人肝癌组织中p53与HSP70相互作用的初步研究

目的 :探讨人肝癌中热休克蛋白 70 (HSP70 )与 p5 3的相互作用。方法 :用免疫组织化学染色法 ,从 12例肝癌组织中筛选HSP70与 p5 3均呈阳性表达的标本 ,并以免疫共沉淀法提取之。然后用SDS PAGE及Westernblot分析双阳性标本中两种蛋白的存在形式。结果 :用免疫组织化学法检测到 12例肝癌组织中有 3例为双阳性 ,用抗HSP70mAb免疫共沉淀的样品 ,可检测到 p5 3蛋白。用抗p5 3mAb免疫共沉淀的样品也可检测到HSP70蛋白。结论 :人肝癌中p5 3与HSP70以复合物的形式而存在 ,此可为肝癌的发病机制及免疫治疗的研究提供新的思路     

Blocking-ELISA for detection of specific antibodies to the glycoproteins of the human immunodeficiency virus type 1 (HIV-1)

A blocking ELISA was developed to confirm the specificity of screening tests for anti-HIV-1 antibodies. A murine monoclonal antibody (McAb) raised against recombinant gp160 was used in combination with a commercial technique (ELA-VIA-1). After determining the optimal experimental conditions, the assay was applied to 92 samples presenting different reactivities by Western blot (WB) analysis. All the sera containing antibodies to gp160/gp120 (53) were positive in our assay. The six patients who sero converted showed a low positivity by ELAVIA-1 (optical density near the cutoff value) reacted by blocking-ELAVIA-1 with an McAb binding inhibition greater than 85%. By contrast, negative samples (29) and specimens that exhibited reactivity only against gag-proteins (10) were not detected (McAb binding inhibition smaller than 15%). This sensitive and specific blocking-ELAVIA-1 represents a convenient alternative to WB as a confirmatory test. The technique is time-saving and inexpensive and can easily be integrated with a screening test for diagnostic or epidemiologic studies on HIV-1 infection.     

中西医结合治疗眼底病的实验和临床研究

眼底病是一种复杂的致盲率很高的眼部疾病 ,包括视网膜脱离、视网膜玻璃体出血和增殖性玻璃体视网膜病变 (PVR)等疾病。本文采用中西医结合的治疗方法对动物模型和 1998～ 2 0 0 0年此类眼病患者进行研究 ,疗效显著 ,使眼底病的治愈率有了很大幅度的提高。中医药对眼底病的治疗常用活血化瘀复方治疗 ,对于从动物实验及临床两方面同时进行中西医结合的系统治疗研究 ,尚未见有报道。     

Tissue-specific distribution of the Goodpasture antigen demonstrated by 2-D electrophoresis and Western blotting

The target of autoantibodies in Goodpasture's disease, the Goodpastureantigen has recently been characterized as the NC1 domain ofthe 3 chain of type IV collagen. In order to study the Goodpastureantigen in different organs, NC1 domains were isolated frombasement membranes (BM) of human glomeruli (GBM), tubules (TBM),alveoli (ABM), placenta (PBM) and aorta (VBM). NC1 preparationswere separated by 2-D electrophoresis, and silver stained orimmunoblotted to determine the subunit structure and antigenicityof different basement membranes. All basement membranes containedmonomeric components of MW 26 kDa and 24 kDa, and associateddimers, corresponding to the 2-D location of 1(IV) and cc2(IV)chains respectively. However, GBM, ABM, and to a lesser extentTBM possessed an extra set of monomeric components of MW 28kDa and associated dimers corresponding to the proposed locationof 3(IV) and 4(IV) chains. 2-D-separated polypeptides were Westernblotted with autoantibodies from patients with Goodpasture'sdisease, a monoclonal antibody to the Goodpasture antigen (P1)and a monoclonal antibody to the bovine 3(IV) chain. The predominantbinding of all these reagents was to cationic 28 kDa monomersof GBM, ABM and TBM, corresponding to the 3(IV) chain, althoughautoantibodies and P1 also bound to neutral 28 kDa monomers,corresponding to the 4(IV) chain. Autoantibodies bound weaklyto more neutral components of PBM and VBM, but neither monoclonalantibody bound to these basement membranes. This study suggeststhat there are two separate type IV collagen networks: one containingthe l(IV) and 2(IV) chains appears to be present in all basementmembranes, and the other containing the 3(IV) and 4(IV) chainshas a tissue specific distribution. The latter network bearingthe Goodpasture antigen is expressed in the basement membranesof both kidney and lung, the organs involved in Goodpasture'sdisease.     

The effects of feeding an Asian or Western diet on sperm numbers, sperm quality and serum hormone levels in cynomolgus monkeys (Macaca fascicularis) injected with testosterone enanthate (TE) plus depot medroxyprogesterone acetate (DMPA)

