抗醛糖还原酶单克隆抗体的制备与特性鉴定

目的:制备抗醛糖还原酶(AR)的单克隆抗体(mAb),并与本室制备的抗醛糖还原酶相似蛋白(ARL-1)mAb进行比较。方法:经RT-PCR获得AR基因,将基因插入pGEX-4T-1(His)6C中,构建重组质粒pGEX-4T-1(His)6C-AR,以重组质粒转化E.coliRosetta诱导表达GST-AR蛋白。以纯化的GST-AR蛋白免疫BALB/c小鼠,采用杂交瘤技术制备mAb。应用间接ELISA和Western blot方法对mAb进行筛选和鉴定。使用Clustalx和Antheprot软件,比较AR与ARL-1的同源性,表达GST-dAR[80~142氨基酸(aa)],与ARL-1差异较大;并分析AR的抗原性,表达GST-dA1(1~79aa)、GST-dA2(80~99aa)、GST-dA3(111~142aa)、GST-dA4(143~316aa)。利用AR全长及截短蛋白,采用Western blot分析制备的抗AR mAb识别AR抗原的部位。结果:获得3株稳定分泌抗AR mAb的杂交瘤细胞系ARB3、AR7B3G4和ARF10。3株抗GST-AR的mAb均为IgG1(κ型),腹水mAb效价为1∶4×105,细胞培养上清mAb效价为1∶1×104,3株mAb均可与胎盘组织中的AR蛋白起反应,而与GST-ARL-1和GST蛋白无交叉反应。它们分别为抗GST-dA1、GST-dA3和GST-dA4蛋白的mAb。结论:成功地制备了3株特异性抗AR mAb,可分别识别AR的1~79、111~142、143~316位氨基酸。将它们与抗ARL-1mAb联合应用,将有助于进一步研究AR与ARL-1蛋白的功能,并为深入探讨AR、ARL-1与相关疾病的关系及进行大规模的流行病学调查提供了有力的工具。     

Diurnal variation of intraocular pressure and the overriding effects of sleep

Intraocular pressure (IOP) was recorded regularly over a 24-h period in 13 normal subjects. All subjects showed a dramatic rise in IOP after sleep, ranging from 37 to 248%. In a second experiment, IOP decreased when 15 subjects remained upright and awake throughout the night. When sleep was not permitted IOP was lowest at 3:00 a.m. (0300); when six of these subjects were permitted to sleep from 0600 to 0800, they showed a rapid and significant increase in IOP of up to 150%, whereas the remaining nine subjects showed (posturally induced) increases of up to 38%.     

rhGM-CSF对小鼠口腔粘膜损伤的防治作用

目的：探讨rhGM-CSF促进口腔腔溃愈合的作用机制。方法：取体外培养的胎儿口腔粘膜成纤维细胞，用rhGM-CSF刺激成纤维细胞后第2、4、6天用细胞计数及^3H－胸腺嘧啶掺入法（^3H-TdR）测定其增殖情况，结果rhGM-CSF对胎儿口腔粘膜成纤维细胞增殖及DNA合成有刺激作用，在80－100ng/ml时，细胞生长达到最佳状态，过高浓度rhGM-CSF的刺激作用反而下降。结论：rhGM-CSF可以通过刺激成纤维细胞增殖及DNA合成促进口腔溃疡愈合。     

肺癌侵犯中央肺动脉继发肺低灌注的CT与DSA对照研究

目的　探讨肺癌侵犯中央肺动脉继发肺低灌注的CT特征及其诊断价值。方法　对2 0例手术及病理证实的肺癌侵犯中央肺动脉 (cancerousinvasionofcentralpulmonaryarteriesinlungcancer,CICPA)患者术前肺血管DSA及增强螺旋CT(含高分辨率CT ,即HRCT)征象行前瞻性对照研究。根据DSA和CT胸膜下末梢肺动脉分布和数量变化 ,即肺血管征、肺毛细血管充盈度和肺实质强化后密度及时间 密度曲线变化 (即肺实质征 ) ,行两者间的盲法对照 ,统计学分析CT与DSA征象及其一致性 ,评估CT对CICPA继发肺低灌注的诊断价值。结果　 2 0例手术病理证实的CICPA病例中 ,共3 0支肺叶动脉受累 ,DSA显示 90 % (2 7/3 0 )出现肺叶低灌注征象 ,CT诊断肺低灌注敏感度为 78%(2 1/2 7) ,特异度 10 0 % (3 /3 ) ,准确度 80 % (2 4/3 0 )。肺低灌注与CPA狭窄程度无明显相关性 (rs=0 40 ,P =0 0 73 )。CT与DSA的肺血管征一致性较好 (Kappa =0 69,P <0 0 0 1) ,而肺实质征一致性稍差 (Kappa =0 3 7,P =0 0 11)。结论　肺低灌注是肺癌侵犯中央肺动脉的常见表现 ,CT对其定性诊断具有重要价值 ,而定量判断有一定限度     

多孔β-TCP/rhBMP-2人工骨在小鼠体内的诱导成骨和早期降解行为的研究

目的 将多孔β—磷酸三钙和重组人的骨形成蛋白—2复合物，植入小鼠肌肉内，观察复合物的骨诱导能力和早期降解行为。方法 将多孔β—磷酸三钙／人骨形成蛋白—2复合物植入25只小鼠右侧大腿后侧肌肉内，单纯多孔β—磷酸三钙植入左侧为对照，分别于72h、1、2、3、4周取材，进行组织学和扫描电镜观察。结果 72h可见幼稚软骨，1周见成熟软骨，2周见编织骨和少量板层骨，3周和4周成熟的板层骨生成，对照组则无骨或软骨生成。2周开始间充质细胞和纤维组织长入材料，3～4周将材料包裹分割，同时可见多核巨细胞反应及吞噬现象，这些是降解早期组织学改变。结论 多孔β—TCP／rhBMP—2复合人工骨显示了良好的生物相容性，有强大的骨诱导能力，有希望作为新型骨移植替代材料用于临床。     

