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检索条件"任意字段=中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议"

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C/EBPβ-regulated Autophagy Facilitates Adipogenesis

C/EBPβ-regulated Autophagy Facilitates Adipogenesis

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者： Liang Guo Jia-Xin Huang Qi-Qun Tang Key Laboratory of Molecular Medicine the Ministry of Education Department of Biochemistry and Molecular Biology Fudan University Shanghai Medical College

Autophagy is a highly conserved self-digestion pathway involved in various physiologicaland pathophysiological *** studies have implicated a pivotal role of autophagyin adipocyte differentiation,but the molecular mechanism for its role and how it isregulated during this process is not ***,we show that CCAAT/enhancer-bindingproteinβ(C/EBPβ),an important adipogenic factor,is required for the activation of autophagyduring 3T3-L1 adipocyte *** autophagy-related gene Atg4b is identifiedas a de novo target gene of C/EBPβand plays an important role in ***,autophagy is required for the degradation of Klf2 and Klf3,two negative regulators of adipocyte differentiation,which is mediated by the adaptorprotein p62/***,the regulation of autophagy by C/EBPβand the role ofautophagy in Klf2/3 degradation and in adipogenesis are further confirmed in *** data describe a novel function of C/EBPβin regulating autophagy and revealthe mechanism of autophagy during adipocyte *** new insights into themolecular mechanism of adipose tissue development provide a functional pathway withtherapeutic potential against obesity and its related metabolic disorders.

关键词： 3T3-L1 preadipocytes Adipogenesis C/EBPβ Autophagy Atg4b

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Forward Genetic Screening for Regulators Involved in Cholesterol Synthesis Using Validationbased Insertional Mutagenesis

Forward Genetic Screening for Regulators Involved in Cholest...

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者： Wei Jiang Bao-Liang Song Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences College of Life Sciences Wuhan University

Somatic cell genetics is a powerful approach for unraveling the regulatory mechanism ofcholesterol ***,it is difficult to identify the mutant gene(s)due to cellsare usually mutagenized chemically or *** identify important genes controllingcholesterol biosynthesis,an unbiased forward genetics approach named validation-basedinsertional mutagenesis(VBIM)system was used to isolate and characterize the 25-hydroxycholesterol(25-HC)-resistant and SR-12813-resisitant *** we report thatfive mutant cell lines were *** which,four sterol-resistant mutants either containa truncated NH2-terminal domain of sterol regulatory element-binding protein(SREBP)-2 terminating at amino acids(aa)400,or harbor an overexpressed SREBP cleavage-activating protein(SCAP).Besides,one SR-12813 resistant mutant was identified tocontain a truncated COOH-terminal catalytic domain of 3-hydroxy-3-methylglutarylcoenzymeA reductase(HMG-CoA reductase).This study demonstrates that the VBIM systemcan be a powerful tool to screen novel regulatory genes in cholesterol biosynthesis.

关键词： forward genetic screen cholesterol SREBP SCAP VBIM HMG-CoA reductase

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miRNA disturbance by loss of Lkb1 in KrasG12D non-small celllung cancer

miRNA disturbance by loss of Lkb1 in KrasG12D non-small cell...

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者： Qian Zhang Huizhong Bao Youxin Jin Institute of Biomedicine and Biotechnology SIAT CAS Institute of Biochemistry and Cell Biology SIBS CAS

Non-small cell lung cancer(NSCLC) accounts for 80–85% of total lung cancer cases withextremely high mortality rates,which can be ameliorated by early diagnosis. Activation ofKRAS and inactivation of tumor suppressor LKB1 are common gene mutations detected inmultiple subtypes of NSCLCs. Concurrence of LKB1 loss with Kras oncogenic mutationsresults in higher malignancy of lung cancer. However, the LKB1- related characteristic microRNAs(miRNA) and their contribution to the malignant progression keeps unknown. Inthis study, miRNAs profiles were elucidated by deep sequencing of both KrasG12 D andKrasG12D Lkb1-/- mouse NSCLC tissues which stand for non-malignant and malignant tumorrespectively, with the lung of wild type C57BL/6 as normal control. Gene function analyses were performed for the targets of significant changed miRNAs to identify the functionof miRNAs, which further confirmed in human NSCLCs, elucidating the diagnostic potentialof miRNAs.

