboratory of Molecular Medicine department of Pathology Drum Tower Hospital Affiliated to Medical SchoolNanjing University department of Oncology Drum Tower Hospital Affiliated to Medical SchoolNanjing University Lujiang Hospital

AIM:To give a comprehensive report of E-cadheringene (CDH1) variations in a population at a high risk for gastric cancer (GC).METHODS:The samples consisted of 178 men and 58 women with a mean age of 62.3 ± 9.4 years and an age range of 30-84 years.A total of 240 cancerfree controls were recruited (mean age of 61.8 ± 10.1 years,age range of 26-82 years).Samples were screened for CDH1 germline mutations by high-resolution melting analysis or directly *** reporter assay,RNA splicing assay and bioinformatic analysis were used to evaluate the effect of ***:Four novel CDH1 sequence alterations were identified in GC patients including a G>T transition 49 bp before the start codon;a three-nucleotide deletion,c.44_46del TGC;one missense mutation,c.604G>A (V202I);and one variation in the intron,c.1320+7A>*** addition,polymorphism frequencies were observed for CDH1-164delT,-161C>A,-73A>C,c.48+6C>T,c.48+62_48+63delinsCGTGCCCCAGCCC,c.894C>T (A298A),c.1224G>A (A408A),c.1888C>G (L630V),c.2076T>C (A692A),and c.2253C>T (N751N) which is similar to the data reported in http://***/projects/SNP/.RNA splicing analysis suggested that the c.1320+7A>G and c.1224G>A variations did not affect exon splicing *** reporter assay demonstrated that the c.-49T variation might be helpful for E-cadherin transcription,though the increase in transcription activity is limited (only 33%).SIFT score and PolyPhen analysis both demonstrated that the L630V missense mutation probably damages protein function,while the V202I variant does ***:This study reveals novel mutations in sporadic GC patients which had been poorly investigated for susceptibility genes.

关键词： Gastric cancer Germline mutation CDH1 Luciferase reporter assay RNA splicing analysis

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Luminal/extracellular domains of chimeric CI-M6PR-C proteins interfere with their retrograde endosome-to-TGN trafficking in the transient expression system

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The Journal of Biomedical Research 2018年第4期32卷 245-256页

作者： Fei Chang Na Li Kang Yan Yumin Huang Hongfei Xu Yongjian Liu Jiangsu Key Laboratory of Xenotransplanation Department of Medical GeneticsNanjing Medical University department of Orthopedics the First Affiliated Hospital of Nanjing Medical University department of Pharmacology & Chemical Biology University of Pittsburgh School of Medicine

The membrane trafficking of cation-independent mannose 6-phosphate receptor（CI-M6PR） between the transGolgi network（TGN） and endosomal compartments is not only critical for maintaining lysosomal function but also a well-known event for understanding molecular and cellular mechanisms in retrograde endosome-to-TGN *** it has been well established in literature that the C-terminus of bovine CI-M6PR determines its retrograde trafficking,it remains unclear whether the luminal domain of the protein plays a role on these sorting *** this study,we found that partial deletion of luminal domain of human CI-M6PR mistargeted the mutant protein to nonTGN ***,replacing the luminal domain of both bovine and human CI-M6PR with that from irrelevant membrane proteins such as CD8 or Tac also altered the TGN targeting of the chimeric *** the other hand,only short sequence from HA fused with the transmembrane domain and C-terminus of the receptor,HA-hCIM6PR-tail,resulted in its preferential targeting to TGN as for the full length receptor,strongly suggesting that sorting of the receptor may be influenced by luminal ***,using this luminal truncated form of HA-hCIM6 PR as a model cargo,we found that the trafficking of the chimeric protein was regulated by the retromer complex through interacting with *** conclusion,our study strongly suggested that the disrupted luminal domain from hCI-M6PR or other irrelevant membrane proteins interfere with the process of membrane trafficking and TGN targeting of CI-M6PR.

