抵抗素对3T3-L1脂肪细胞葡萄糖摄取的影响

【目的】构建载有小鼠抵抗素（resistin）基因及其反义核酸的重组真核表达质粒，探讨抵抗素对3T3-L1脂肪细胞葡萄糖摄取的影响。【方法】用RT-PCR技术从小鼠脂肪组织扩增抵抗素基因cDNA，A／T克隆于pGEM-T载体，再亚克隆于pcDNA3．1（＋）和pcDNA3．1（-）真核表达载体，构建载有小鼠抵抗素（resistin）基因及其反义核酸的重组真核表达质粒pcDNA3．1^（＋）resistin和pcDNA3．1^（-）-resistin^（-）。将pcDNA3．1^（-）resistin^（-）转染小鼠3T3-L1脂肪细胞，通过G418筛选稳定表达的细胞株。3T3-L1脂肪细胞分为对照、瞬时、载体和稳定4个细胞组（每组n=6），用^3H-2-脱氧葡萄糖进行非胰岛素和胰岛素介导的葡萄糖摄取试验。【结果】插入pcDNA3．1^（＋）的DNA片段的核苷酸序列与小鼠抵抗素基因mRNA编码区序列完全一致．且方向正确：插入pcDNA3．10^（＋）的DNA片段的核苷酸序列与抵抗素基因mRNA编码区核苷酸序列完全一致，且方向相反。对照组、瞬时组、载体组和稳定组的3T3-L1脂肪细胞非胰岛素和胰岛素介导的葡萄糖摄取率的无显著性差别（P〉O．05）。【结论】成功构建了载有抵抗素基因和载有抵抗素基因反义核酸的重组真核表达质粒。抵抗素对3T3-L1脂肪细胞的葡萄糖摄取均无明显影响

Preparation and Characterization of Sinter-Resistant RhSm2O3/SiO2 Catalyst and Its Performance for Partial Oxidation of Methane to Syngas

以乙酰丙酮铑(Rh(acac)_3)和乙酰丙酮钐(Sm(acac)_3)为前驱体,用浸渍法制备了Rh/SiO_2和Rh-Sm_2O_3/SiO_; 2催化剂。采用原位红外光谱、热重分析、低温N_2吸附、X射线粉末衍射、高分辨透射电子显微镜、H_2-程序升温还原和X射线光电子能谱等实验技术对催; 化剂的制备过程,比表面积和物相以及Rh与Sm_2O_3间的相互作用进行了表征,并以甲烷部分氧化制合成气为目标反应对催化剂的稳定性进行了考察。研究; 表明:以Rh(acac)_3和Sm(acac)_3为前驱体采用简单的浸渍法即可制备出Rh平均粒径为2.3; nm且具有良好抗烧结性能的Rh-Sm_2O_3/SiO_2催化剂。在浸渍过程中乙酰丙酮化合物通过与SiO_2表面羟基形成氢键而负载于载体表面。S; m(acac)_3在SiO_2表面的单层负载量(质量分数)约为31%,对应于Sm_2O_3的质量分数约为15%,只要Sm(acac)_3的质量分; 数低于这一阈值,均可保证分解后生成的Sm_2O_3以高分散形式负载于SiO_2上,且不会因高温(800; °C)焙烧而团聚。高分散于SiO_2表面的Sm_2O_3与Rh之间存在强的相互作用,可显著提高Rh的分散度,防止其在高温反应条件下烧结,进而使低; Rh负载量的催化剂表现出良好的甲烷部分氧化制合成气反应活性和稳定性。Rh/SiO2 and Rh-Sm2O3/SiO2 catalysts were synthesized by the conventional impregnation method using rhodium acetylacetonate (Rh(acac)(3)) and samarium acetylacetonate (Sm(acac)(3)) as precursors. The preparation and catalytic properties, as well as the interaction between Rh and Sm2O3, were characterized in detail by in situ infrared spectroscopy (IR), thermogravimetric analysis (TG), N-2 physisorption (Brunauer-Emmett-Teller (BET) method), X-ray powder diffraction (XRD), transmission electron microscopy (TEM), temperature-programmed reduction (H-2-TPR) and X-ray photoelectron spectroscopy (XPS). The performance of the catalysts for the partial oxidation of methane (POM) to syngas was also investigated. The results showed that a sinter-resistant Rh-Sm2O3/SiO2 catalyst with an average Rh particle size of similar to 2.3 nm could be synthesized using the conventional impregnation method with Rh(acac)(3) and Sm(acac)(3) as precursors. The surface silanol groups of SiO2 acted as the centers to interact with M(acac)(3) (M=Rh, Sm) molecules when SiO2 was impregnated in the M(acac)(3) solution, leading to the formation of a hydrogen-bonded M(acac)(3) layer on the SiO2 surface. In this experiment, the monolayer coverage of Sm(acac)(3) on the SiO2 surface was equal to a Sm(acac)(3) loading (mass fraction) of approximately 31%, which in turn corresponded to a Sm2O3 loading of approximately 15%. When a Sm(acac)(3)/SiO2 sample with Sm(acac)(3) loading below 31% was heated in air to approximately 360?, the monolayer Sm(acac)(3) species decomposed into highly dispersed Sm2O3 species on the SiO2 surface, which displayed superior stability against sintering at high temperature. No aggregation of the Sm2O3 species was observed even when the sample was heated to 800 degrees C in air. The strong interaction between the highly dispersed Sm2O3 and Rh plays a key role in increasing the dispersion of Rh species in the catalyst and preventing the Rh species from sintering under high temperature conditions. This factor should also be responsible for the superior activity and stability of the Rh-Sm2O3/SiO2 catalyst with extremely low Rh loading for the catalytic partial oxidation of methane to syngas.国家重点基础研究发展规划项目（2013CB933102）,国家自然科学基金（21473144,21373168）,国家基础科学人才培养基金项目（J1310024）及教育部创新研究团队项目（IRT_14R31）资