关键词： NSCLC Lkb1 Kras miRNA

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Sonchus asper extract inhibits LPS-induced oxidative stress andpro-inflammatory cytokine production in RAW264.7 macrophages

Sonchus asper extract inhibits LPS-induced oxidative stress ...

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者：王兰 Colloge of Life Science Henan Normal University

Sonchus asper is an herb that is used extensively to treat bronchitis, asthma, wounds,burns, and cough. To determine its mechanism of action, we examined the effects of anethyl acetate fraction(EAF) of S. asper on nitric oxide(NO) production and prostaglandin(PG)-E2 levels in lipopolysaccharide(LPS)-stimulated RAW264.7 macrophages. We observedsignificant inhibition associated with suppression of oxidative stress as evidencedby increased endogenous superoxide dismutase(SOD) activity and intracellular glutathione(GSH) levels, decreased reactive oxygen species(ROS) generation and lipid peroxidation,and restoration of the mitochondrial membrane potential. To confirm its antiinflammatoryeffects, expression of inducible NO synthase(iNOS), cyclooxygenase-2(COX-2), tumor necrosis factor(TNF)-α, and the anti-inflammatory cytokines IL-1β and IL-6 wereanalyzed by semi-quantitative RT-PCR. EAF treatment significantly reduced the expressionof all of these factors in a dose-dependent manner, and enhanced expression of the antioxidantsMnSOD and heme oxygenase-1(HO-1). Additionally, HPLC fingerprint resultssuggest that rutin, caffeic acid, and quercetin may be the active ingredients in EAF. Takentogether, these findings imply that the anti-inflammatory effect of EAF on LPS-stimulatedRAW264.7 cells is mediated by suppression of oxidative stress.

关键词： oxidative stress anti-inflammatory

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Inhibition of the IgE-mediated activation of RBL-2H3 cells by TIPP,a novel thymic immunosuppressive pentapeptide

Inhibition of the IgE-mediated activation of RBL-2H3 cells b...

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者： Qianqian Lian Yanna Cheng Fengshan Wang School of Pharmaceutical Sciences Shandong University

TIPP is a novel thymic immunosuppressive pentapeptide originally obtained from calfthymic immunosuppressive extract. The present study was aimed to investigate the inhibitoryactivity of TIPP on IgE-mediated activation of RBL-2H3 cells. Release of β-hexosaminidase, intracellular calcium assay, membrane ruffling, mRNA levels of cytokines,COX-2 expression and phosphorylation of mitogen-activated protein kinase(MAPK) weredetermined by colorimetric assay, fluorescence spectrophotometer, confocal fluorescencemicroscope, quantification PCR, and western blot respectively. The results showed thatTIPP significantly inhibited the degranulation in IgE-antigen complex-stimulated RBL-2H3 cells. It suppressed the increase in intracellular calcium and rearrangement of F-actin, andalso signicantly attenuated the transcription of proinammatory cytokines, such as IL-3, IL-6 and MCP-1. Western blot analysis showed that TIPP has an inhibitory activity on thephosphorylation of ERK, but shows no effect on phosphorylation of JNK and p38. Our datasuggest that TIPP effectively suppresses IgE-mediated activation of RBL-2H3 cells viablocking ERK MAP kinases signalling pathway.

关键词： allergy immunosuppressive peptide

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Structural insights into Paf1 complex assembly and histone binding

Structural insights into Paf1 complex assembly and histone b...