关键词： CI-M6PR TGN targeting retrograde trafficking SNX5

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Association of eleven single nucleotide polymorphisms with refractive disorders from Eskisehir,Turkey

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International Journal of Ophthalmology(English edition) 2021年第6期14卷 812-817页

作者： Nadir Unlu Ebru Erzurumluoglu Gokalp Serap Arslan Oguz Cilingir Muzaffer Bilgin Engin Yildirim Huseyin Gursoy department of Ophthalmology Eskisehir Osmangazi University Medical FacultyEskisehir 26040Turkey department of medical genetics Eskisehir Osmangazi University Medical FacultyEskisehir 26040Turkey department of Biostatistics Eskisehir Osmangazi University Medical FacultyEskisehir 26040Turkey department of Pharmacology Eskisehir Osmangazi University Medical FacultyEskisehir 26040Turkey

AIM:To investigate relationship between refractive errors and eleven single nucleotide polymorphisms(SNPs)in HGF,GC,MFN1,GNB4,and VDR genes in Turkish ***:A group of 212 participants with myopia(n=91),hyperopia(n=45),and emmetropia(n=76)were investigated in this *** in HGF,GC,MFN1,GNB4 and VDR genes were studied by Snap Shot ***:The patients in this study consists of 47 female/44 male(age:23.47±4.30)patients with myopia,20 female/25 male(age:31.20±8.02)with hyperopia and 33 female/43 male(age:25.22±6.60)with *** genotype distribution of the rs7618348 polymorphism,which was the only statistically significant one between myopia and emmetropia *** genotype distribution of the rs3819545,rs3735520,rs7041,and rs2239182 polymorphisms,which were statistically significant between hyperopia and emmetropia ***:The importance of genetic predisposition to refractive errors with respect to etiology of the disease is *** is known that polymorphism studies may differ because of genetic diversity among populations so larger cohort studies are required in different populations to enlighten the etiology of the refractive errors.

关键词： refractive disorders myopia hyperopia genetics single nucleotide polymorphisms Turkey

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Therapeutic potential of mesenchymal stem cells in the treatment of acute liver failure

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World Journal of Gastroenterology 2022年第28期28卷 3627-3636页

作者： Carl Randall Harrell Dragica Pavlovic Valentin Djonov Vladislav Volarevic Regenerative Processing Plant LLC Palm HarborFL 34176United States department of genetics Faculty of Medical SciencesUniversity of KragujevacKragujevac 34000Serbia Institute of Anatomy University of BernBern 3012Switzerland department of medical genetics and Microbiology and Immunology Faculty of Medical SciencesUniversity of KragujevacKragujevac 34000Serbia

Acute liver failure(ALF)is a severe and life-threatening condition in which rapid deterioration of liver function develops in a patient who has no preexisting liver *** stem cells(MSCs)are immunoregulatory stem cells which are able to modulate phenotype and function of all immune cells that play pathogenic role in the development and progression of *** in juxtacrine and paracrine manner attenuate antigen-presenting properties of dendritic cells and macrophages,reduce production of inflammatory cytokines in T lymphocytes,suppress hepatotoxicity of natural killer T(NKT)cells and promote generation and expansion of immunosuppressive T,B and NKT regulatory cells in acutely inflamed *** to their nano-sized dimension and lipid envelope,intravenously injected MSC-derived exosomes(MSC-Exos)may by-pass all biological barriers to deliver MSC-sourced immunoregulatoy factors directly into the liver-infiltrated immune cells and injured *** obtained by us and others revealed that intravenous administration of MSCs and MSC-Exos efficiently attenuated detrimental immune response and acute inflammation in the liver,suggesting that MSCs and MSC-Exos could be considered as potentially new remedies in the immunotherapy of *** this review,we emphasize the current knowledge about molecular and cellular mechanisms which are responsible for MSC-based modulation of liver-infiltrated immune cells and we discuss different insights regarding the therapeutic potential of MSCs in liver regeneration.

关键词： Mesenchymal stem cells Acute liver failure Therapy Immunomodulation Regeneration

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