Expression and activity characterization of thrombosis targeting protein hu3D3V_H-tTF to blood vessel of lung cancer

目的制备一种用于肺癌血管靶向栓塞治疗的融合蛋白hu3D3VH-tTF,并鉴定其生物学活性。方法利用重叠PCR技术构建tTF与hu3D3VH的融合基因,克隆至表达载体pET22 b(+),在E.coliBL21(DE3)中表达,镍亲和色谱柱纯化目的蛋白。ELISA检测融合蛋白hu3D3VH组分与肺腺癌细胞A549选择性结合活性,凝血实验和FⅩ活化实验鉴定融合蛋白tTF组分的促凝血活性。结果获得序列正确的hu3D3VH/tTF/pET22 b(+)重组子,融合蛋白在E.coliBL21(DE3)中高效表达。纯化后的融合蛋白与肺腺癌细胞A549具有选择性结合活性,并能活化FⅩ、有效促发血液凝固。结论成功构建hu3D3VH/tTF/pET22 b(+)重组子,hu3D3VH/tTF融合蛋白具有hu3D3VH的选择性结合能力同时具有TF的促凝血活性,为开展选择性肺肿瘤血管血栓性栓塞研究奠定了基础。 【英文摘要】 Purpose To prepare the fusion protein of hu3D3VH-tTF for thrombosis targeting therapy of lung cancer and to analyze its biological activities.Methods Fusion gene hu3D3VH-tTF was constructed by overlap PCR,cloned into expression vector pET22 b(+),and expressed in E.coli BL21(DE3).The fusion protein hu3D3VH-tTF was purified through Nickel-affinity chromatography column.The selective binding activity of the hu3D3VH moiety of fusion protein was analyzed using ELISA and the coagulation activity of the tTF moiety...福建省自然科学基金项目(C0410004);; 厦门大学科技创新项目(XDKJCX20053026

Improvement of column of machine tool based on modal analysis

机床在工作时,由于要承受各种变载荷而产生振动,其精度和寿命会受到影响,因此有必要对机床进行模态分析,了解其动态特性,以便进一步分析和改进。采用3d建模软件对机床进行建模和装配,将得到的模型导入HyPErMESH建立有限元模型,再将有限元模型导入AnSyS进行模态分析,得到机床在两种工作情况下的固有频率和振型。最后根据模态分析结果对机床立柱进行优化,得出优化结构。When the machine tool is in operation,it will undergo a variety of dynamic loads which as a consequence incur vibrations and degrade machining precision.Therefore,modal analysis is highly demanded to gain an insight into the dynamics of machining system and direct further analysis and optimization.In such a circumstance a corresponding machine tool is established via 3D CAD software and subsequently preprocessed by HYPERMESH.The finite element model produced by HYPERMESH is eventually analyzed by ANSYS to receive the natural frequencies and mode shapes of the target machine tool.An optimization design of the column of the target machine tool is thus developed based upon the modal information.国家自然科学基金资助项目(70772093);福建省高等学校新世纪优秀人才支持计划资助项目;厦门市科技计划项目(3502Z20090003

Studies of preparation and properties of RGD-tTF water-based ferrofluids

目的探讨RGD-tTF水基磁流体通过磁场和RGD多肽在体外对内皮细胞双靶向的功能。方法通过化学沉淀法以柠檬酸钠为表面活性剂制备水基磁流体(MnFe2O4),弱酸改性后与重组的RGD-tTF融合蛋白结合,利用H-600透射电镜观测纳米粒径,以SUQID鉴定磁性,用MTT法、因子X活化检测和流式细胞仪检测RGD-tTF磁流体生物活性。结果成功制备出的水基磁流体能在磷酸盐缓冲液中稳定分布且具有生物兼容性,实验表明RGD-tTF与水基磁流体结合后对RGD和tTF生物活性均无显著影响,并证实在磁场的作用下能实现了水基磁流体对RGD-tTF的定位作用。结论一种具有内皮细胞双靶向功能的RGD-tTF水基磁流体已制备成功。 【英文摘要】 Purpose To study the property of RGD-tTF water-based ferrofluids double targeting EC304 cells in vitro by magnet and RGD peptide.Methods Water-based ferrofluids(MnFe2O4) were prepared by chemical precipitation method using citrate as surfactant, dispersed in weak acid to create surface charges,and coated with recombinant RGD-tTF protein.Proteins coated ferrofluids were characterized by H-600 transmission electron microscopy(TEM)and Superconducting Quantum Interference Device(SQUID),and its biological activi...教育部和厦门大学出国留学人员启动基金资

pJAK2 polypeptide,an antagonist of suppressors of cytokine signaling-1,can enhance the antitumor effect of dendritic cells