The aim of this study was to elucidate the effects of an Asian diet compared to a Western diet on sperm numbers and quality, and serum hormones in cynomolgus monkeys (Macaca fascicularis) injected with testosterone enanthate (TE) plus depot medroxyprogesterone acetate (DMPA). Thirty male monkeys were divided into three groups of ten animals each. The first group (control) was fed with standard diet 'monkey chow' (9% fat, 13% protein, 78% carbohydrate); the second group was fed an 'asian' diet (15% fat, 15% protein, 70% carbohydrate); the third group was fed a 'Western' diet (35% fat, 25% protein, 40% carbohydrate). These diets were administered from the beginning (adaptation) until the experiment was terminated. Three months after the adaptation period, all groups were injected with 20 mg TE (once per week) and 25 mg DMPA (once every 6 weeks) for 18 weeks, while TE injections were continued for another 6 weeks. There were no differences in sperm numbers and quality, or in hormone levels between the first and second groups. In both of these groups azoospermia was achieved in 100% of animals, while in the third group only 70% achieved azoospermia. In all 3 groups, spermatozoa were once again detectable by week 33. However, by the end of the study at week 39, sperm numbers in the first and second groups reached only severe oligozoospermia (two animals remained azoospermic in the first group) while in the third group, numbers had returned to normozoospermia. The quality of spermatozoa during and after the treatment in the third group was better than in the first and second groups. Hormone concentrations decreased more rapidly in both the first and second groups, compared to the third group, while the recovery period was slower in the first and second groups, compared to the third group. It is concluded that different formula diets result in differential decreases in sperm numbers and quality, and in hormone concentrations in M. fascicularis injected with TE in combination with DMPA. Animals fed with either monkey chow or an Asian diet exhibited more severe and prolonged decreases in these parameters than did animals fed with a Western diet.     

Insulin-like growth factor (IGF)-I,-II and IGF-binding proteins in the cyst fluid of a patient with astrocytoma

Insulin-like growth factor (IGF)-I,-II and IGF-binding proteins (IGFBPs) were demonstrated in the cyst fluid of a patient with a hypothalamic astrocytoma. The astrocytoma cyst fluid was subjected to gel chromatography at low pH and the IGF-I and IGF-II levels were measured by specific radioimmunoassays. Immunoreactive IGF-I and IGF-II levels were 19 ng/ml and 78 ng/ml respectively. Several-fold higher IGF-II values were obtained when cyst fluid was not extracted or was extracted with acid ethanol before radioimmunoassay analysis. The immunoreactive IGFBP-1 concentration was 26 ng/ml. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and subsequent Western ligand blotting with [¹²⁵I]IGF-II revealed bands at 200, 34.5, 29.5, 24 and 21 kD as visualized by autoradiography. Binding studies demonstrated that these binding proteins bind specifically [¹²⁵I]IGF-I and [¹²⁵I]IGF-II. These observations suggest that IGFs as well as IGF-binding proteins are produced by astrocytoma cells and may act in a paracrine or autocrine fashion capable of modulating the growth of astrocytoma tumours.     

子宫内膜癌细胞中孕激素受体的多因素调节

目的　通过体外实验研究雌激素、孕激素、胰岛素 /胰岛素样生长因子Ⅰ (Insulin -likegrowthfactorⅠ ,IGF -Ⅰ )对内膜癌细胞中孕激素受体 (Progesteronereceptor,PR )亚型表达的调节 ,探讨内膜癌细胞中PR亚型表达的意义。方法　体外培养高分化子宫内膜癌细胞Ishikawa ,按实验目的分别加入雌激素、孕激素、雌激素孕激素联合、胰岛素及IGF -Ⅰ刺激 2 4h、 4 8h后 ,采用Westernblot方法测定各个条件下内膜癌细胞中两种孕激素受体亚型的表达。结果　内膜癌细胞Ishikawa在自然状态下 ,PR -A和PR -B均阳性表达 ;细胞经血清限制后 ,总PR及两种亚型都明显下调。雌激素可以上调细胞中PR -A和PR -B表达水平 ,但在高血清时明显。孕激素单独或雌、孕激素联合应用在 4 8h时都可以降低Ishikawa细胞中PR表达。胰岛素 /IGF -Ⅰ在 2 4h时可以上调PR -B水平 ,4 8h时此上调作用消失。结论　①雌激素对PR的上调作用与血清浓度有关 ,血清中可能含有雌激素作用所需要的某些因子 ;②孕激素对内膜癌细胞Ishikawa中PR有下调作用 ;③胰岛素 /IGF -Ⅰ可以上调内膜癌细胞中PR表达 ,主要是PR -B表达 ,但此作用持续时间短     

金黄色葡萄球菌NorA蛋白免疫检测方法的建立

目的建立金黄色葡萄球菌耐药转运蛋白NorA免疫检测方法。方法利用琼脂稀释法测定诺氟沙星（NFLX）对临床分离的6株金葡菌和标准菌株ATCC25923的MIC。对norA基因进行克隆、表达，提取表达产物包涵体，并进行了纯化。将纯化的表达蛋白作为免疫原免疫家兔，获取NorA蛋白的抗体血清。利用制得的抗体通过Western blotting检测临床分离的6株金葡菌的NorA蛋白表达水平。结果氟喹诺酮类药物耐药水平较高的临床分离金葡菌菌株的NorA蛋白表达水平一般较高，敏感菌株的耐药水平较低且接近。结论免疫检测方法可用于检测金葡萄菌耐药转运蛋白NorA的表达水平。     

中西文化差异与旅游资料的翻译

目的:提高旅游资料翻译的质量,促进中国旅游业的进一步发展.方法:通过分析中西文化的主要差异和旅游资料翻译的特点,提出旅游资料翻译中的文化处理原则和具体的翻译技巧.结论:在具体的旅游翻译实践中,运用添加解释性语境信息、删减冗余信息、类比、再创造等方法,缩小文化差异,可成功地进行文化交流.