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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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RhoGDIβ switches cell fate between smooth muscle cell versus adipocyte via regulating actin filament formation

RhoGDIβ switches cell fate between smooth muscle cell versu...

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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：黄海艳张文婷蒋文艳陈素贞汤其群复旦大学上海医学院生化系

Mesenchymal stem cells can differentiation into multiple mesoderm cell types under differentinducing conditions and the inducers of the differentiation along one lineage oftenrepresses the differentiation toward alternative lineages, but the extracellular signals integrationinvolved in the decision of commitment fate is largely unknown. Here, our iTRAQbasedproteomics profiling studies identify RhoGDIβ, an inhibitor of small G-protein Rhofamily, as a critical component of the switch in the decision of adipocyte fate versussmooth muscle cell fate, in response to BMP4 exposure. RhoGDIβ is notably downregulatedduring BMP4-induced adipocyte lineage commitment, which is involved with cytoskeleton-associated protein lysyl oxidase. Introduction of excessive RhoGDIβ completelyprevents BMP4-induced adipocyte lineage commitment and simultaneously stimulatesearly smooth muscle commitment via suppressing the activation of Rac1. Overexpressionof RhoGDIβ induces the stress fibers of F-actin in phospho-MLC dependent manner, indicatingthat the cytoskeletal tension regulated by RhoGDIβ contributes to the determinationof adipocyte versus myocyte commitment. Furthermore, the overexpression of RacV12(Rac1 active form) totally rescues the inhibition of RhoGDIβ on adipocyte commitment,simultaneously preventing the formation of the smooth muscle phenotype and disruptingthe stress fiber in cells overexpressing RhoGDIβ. Collectively, these results indicate thatRhoGDIβ functions as a novel BMP4 signaling node that directs the commitment towardadipocyte versus myocyte lineages. This study will shed a light on clarifying the molecularbasis of the switch between adipogenesis and myogensis.

关键词： RhoGDIβ smooth muscle cells adipocytes Rac1 Mesenchymal stem cells

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The search of the degradation pathway of dysbindin

The search of the degradation pathway of dysbindin

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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：朱春燕杨微许琪中国医学科学院基础医学研究所

Background The correct degradation of proteins is vital for the homeostasis of neuronal system. DISC1,a suspectable gene for schizophrenia, has been reported to be aggregated in patients ofschizophrenia. DTNBP1 is another most studied susceptible gene of schizophrenia. Dysbindin-B, lacking the tenth extron, differs from whole length A due to the selective splicingof DTNBP1. The altered level of dysbindin expression has been reported in both schizophreniapatients and sandy mouse. But the degradation pathways for both dysbindin-Aand dysbindin-B are not clear. To search the possible degradation pathways,we conducteda yeast two-hybrid *** Firstly, as the bait, the sequence of dysbindin-A, dysbindin-B, pest of dysbindin-A wereconstructed to pGB plasmids. Secondly, the result of self-activation indicated only pest ofdysbindin-A can be used in the following experiment. Thirdly, the prey plasmids wereamplified and collected(human fetal brain library). Then, the prey and bait plasmids wereco-transformed to Y190 yeast line. The positive clones were ***23 positive clones on Leu-Trp-His-SD+3AT plates were found. All of the positive cloneswere proved to be negative by β-galactosidase detection. All of the positive clones on Leu-Trp-His-SD+3AT plates were sequenced. Only one full length and code-matchedgene(TUBB4B)was *** TUBB4 B encodes tubullin β-4B which is a number of microtube. No evidence indicates tubullinβ-4B is related to any degradation pathway. In our experiment, only pest of dysbindin-A do not stimulate self-activation and can be used as bait. Self-activation is one of themost obvious limits which contribute to the shortage of yeast two-hybrid.

关键词： dysbindin protein degradation

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Overexpression of miR-34a delays irradiation-induced H2AXactivation and DSBs repair in lung cancer cells

Overexpression of miR-34a delays irradiation-induced H2AXact...

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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： Deling Lin Wan Yao Yi Song Department of Biochemistry and Molecular Biology Beijing Institute of Radiation Medicine

