PAI－1糖基化位点突变体的构建及其在哺乳动物细胞中表达

Ⅰ型纤溶酶原激活物抑制剂（ＰＡＩ－１）是一种Ｍｒ为５０×１０３的单链糖蛋白，有３７９个氨基酸残基，３个Ｎ型糖基化位点。构建ＰＡＩ－１糖基化突变体，以便研究糖链的功能。用寡核苷酸定位突变技术将３个糖基化位点２０９，２６５，３２９位进行突变，把３个糖基化位点都发生了突变的ＰＡＩ－１ｃＤＮＡ组装到真核表达载体ｐＳＶ２中，得到真核表达质粒ｐＺＨ－ｐ１－Ｍ３Ｅ；在二氢叶酸还原酶缺陷型中国仓鼠卵巢细胞（ＣＨＯｄｈｆｒ－）中进行短暂表达，用发色底物法和夹心ＥＬＩＳＡ方法检测培养液中ＰＡＩ－１的活性和含量。结果：糖基化位点突变的ＰＡＩ－１能在ＣＨＯ细胞中表达，但表达水平及活性较低。非糖基化ＰＡＩ－１的活性和抗原分别为４．３４ＩＵ／ｍｌ和３．１５μｇ／Ｌ结论：用寡核苷酸定位突变方法获得了ＰＡＩ－１糖基化突变体，并且在ＣＨＯ细胞中得到表达。     

Lung function, smoking and survival in severe alpha 1-antitrypsin deficiency, PiZZ

Lung function, smoking, age and mortality data in 158 adult severe alpha₁-antitrypsin deficient, PiZZ individuals, followed from 1963 to 1982 were analyzed. Low initial FEV₁ value was significantly associated with increased mortality (p < 0.005). A 3 yr mortality rate of 40% was found in individuals whose initial FEV₁ values were less than 30% of that predicted. In contrast, the corresponding 3-yr mortality among those whose initial FEV₁ values were between 30 and 65% of that predicted was only 7%. Smokers were found to have significantly lower FEV₁ levels (p = 0.008) and higher mortality (p < 0.005) than non-smokers. The difference between current and ex-smokers in mortality and FEV₁ level were not statistically significant (p = 0.9 and p>0.25, respectively). Cross-sectional analysis of the initial FEV₁ values indicated a significant decline (p < 0.005) of FEV₁ with increasing age. This decline was greater among smokers than non-smokers. Longitudinal analysis of FEV₁ rates of decline in 80 cases with follow-up FEV₁ measurements failed to detect any significant differences between smokers and non-smokers, but was performed late in the disease process. The application of these results to the planning of studies on replacement therapy, smoking intervention strategy and longitudinal follow-up is discussed.     

Chromosomal Mapping of Genetic Locus Associated with Thymus-size Enlargement in BUF/Mna Rats

The thymoma-prone rat of the BUF/Mna strain is a useful model for human thymoma. In this strain thymoma development is regulated by a single autosomal susceptible gene, Tsr-1. At pre-thymoma age, BUF/Mna rats have extremely large thyrauses, when compared to those of other strains of rats. Genetic studies in crosses between BUF/Mna rats with large thymuses and WKY/NCrj rats with small thymuses suggested the presence of a major autosomal gene, Ten-1 , which contributes to thymus enlargement in a backcross population. Linkage studies between Ten-1 and microsatellite markers in backcross rats of (WKY/NCrj×BUF/Mna)Fl×BUF/Mna have led to the localization of Ten-1 in chromosome 1. This result may provide an approach to clone Tsr-1 , which could be allelic to Ten-1.     

角质层及其类脂对雌二醇经皮渗透的作用

采用扩散装置和放射性同位素标记物测定,辅以FTIR和DSC技术,研究了雌二醇(OE)在角质层、去脂角质层、表皮以及活性表皮中的渗透性质和分配性质,分析角质层及其类脂在亲脂性药物经皮渗透中的作用。实验表明角质层是该类药物渗透的屏障,OE在其中的渗透速度受到角质层通透性和药物在角质层分配系数的双重控制,而OE从角质层转运至活性表皮的过程不成为限速因素。角质层的屏障性质仅部分来源于角质层类脂,而角质层细胞膜产生的阻力则是角质层屏障性质的重要组成部分。角质层类脂同时又可能是OE扩散的途径,该类物质通过对亲脂性药物增溶、提高后者在角质层中的分配产生影响。促进剂1,8-桉油精(1,8-CN)对OE的促渗作用也部分地依赖于角质层类脂的存在,通过一种可逆的物理相互作用改变类脂途径的通透性。     

