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71.
腺病毒介导的HSV—tk基因治疗大鼠脑胶质瘤实验研究 总被引:4,自引:0,他引:4
目的:带有HSV-tk基因的重组腺病毒(AdHCMV-tk)结合核苷类似物(NA)治疗大鼠C6脑胶质瘤。方法:用X-gal染色测定AdHCMV-lacZ转染大鼠C6胶质瘤细胞的效率。用AdHCMV-tk/ACV、GCV离体及活体治疗大鼠C6胶质瘤。结果:AdHCMV-lacZ感染C6细胞效率达100%,AdHCMV-tk感染C6细胞,在病毒感染复数为1000时,GCV和ACV半致死剂量分别为3μg/ml和20μg/ml,Ad-HCMV-tk/ACV治疗大鼠C6胶质瘤模型,大鼠生存期超过90天,而对照组分别为17.0±1.6天(生理盐水组)、14.5±1.3天(AdHCMV-lacZ组),P<0.001。结论:重组腺病毒对靶细胞感染效率可达100%,AdHCMV-tk用GCV的杀伤C6胶质瘤细胞比ACV强,而HSV-tk/ACV用腺病毒介导治疗大鼠脑肿瘤疗效显著。 相似文献
72.
73.
JOHN A. CARVER KERRIE A. NICHOLLS ANDREW J. AQUILINA ROGER J.W. TRUSCOTT 《Experimental eye research》1996,63(6):639-647
The high-molecular-weight (HMW) protein from the lens is composed mostly of α-crystallin in a highly aggregated state. Bovine HMW protein was carefully separated from α-crystallin by size-exclusion chromatography. α-Crystallin has chaperone-like ability whereby it stabilizes other proteins under conditions of stress (e.g. heat). Comparison of bovine HMW protein and α-crystallin shows that the HMW protein has a markedly reduced chaperone ability compared to α-crystallin. However, in contrast to the results of other workers, we observe no alteration with age in the ability of α-crystallin to act as a chaperone. Using electrospray ionisation mass spectrometry, changes in the phosphorylation of the α-crystallin subunits with age have been quantified. Phosphorylation of α-crystallin occurs early in life but does not alter in proportion after about three years of age. In addition, phosphorylation of the A subunit of α-crystallin has little effect on its chaperone ability. As is found in the artificially prepared HMW complex of α- and γ-crystallin, NMR spectroscopy shows that in the naturally occurring HMW protein, the short C-terminal extension of the αBsubunit has lost its flexibility whereas the αAsubunit extension is still flexible. Post-translational modifications therefore seem to have little effect on the chaperone action of α-crystallin, but alterations in the quaternary structure of α-crystallin via incorporation into the HMW aggregate, lead to major changes in the chaperone ability of the protein. The results are consistent with the notion that one of the contributing factors to cataract formation in the lens is the depletion of α-crystallin with age as it is converted into the HMW protein. 相似文献
74.
What is the value of bcl-2 protein detection for histopathologists? 总被引:15,自引:0,他引:15
75.
从包涵体中纯化重组人幽门螺杆菌热休克蛋白A亚单位 总被引:3,自引:1,他引:2
目的:建立一种有效从包涵体中纯化重组人幽门螺杆菌热休克蛋白A亚单位HspA的方法。方法:重组基因工程大肠杆菌发酵后,表达的Hp r HspA包涵体经洗涤,变性,复性,采用Q Sepharose High Performance阴离子交换层析和Superdex75凝胶过滤层析分离纯化,使用SDS-PAGE和HPLC检测纯度,选用ELISA和动物实验对纯化蛋白的免疫学活性和生物学活性进行鉴定。结果:Hp的HspA包涵经洗涤和溶解后,Hp的HspA的纯度>60%,包涵体溶解液经阴离子交换层析和凝胶过滤层析后纯度超过95%,Hp的HspA纯品经检测具有良好的免疫学活性和生物学活性。结论:本研究建立的分离纯化方法可从包涵体中获得高纯度的重组HspA蛋白,为进一步的动物实验研究奠定了基础。 相似文献
76.
INTRODUCTIONThemicrotubule-associatedprotein蚲ishyperphos-phorylatedandglycosylatedinAlzheimerdisease(AD),andtheseabnormalmodificationsformedthebasisofprogressivelyretrogradeneurofibrillarydegenerationseeninADbrainandtherebythedementia(1,2).ADab-normallyphosphorylated蚲notonlyismicrotubuleas-semblyincompetent,butalsoinhibitsassemblyanddis-assemblesthepreassembledmicrotubulesinvitro(3).Inthetangle-bearingneuronsinADbrain,thenormalcytoskeletonisdisruptedandreplacedwi… 相似文献
77.
O. GAROFALO P. G. E. KENNEDY M. SWASH† J. E. MARTIN† P. LUTHERT B. H. ANDERTON P. N. LEIGH 《Neuropathology and applied neurobiology》1991,17(1):39-45
The expression of two heat shock proteins, HSP72 and p57, in addition to ubiquitin, has been studied immunocytochemically in nine amyotrophic lateral sclerosis (ALS) cases and 10 age-matched controls. HSP72 and p57 antibodies did not identify the characteristic ubiquitin-immunoreactive inclusions present in anterior horn cells in ALS spinal cord. Antibodies to HSP72, but not to p57 or ubiquitin, strongly labelled structures corresponding to polyglucosan bodies in spinal grey matter. Such immunoreactive profiles were more abundant in ALS cases, although they were also present in control material. They were sometimes identified by haematoxylin and eosin and periodic acid Schiff reaction, but were not labeled by phosphotungstic acid haematoxylin or by antibodies to glial fibrillary acidic protein. Although ubiquitin, HSP72 and p57 are stress-induced proteins, they are expressed differently and might therefore have different significance in neuronal degeneration. 相似文献
78.
Expression of the neurofilament protein NF-H in L cells. 总被引:1,自引:0,他引:1
79.
80.
目的:在原核系统中表达并纯化大肠杆菌胞嘧啶脱氨酶(cytosine deaminase,CD),制备鼠抗大肠杆菌CD多克隆抗体,方法:亚克隆CD基因到原核表达载体pMAL-c2和pBV222中,并转化入大肠杆菌DH5α内,诱导表达并纯化MBP-CD和6his-CD融合蛋白,用纯化的MBP-CD融合蛋白免疫小鼠制备多克隆抗体。结果:通过重组质粒的酶切筛选出重组阳性克隆,成功地表达和纯化出MBP-CD和6his-CD融合蛋白,用纯化的MBP-CD成功制备了鼠抗CD多克隆抗体,并用6his-CD和GST-CD重组蛋白进行Western印迹分析,证实了抗体的正确性,结论:应用多克隆抗体可以检测体内外CD基因的表达,为临床前和临床上深入开展CD基因的生物治疗研究提供重要的实验材料。 相似文献