目的:探讨细胞因子信号转导抑制因子-1(SuPPrESSOrS Of CyTOkInE SIgnAlIng1,SOCS1)拮抗物PJAk2多肽(氨基酸序列号为1001-1013)参与树突状细胞(dEndrITIC CEllS,dCS)的体外诱导培养后对dCS抗肿瘤作用的影响。方法:采集健康人外周血,离心获得单个核细胞,用重组人粒细胞巨噬细胞集落刺激因子(rECOMbInAnT HuMAn grAnulOCyTE-MACrOPHAgE COlOny STIMulATIng fACTOr,rHgM-CSf)及重组人白细胞介素-4(rECOMbInAnT HuMAn InTErlEukIn-4,rHIl-4)诱导dCS,第5天分为4组:单纯dCS培养(对照)组、抗原负载(lySATE-dCS)组、多肽修饰(PJAk2-dCS)组和抗原与多肽共培养(lySATE+PJAk2-dCS)组,第6天各组加入肿瘤坏死因子-α(TuMOr nECrOSIS fACTOr-AlPHA,Tnf-α)促成熟。倒置显微镜下观察dCS形态;fCM法检测dCS表型;乳酸脱氢酶(lACTATE dEHydrOgEnASE,ldH)细胞毒实验检测各组细胞毒性T淋巴细胞(CyTOTOXIC T lyMPHOCyTE,CTl)对胃癌细胞bgC-823的靶向杀伤作用;ElISA法检测白细胞介素-12(InTErlEukIn-12,Il-12)和γ干扰素(InTErfErOn-γ,Ifn-γ)的水平。结果:与未加入诱导剂组相比,各组均成功诱导出成熟dCS,均高表达Cd80、Cd83、Cd86和人类白细胞dr抗原(HuMAn lEukOCyTE AnTIgEn dr,HlA-dr),但以lySATE+PJAk2-dCS组的表达水平最高。在10:1~30:1的效靶比范围内,CTl杀伤作用与效靶比呈正相关。当效靶比为30:1时,对照组的CTl杀伤率达(19.77±2.34)%,低于其他3组(P<0.01),而lySATE+PJAk2-dCS组较lySATE-dCS组及PJAk2-dCS组都高(P<0.05)。lySATE+PJAk2-dCS组培养上清液中Il-12及Ifn-γ的分泌水平明显高于对照组(P<0.01)。结论:SOCS1拮抗物PJAk2多肽(1001-1013)可增强dCS对胃癌细胞的抗原递呈及特异性抗肿瘤作用。Objective:To investigate the effect of pJAK2 polypeptide,an antagonist of SOCS1(suppressors of cytokine signaling 1),on antitumor effect of in vitro cultivation-induced DCs(dendritic cells).Methods:Peripheral blood was collected from the healthy volunteers,and the PBMCs(peripheral blood mononuclear cells)were isolated.DCs were induced by rhGM-CSF(recombinant human granulocyte-macrophage colony-stimulating factor)and rhIL-4(recombinant human interleukin-4).On the fifth day,DCs were divided into four groups:control group,Lysate-DCs group,pJAK2-DCs group,and Lysate + pJAK2 DCs group.On the sixth day,TNF-α(tumor necrosis factor-alpha)was added into each group.The morphological features of DCs were observed under an inverted microscope;the phenotypes were detected by FCM(flow cytometry);the killing effect of CTLs(cytotoxic T lymphocytes)on gastric cancer BGC-823 cells was evaluated by LDH(lactate dehydrogenase)cytotoxicity test;the concentrations of IL-12(interleukin-12)and IFN-γ(interferon-γ)were detected by ELISA(enzyme-linked immuno sorbent assay).Results:Mature DCs presented typically morphological and phenotypic features;the DCs in Lysate + pJAK2-DCs group had the highest expression levels of CD80,CD83,CD86 and HLA-DR(human leukocyte antigen DR).When the ratio of effectors to target cells ranged from 10:1 to 30:1,the killing activity of CTLs had a positive correlation with the ratio.When the ratio of effectors to target cells was 30:1,the killing activity of CTLs in the control group was(19.77±2.34)%,which was lowest as compared with the other groups(P < 0.01),meanwhile the killing activity of CTLs in Lysate + pJAK2-DCs group was higher than those in Lysate-DCs and pJAK2-DCs groups(P < 0.05).The levels of IL-12 and IFN-γ secretion in Lysate + pJAK2-DCs group were apparently higher than those in the control group(P < 0.01).Conclusion:An antagonist of SOCS1,pJAK2 polypeptide,can enhance the ability of antigen presentation and specific antitumor effect of DCs on gastric cancer cells.南京军区医学科技创新课题资助项目(编号:10MA068); 福建省自然科学基金面上项目(编号:2010D013); 厦门市科技计划创新项目(编号:3502z20104014

