树舌多糖GF对小鼠HepA癌细胞PTEN蛋白表达的影响

目的 探明树舌多糖(GAPS)GF对小鼠HepA癌细胞PTEN蛋白表达的影响.方法 昆明种小鼠随机分肿瘤组和树舌多糖组.接种后各组肌肉注射给药,分别给予生理盐水和树舌多糖GF.连续10 d,于末次给药24 h,处死小鼠,剥离肿瘤组织.Dot blot技术检测各组样品PTEN蛋白的表达量.结果 肿瘤组检测结果为阴性,树舌多糖组检测结果为阳性.结论 树舌多糖GF抗肿瘤的作用靶点之一可能为抑癌基因PTEN.

小鼠HepA癌细胞PTEN蛋白表达及树舌多糖GF的影响

目的:探明小鼠HepA癌细胞PTEN蛋白表达及树舌多糖GF的影响.方法:昆明种小鼠随机分为三组:肿瘤组、树舌多糖组和阴性对照组.肿瘤组、树舌多糖组接种后各组肌肉注射给药,正常组和肿瘤组给与生理盐水,树舌多糖组给与树舌多糖GF;采用超速离心技术制备、SDS-PAGE分离细胞可溶性蛋白;Western blot和Dot blot技术检测各组样品PTEN蛋白的表达量.结果:树舌多糖组检测PTEN结果为阳性,肿瘤组检测PTEN结果为阴性.结论:树舌多糖GF抗肿瘤的作用靶点之一可能为抑癌基因PTEN.

AIM: Expression of bcl-2 in Marek's disease (MD) tumor was analyzed and relationship between bcl-2 gene and lymphomagensis clarified. METHODS: Model of MD tumor was reproduced on SPF White Leghorn chickens, collected tumor and normal tissues, conserved in liquid nitrogen. Total RNA in the tissues was obtained by AGPC method, detected their concentration and analyzed by 1% denatured agrose gel containing formaldehyde. A plasmid (pTTCB1) that contains chicken bcl-2 1.2 kb fragment was amplified, extracted and digested with restriction endonuclease (RE), the 1.2 kb fragment was recoveried and puarified with purigene kit, labeled probes with [α-32P]. Expression of bcl-2 was analyzed with Dot blot and Northern blot hybridization. RESULTS: (1) There are bcl-2 transcripts in MD tumor and relevent normal tissues; (2) Expression of bcl-2 in MD tumor is much higher than that in normal tissues. CONCLUSION: Our findings indicated that expressive products of chicken bcl-2 promotes lymphomagenesis of MD by blocking apoptotic cell death.

外加电场固定液相分子快速斑点免疫分析

目的建立一种广泛适用于临床上筛选抗体进行疾病诊断和在分子生物学研究中测试各种蛋白表达的快速、简便的检测方法.方法用一种特殊的多孔滤膜装置同时给多份样品施加电场,将其吸附于NC膜,进行免疫测试,称之为外加电场固定液相分子快速斑点免疫分析.分别进行线性、灵敏度和干扰因素测试.用其筛选自身免疫性疾病患者抗豚鼠内耳抗体和豚鼠内耳bFGF、FGFR、NFκB、IGF1R分子的表达.结果呈色反应的灰度值与抗体稀释倍数的对数之间具有很好的线性关系,相关系数r=0.995,P＜0.0001.6.4%SDS不影响检测结果,灰度值与SDS浓度之间的相关系数r=0.455,P＞0.05.其检测抗原灵敏度为23 ng,当待检样品中含99.86%杂蛋白时(待检蛋白0.149%),不能检出该分子.抗内耳抗体,31%(5/16)系统性红斑狼疮患者阳性,5%(1/19)类风湿病和强直性脊柱炎阳性,对照组2%(1/48)阳性.豚鼠内耳表达bFGF、FGFR、NFκB和IGF-IRα等4种分子.结论外加电场固定液相分子快速斑点免疫分析是一种抗表面活性剂干扰的高效抗原抗体检测方法.