人骨髓间充质干细胞分离培养方法的改进

背景:当前分离培养骨髓间充质干细胞方法的有多种,但在外科手术中进行获取的方法仍然较少,而外科分离所得细胞对于相应疾病的具有揭示原因和提供研究线索的价值。目的:在骨科手术中以直接贴壁差速贴壁分离培养纯化与鉴定骨髓间充质干细胞。方法:在骨科关节置换等大手术中吸取少量人骨髓血,采用直接贴壁法分离骨髓间充质干细胞,在贴壁后36～48h行洗盘处理,并通过长期体外培养,特定培养基自身的筛选作用对细胞进行筛选,当细胞生长达到足量时进行鉴定。结果与结论:采用直接贴壁法能够在长骨髓腔内混合血中分离培养骨髓间充质干细胞并形成集落,其形态学表现和表面分子经鉴定符合骨髓间充质干细胞特点,能够获得符合要求的人骨髓间充质干细胞,从而进行后续实验

Preparation and Application of Soluble Human Squamous Cell Carcinoma Antigen Expressed by Escherichia coli

旨在建立基于大肠杆菌表达系统的高效可溶性表达人鳞状上皮细胞癌抗原(SCCAg)方法,获得具有较好活性的重组SCCAg抗原并应用于建立抗原检测方法; 。基于pGEX-6P-l载体和大肠杆菌E. coli; ER2566菌株开展重组SCCAg抗原可溶性表达纯化方法研究,评价纯化抗原活性,筛选特异性单克隆抗体,初步建立并评价SCCAg抗原检测方法。结果; 显示,pGEX-6P-l载体和E coli; ER2566菌株可用于建立较高效的可溶性表达和纯化SCCAg抗原的方法,获得了具有较高纯度和活性的重组SCCAg抗原,筛选获得特异性单克隆抗体并; 初步建立了 SCCAg管式化学发光检测方法。建立了有效的基于大肠杆菌表达系统的可溶性表达和纯化SCCAg的方法。The aims of this study are to establish a method for efficient soluble; expression of human squamous cell carcinoma antigen(SCCAg ) based on; Escherichia coli expression system and obtain the recombinant SCCAg; antigen in fine activity, then apply it in the detection method; establishment of antigen. The study on the method of soluble expression; and purification of recombinant SCCAg antigen was conducted based on; pGEX-6P-l vector and E. coli ER2566 strain. The activity of the purified; antigen was evaluated by Abbott Kit and the specific monoclonal antibody; was screened by indirect ELISA. It was proved that PGEX-6P-1 vector and; E. coli strain ER2566 could be used to establish efficient soluble; expression and purification method for recombinant SCCAg antigen.; Moreover, the recombinant SCCAg antigen was proved to be in high purity; and activity. Thus,the SCCAg detection method of chemical luminous tube; was established with the specific monoclonal antibodies. In conclusion,; an effective method for the expression and purification of SCCAg, which; is based on the E. coli expression system, is established.国家高技术研究发展计划(863计划

Effects of Serum Medicated with Duhuo Jisheng Decoction on the Induced Degeneration of Articular Chondrocytes in Vitro Culture Based upon “caveolin-p38MAPK” Signal Pathway