RE-X二元合金相图的热力学数据库

利用CALPHAD方法,采用亚正规溶体模型、亚点阵模型以及理想气体模型来描述RE-X(Ag,Bi,Cr,Mn,Mo,V,Zn)中二元系各相的Gibbs自由能,并结合相平衡及热力学性质的实验结果,对Ag-RE(RE:Sc,Y,Nd,Sm,Gd,Tb,Ho,Er)、Bi-RE(RE:Nd,Tm,Er,Ho,Pr,Gd)、Cr-RE(RE:Ce,Nd,Sm,Lu)、Mn-RE(RE:Pr,Nd,Sm,Eu,Tb,Dy,Ho,Er,Tm,Yb,Lu)、Mo-RE(RE:Sc,Y,La,Ce,Pr,Nd,Sm,Eu,Tb,Dy,Ho,Er,Tm,Yb,Lu)、V-RE(RE:La,Ce,Pr,Nd,Ho,Lu)和Zn-RE(RE:Y,Ce,Pr,Nd,Sm)各二元系相图进行热力学优化与计算。计算结果与实验数据取得很好的一致性,并结合其他相关稀土二元系相图热力学计算,初步建立部分稀土二元合金相图的热力学数据库。该热力学数据库可以提供相平衡及热力学性质等多种信息,为外推计算稀土多组元体系的相平衡提供理论基础,并为高性能稀土合金材料的设计及制备提供重要的理论指导

Effect of Helicobacter pylori on the function of peripheral blood monocyte-derived dendritic cells in gastric cancer patients

分析HP阳性和HP阴性的胃癌患者外周血单核细胞来源树突状细胞(MOdCS)功能的差异性及其临床意义。方法:用尿素14C呼气试验对解放军第一七四医院2011年1月至2012年10月收治的84例胃癌患者进行HP感染状态鉴定,分别采集HP阳性和阴性胃癌患者外周血,分离外周血单个核细胞,采用经典方法(rHgM-CSf、rHIl-4联合rHTnf-α)诱导产生dCS,采用流式细胞仪检测dCS表型,采用ldH释放法检测dCS致敏T细胞对胃癌细胞的毒性杀伤作用,采用ElISA方法检测细胞因子Il-12、Ifn-γ的分泌水平。结果:两组MOdCS成熟过程形态变化无差异,HP阳性组MOdCS表面标记分子Cd1A、Cd80、Cd83、Cd86和HlA-dr平均表达百分率均高于HP阴性组,其中Cd80、Cd83、Cd86的表达水平差异有统计学意义,Cd1A、HlA-dr差异无统计学意义。HP阳性组dCS致敏T淋巴细胞对胃癌细胞杀伤率和Il-12、Ifn-γ的分泌水平均高于HP阴性组(P<0.05)。结论:HP感染状态不影响胃癌患者外周血MOdCS成熟过程形态变化,HP感染的胃癌患者MOdCS成熟和活化水平更高。Objective:This study aimed to compare and analyze the functional differences between peripheral blood monocyte-derived dendritic cells(DCs) of Helicobacter pylori-positive and H.pylori-negative patients with gastric cancer.Methods:H.pylori infection was detected in 84 patients with gastric cancer in our hospital from January 2011 to October 2012 by the14C-urea breath test.DCs were generated from monocytes isolated by an adherent method from the two groups of patients and cultured in the presence of rhIL-4,rhGM-CSF,and rhTNF-α.Furthermore,the expression of surface marker molecules was determined by fluorescence-activated cell sorting analysis.The cytotoxicity of DCs pulsed T cells against gastric carcinoma cell was assessed by the lactate dehydrogenase-releasing assay.The secretion of IL-12 and IFN-γ in the supernatant was determined by enzyme-linked immunosorbent assay.Results:No difference was observed in the morphological change of the maturation process.The mean expression of CD1a,CD80,CD83,CD86,and HLA-DR molecules in DCs of H.pylori-infected patients was higher than that in DCs of H.pylori-negative group,and the differences were statistically significant except for CD1a and HLA-DR.The cytotoxicity activities,IL-12 release,and IFN-γ release in the H.pylori-positive group were significantly higher than those in the H.pylori-negative group(P<0.05).Conclusion:H.pylori infection has no effect on the morphological change of the maturation process of monocyte-derived DCs.These data clearly demonstrate that monocyte-derived DCs of H.pylori-infected patients with gastric cancer can induce stronger maturation and activation than those of H.pylori-negative patients.南京军区医学科技创新课题(编号:10MA068); 福建省自然科学基金资助项目(编号:2010D013); 厦门市科技计划创新项目(编号:3502z20104014)资助~

