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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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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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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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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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The inhibitory role of Co2+ on □-glucosidase: inhibition kinetics and molecular dynamics simulation integrating study

The inhibitory role of Co2+ on □-glucosidase: inhibition ki...

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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： Xuan Li Zhi-Rong L(u) Dong Shen Yi Zhan Jun-Mo Yang Yong-Doo Park Hai-Meng Zhou Qing Sheng Jinhyuk Lee College of Life Sciences Zhejiang Sci-Tech University Hangzhou 310018 P.R.China Zhejiang Provincial Key Laboratory of Applied Enzymology Yangtze Delta Region Institute of Tsinghua University 705 Yatai Road Jiaxing 314006 P.R.China Zhejiang Provincial Key Laboratory of Applied Enzymology Yangtze Delta Region Institute of Tsinghua University705 Yatai Road Jiaxing 314006 P.R.China School of Life Science Tsinghua University Zhongguancun Street Beijing 100084 P R.China Department of Dermatology Sungkyunkwan University School of Medicine Samsung Medical CenterSeoul 135-710 Korea Korean Bioinformation Center (KOBIC) Korea Research Institute of Bioscience and BiotechnologyDaejeon 305-806 Korea Department of Bioinformatics University of Sciences and Technology Daejeon 305-350 Korea

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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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The changes of telomere length of the Culture Human Lymphocytes

The changes of telomere length of the Culture Human Lymphocy...

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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： ruifeng ruifeng zhang yuefeng guo 中国辐射防护研究院

Objective To investigate the effect of 12C heavy ion irradiation on telomerelength in culture human *** Culture human lymphocyte line Peng-EBVwere exposed to 12C heavy ion at the irradiation doses of 0.1 Gy,0.5 Gy,1.0 Gy,2.0 Gyand the dose rate was 0.3-0.5 Gy/min.,*** cells were collected at 24,48,72 hafter *** DNA was *** telomere length was measured byfluorescence quantitative PCR,in order to estimate time and dose-response of change intelomere length induced by 12C heavy ion irradiation in culture human *** relative telomere length after exposure all significantly increased(except for 0.5,2.0 Gy exposed cells at 24 h);The relative telomere length of 1.0 Gy exposed cells and at72 h post-irradiation all highly significantly increased;1.0 Gy exposed cells entended moreobviously than that were exposed of 0.1 Gy,0.5 Gy and 2.0 Gy;The relative telomerelength of every dose group increased with prolongation of exposure *** 12Cheavy ion irradiation at 0.1,0.5,1.0and 2.0Gy may all result in telomere extension in culturehuman lymphocytes.

关键词： Telomere length 12C Fluorescence quantitative PCR

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CDT1 Gene Regulation of Genomic Instability Cell ThroughApoptosis and Cell Cycle

CDT1 Gene Regulation of Genomic Instability Cell ThroughApop...

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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：原雅艺党旭红左雅慧中国辐射防护研究院

Objective. To overexpress CDT1 gene and investigate its effect on apoptosis andcell cycle in human liver cells that were genomic instability induced by radiation(GIR)Methods Lentivirus-mediated CDT1 gene overexpression system was constructedand verified its validity and infectious efficiency. The apoptosis was detected byAnnexin Ⅴ-APC flow cytometry(FCM) and the cell cycle was determined by FCM withsingle PI staining. And the expression changes of p53, ATM, ATR, Bcl-2, Caspase3, Cyclin Egene were assayed by real time fluorescence quantitative PCR technology. ResultsThe successfuloverexpression of CDT1 gene in GIR was confirmed using quantitative polymerasechain reaction(qPCR) and FCM. Compared with negative control group, cell apoptosis wasremarkably induced in overexpression of CDT1 gene group. Meanwhile, overexpression ofCDT1 gene was able to significantly suppress p53 and Bcl-2 expression. Conclusion These results suggest that CDT1 play a critical role in the regulation of genomic instability through increase the apoptosis rate, and decrease the expression of p53 and Bcl-2 gene.

关键词： CDT1 gene genomic instability overexpression Cell cycle apoptosis

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Crystal structure of Rot, a global regulator of virulence genes in Staphylococcus aureus

Crystal structure of Rot, a global regulator of virulence ge...

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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：朱玉威范晓娇滕脉坤李旭中国科学技术大学

Staphylococcus aureus is a highly versatile pathogen that can infect the tissue ofhuman by producing a large arsenal of virulence factors,which was tightly regulated by acomplex regulatory ***,sharing sequence similarity to SarA homologues,is aglobal regulator that regulates numerous virulence ***,the recognition modelof Rot to the promoter region of target genes and the putative regulation mechanism *** this study,we report the 1.77resolution X-ray crystal structure of *** structure reveals that two Rot molecules form a compact homodimer,each of whichcontains a typical helix-turn-helix module and aβ-hairpin motif connected by a *** fluorescence polarization results indicate that Rot preferentially recognizes ATrichdsDNA with≈30 base pair nucleotides,and the conserved positively charged residueson the winged-helix motif are vital for binding to the AT-rich *** propose that theDNA recognization model of Rot may be similar to that of SarA,SarR and SarS,in whichthe helix-turn-helix motifs of each monomer interact with the major grooves of targetdsDNA and the winged motifs contact the minor ***,the structureshows that Rot adopts a novel dimerization model differing from that of other SarA *** expected,perturbation of the dimer interface abolishes the dsDNA binding abilityof Rot,suggesting that Rot functions as a *** addition,we further confirm our resultsin vivo by measuring the transcriptional regulation ofα-toxin,a major virulence factorsproduced by most *** strains.

关键词： Rot virulence genes Staphylococcus aureus

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The M-T Hook Structure Dramatically Increases the Potency of HIV-1 Fusion Inhibitors to Overcome Drug-resistance

The M-T Hook Structure Dramatically Increases the Potency of...

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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：姚雪崔胜医科院病原所

Peptides derived from the C-terminal heptad repeat region(CHR) of HIV-1 gp41 are potentfusion inhibitors by targeting the N-terminal heptad repeat region(NHR). We have recentlyidentified that the unique M-T hook structure preceding the pocketbinding motifof CHR peptides can greatly enhance the binding of inhibitors to the NHR pocket thus improvingthe anti-HIV activity. Here, we incorporated the M-T hook to the N-terminus of Sifuvirtide(SFT), a potent CHR-derived inhibitor currently under phase III clinical trials inChina. The high-resolution crystal structure of the resulting peptide MT-SFT confirmed ahighly conserved M-T hook conformation. Compared to SFT, MT-SFT exhibited significantlyimproved activities to bind to the NHR and to block the formation of six-helix bundle(6-HB) and to inhibit HIV-1 Env-mediated cell fusion, viral entry and replication. Importantly,MT-SFT inhibited SFT- and Enfuvirtide(T20)-resistant HIV-1 variants more efficientlyand had a dramatically increased genetic barrier to induce drug resistance. Therefore, ourstudies have verified that the M-T hook structure offers a general strategy for designingnovel HIV-1 fusion inhibitors and provide new insights into viral entry and inhibition.

关键词： HIV fusion inhibitor

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