抵抗素诱导脐静脉内皮细胞功能异常

【目的】研究抵抗素（resistin）对脐静脉内皮细胞功能的影响，以探讨抵抗素在粥样硬化性疾病中的作用及其机制。【方法】原代培养人脐静脉内皮细胞，不同浓度人抵抗素（0、50、100ng／mL）培养24h。流式细胞检测抵抗素对内皮细胞间黏附分子（ICAM-1）、血管细胞间黏附分子（VCAM-1）与活性氧簇（ROS）表达的影响；RT-PCR检测抵抗素对内皮素（ET-1）、内皮型一氧化氮合酶（eNOS）、诱导型一氧化氮合酶（iNOS）mRNA表达的影响。【结果】人脐静脉内皮细胞经人抵抗素（50ng／mL、100ng／mL）处理24h后ICAM-1和ET-1mRNA表达显著增高，而各组间VCAM-1表达、ROS生成，以及eNOS和iNOS mRNA表达无明显差别。【结论】抵抗素可通过增加ICAM-1表达，上调ET-1表达直接促进内皮细胞激活，提示脂肪细胞与内皮细胞的相互作用可能是动脉粥样硬化性疾病的发病因素之一

新诊断2型糖尿病患者血清炎症因子及外周血单个核细胞中核因子κB活性变化

【目的】探讨新诊断2型糖尿病患者血清炎症因子超敏-C反应蛋白（hs-CRP）、肿瘤坏死因子α（TNF-α）、白介素-6（IL-6）水平及外周血单个核细胞（PMNC）中NF-κB的活性变化。【方法】选择健康成人66例，新诊断2型糖尿病患者92例，分为正常对照组和2型糖尿病组，分析比较两组血清炎症因子及PMNC中NF-κB活性水平。血清hs-CRP、TNF-α、IL-6采用酶联免疫吸附法测定。免疫印迹分析检测PMNC中核因子KBp65（Set^536）的磷酸化水平，β-actin作为内参。【结果】2型糖尿病组患者的血清CRP、IL-6、INF-α水平均显著高于正常对照组（P〈0．001）；糖尿病组患者PMNC中NF-κB P65（Ser^536）磷酸化水平高于正常对照组（0．85±0．38vs0．47±0．25，P〈0．05）；在糖尿病组，log（CRP）与质量指数（BMI）、腰围、腰臀比、稳态模型法估计的胰岛素抵抗指数（HOMA-IR）、log（IL-6）、log（TNF-α）、糖基化血红蛋白（hemoglobin，Alc）呈正相关。【结论】结果提示2型糖尿病患者处于炎症状态，血清炎症因子水平及PMNC中NF-κB活性的升高可能与胰岛素抵抗和动脉粥样硬化的形成有关