p38MAPK induces apoptosis of glioma cell

目的　研究 p38MAPK基因转染大鼠胶质瘤细胞系C6后对其生物学特性的影响 .方法　利用脂质体介导法将p38MAPK基因导入大鼠胶质瘤细胞系 C6中 ,用免疫细胞化学染色检测其在细胞转染前后的表达情况 ,用 HE染色、流式细胞仪等方法研究其对细胞形态、粘着状况和生长周期的影响 .结果　转染 p CMV5 - p38MAPK质粒组 p38MAPK蛋白表达阳性 ,细胞形态发生变化 ,贴壁性降低 ,出现大量凋亡细胞 .结论　转染 p38MAPK基因可诱导胶质瘤细胞凋亡 【英文摘要】 AIM To study the effect of p38MAPK transfection on the biological characteristics of glioma cell C6. METHODS p38MAPK was transfected into glioma cell C6 by lipofectin. Expression of p38MAPK was detected by immunocytochemistry. HE staining and flow cytometry were adopted to measure the cell morphology, adhesion and cell cycle. RESULTS p38MAPK was expressed in transfected glioma cells, with cell biological characteristics changing and apoptotic cells emerging. CONCLUSION Apoptosis of glioma cell could be ...高等学校骨干教师计划资

p38MAPK induces apoptosis of glioma cell

目的　研究 p38MAPK基因转染大鼠胶质瘤细胞系C6后对其生物学特性的影响 .方法　利用脂质体介导法将p38MAPK基因导入大鼠胶质瘤细胞系 C6中 ,用免疫细胞化学染色检测其在细胞转染前后的表达情况 ,用 HE染色、流式细胞仪等方法研究其对细胞形态、粘着状况和生长周期的影响 .结果　转染 p CMV5 - p38MAPK质粒组 p38MAPK蛋白表达阳性 ,细胞形态发生变化 ,贴壁性降低 ,出现大量凋亡细胞 .结论　转染 p38MAPK基因可诱导胶质瘤细胞凋

Self-healing of fractured one dimensional brittle nanostructures

Recent experiments have shown that fractured GaAs nanowires can heal spontaneously inside a transmission electron microscope. Here we perform molecular-dynamics simulations to investigate the atomic mechanism of this self-healing process. As the distance between two fracture surfaces becomes less than 1.0 nm, a strong surface attraction is generated by the electrostatic interaction, which results in Ga–As re-bonding at the fracture site and restoration of the nanowire. The results suggest that self-healing might be prevalent in ultrathin one-dimensional nanostructures under near vacuum conditions

Self-healing in fractured GaAs nanowires

Molecular dynamics simulations are performed to investigate a spontaneous self-healing process in fractured GaAs nanowires with a zinc blende structure. The results show that such self-healing can indeed occur via rebonding of Ga and As atoms across the fracture surfaces, but it can be strongly influenced by several factors, including wire size, number of healing cycles, temperature, fracture morphology, oriented attachment and atomic diffusion. For example, it is found that the self-healing capacity is reduced by 46% as the lateral dimension of the wire increases from 2.3 to 9.2 nm, and by 64% after 24 repeated cycles of fracture and healing. Other factors influencing the self-healing behavior are also discussed