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者： Xinlei Chu Xiaohong Qin Huisha Xu Lei Li Zheng Wang Fengzhi Li Hao Zhou YuequanShen Jiafu Long Nankai University

The highly conserved Paf1 complex(PAF1C)plays critical roles in RNA polymerase II transcriptionelongation and in the regulation of histone *** has also been implicatedin other diverse cellular activities,including posttranscriptionalevents,embryonic developmentand cell survival and maintenance of embryonic stem cell ***,we reportthe structure of the human Paf1/Leo1 subcomplex within *** overall structurereveals that the Paf1 and Leo1 subunits form a tightly associated heterodimer throughantiparallel beta-sheet *** biochemical experiments indicate that Leo1binds to PAF1C through Paf1 and that the Ctr9 subunit is the key scaffold protein in ***,we show that the Paf1/Leo1 heterodimer is necessary for itsbinding to histone H3,the histone octamer,and nucleosome in *** results shed lighton the PAF1C assembly process and substrate recognition during various PAF1Ccoordinatedhistone modifications.

关键词： Transcription elongation RNA polymerase II Paf1 complex Histone modification

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hCINAP Negatively Regulates NF-κB Signaling by Recruiting thephosphatase PP1 to the IKK Complex for Deactivation

hCINAP Negatively Regulates NF-κB Signaling by Recruiting t...

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者： Linglong Qu Yapeng Ji Xiaofeng Zheng School of Life Sciences Peking University

The transcription factor NF-κB acts as a critical regulator in innate and adaptive immunesystem in response to a variety of stimuli, therefore both activation and inactivation of NF-κB signaling needed to be tightly controlled to maintain homeostasis. Despite extensiveand deep studies on the mechanisms of IKK activation, the molecular mechanisms underlyingIKK deactivation remain poorly understood. Here we demonstrated that human coilin-interacting nuclear ATPase protein(hCINAP) is a novel negative regulator in NF-κB signalingby targeting IKK. hCINAP associates with IKKα and IKKβ and inhibits IKK *** knock-down and knock-out of hCINAP markedly enhances IKK phosphorylation,NF-κB activation and NF-kB-responsive cytokines transcription. Futhermore,hCINAP directly interacts with catalytic subunits of serine/threonine protein phosphatase 1(PP1) and recruits PP1 to IKK for dephosphorylation. Taken together, our study identify apreviously unrecognized role for hCINAP in negative regulation of NF-κB signalingthrough inhibiting IKK phosphorylation by recruiting PP1, which provides new insight intothe mechanism of IKK deactivation and the negative regulation of inflammatory reactions.

关键词： hCINAP NF-κB IKK PP1

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Expression, purification and characterization of a recombinant humanized bispecific antibody targeting PD-1 and c-MET

Expression, purification and characterization of a recombina...

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者：吴毅于敏 The Key Laboratory of Molecular Medicine he Ministry of Education Department of Biochemistry and Molecular Biology Shanghai Medical College Fudan University

A recombinant humanized bispecific antibody（rh BsAb）, recognizing programmed celldeath protein 1（PD-1） and hepatocyte growth factor receptor（HGFR or c-MET） was constructedwith two monoclonal antibodies（mAbs）, which were obtained from hybridomasand modified by aligning with human antibody database and screening through DiscoveryStudio software. The rhBsAb was expressed by HEK 293 cell, then purified by Protein A affinitychromatography. SDS-PAGE and Western blotting analysis of the purified rhBsAb indicatedthat the rhBsAb was successfully expressed as a 50kDa protein with yields of approximately150mg/L of HEK 293 cell culture supernatant. The purity of the rhBsAb was upto 95%. The rhBsAb was further confirmed by N-terminal sequencing, MALDI-TOF-MS andindirect flow cytometry. PD-1 blocked by the rhBsAb improved T cell cytokine（IL-2 andIFN-γ） secretion by 3-6 fold. The phosphoration level of c-MET, AKT and ERK1/2 wasdownregulated by blocking c-MET with the rhBsAb.

关键词： bispecific antobody PD-1 c-MET HEK 293

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Screening of chemoeffectors for Chemoreceptor Tsr to disclose the possible molecular mechanism of repellents

Screening of chemoeffectors for Chemoreceptor Tsr to disclos...