Recruitment of cyanobacteria from sediment of the Shanzai Reservoir

采用正交试验考察了温度、光照、营养盐和物理扰动4个因素对山仔水库冬季沉积物中蓝藻复苏的影响,每个因素设置两个水平,培养周期为6d,并以蓝藻复苏量为考察指标.结果表明,温度和光照为蓝藻复苏的主要影响因子,上覆水体的营养盐、物理扰动对沉积物中蓝藻门复苏的影响作用不显著,不同的蓝藻种属对温度和光照条件的响应程度不完全一致,蓝藻门微囊藻属(MICrOCySTIS)对温度和光照的复苏响应显著,颤藻属(OSCIllATOrIA)仅对温度的复苏响应显著.同时,通过设置6.0~16.0℃之间6个温度梯度及50和2000lX两个光照梯度,进行了沉积物柱状样复苏模拟实验.结果显示,山仔水库冬季沉积物微囊藻属和颤藻属在10℃左右开始复苏,微囊藻属对光的敏感性使其更容易处于优势地位.The effects of four environmental factors,including temperature,light,nutrient and physical disturbance,on the cyanobacteria recruitment from the winter sediment in the eutrophic Shanzai Reservoir were investigated using an orthogonal experiment.Two levels were designed for each factor and the experiments lasted for six days.The results showed that temperature and light were the most important for the recruitment of Microcystis and Oscillatoria.Increasing temperature would promote the recruitment of Microcystis and Oscillatoria which would make the cyanobacteria to dominate.The effects of the overlying water nutrient status and the physical disturbance on the recruitment of Microcystis and Oscillatoria from sediments were not obvious.Based on the results,different temperatures and light intensities were designed in the simulation experiment.The incubation temperatures were increased from 6.0 to 16.0 ℃ with six levels while each temperature level was kept for four days under 2000 and 50 lx light intensities,respectively.Recruitment was calculated by the diminution of benthic cyanobacteria abundance,and moreover,by the increase of cyanobacteria abundance in the water column.It is shown that the recruitment of Microcystis and Oscillatoria started at about 10 ℃.It was also demonstrated that the migration of Microcystis from the sediments was more pronounced at higher light intensity(2000 lx) than in dark(50 lx) treatments.国家自然科学基金项目(No.41101060);福建省自然科学基金(No.2010J01250);福建省教育厅科学研究基金(No.JA10085);厦门大学近海与海洋环境国家重点实验室青年学者访问基金项目(No.MELRS1103)---

HIV-1 gp160截短蛋白在酿酒酵母中的表达

利用PCR技术从pNL-43上扩增出截短的编码gp160蛋白的基因片断,克隆到酿酒酵母表达载体YEpFLAG-1上构建表达质粒YEp-gp160Δ12,电转化到酿酒酵母中,用缺色氨酸的SC培养基筛选出阳性克隆,重组子经YP培养基诱导后进行全菌蛋白的SDS-PAGE和Western Blotting分析,筛选出高表达菌株.纯化后的重组gp160Δ12(rgp160Δ12)蛋白经ELISA鉴定显示具有良好的生物活性

重组毕赤酵母高密度发酵表达H5N1禽流感病毒糖蛋白

在10L发酵罐中,对高致病性禽流感病毒H5N1糖蛋白HA1在重组毕赤酵母中的表达发酵工艺进行了研究。通过分批补料培养方法探讨不同培养温度、诱导温度、补料方式、微量元素等因素对菌体的生长以及重组蛋白表达和活性的影响。结果表明,菌种培养和诱导温度均为25oC时,菌体的生长、分泌表达量和与广谱中和抗体的反应活性较好;微量元素是影响重组HA1蛋白生物活性的重要因素;通过优化高密度发酵工艺,H5N1病毒糖蛋白HA1在发酵罐中的表达量比摇瓶培养提高10.5倍,达到约120mg/L,为大规模制备高致病性禽流感病毒的HA1蛋白奠定了基础

抗H5亚型禽流感病毒单链抗体在毕赤酵母中的分泌型表达和生物活性分析

在本实验室研制出的多株针对H5N1亚型禽流感血凝素单抗中,13D4单抗对所有H5亚型病毒均有血凝抑制和中和活性,具有特异性高、反应性强和识别广的特点,且在小鼠实验中显示了对各种代表株禽流感的感染和发病均具有良好的治疗效果。在此研究基础上,本实验通过基因工程构建含有13D4单链抗体(scFv)基因的毕赤酵母表达载体,实现目的蛋白的分泌性表达和纯化。经过竞争法和血凝抑制检测其活性,表明获得的单链抗体具有与原始鼠源抗体相近的反应活性和相同的识别表位。H5N1广谱中和单抗13D4的单链抗体的成功构建,为进一步研制针对H5N1禽流感病毒的治疗性抗体奠定了基础