Methods of detecting mature human Ia-like antigen and their metabolic precursors

Ia antigens were shown to be present in the cell almost exclusively as mature alpha beta dimers which split into separate alpha and beta chains after boiling in SDS. In contrast metabolically labelling the cells with [35S]methionine resulted in only free alpha and beta chains being labelled. It is concluded that this widely used type of labelling, although useful for studying intermediate synthesis, should not be used for labelling mature cell surface molecules.     

血栓闭塞性脉管炎患者血栓素、前列环素及血液流变学指标的检测分析

目的 :探讨血栓闭塞性脉管炎患者血栓素B2 、前列环素与血液流变学指标的变化及其与发病机制的关系。方法 :用放射免疫法测定TXA2 和PGI2 的代谢产物血栓素B2 (TXB2 )、6 酮 前列腺素F1α( 6 K PGF1α) ,LBY N6A自清洗旋转式粘度计测定血液流变学指标 ,并与对照组对比分析。结果 :血栓闭塞性脉管炎患者TXB2 ( 53.59± 83.2 1ng/L)、6 K PGF1α( 14 .50± 3.4 5ng/L) ,与对照组相比差异显著 ;全血高切粘度、低切粘度、血浆粘度及纤维蛋白原疾病组均高于对照组 ,差异具有统计学意义。结论 :血栓闭塞性脉管炎患者存在TXA2 /PGI2失衡 ,并与血液流变学的改变有密切关系     

EB病毒LMP1在鼻咽癌细胞中通过IκBα的降解活化NF-κB

目的　阐明鼻咽癌细胞中EB病毒编码的潜伏膜蛋白 1(LMP1)活化核转录因子NF κB的机制。方法　利用强力霉素Dox诱导表达LMP1的鼻咽癌细胞株Tet on LMP1 HNE2为实验模型 ,首先应用免疫印迹方法测定Dox诱导后不同时相LMP1的表达动力学以及IκBs蛋白量及功能的改变。进而用间接免疫荧光法检测NF κB的亚细胞定位。最后采用瞬间共转染及报道基因活性分析分析NF κB的活性。结果　在鼻咽癌细胞Tet on LMP1 HNE2中 ,Dox处理 15分钟后LMP1的表达迅速升高并维持与较高水平直至 12 0分钟。LMP1的诱导性表达导致IκBα的磷酸化并降解 ,但IκBα蛋白总量无改变。继IκBα的磷酸化并降解 ,NF κB(P6 5 )自胞浆易位至胞核且活性升高。IκBα的显性负性突变子抑制NF κB(P6 5 )的核易位及报道活性。LMP1的诱导性表达并未引起IκBβ蛋白水平变化。结论　在鼻咽癌细胞Tet on LMP1 HNE2中 ,EB病毒LMP1通过IκBα的磷酸化并降解激活核转录因子NF κB的活性 ,并且 ,LMP1诱导的NF κB活性能被IκBα的显性负性突变子完全抑制。IκBβ在此信号传导途径中无改变。LMP1表达前后IκBα蛋白总量维持恒定可能是由于NF κB的活化迅速启动了IκBα的重头合成这一自身调节环路所致。     

氧化低密度脂蛋白功能性调节人内皮细胞的生长调节致癌基因α

目的　探讨oxLDL对人内皮细胞的CXC亚家族趋化因子GROα的调节作用及生理意义。方法　超高速离心得LDL ,氧化后得oxLDL。逆转录PCR分析人内皮细胞系ECV30 4细胞GROαmRNA。GAPDH被用作PCR中正常改变的对照物。酶联免疫检测仪测定ECV30 4细胞表面连接及分泌到溶液中的GROα蛋白。静态细胞粘附试验测定ECV30 4细胞表面连接的GROα蛋白的生理意义。结果　正常ECV30 4细胞不表达GROαmRNA ;OxLDL显著调节其表达 ,诱导效应首先出现在处理后 1h ,最大在 2h ,在 4h时下降。相比较 ,LDL对其mRNA表达没有影响。正常ECV30 4细胞低水平表达细胞表面的GROα蛋白。OxLDL呈浓度依赖性、时间依赖性地上调其表面的GROα蛋白。相比较 ,LDL对其表面表达的GROα蛋白无调节作用。OxLDL对ECV30 4细胞分泌到细胞溶液中GROα蛋白很少有影响。 4 0 μg/mloxLDL处理ECV30 4细胞 2 4h ,造成显著的人单核细胞系U937细胞粘附到ECV30 4细胞数目的增加。用GROα抗体预处理oxLDL刺激的ECV30 4细胞 ,可显著减低U937细胞粘附到ECV30 4细胞的U937细胞数目 (大约 2 / 3)。结论　oxLDL可能功能性上调内皮细胞GROα的表达。     