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者：陈曦马晓旻来鲁华北京大学前沿交叉学科研究院定量生物学中心北京大学化学与分子工程学院

Chemotaxis is a phenomenon that motile bacteria move directionally alone chemicalgradients in the *** ability depends on the chemoreceptors which can detectminute concentration changes in the chemical environment,and generate a highlyamplified signal that modulates the cell’s flagella motor *** aretransmembrane proteins including periplasm domain,transmembrane domain and *** exist as trimer of dimers in *** are five types of chemoreceptorsin ***:Tar for aspartate,Tsr for serine,Tap for peptides,Trg for ribose and galactoseand Aer for *** cytoplasm domains of these receptors are highly conserved,andthe ligand selectivity depends on the periplasm *** Bi et *** discoveredsix novel attractants and two antagonists for the Escherichia coli chemoreceptorTar and revealed the molecular mechanisms for attractants binding and ***,no repellent molecules have been discovered,and the molecular mechanism ofrepellent remains *** research focus on another chemoreceptor,*** want todiscovery the chemoeffectors of Tsr and explore the molecular mechanism of repellentbinding and ***,Autodock was used to virtually screen compounds that bind toTsr ligand binding *** the potential chemoeffectors virtually screened out,fourattractants and four repellents were discovered in microfluidic *** bindingaffinity of these molecule with Tsr periplasm domain was measured by Isothermal titrationcalorimetry(ITC)and Microscale Thermophoresis(MST).Then we will verify the responseof the chemotaxis signal pathway to these chemoeffectors and explore the molecularmechanisms of the chemoeffectors,especially repellents,binding and signaling.

关键词： chemoreceptor Tsr molecular mechanism

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酶解法制备南极磷虾油的研究

酶解法制备南极磷虾油的研究

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者：徐晓斌王世立山东省医药生物技术研究中心

背景南极磷虾油提取自南极磷虾,其富含的磷脂、多不饱和脂肪酸和虾青素等营养物质在预防心脑血管疾病,促进大脑发育、延缓衰老、缓解痛风和类风湿关节炎等方面具有显著功效。酶解提油法是最近几年来研究和利用的新提油工艺,其原理是通... 详细信息

背景南极磷虾油提取自南极磷虾,其富含的磷脂、多不饱和脂肪酸和虾青素等营养物质在预防心脑血管疾病,促进大脑发育、延缓衰老、缓解痛风和类风湿关节炎等方面具有显著功效。酶解提油法是最近几年来研究和利用的新提油工艺,其原理是通过酶解作用使脂类与复合的蛋白质、多糖分离开,从而促使油脂的释放。酶解法在制备鱼油上应用较广,而用酶解法制备南极磷虾油鲜有报道。目的优化南极磷虾油酶解法制备工艺,包括优化其酶解条件及萃取条件。方法酶解法制备南极磷虾油的基本流程是:将冷冻鲜虾在室温中解冻,用组织捣碎机打成匀浆;加适量复合蛋白酶在适宜温度下酶解一定时间;用有A液和B液将虾油萃取出来(A液和B液为常用萃取剂的一种或几种配比);将萃取液真空干燥获得南极磷虾油,并以提取率为指标进行以下实验。酶解条件的优化:设置25℃、35℃、45℃、55℃、65℃五个温度梯度进行单因素试验,设置1h、2h、3h、4h、5h、6h六个时间梯度进行单因素试验,设置0、0.05%、0.1%、0.15%、0.2%五个酶量梯度进行单因素试验;酶解条件的三个因素酶解温度、酶解时间和酶量进行响应面分析。萃取条件的优化:A液和B液添加顺序的比较;设置匀浆中含水量的0.25倍、0.5倍、0.75倍、1倍四个A液量梯度进行单因素试验,设置A液量的0.5倍、1倍、2倍、3倍四个B液量梯度进行单因素试验,设置5min、20min、35min、50min、65min五个时间梯度进行单因素试验;萃取条件的三个因素A液量、B液量和萃取时间进行响应面分析。结果单因素试验及响应面分析结果显示,添加0.14%的复合蛋白酶在45℃下酶解4.9小时,加入匀浆含水量的0.78倍A液搅拌混匀,再加入2倍A液量的B液搅拌混匀,搅拌萃取40min,可保证虾油品质的同时获得最大提取率95%。所得虾油磷脂含量为28%。

关键词：南极磷虾油酶解法

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