Objective: MiR-34 a was a member of the microRNAs that induced by irradiation. In thisstudy, we pay attention to the function of miR-34 a in H2 AX activation, DNA double strandbreaks repair and tumor radio-sensitivity after irradiation in A549 lung cancer cells. Methods:MiR-34 a was over-expressed in the NSCLC A549 cell lines, and then double strandsbreaks and repair were measured by micronucleus experiment and single cell gel electrophoresistest in vitro. Western-blot was used to detect the expression of H2 AX, γH2AX, andCCND1. Then γH2AX foci were observed by immunofluorescence. Cell cycle distributionand cell proliferation were detected by flow cytometry and CCK8 analysis. Results: After 10 Gy irradiation, micronueleatede cells were increased in the miR-34 a over-expressed A549 cells. Over-expression of miR-34 a delays the activation of H2 AX after irradiation *** addition, miR-34 a inhibited CCND1 expression and reduced G1 cells. AccumulationA549 cells at the G1 stage can also delay he activation of H2 AX after irradiation. Conclusion:Our data suggest that over-expression of miR-34 a in iR-34 a deficient A549 cells inhibitedCCND1 expression and G1 cell cycle arrest, which delayed activation of H2 AX afterirradiation. These effects proceed to impact on the DNA damage repair and finally have aninfluence on radio-sensitivity of tumor cells.

关键词： miR-34a Ionizing Radiation γH2AX

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Study on serum proteoics in rats irradiated by 137Cs γ-ray

Study on serum proteoics in rats irradiated by 137Cs γ-ray

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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： Yahui Zuo Xuhong Dang Huifang Zhang Zhikai Duan China Institute for Radiation Protection

Objective To investigate the changes of proteomics in serum of Sprague-Dawley(SD)rats after accumulated irradiation by 137 Cs γ ray. Methods Thirty matureSD rats were randomly divided into three groups:0.2Gy group, 2Gy group and healthy controlgroup. Rats were irradiated at dose rate of 0.336mGy/min for 10 days and 20 dayscontinuously. Isobaric tags for relative and absolute quantitation(iTRAQ) was used to analyzethe different protein expression in serum of irradiated rats. Gene Ontology, KEGGpathway and protein-protein interaction network analysis was conducted using *** In total, 363 protein spots were identified. 29 proteins were differentially expressedboth in two groups compared with control, of which 10 were up-regulated and 19down-regulated. Based on softwares about classtification of biological process categories,classtification of cellular component and classtification of molecular function, differentiallyexpressed proteins may play important role in metabolic process, regulation of biologicalprocess, protein binding, catalytic activity. Analysis with the PAJEK software demonstratedthat those 16 differentially expressed proteins could form a complicated interaction networkwhere Glutathione S-transferase P1(GSTP1), Phosphoglycerate kinase 1(PGK1) andProtein disulfide-isomerase(PDI) might be key *** Accumulated irradiationcan induce differentially expressed proteins in serum of irradiated rats. Analysis on functional roles of the screened proteins GSTP1, PGK1 and PDI may provide insight intofurther mechanistic investigations and underlying molecular biomarkers induced by accumulated irradiation.

关键词： Accumulated irradiatin Serum proteomics Isobaric tags for relative and absoluteq uantitation(iTRAQ) Interaction network

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Inhibitive effect of dexamethasone loaded layered doublehydroxide on Th2 cell development in a rat asthma model

Inhibitive effect of dexamethasone loaded layered doublehydr...

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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： Wenrui wang Rongrong Zhu Hui Liu ShiLong Wang Research Center for Translational Medicine at East Hospital School of Life Sciences and Technology Tongji University Eastern Hepatobiliary Surgery Hospital Second Military Medical University

Although inhaled steroids are the treatment of first choice to control asthma, frequentcorticosteroid steroid administration is associated with many side effects such as adrenocorticalsuppression, Cushing's syndrome and osteoporosis. The purpose of this study wasto develop a drug delivery system to overcome the limitation of corticosteroid and improvethe therapeutic efficacy in an ovalbumin(OVA)-induced allergic rats model ofasthma. Dexamethasone Sodium Phosphate(Dexa) was incorporated into layered doublehydroxides(LDHs) through a co-precipitation method with an average particle size of approximately100 nm. The X-ray diffraction patterns and FT-IR spectra of the Dexa-LDHs indicatedthat the Dexa molecules were successfully intercalated into LDH gallery via electrostaticinteraction. A gradual and biphasic in vitro release behavior of the drug from LDH inpH=7.4 phosphate buffered saline(PBS) was observed, and the parabolic diffusion modelwas used to simulate the release kinetics of Dexa from the LDHs. The potential of Dexa-LDHs as an anti-asthmatic agent was evaluated using a murine model of ovalbumin-challenged allergic asthma. The numbers of inflammatory cells in bronchoalveolar lavage(BAL) fluid were significantly reduced when the rats was administrated with Dexa-LDHs. Furthermore, Dexa-LDHs significantly inhibited the increase in T-helper-2-type cytokinessuch as IL-4 and IL-13 in bronchoalveolar lavage fluid(BALF), and also effectivelysuppressed airway hyperresponsiveness. In conclusion, these results suggested that Dexa-LDHs had great potential for the treatment of airway inflammatory diseases such asasthma.