目的:观察独活寄生汤含药血清对兔退变软骨细胞“CAVEOlIn-P38MAPk“信号通路的调控作用,探讨独活寄生汤治疗骨关节炎的作用机制。方法:将20只3月龄新西兰兔随机分为生理盐水组(空白血清组)和独活寄生汤组(含药血清组),每组10只。分别在24 H、36 H、48 H不同采血时间点采集独活寄生汤含药血清和空白血清,将5%、10%、15%、20%不同浓度两种血清作用于体外培养第2代软骨细胞,确定含药血清最佳干预条件;建立体外退变软骨细胞模型,分别给予独活寄生汤含药血清(含药血清组)和空白血清(空白血清组)干预36 H,收集软骨细胞,运用WESTErn blOT法检测血清干预后退变软骨细胞CAVEOlIn-1、P38、P-P38蛋白表达,rT-PCr法检测血清干预后退变软骨细胞白细胞介素(Il)-1β、肿瘤坏死因子(Tnf)-α、基质金属蛋白酶(MMP)-3、MMP-13、CAVEOlIn-1 M rnA表达。结果:浓度为15%的36 H采血时间点含药血清的促增殖作用最明显;退变软骨细胞中存在“CAVEOlIn-P38MAPk“信号通路的激活,独活寄生汤含药血清可抑制CAVEOlIn-1、P-P38蛋白表达及Il-1β、Tnf-α、MMP-3、MMP-13、CAVEOlIn-1 M rnA的表达,差异有统计学意义(P<0.05)。结论:独活寄生汤能通过抑制“CAVEOlIn-P38MAPk“信号通路的激活及其下游效应分子,从而有效抑制软骨细胞凋亡。Objective:To observe the effects of serum medicated with Duhuo Jisheng Decoction on the induced degeneration of articular chondrocytes in vitro culture based upon "caveolin-p38MAPK" signal pathway and probe the mechanism of Duhuo Jisheng Decoction in the treatment of osteoarthritis.Methods:20 March old New Zealand rabbits were randomly divided into normal saline group(blank serum group)and Duhuo Jisheng Decoction group(drug containing serum group),10 rats in each group.The second generation of cartilage cells was cultured in vitro using different time points of sample collection such as 24 h,36 h and 48 h,serums medicated with 5%,10%,15% and 20% concentrations of Duhuo Jisheng Decoction and blank serums.The best intervention time of medicated serum was determined,which was then used to deal with the degenerating chondrocytes.The degenerated cartilage cell models in vitro were established,which were intervened by the serum medicated with Duhuo Jisheng Decoction(drug containing serum group)and the blank serum(blank serumgroup)after 36 h,collecting chondrocytes.The Western Blot method was used to detect the expression of degenerated cartilage cells caveolin-1 and p-p38 protein after serum intervention.The RT-PCR method was usedto detect the expression of degenerated cartilage cells IL-1β,TNF-α,MMP-3,MMP-13 and caveolin-1 m RNA after serum intervention.Results:The concentration of 15% and 36 h sampling time point serum on the proliferation;the activation of "caveolin-p38MAPK" signaling pathway in the degeneration of cartilage cell,Duhuo Jisheng decoction containing serum can inhibit the expression of caveolin-1,p-p38 protein expression and IL-1β,TNF-α, MMP-3,MMP-13,caveolin-1,m RNA,the difference was statistically significant(P < 0.05).Conclusion:Duhuo Jisheng decoction can inhibit the "caveolin-p38MAPK" signal pathway and its downstream effector molecules, which can effectively inhibit the apoptosis of cartilage cells.国家自然科学基金资助项目(81302986); 福建省卫生厅青年科研课题资助计划项目(2012-2-69); 福建省自然科学基金(2013J01389); 福州市卫生系统科技计划(2013-S-wq6

Capturing the labile fullerene[50] as C50Cl10

地址: 1. Xiamen Univ, State Key Lab Phys Chem Solid Surfaces, Xiamen 361005, Peoples R China 2. Xiamen Univ, Dept Chem, Xiamen 361005, Peoples R China 3. Chinese Acad Sci, Inst Chem, Beijing 100080, Peoples R China 4. Chinese Acad Sci, Wuhan Inst Phys & Math, State Key Lab Magnet Resonance & Mol Phys, Wuhan 430071, Peoples R China 电子邮件地址: [email protected]

装配误差和齿面修形对曲线圆柱齿轮传动误差的影响

基于齿轮啮合理论，推导了带抛物线修形的曲线圆柱齿轮齿面方程，研究了齿轮副有限元模型初始接触位置的求解方法；利用齿轮参数化有限元技术，建立了齿轮副的有限元模型，从有限元分析结果中提取了齿轮载荷传动误差；通过20余组算例分别研究了齿轮副的装配误差、修形量、刀盘曲率半径和修形曲线偏置量对载荷传动误差的影响。结果表明，装配误差可以降低载荷传动误差的波动幅度；较大的齿面修形量可减小载荷传动误差波动幅度；较大的刀盘曲率半径可减小载荷传动误差值和误差波动幅度；齿面修形曲线的偏置对载荷传动误差的影响较小。总的来说，采用较大曲率半径的修形刀盘可加工出传动性能更好的曲线圆柱齿轮