Nasal lavage leukotrienes in infants with RSV bronchiolitis

Respiratory syncytial virus (RSV) bronchiolitis is a very common infection in infants and, after the acute phase, a number of patients develop a reactive airway disease that lasts for years. Although the pathogenesis of the lung damage after RSV bronchiolitis is still largely unknown, previous studies suggest that leukotrienes may play an active part in it. The aim of this study was to measure leukotriene levels in the nasal lavage fluid (NLF) collected in infants during RSV bronchiolitis and 1 month later. Cysteinyl leukotrienes (Cys-LTs) and leukotriene B(4) (LTB(4)) were measured in the NLF of 22 infants with their first episode of RSV bronchiolitis and 16 healthy infants. A second NLF sample was collected to measure leukotriene levels 1 month after the acute disease. NLF Cys-LT levels were significantly higher in infants with RSV bronchiolitis than in healthy controls [950 pg/ml (285.5-2155.9) vs. 110.5 pg/ml (66.5-451.3), p = 0.01], and they remained so a month after the acute infection (p = 0.02). A subanalysis showed no difference in Cys-LTs concentrations, either between bronchiolitis infants with and without a family history of atopy, or between those with and without passive exposure to cigarette smoke. No significant difference was found between the LTB(4) levels measured in the bronchiolitis cases and the control children. Cys-LTs are significantly increased in the NLF of infants with acute RSV bronchiolitis, and remain so at 1-month follow-up, suggesting a possible role of these eicosanoids in the pathogenesis of the disease.     

铁道车祸危险因素与行车安全经济效益的探讨

作者以1380名机车乘务员为研究对象,测查其生理心理指标,探讨发生车祸的危险因素,分析34年车祸流行的规律,并对行车安全的社会经济效盖进行了估算。研究结果显示,社会安定是减少车祸、保证行车安全的重要因素。通过多因素分析,揭示车祸的危险因素是低文化程度、过度疲劳、睡眠不足、视运动反应迟钝和弱神经类型,其间存在交互作用。实践表明,推行安全行车千日活动,可获巨大社会经济效益(1∶7)。     

高血压病患者血清瘦素含量与胰岛素抵抗水平的相关性研究

目的：探讨高血压病患者血清瘦素水平与胰岛素抵抗的关系。方法：测定217例高血压病患者(男92例，女125例)的空腹血清瘦素含量、空腹血糖、胰岛素、收缩压、舒张压和体质量指数(BMI)，稳态模式评估法计算胰岛素抵抗指数(HOMA-IR)。分析瘦素与其他各项参数的相关性。结果：以HOMA—IR的25％位点，作为判断胰岛素抵抗的切割点，把高血压病患者分为胰岛素抵抗组(IR)和胰岛素敏感组(IS)，血清瘦素浓度IR组显著高于IS组(P＜0．05)。血清瘦素浓度与HOMA—IR呈显著正相关(男性r＝0．407，P＜0．01；女性r＝0．254，P＜0．01)；校正年龄和BMI后，男性组两者仍呈显著正相关(r＝0．219，P＜0．05)，女性组两者无相关；逐步回归分析显示，男性组HOMA—IR为血清瘦素浓度的独立预测因素。结论：男性高血压病患者血清瘦素浓度与胰岛素抵抗直接相关，女性则无直接相关性。性别差异的机制有待进一步探讨。     

Porins and lipopolysaccharide stimulate platelet activating factor synthesis by human mesangial cells.

Porins, a family of hydrophobic proteins located in the outer membrane of the cell wall of gram-negative bacteria and lipopolysaccharide (LPS), were shown to stimulate the synthesis of platelet activating factor (PAF), a phospholipid mediator of inflammation and endotoxic shock, by cultured human glomerular mesangial cells (MC). The synthesis of PAF induced by porins was rapid (peak at 20 min) and independent either from contamination by LPS or from generation of an endotoxin-induced cytokine such as tumor necrosis factor (TNF) since it was not prevented by cycloheximide, an inhibitor of protein synthesis or anti-TNF blocking antibodies. LPS also stimulated PAF synthesis by MC. However, the kinetic of PAF synthesis induced by LPS was biphasic with an early and transient peak at 10 minutes and a second and sustained peak at three to six hours. This second peak required an intact protein synthesis and was prevented by anti-TNF antibodies, suggesting the dependency on LPS-induced synthesis of TNF. Experiments with labeled precursors demonstrated that in MC, either after stimulation with porins or LPS, PAF was synthesized via the remodeling pathway that involves acetylation of 1-0-alkyl-sn-glyceryl-3-phosphorylcholine (2-lyso-PAF) generated from 1-0-alkyl-2-acyl-sn-glyceryl-3-phosphorylcholine by phospholipase A2 (PLA2) activity. Porins and LPS, indeed, induced PLA2-dependent mobilization of [14C]-arachidonic acid that was inhibited by p-bromodiphenacylbromide (PBDB). PBDB, an inhibitor of PLA2, also blocked PAF synthesis by preventing the mobilization of 2-lyso-PAF, the substrate for PAF-specific acetyltransferase.(ABSTRACT TRUNCATED AT 250 WORDS)