Agilent 2100 Bioanalyzer在基因差异表达研究中的应用

目的观察Agilent 2100 Bioanalyzer 芯片分析系统(以下简称Bioanalyzer)在基因差异表达研究中的应用。方法应用限制性显示技术分别从正常和热休克处理后的酿酒酵母细胞中分离出cDNA片段,然后再用Bioanalyzer和传统的琼脂糖凝胶电泳技术对RD-PCR产物进行检测分析。结果Bioanalyzer能更快速、敏感地分离和显示差异表达的基因片段,并且通过对差异片段进行定量比较,发现了数个表达有明显差异的基因片段。结论Bioanalyzer在基因差异表达研究中具有重要的应用价值。     

日本血吸虫成虫67kD蛋白的分离与鉴定

目的 :分离日本血吸虫感染及免疫血清识别的成虫抗原 (AWA)中的特异蛋白带 ,为血吸虫病免疫诊断提供新的抗原分子。方法 :免疫印迹法分析AWA的特异蛋白带 ,电泳层析法分离靶抗原。结果 :获得了感染血清和免疫血清识别的 6 7kD蛋白。结论 :电泳层析法是一种分离血吸虫抗原的有效方法     

等电聚焦电泳分析红细胞Gd(-)壮族富宁型〔Gd(-)Zhuang-Funing〕

本文采用聚丙烯酰胺凝胶等电聚焦电泳(IEF),借助园盘电泳仪的简易装置,对正常 B 型G6PD 及变异型 G6PD 进行分析,获得了满意效果。     

Identification of differentially expressed proteins in human stage I lung adenocarcinoma and tumor-adjacent tissues with two-dimension gel electrophoresis profiling

Objective： The aim of this study was to establish reproducible two-dimensional electrophoretic assay used for profiling and identification of differentially expressed proteins in human stage I lung adenocarcinoma and paired normal tumor-adjacent tissue. Methods： The proteins from 12 human stage I lung adenocarcinoma tissues and normal tumor-adjacent tissues were separated using isoelectric focusing electrophoresis （the first dimension） and the subsequent homogeneous SDS-polyacrylamide gel electrophoresis （SDS-PAGE） （the second dimension）. The differentially expressed proteins were determined with PDQuest image analysis software, and identified using matrix-assisted laser desorption/ionization time of flight mass spectrometry （MALDI-TOF-MS） and database searching. Results： The well-reproducible 2-DE gel patterns of human stage I lung adenocarcinoma and normal tumor-adjacent tissues were profiled and 26 differentially expressed proteins uncovered. Nine of these 26 protein spots were cut out from the preparation gels and determined with MALDI-TOF-MS. Searching against the protein database, four candidate proteins were identified. They were 60S acidic ribosomal protein P2, Cathepsin B1, Apolipoprotein A-I precursor, and La 4.1 protein. Conclusion： In this study, high reproducible 2-DE gel protein images of human stage I lung adenocarcinoma and paired normal tumor-adjacent tissues were achieved successfully, and 4 differentially expressed proteins were revealed. These data will be helpful for screen of early biomarker and study of molecular mechanisms of human lung adenocarcinoma.     

人血清载脂蛋白A-I的分离、纯化及其测定

用超离心法制备人血清高密度脂蛋白(HDL);甲醇/氯仿脱脂;经Sephadex G-150层析及DEAE-纤维素(DE-52)离子交换得纯化载脂蛋白A-I(Apo A-I),免疫新西兰兔,获得效价为1:32的特异性抗人Apo A-I抗血清。用本室建立的单向火箭免疫电泳法测定了158名健康中年人与9名老人的血清Apo A—I含量,结果发现:β-脂蛋白<5g/L,总胆固醇<2mmol/L的人血清Apo A-I含量男女分别为2.06±0.16g/L和2.07±0.15g/L;而β脂蛋白≥5g/L,总胆固醇≥2mmol/L的人血清Apo A-I含量男女分别为1.75±0.15g/L和1.82±0.1g/L,由此可见,血脂正常的男性和女性的Apo A—I含量均显著高于血脂高的男性和女性。80岁以上老年人的血清Apo A-I含量为2.1±0.1g/L,也在正常范围。与文献报道比较,我国人的血清Apo A-I含量高于外国人,这与我国人HDL水平较外国人高一致。     

大肠侧向发育型肿瘤细胞株双向电泳技术成功建立

目的:建立侧向发育型肿瘤细胞(LST)株双向电泳技术.方法:提取LST细胞株总蛋白质,采用不同pH范围IPG胶条进行LST细胞株总蛋白质双向电泳,并对实验步骤进行一系列的调整优化.结果:获得了较为清晰的LST细胞株双向电泳图谱,采用pH 3～10线性IPG胶条分离出1356±43个蛋白质斑点;pH 4～7 IPG胶条两种上样量(250μg和150μg)分别分离出了1285±51和989个蛋白质斑点.结论:通过相关条件的调整优化,成功建立了双向电泳技术,为研究LST特殊性生物学行为相关蛋白质奠定了坚实的基础.     

血管通透性的调节和游离微血管技术在其研究中的应用

Vascular hyperpermeability is a cardinal feature of inflammation or bum in which an array of inflammatory mediators can cause such changes in the microvessels. The fimctional measures of microvascular exchange that represent the properties of microvascular wails are the permeability coefficients which have been reported from measurements on intact whole organisms (including human subjects}, on perfused tissues and organs, on single perfused microvessels, and on monolayers of cultured microvascular endothelial cells. In this review, we summarize some experiments of vascular permeability in individually isolated perfused microvessels.     

毛细管气相色谱法测定人体血液中的游离脂肪酸和总脂肪酸

采用石英毛细管气相色谱法测量人体血液中的游离脂肪酸（FFA）和总脂肪酸（TFA），方法灵敏，可分离血浆中近20种脂肪酸，其中油酸（C18：1）、软脂酸（C16：0），亚油酸（C18：2）、硬脂酸（C18：0）和软脂油酸（C16：1）约占血浆中FFA总量的90％。本方法所需血量少，尤其对采血较困难的婴儿进行脂肪营养研究适用。     

~(125)碘-绒毛膜促性腺激素的电泳纯化及分析

标记抗原在贮存过程中,由于受幅射分解和脱碘影响而使免疫活性和生物活性降低。我们对自制的~(125)碘-绒毛膜促性腺激素进行了实验观察。结果显示:贮存1月后,其免疫活性和生物活性分别降低了33.1％和44.0％;经电泳纯化后的标记物的免疫活性和生物活性明显提高,分别达到了最初的90.5％和95.3％。