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目的探讨全反式维甲酸(ATRA)对人胃癌细胞株SGC7901的增殖及对胞膜蛋白caveolin-1表达和定位的影响。方法 MTT法检测不同浓度(5、10、25μmol/L)ATRA对SGC7901细胞增殖的影响;免疫荧光法检测cavolin-1在细胞中的定位,Western blot检测caveolin-1的表达。结果 ATRA可明显抑制人胃癌细胞株SGC7901的增殖。ATRA处理3 d后,SGC7901细胞caveolin-1的表达无明显变化,但caveolin-1在胞浆中的分布明显降低,胞膜上分布明显增加。结论 ATRA可明显抑制SGC7901的增殖,促进caveolin-1在细胞膜上定位。 相似文献
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许多学者基于巨噬细胞的贴壁特性曾使用多种方法从外周血分离人的巨噬细胞。但目前使用的一些方法尚有不足之处,可归纳为以下三点: 1.细胞的收获量和活力往往不稳定; 2.或者需要预先用小牛血清处理塑料物; 3.或者需要使用以前贴壁的细胞株的微量渗出物包被塑料物。 1978年Rinehart等虽用利多卡因(Lidoeathe)从未经处理过的塑料表面分离过人的单核细胞,但用他这种方法收获的细胞量又少又不稳定,细胞的活力也较差。本文作者所使用的低pH值利多卡因改良法可以经常产生功能好的人单核细胞,并与下列几种贴壁分离法作了比较:(1)用橡皮头(rubber policeman)进行手工刮除法;(2)用微量渗出物包被塑料物;(3) 相似文献
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目的:利用毕赤酵母表达系统表达两种融合形式的乙肝病毒表面抗原(HBsAg-s和s-HBsAg),并对融合蛋白的免疫原性进行研究。方法:用毕赤酵母分泌型表达质粒pPIC9k为载体,通过PCR方法引入(Gly4Ser)3连接肽的编码基因将GM-CSF融合到HBsAg基因的3′端或5′端进行分泌表达,表达产物以SDS-PAGE及Western blot进行分析并通过DEAE-Sepharose Fast Flow离子交换柱进行蛋白纯化。将纯化的两种融合蛋白与单纯的HBsAg蛋白分别免疫小鼠,ELISA方法检测HBsAg特异的抗体反应水平。结果:两种HBsAg/GM-CSF融合蛋白均可在毕赤酵母菌株GS115中获得了分泌表达,Western blot分析表明表达产物能够与抗GM-CSF抗体及抗HBsAg抗体发生特异性结合。免疫后的血清ELISA结果显示,相比单纯的HBsAg蛋白,两种融合蛋白可以诱导出更高的抗体水平(P<0.05),其中将GM-CSF蛋白融合在HBsAg蛋白的C端效果要好于融合在N端。结论:通过与GM-CSF蛋白的融合可以显著增强HBsAg的免疫原性,这为提高乙肝疫苗的免疫效果提供了新的思路与方法。 相似文献
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特异性刺激物的化学性质的近代研究表明,花粉粒含有一定的油质和多醣物,它可明显地被鼻粘膜分泌物迅速地消化并随后释放出大约13种可溶性抗原性蛋白质。King 相似文献
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多数肱骨外科颈骨折经闭合整复 ,用超肩石膏托或夹板、外展支架等外固定后能取得满意效果。除了常见的内固定方法外 ,我们自 1 996年以来 ,用三叶接骨钢板内固定治疗肱骨外科颈骨折 1 8例 ,优良率达 90 %以上 ,现报告如下。1 临床资料1 .1 一般资料 1 8例中男 1 1例 ,女 7例。年龄 30~ 70岁 ;病程为伤后 1~ 1 0天 ;左肩 8例 ,右肩 1 0例 ;内收型骨折 5例 ,外展型骨折 1 3例 ;合并肩关节前脱位 2例。所有骨折均经闭合整复后因对位不佳而转手术治疗。1 .2 治疗方法 ( 1 )手术方法 :在全麻或高位臂丛麻醉下取仰卧位且伤肩垫高 ,麻醉阻… 相似文献
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BACKGROUND: Transplantation of cell lines expressing neurturin (NTN) has been used to treat animal models of Parkinson's disease. However, gone homology between humans and rats varies greatly, so experimental results are not entirely suitable for understanding cellular transplantation in humans. OBJECTIVE: To explore expression of NTN in African green monkey kidney cells (Vero cells); to obtain a stably NTN expressing cell line. DESIGN, TIME AND SETTING: An observation study of gone engineering and cellular biology was performed at the Institute of Medical Biology, Chinese Academy of Medical Science and Peking Union Medical College between 2005 and 2008. MATERIALS: Human embryonic hepatic tissues, expression vector pcDNA3, and Vero cells were prepared in this laboratory. Primers were synthesized by TaKaRa Biotechnology, Dalian, China; RNA extraction kit and plasmid extraction kit were purchased from Shanghai Watson Bioengineering, China; G418 and MTT were purchased through Sigma, USA; Lipofectamine2000 was a product of Invitrogen, USA; mice anti-human NTN antibody and fluorescent labeling goat anti-mouse IgG antibody were provided by Jingmei Biotech, China. METHODS: Total RNA was harvested from human embryonic hepatic tissues, and NTN cDNA was cloned by RT-PCR method, followed by subcloning into the pcDNA3 eukaryotic expression vector. The obtained pcDNA3/hNTN was stably transfected into Vero cells using Lipofectamine 2000, and stably expressing clones were selected using G418. MAIN OUTCOME MEASURES: NTN mRNA and protein expressions were respectively identified by RT-PCR and immunofluorescence. The morphology of transfected cells was observed under inverted microscopy, and the growth characteristics of those cells were determined using MTT method. RESULTS: A clonal cell line, stably expressing human NTN mRNA and protein, was obtained through stable transfection of pcDNA3/hNTN into Vero cells. The transfected Veto cells exhibited irregular morphology, rather than a spindle shape. The growth retardation phase was prolonged, but the number of cells was identical to non-transfected cells. CONCLUSION: Vero cell lines, which stably expressed human NTN protein, were obtained, and expression patterns of these cell lines were acceptable. 相似文献
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关于Ⅰ型变态反应与肿瘤的关系,自50年代就已引起很多学者的重视。目前多数人认为这两者之间存在着一定的关系,少数人则持否定的态度。因此弄清这两者的关系,有利于我们开展肿瘤的防治工作。一、临床观察八十年代前后仍有很多学者认为变态反应状态有预防肿瘤作用的可能,这一概念已得到不少临床工作者的支持。例如1983年 相似文献