A Novel Method for Quantification of Circulating DNA in Serum by Capillary Zone Electrophoresis

In this paper, we first presented a novel method for quantification of circulating DNA in human serum based on capillary zone electrophoresis with laser-induced fluorescence detection (CZE-LIF). The serum was digested by proteinase to release free DNA, and then CZE-LIF system was used for the quantification of total circulating DNA. This method was successfully used to quantify the circulating DNA levels in sera from healthy individuals and certain cancer patients.We found the significantly elevated circulating DNA levels in certain prostate cancer patients. Our results demonstrated that CZE-LIF system has good linearity, excellent sensitivity (0.5 ng/mL DNA),satisfactory reproducibility (RSDs in one day and between days were both less than 5%) and reliability, and is well suitable to the quantification of the circulating DNA in human serum or plasma.     

动力学涂层毛细管电泳分离双链脱氧核糖核酸片段

以异丙醇为聚合反应链转移试剂，水相法合成了短链聚N，N－二甲基丙烯酰胺（PDMA），研究表明，该聚合物能在毛细管内壁形成稳定的动力学涂层，从而有效地抑制电渗流和毛细管内壁与DNA的作用。这种介质被成功地应用于DNA片段的高效分离。     

藻胆蛋白的Langmuir-Blodgett膜——Ⅰ.R-藻红蛋白单分子膜及蛋白构象变化的研究

测定了R-藻红蛋白(R-PE)在气/液界面上的表面压-面积曲线(tt-A等温线),结果表明R-PE具有很好的成膜性能,且由n-A等温线求得的单分子占有面积与R-PE以盘状形态“平躺”于液面上时的分子占有面积相同。R-PELB膜的透射电子显微镜(TEM)图象和椭圆偏振关于R-PE单分子膜厚度的测定,说明R-PE分子在基片上的取向是其盘平面与基片表面平行。根据这些依据,提出了R-PE单分子膜的结构。将R-PE在气/液界面上形成的单分子膜在适当表面压下用LB技术转移到基片上,测得了蛋白累积单分子膜的吸收及荧光光谱,R-PE单分子膜的吸收和荧光光谱与其在水溶液中无显著差别。膜的CD光谱与R-PE水溶液的CD光谱比较说明,当R-PE成膜后其二级结构发生了变化,即蛋白的p-折叠片构象组分增加。     

毛细管电泳的原理及应用（第二讲）毛细管电泳的原理及应用

 毛细管电泳的原理及应用（第二讲）毛细管电泳的原理及应用罗国安，王义明（清华大学生命科学与工程研究院，清华大学化学系北京１０００８４）１概述毛细管电泳（ＣＥ）除了比其它色谱分离分析方法具有效率更高、速度更快、样品和试剂耗量更少、应用面同样广泛等优点外，其仪器结构也比高效液相色谱（ＨＰＬＣ）简单。ＣＥ只需高压直流电源、进样装置、毛细管和检测器。前三个部件均易实现，困难之处在于检测器。     

THE PREPARATION AND STUDY ON THE NANO-TiO2/SILK FIBROIN COMPOSITE FILMS BY THE SOL-GEL METHOD

Based on the sol-gel technique using butyl titanate as oxide precursor, the regenerated SF (silk fibroin)/nano-TiO2 composite films were synthesized. Different amounts of butyl titanate to SF were used to verify this effect on the characteristics of the formed materials. Samples were characterized by thermogravimetric analysis, X-ray diffractometry, UV, AFM and FT-IR spectroscopy.The experimental results reveal that, compared to the pure silk fibroin films, the mechanical strength of these regenerated SF/nano-TiO2 composite films were increased and the dissolubility in water of SF/nano-TiO2 composite films in aqueous solution were decreased. The diameter of nano-TiO2 particle films was about 80nm through UV and AFM. The nano-TiO2 particles were well dispersed in the regenerated silk fibroin. It was found that the crystal structures of the composite films were transited from typical Silk Ⅰ to typical Silk Ⅱ by the XRD and FTIR. Furthermore, the crystallinity of the composite films was obviously improved. Through the TGA, it was demonstrated that the heat transition temperature of composite films was also enhanced.     

Ion Recognition and Analytical Application of a Fibroin Modified Electrode

A novel fibroin modified electrode with ion recognition was reported. The membrane with isoelectric point of pH 4.5, was modified on graphite and carbon fiber electrodes. The pH-responsive ion recognition of the modified electrode was investigated by use of some neurocompounds. The fibroin carbon fiber electrode has been used for in-vivo determination.     

藻胆体核心复合物结构与功能的研究（Ⅱ）

从螺旋藻藻胆体中分离出４种不同种结构和光谱形式的变藻蓝蛋白复合物ＡＰＩ、ＡＰⅡ、ＡＰⅢ和ＡＰＢ，利用吸收光谱、荧光光谱比较了三聚体和单体的光谱特性，通过对吸收光谱的光谱解叠以及各组分的归属，研究了变藻蓝蛋白复合物内各色团间相互作用的性质和在能量传递中的功能，结果表明，复合物内色团间的作用关系可以用Ｆｏｒｓｔｅｒ偶极－偶极作用机制来解释，由于妆蛋白和同源亚基的存在影响其结构的对称性，进而影响各色团间     

毛细管电泳

毛细管电泳（ＣＥ）是当前分析化学前沿领域及研究热点之一，本文介绍了ＣＥ基本原理和特点；ＣＥ的六种分离模式和分离机理；ＣＥ的关键技术之一－检测器以及ＣＥ的一些应用。根据ＣＥ特色，介绍了ＣＥ作为一种纳技术在生命科学中的应用，ＣＥ的激光诱导荧光检测器的广泛使用，ＣＥ和质谱联用成为生物分子分析的手段及ＣＥ在小分子及离子方面的应用。     

非水体系毛细管电泳-紫外检测法测定苯甲酸和苯甲醛

建立了用非水相体系高效毛细管电泳-紫外检测法同时测定苯甲酸和苯甲醛的新方法，考察了运行电压、非水相介质和电解质等因素的影响，在25℃下，以V（乙腈）：V（碳酸丙烯酯）=1：1的混合液为溶剂，缓冲体系中含15mmol／L十六烷基三甲基溴化铵体积分数1％乙酸，重力进样30S，运行电压20kV，毛细管总长45cm有效长度30cm，φ75μm，检测波长285nm。苯甲酸线性范围为5～40μg／mL，线性方程为：Y=13．473ρ＋13．336，相关系数r=0．9985，检出限为0．92μg／mL，RSD为3．8％。苯甲醛的线性范围75～1125μg／mL线性方程为：Y=5．2449ρ＋564．01，相关系数r=0．9997，检出限为15．60μg／mL，RSD为3．5％。已用于经空气氧化后的苯甲醛中苯甲酸和苯甲醛的测定。     

大鼠肝脏金属硫蛋白的分离纯化和金属硫蛋白亚型-Ⅱα结构域片段的制备及其核磁共振鉴定

Wistar雄性大鼠经腹腔注射CdCl_2诱导其肝脏合成金属硫蛋白。取鼠肝,先经匀浆、热变性、乙醇-氯仿萃取、丙酮沉淀等步骤,后由Sephadex G-75、DEAE-52柱层析,得金属硫蛋白的两亚型Ⅰ和Ⅱ。经原子吸收光谱测定每蛋白分子含有5个镉和2个锌。用枯草杆菌酶割法制备并分离了金属硫蛋白亚型Ⅱ的α结构域片段。还用NMR研究了金属硫蛋白的两个亚型及所得的α结构域片段。