尿毒症及血透对血清一氧化氮浓度的影响及其临床意义

为研究尿毒症及血液透析对血清一氧化氮 (NO)浓度的影响及其临床意义 ,应用硝酸盐还原酶法检测了 1 0例尿毒症未透析患者、1 4例维持性血液透析患者透析前后及 1 0例正常对照者血清NO浓度。结果 :尿毒症未透析及血透患者NO水平均较正常对照显著升高 (P <0 .0 5) ,且尿毒症未透析患者血NO浓度与肿瘤坏死因子 α水平呈显著性正相关 (r =0 .858,P <0 .0 0 1 )。血液透析后NO浓度较透析前显著降低 (P <0 .0 5) ,但透析间期又会导致其蓄积。结果提示 :NO的蓄积对尿毒症的临床表现可能有一定影响。     

GNAS gene mutations affecting XLαs and bone health: A long neglected relationship

The GNAS locus is an imprinted site. The α-subunit of the stimulatory G protein (Gsα) and extralarge variant (XLαs) are the two important products of the GNAS locus. The abnormal expression of Gsα is associated with pseudohypoparathyroidism (PHP) and related disorders, including Albright hereditary osteodystrophy (AHO), pseudopseudohypoparathyroidism (PPHP), and progressive osseous heteroplasia (POH). XLαs protein can mimic the catalytic intracellular synthesis of cyclic adenosine monophosphate (cAMP) by Gsα in response to parathyroid hormone (PTH) stimulation, which may be involved in the pathogenesis of PPHP and POH in patients with paternal GNAS defects. A paternally inherited nonsense variant in the first exon of XLαs in an adult patient may be associated with fractures and osteopetrosis. The relationship between the XLαs product of the GNAS locus and bone remodeling may have been overlooked. Here, we summarize the phenotypes of genetic mouse models and clinical cases of XLαs variations and suggest that the abnormal paternal expression of XLαs may be associated with the development of POH and affect osteoblast and osteoclast differentiation.     

A high-yield and simplified procedure for the synthesis of α-[C]Methyl-l-tryptophan

Alpha-[¹¹C]methyl-l-tryptophan (AMT) has been synthesized by stereoselective methylation with [¹¹C]methyl iodide of the lithium-enolate generated by treating dimethyl 2(S),3a(R),8a(S)-(+)-hexahydro-8(phenylsulfonyl) pyrrolo[2,3-b]indole-1,2-dicarboxylate (2) with lithium diisopropyl amide (LDA) at −55 °C, followed by ring opening using trifluoroacetic acid and alkaline hydrolysis of the protecting groups. The crude product was purified by a simple reverse-phase C-18 Sep-Pak procedure. The purified product was isolated with an average radiochemical yield of 53 ± 12% (decay corrected) in 30–35 min from [¹¹C]methyl iodide. At end of synthesis (EOS), 138 ± 35 mCi (n = 24) of product was collected with a specific activity of ca. 1–1.3 Ci/μmol (EOS) (4–5 Ci/μmol @ EOB) starting from 1.5 Ci (EOB) of [¹¹C]CO₂.     

Further studies of the turnover of dog antithrombin III. Study of 131I-labelled antithrombin protease complexes

Fresh plasma containing 131I-antithrombin III (*I-AT) was coagulated and incubated at 37 degrees C for 2 hr. A "complex peak," separated on heparin-agarose contained AT and *I-AT antigen but no heparin cofactor activity. Crossed immunoelectrophoresis showed only AT complexes. SDS PAGE showed 80% of the *I-AT in a major band (approximately 80,000 daltons), 15% in a minor band (approximately 100,000 daltons) and the rest in trace bands (approximately 60,000 and/or 115,000 daltons). Ammonia treatment of the complex peak released alpha-thrombin. After i.v. injection 80% of the complexed *I-AT, chiefly as the major band, left the plasma with t 1/2 approximately 15 min and was almost immediately catabolized to low molecular weight breakdown products. A major catabolic site was the liver. A simple kinetic model describes the findings approximately.     

5α-还原酶抑制剂对过度训练大鼠雄激素作用的影响

 目的探讨非甾体类5α-还原酶抑制剂DQI对大运动量游泳训练大鼠血清雄激素水平、EPO水平及肾脏组织EPO合成能力的影响。方法以渐增游泳训练为大鼠运动应激模型，运用酶免和放免分析法观察了血睾、血清EPO及血清5α-双氢睾酮的含量，以及运用RT PCR法观察了肾脏组织EPOmRNA的表达水平。结果本运动模型可提高靶组织睾酮向5α-双氢睾酮的转化，而DQI可以显著抑制这一过程，并可以提高肾脏组织EPO的合成能力及血清EPO含量。结论非甾体类5α-还原酶抑制剂DQI对维持及恢复运动能力有一定作用。其机制与整合内源性睾酮及增加内源性EPO有关。