关键词： airway hyperresponsiveness Dexamethasone Sodium Phosphate solid lipidn anoparticles

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Studies on the purification of polysaccharides from Mutant Irpexlacteus Fr.(ILN10) and their anti-nephritic effect

Studies on the purification of polysaccharides from Mutant I...

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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： Di Wang Jiahui Lu Qingfan Meng Lirong Teng Jilin Universtiy

Irpex lacteus, one of white-rot basidiomycetes, has been widely used as a traditionalChinese medicine which displays a remarkable therapeutic effect on chronic *** present study was aimed to investigate the antinephritis activity of purified polysaccharideextracted from Irpex lacteus in both in vitro and in vivo experiments. Two water-soluble polysaccharide fractions(ILN3A, ILN3B) were isolated from Irpex lacteus via deproteinization,decolorization, and purification using dicthy laminoethyl(DEAE)-celluloseion exchange and sephadex G100 chromatographies. The average molecular weights andmonosaccharide composition of the two polysaccharide fractions were further analyzed. Invitro data show that ILN3 A exhibited more significant inhibitory effects on murine mesangialcells(HBZT-1) than that of ILN3 B with an IC50 value of 97.28 μg/*** nephritis rats induced by cationic bovine serum albumin(C-BSA) were applied to confirmthe therapeutic effects of ILN3 A on glomerulonephritis. ILN3 A significantly decreasedthe urine protein(UP), total cholesterol(TC), triglycerides(TG) and serum creatinine(Scr)level in nephritis rats. ILN3 A also down-regulated the release of IL-2, IL-6 and TNF-α,and upregulated IL-2R, thus inhibiting the inflammatory reaction. ILN3 A mediated therapetuticeffects on glomerulonephritis may be attributable to the inhibition of NF-κB *** results indicated that ILN3 A polysaccharide fraction can effectively improve thenephritis symptoms, alleviate glomerular immune inflammation and reduce the kidneydamage of nephritis rats, thus have great potential for industrial application.

关键词： Irpex lacteus Fr.Anti-nephritic effect

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Low micromolar zinc enhances abnormal aggregation of humanTau in an inducible cell model

Low micromolar zinc enhances abnormal aggregation of humanTa...

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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：胡吉英刘晓灵程晓庆陈杰梁毅武汉大学生命科学学院病毒学国家重点实验室

Abnormal aggregation of human Tau protein leads to the generation of neurofibrillarytangles, a key neuropathological feature in tauopathies including Alzheimer *** factors such as labile metal ions within neurons are likely to play a role in thedevelopment of tauopathies. We have previously demonstrated that zinc, an essential elementthat is highly concentrated in brain, directly accelerates the fibrillization of humanTau in vitro. However, how dose zinc influences intracellular Tau aggregation has not beenstudied yet. Herein, by employing an aggregation cell model of a Tau pathological mutant,full-length Tau-ΔK280 or Tau244-372-ΔK280, as an inducible cell model, we have investigatedthe effects of zinc on abnormal aggregation of human Tau and its cysteine mutants inSH-SY5 Y neuroblastoma cells. As evidenced by confocal laser scanning microscopy, Sarkosyl-insoluble SDS-PAGE, and Western blot, such a Tau pathological mutant forms amyloidfibrils in SH-SY5 Y cells, and 10-50 M zinc strongly enhances abnormal aggregation of Tau-ΔK280 in such an inducible cell model. Our in vitro experiments indicate that 10-50 M zincstrongly accelerates the fibrillization of Tau-ΔK280 via bridging of Cys-291 and ***, 10-50 M zinc significantly promotes hyperphosphorylation of full-length Tau-ΔK280 in SH-SY5 Y cells. Our data demonstrate that zinc directly accelerates the intracellularaggregation of human Tau, providing clues to understanding the relationship betweenzinc dyshomeotasis and the etiology of tauopathies.

关键词： confocal microscopy electron microscopy(EM) protein aggregation proteinp hosphorylation neurodegenerative disease Tau protein

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H3R2 site is involved in DNA damage response and repair

H3R2 site is involved in DNA damage response and repair

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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：何明洪周金秋李宏韬钟蔚丽 SIBCB

DNA damage is a serious threat to genome integrity, which can result in many *** have evolved multiple pathways to respond and repair the break, especially thealterations in chromatin structure. Meanwhile, DNA damage induced histone degradationpartially depends on histone acetylation, which suggests that histone modifications playvery important roles in DSBs response and repair. Here, we show that H3R2 site is importantfor DNA damage response and repair. H3R2 A mutation is sensitive to MMS treatment,which can be suppressed by H4K16 R mutation. H4K16 acetylation is involved in DNA damage induced histone degradation, which prompts us to detect histone degradation inH3R2 A mutant. The result shows that the histone degradation in H3R2 A mutant is faster than that in wild type yeast cells when DNA damage occurs. Additionally, H3R2 A mutation reduces efficiency of non-homologous end joining(NHEJ) and(or) homologous recombination(HR). Further more, H3R2 methylation levels are increased after DNA damage occurs,suggesting that H3R2 methylations may be involved in DNA damage response and *** together, our work implicates that H3R2 site may be involved in DNA damage response and repair by affecting histone degradation, NHEJ and(or) HR. H3R2 methylations may play important roles in these processes.

