IGF-Ⅰ及其受体在牛体外受精早期胚胎中的表达

采用成分明确的体外培养系统产生牛体外受精胚胎,结合荧光免疫定位的方法,利用激光共聚焦成像系统研究了在牛卵母细胞及不同发育阶段早期胚胎中IGF-Ⅰ及其受体的表达.结果表明,IGF-Ⅰ及其受体从未成熟的卵母细胞、成熟的卵母细胞到早期胚胎发育的各个阶段均有表达,在COC的颗粒细胞中也有表达.IGF-I在成熟卵母细胞中主要定位于细胞膜附近,在未成熟卵母细胞、2细胞期胚胎中分布于细胞膜及膜下区域,在8细胞期胚胎和桑椹胚中分布于整个细胞质中,在囊胚阶段主要位于滋胚层,内细胞团细胞有少量的表达.IGF-IR在未成熟卵母细胞和成熟卵母细胞以及2细胞、8细胞期胚胎中集中分布于卵裂球远离胚胎中心的细胞膜上,在桑椹胚期时遍布于整个胚胎细胞的细胞质中,在囊胚期时主要分布在滋胚层,内细胞团细胞有少量表达.     

牦牛血清转铁蛋白对人红细胞膜氧化损伤的保护作用

牦牛血清中分离纯化的转铁蛋白对人红细胞膜的抗氧化具有保护作用 .方法 :采用黄嘌呤 /黄嘌呤氧化酶、Fe2 + /H2 O2 两个体系产生的O·2 -、OH·对红细胞膜造成氧化损伤 ,通过丙二醛含量、超氧化物歧化酶和谷胱甘肽过氧化酶活性的测定研究牦牛转铁蛋白在抗氧化方面的作用与机制 .结果 :O·2 -和OH·可以造成红细胞膜丙二醛量的显著增加 ,超氧化物歧化酶和谷胱甘肽过氧化酶活性的下降 .当适量不同浓度牦牛血清转铁蛋白在黄嘌呤 /黄嘌呤氧化酶、Fe2 + /H2 O2 两个体系前处理可以抑制丙二醛的产生 ,保护红细胞膜免受丙二醛的氧化损伤并且使超氧化物歧化酶和谷胱甘肽过氧化酶活性升高 .提示 ,牦牛血清转铁蛋白对O·2 -和OH·自由基引起的氧化损伤有保护作用     

银纳米粒子与血清白蛋白相互作用的滞后效应

用圆二色光谱,紫外-可见光谱研究银纳米粒子与血清白蛋白的相互作用及其相关效应.拟合计算圆二色光谱数据发现,与银纳米粒子作用后的血清白蛋白中α-螺旋含量减少,而α-折叠、转角和无规卷曲等结构含量增加,其构象在作用后变得更为松散和伸展.通过时间扫描吸收光谱跟踪研究发现,在血清白蛋白与银纳米粒子相互作用的过程中,银纳米粒子的包覆与聚集、血清白蛋白的构象变化具有双重滞后效应.     

基于硅胶载体表面分子印迹聚合物对牛血清蛋白识别性能

以硅胶作为载体,丙烯酰胺和2-丙烯酰胺基-2-甲基丙磺酸为双功能单体,牛血清蛋白质作为模板分子,制备了牛血清蛋白表面分子印迹聚合物,并对其制备方法、性状和性质进行了检测。结果证明制备的分子印迹聚合物大小在400 nm左右,当功能单体的配比为5∶1时为最优配比,分子印迹聚合物对牛血清蛋白质的结合在60 min时达到平衡,并且在浓度为0. 6mg·m L~(-1)的情况下,结合率达到饱和。特异性检测可知分子印迹聚合物对牛血清蛋白的结合率最高并且其印迹因子(IF)高达1. 58,说明了它对目标蛋白具有良好的选择识别能力,且重复性能好,重复检测后吸附量在80%左右。     

硼酸缓冲液应用于电泳实验的效果分析

血清蛋白醋酸纤维薄膜电泳是医学院校生物化学实验中的重要实验,传统巴比妥缓冲液具有毒性且不经济,该文旨在研究硼酸缓冲液的效果。采用DY—l型电泳仪配套水平电泳槽进行醋酸纤维薄膜电泳,控制电流时间和室内温度。显示硼酸缓冲液电泳可以分离出5条清晰的蛋白区带,并且电泳10次后仍然效果良好。由此得出离子强度为0.08的硼酸缓冲液完全可以替代巴比妥,无毒且经济,值得推广应用。     

荒漠沙蜥血清蛋白含量的年周期变动

对荒漠沙蜥（Phrynocphalus przewalskii)血清蛋白仿及血清蛋白组分的周期变化进行了测定,测定时间为4－10月的每月中旬及1月中旬（进行人工冬眠的蜥蜴）,血清总蛋白含量采用紫外分光光度法测定,血清蛋白组合组分采用醋酸纤维素薄膜电泳分离,然后用双波长色谱扫描仪扫描,计算相对含量,测定结果表明,荒漠沙蜥的血清总蛋白含量在深冬眠期的1月最高,为81．45g/L,4月出 一进入繁殖季节,其含量开始下降,夏季（6－8月）降为43．81g/L,其中7月最低,为38．07g／L,秋季的含量略低于春季的含量,但差异不显著（P＞0．05）,在冬 眠前（10月下旬）及冬眠期（1月中旬）,雌性的含量高于雄性,且差异显著（P＜0．05）,在出眠后的繁殖季节雄性的含量高于雌性,在冬眠季节清蛋白的比例相对增加,而球蛋白的比例相对减少。     