MPTP对鼠肝脂质过氧化作用的影响

本文报道给C57小鼠腹腔注射不同剂量1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)后,发现组3(MPTP35 mg/kg,每天一次,共7天)鼠肝匀浆、线粒体和微粒体的膜丙二醛含量明显增加,与对照组相比分别增加70.5%,67%和51.4%(P<0.01),而组1(MPTP 35mg/kg,每4小时一次,共3次)和组2(MPTP35mg/kg,每天一次,共4天)鼠肝丙二醛含量与对照组相近。结果表明MPTP有明显地促进鼠肝脂质过氧化的作用,并与其剂量有关。     

Interleukin 1 receptor antagonist (IL1RA) in acute leukaemia: IL1RA is both secreted spontaneously by myelogenous leukaemia blasts and is a part of the acute phase reaction in patients with chemotherapy- induced leucopenia

Abstract: Blast cells derived from peripheral blood of patients with acute myelogenous leukaemia (AML) were cultured in vitro and interleukin 1 receptor antagonist (IL1RA) concentrations determined in culture supernatants. AML blasts derived from patients classified as AML-M4 and AML-M5 subtype showed an increased release of IL1RA. IL1α and IL1β caused a similar increase in AML blast release of IL1RA, and addition of anti-ILl antibodies decreased IL1RA release. IL1RA release from AML blasts was also increased by stem cell factor, tumour necrosis factor α (TNFα), granulocyte-macrophage colony-stimulating factor and macrophage colony-stimulating factor, whereas interleukin 3, interleukin 6, leukaemia inhibitory factor and granulocyte colony- stimulating factor did not significantly alter IL1RA release. When investigating IL1RA serum levels, serum concentrations were decreased in acute leukaemia patients with chemotherapy-induced cytopenia compared with healthy controls. Serum levels of both IL1RA as well as IL1β and soluble TNFα receptors increased when the leucopenic patients developed complicating bacterial infections.     

电针改善硬膜外吗啡用于术后镇痛所引起的免疫抑制

为观察硬膜外吗啡和电针对术后患者免疫功能的影响，检测自然杀伤细胞（ＮＫｃｅｌｌ）活性和ＰＨＡ诱导白细胞介素２（ＩＬ－２）水平在单纯胆囊切除术患者术前和术后第１、３、７天的动态变化情况。结果吗啡组ＮＫ活性在术后第１、３、７天抑制，手术组仅在术后第１、３天出现抑制，而抑制率低于同天的吗啡组，电针可拮抗吗啡引起的ＮＫ活性抑制加深状况。在术后第１天，手术组和吗啡组ＩＬ－２水平均下降，吗啡＋电针组无明显变化，术后第７天吗啡＋电针组ＩＬ－２升高接近正常人水平。表明电针能改善硬膜外吗啡引起的免疫抑制，促进术后机体的恢复。硬膜外吗啡结合电针是值得推荐的术后镇痛方法。     

115 BRCA1基因与乳腺癌研究进展

乳腺癌易感基因BRCA1作为抑癌基因,其编码产物在细胞周期调控、DNA损伤修复以及肿瘤细胞的凋亡中扮演着重要角色.大量研究证明,BRCA1基因结构、功能异常与乳腺癌的发生发展有十分密切的联系.本文拟就BRCA1与乳腺癌相关研究的最新进展作一综述.     

Molecular analysis and polymorphism of the DLA-DQB genes

Abstract: Partial-length cDNA clones and full-length genomic clones corresponding to a complete canine DQB class It gene were isolated. Southern analyses suggested the presence of two DQB genes - one of which appeared to be a pseudogene lacking exon 2 called DQB2. The other DQB gene, called DQB1, was isolated from a genomic phage clone and contained six exons. The DQB1 clone was restriction mapped, and exon 2 was sequenced from 70 dogs. Twenty alleles were found. Most of the amino acid substitutions occurred at putative positions in the peptide binding site. Inheritance of these sequences showed Mendelian segregation with one or two alleles per dog. Cluster analysis of the nucleotide and predicted amino acid sequences subdivided the canine DQB1 alleles into four major allelic groups. The number of nonsynonymous changes was higher than the number of synonymous changes in the putative antigen recognition sites suggestive of positive selection.