关键词： H3R2 DNA damage

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Identification of Novel Surface-Exposed Proteins of Rickettsiarickettsii by Affinity Purification and Proteomics

Identification of Novel Surface-Exposed Proteins of Ricketts...

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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：龚文平熊小路齐永焦俊段长松温博海军事医学科学院

Rickettsia rickettsii, the causative agent of Rocky Mountain spotted fever, is the mostpathogenic member among Rickettsia spp. Surface-exposed proteins（SEPs） of R. rickettsiimay play important roles in its pathogenesis or immunity. In this study, R. rickettsii organismswere surface-labeled with sulfo-NHS-SS-biotin and the labeled proteins were affinity-purified with streptavidin. The isolated proteins were separated by two-dimensionalelectrophoresis, and 10 proteins were identified among 23 protein spots by electrosprayionization tandem mass spectrometry. Five（OmpA, OmpB, GroEL, GroES, and a DNAbindingprotein） of the 10 proteins were previously characterized as surface proteins of ***. Another 5 proteins（Adr1, Adr2, OmpW, Porin, and TolC） were first recognizedas SEPs of R. rickettsii herein. The genes encoding the 5 novel SEPs were expressed in Escherichiacoli cells, resulting in 5 recombinant SEPs（rSEPs）, which were used to immunizemice. After challenge with viable R. rickettsii cells, the rickettsial load in the spleen, liver, orlung of mice immunized with rAdr2 and in the lungs of mice immunized with other rSEPsexcluding rTolC was significantly lower than in mice that were mock-immunized with *** in vitro neutralization test revealed that sera from mice immunized with rAdr1, rAdr2,or rOmpW reduced R. rickettsii adherence to and invasion of vascular endothelial cells. Theimmuno-electron microscopic assay clearly showed that the novel SEPs were located in theouter and/or inner membrane of R. rickettsii. Altogether, the 5 novel SEPs identified hereinmight be involved in the interaction of R. rickettsii with vascular endothelial cells, and all ofthem except TolC were protective antigens.

关键词： Rickettsia rickettsii surface-exposed proteins protective antigens pathogenesist ransmission electron microscopy

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D-ribose,an overlooked player in type 2 diabetes mellitus?

D-ribose,an overlooked player in type 2 diabetes mellitus?

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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： Tao Tao Su Rongqiao He Institute of Biophysics Chinese Academy of Sciences

Type 2 diabetes mellitus is a metabolic disorder that is characterized by high blood glucose due to either insulin resistance or insulin deficiency. A direct correlation between Dglucose and diabetic complications has long been established, and is the focus of most research in this field. In contrast, D-Ribose has been overlooked so far as a potential risk player in the development of *** published reports found abnormally high levels of D-ribose in the urine of diabetic patients. Elsewhere, intravenous administration of D-ribose resulted in a decrease in blood D-glucose, concomitantly with an increase in the level of glycated serum proteins. It is known that administration of high dose of D-ribose rapidly glycates proteins in vivo and in vitro, which results in the production of high levels of advanced glycation end products.D-glucose, in contrast, cannot be used to the same effect, even under identical ***, as we hypothesized, dysfunction of the metabolism of D-ribose may play a roleinvolving the development of complications of type 2 diabetes mellitus, and it is feasible that D-ribose could be used as a biomarker for type 2 diabetes mellitus. That is, diabetics suffer from dysfunction of energy metabolism, involving not only D-glucose, but also *** viewpoint for D-ribose in diabetes is based on these observations:(i) Diabetics display abnormal increases in the level of uric D-ribose;(ii) Raised levels of D-ribose result in increased ribosylation activity, in the result of which more advanced glycation end products accumulate in cells;and(iii) D-ribose could be used as a new biomarker for type2 diabetes mellitus, as it is easily measured in the urine of diabetic *** suggest that this novel view of type 2 diabetes mellitus deserves further and detailed in vestigation, both at the level of basic and clinical research, to clarify whether and how Dribose and its metabolism are related with diabetic complications.

关键词： ribosylation type 2 diabetes mellitus

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