Carnosic Acid Encapsulated in Albumin Nanoparticles Induces Apoptosis in Breast and Colorectal Cancer Cells

Carnosic acid (CA) is a natural phenolic compound with several biomedical actions. This work was performed to study the use of CA-loaded polymeric nanoparticles to improve the antitumor activity of breast cancer cells (MCF-7) and colon cancer cells (Caco-2). CA was encapsulated in bovine serum albumin (BSA), chitosan (CH), and cellulose (CL) nanoparticles. The CA-loaded BSA nanoparticles (CA-BSA-NPs) revealed the most promising formula as it showed good loading capacity and the best release rate profile as the drug reached 80% after 10 h. The physicochemical characterization of the CA-BSA-NPs and empty carrier (BSA-NPs) was performed by the particle size distribution analysis, transmission electron microscopy (TEM), and zeta potential. The antitumor activity of the CA-BSA-NPs was evaluated by measuring cell viability, apoptosis rate, and gene expression of GCLC, COX-2, and BCL-2 in MCF-7 and Caco-2. The cytotoxicity assay (MTT) showed elevated antitumor activity of CA-BSA-NPs against MCF-7 and Caco-2 compared to free CA and BSA-NPs. Moreover, apoptosis test data showed an arrest of the Caco-2 cells at G₂/M (10.84%) and the MCF-7 cells at G2/M (4.73%) in the CA-BSA-NPs treatment. RT-PCR-based gene expression analysis showed an upregulation of the GCLC gene and downregulation of the BCL-2 and COX-2 genes in cells treated with CA-BSA-NPs compared to untreated cells. In conclusion, CA-BSA-NPs has been introduced as a promising formula for treating breast and colorectal cancer.     

Human Serum Amyloid a Impaired Structural Stability of High-Density Lipoproteins (HDL) and Apolipoprotein (Apo) A-I and Exacerbated Glycation Susceptibility of ApoA-I and HDL

Human serum amyloid A (SAA) is an exchangeable apolipoprotein (apo) in high-density lipoprotein (HDL) that influences HDL quality and functionality, particularly in the acute phase of inflammation. On the other hand, the structural and functional correlations of HDL containing SAA and apoA-I have not been reported. The current study was designed to compare the change in HDL quality with increasing SAA content in the lipid-free and lipid-bound states in reconstituted HDL (rHDL). The expressed recombinant human SAA1 (13 kDa) was purified to at least 98% and characterized in the lipid-free and lipid-bound states with apoA-I. The dimyristoyl phosphatidylcholine (DMPC) binding ability of apoA-I was impaired severely by the addition of SAA, while SAA alone could not bind with DMPC. The recombinant human SAA1 was incorporated into the rHDL (molar ratio 95:5:1, 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC): cholesterol: apoA-I) with various apoA-I:SAA molar ratios from 1:0 to 1:0.5, 1:1 and 1:2. With increasing SAA1 content, the rHDL particle size was reduced from 98 Å to 93 Å, and the α-helicity of apoA-I:SAA was decreased from 73% to 40% for (1:0) and (1:2), respectively. The wavelength maximum fluorescence (WMF) of tryptophan in rHDL was red-shifted from 339 nm to 345 nm for (1:0) and (1:2) of apoA-I:SAA, respectively, indicating that the addition of SAA to rHDL destabilized the secondary structure of apoA-I. Upon denaturation by urea treatment from 0 M to 8 M, SAA showed only a 3 nm red-shift in WMF, while apoA-I showed a 16 nm red-shift in WMF, indicating that SAA is resistant to denaturation and apoA-I had higher conformational flexibility than SAA. The glycation reaction of apoA-I in the presence of fructose was accelerated up to 1.8-fold by adding SAA in a dose-dependent manner than that of apoA-I alone. In conclusion, the incorporation of SAA in rHDL impaired the structural stability of apoA-I and exacerbated glycation of HDL and apoA-I.     

一步法提取鸭血清转铁蛋白

一步法提取鸭血清转铁蛋白是利用热变性沉淀去除鸭血清中的杂蛋白，在微碱性（pH值为8．0）条件下，75℃保温25min，转铁蛋白的纯化倍数为1．11，收率为96％，而硫酸铵分级沉淀法的纯化倍数为1．03．收率为51％。一步法与硫酸铵分级沉淀法相比，方法简单，成本低廉，适宜工业化生产。     

黄粉虫对血清总胆固醇及甘油二酯的作用

利用黄粉虫的幼虫含较多亚油酸成分，喂正常迪卡公鸡，并对它的血清总胆固醇及甘油三酯的作用进行研究．试验动物随机分成３组，Ｉ组为对照组，Ⅱ组为黄粉虫幼虫组，６ｇ／ｄ，Ⅲ组为复方亚油酸乙酯丸组，１丸（含亚油酸乙酯３００ｍｇ）／ｄ，实验期１８ｄ．结果表明，血清总胆固醇，Ⅱ组为（２．７３±０．２６）ｍｍｏｌ／Ｌ，Ⅲ组为（２．８３±０．３１）ｍｍｏｌ／Ｌ，均明显低于对照组（３．４０±０．６）ｍｍｏｌ／Ｌ，Ｐ＜０．０１）．血清甘油三酯，Ⅱ组为（０．４０±０．１３）ｍｍｏｌ／Ｌ，Ⅲ组为（０．３６±０．１１）ｍｍｏｌ／Ｌ，均明显低于对照组（（０．９±０．３）ｍｍｏｌ／Ｌ，Ｐ＜０．０１）．试验说明用黄粉虫的幼虫喂迪卡公鸡有显著降低血清总胆固醇和甘油三酯作用．