动脉粥样硬化斑块中内皮素生成及分布的免疫组织化学观察

应用免疫组化方法对人动脉粥样硬化（ＡＳ）斑块中内皮素（ＥＴ）进行分析，发现除内皮细胞外，增生的平滑肌细胞（ＳＭＣ）中也含有大量的内皮素；在内皮剥脱的大鼠胸主动脉，增生的内膜ＳＭＣ能产生丰富的内皮素。内皮素放射免疫测定证实ＳＭＣ增生的活跃程度与内皮素量呈正比。提示内皮素合成增多与ＡＳ斑块内ＳＭＣ增生关系密切。     

阿糖胞苷纳米粒的制备及其释药规律研究

采用乳化聚合法制备阿糖胞苷纳米粒，研究其体内外释药特性。结果表明阿糖胞苷纳米粒体外释药规律符合双指数方程，有明显的缓释作用。在家兔体内的药物动力学过程符合二室模型，与阿糖胞苷注射剂相比，t1／2β和MRT延长，CL降低，表明阿糖胞苷纳米粒可显著延长阿糖胞苷在体内存留时间，具有明显的缓释特征。     

巢蛋白和阶段特异性胚胎抗原-1在大鼠2型星形胶质细胞中的表达

目的 观察1型和2型星形胶质细胞(T1A、T2A)是否表达神经干细胞的标志物、是否具有神经干细胞的特性.方法 取新生大鼠脑皮质,体外培养纯化的O-2A祖细胞、T1A和T2A,应用激光共焦双重免疫荧光标记技术检测巢蛋白和阶段特异性胚胎抗原-1(SSEA-1)的表达;观察O-2A祖细胞、 T1A和T2A在碱性成纤维生长因子(bFGF)和表皮生长因子(EGF)的培养液中生长方式的改变.结果 巢蛋白在O-2A祖细胞和T2A中表达,T1A不表达;SSEA-1仅在T2A中表达.在干细胞培养基中培养10d,T2A形成能增殖和连续传代的细胞球,细胞球巢蛋白标记阳性,贴壁后分化细胞具有神经元、星形胶质细胞和少突胶质细胞样形态;但相同培养条件下的O-2A祖细胞和T1A生长方式无改变.结论 巢蛋白和SSEA-1在两型星形胶质细胞中的表达存在差异,T2A具有神经干细胞的某些生物学特性.     

钴60放射法制备免疫功能抑制大鼠模型

目的通过系统的钴60放射计划,对大鼠免疫功能抑制的合适剂量进行评估,为干细胞移植研究提供合适的的动物模型。方法SD大鼠随机分为12.5Gy、10Gy、7.5Gy、5Gy4个剂量组,10只/组。计算放射后2个月的死亡率。检测白细胞数量,评价各剂量组动物的白细胞参考值范围。结合死亡率和白细胞参考值范围,确定合适的放射剂量。通过对外周血中性粒细胞(PMN)吞噬实验和外周血白细胞移行抑制实验(MIT),对细胞免疫功能进行检测。结果行12.5Gy照射的大鼠,3d内全部死亡;10Gy放射后的大鼠在1个月内全部死亡;7.5Gy照射的大鼠2个月内死亡1只;5Gy照射的大鼠没有死亡情况发生。设定95%作为可信区间,计算白细胞数量参考值范围(109个/L),未放疗组:16.978+1.96×6.46;5Gy放疗组:4.93+1.96×0.72;7.5Gy放疗组:2.313+1.96×0.782;10Gy放疗组:1.03+1.96×0.507。t检验表明,随着放疗剂量的增加,各实验组的白细胞数量显著减少。对于细胞免疫功能的检测,外周血中性粒细胞(PMN)吞噬实验结果表明,机体全身免疫功能降低;外周血白细胞移行抑制实验(MIT)表明,机体T细胞免疫功能降低。结论7.5Gy可以一定程度上抑制大鼠的免疫功能,为合适的放疗剂量。     

Detection of turkey rhinotracheitis virus in turkeys using the polymerase chain reaction

Six-week-old turkey poults were infected with the virulent UK/3B/85 strain of TRTV. Tracheal and oesophageal swabs were made every 2 to 3 days from groups of five poults and the RNA extracted. The TRTV RNA was then reverse-transcribed into complementary DNA (cDNA) using an oligonucleotide complementary to the 3' end of the fusion protein (F) mRNA. The cDNA was then used in a polymerase chain reaction (PCR) with an upstream primer to generate a product of approximately 0.5 kbp which was detected by ethidium bromide staining after electrophoresis. In this way, TRTV was detected in both types of swab for 17 to 19 days post-infection, nearly 2 weeks after the peak titres of infectious virus. Swabs which were allowed to dry completely before RNA extraction were as successful as swabs kept wet and extracted almost immediately, useful for when samples are collected in the field. The oligonucleotides amplified the 0.5 kbp product from TRTV strains isolated in six countries over a 13-year period, indicating that they might be usable as 'universal' oligonucleotides for TRTV detection.     

Significance of cytogenetic and fluorescence in situ hybridization analysis in evaluating antichronic myeloid leukemia efficacy of different immune effector cells

We report on the antileukemia effect of interleukin 2 (IL2) on different immune cells from 22 patients with chronic myeloid leukemia (CML). Bone marrow cells from these patients were first cultured in modified long-term bone marrow culture medium for several days, then separately cultured with lymphokine activated killer cells (LAK), cytokine-induced killer cells (CIK), and dendritic cell cocultured CIK (DC-CIK) for another 1-2 days. They were then detected for presence of the Philadelphia chromosome (Ph) by cytogenetic analysis and fluorescence in situ hybridization (FISH). The percentage of Ph-chromosome-positive cells in the bone marrow mononuclear cells after culturing with CIK and DC-CIK was significantly lower than that after culturing with IL2 or LAK. Our results demonstrate that cytogenetics and FISH are useful techniques for the evaluation of the anti-CML effect of immune cells and that CIK or DC-CIK can be appropriate candidates for adoptive immune cell therapy in vivo or for leukemia cell purging ex vivo.     

路氏乳杆菌JCM1081黏附相关蛋白的初步鉴定

目的研究路氏乳杆菌(Lactobacillus reuteri,也称罗伊氏乳杆菌)JCM1081菌体表面蛋白对其黏附HT-29细胞的影响。方法将路氏乳杆菌JCM1081菌体进行胰蛋白酶、蛋白酶K处理;用氯化锂和盐酸胍对乳杆菌表面的蛋白进行抽提,进行SDS-PAGE后与黏蛋白受体进行Western blot,并对杂交阳性蛋白进行质谱分析鉴定。结果路氏乳杆菌JCM1081菌体经胰蛋白酶、蛋白酶K处理后,其对HT-29细胞的黏附力显著下降(P<0．01);用氯化锂去除路氏乳杆菌JCM1081菌体外表面的S层蛋白后,路氏乳杆菌JCM1081对HT-29细胞的黏附力无显著变化;Western blot结果显示相对分子质量(Mr)为29×103和14×103的两种菌体表面蛋白与黏蛋白受体杂交中出现了强阳性;质谱分析结果显示29×103蛋白与路氏乳杆菌ATCC55730的h0793蛋白相似性高达71．1％。结论路氏乳杆菌JCM1081菌体表面的蛋白参与了乳杆菌的黏附,其中29×103和14×103的两种胞壁表面蛋白能够特异地识别黏蛋白受体并与之结合,29×103蛋白属ABC转运蛋白家族。     

Deletion of stem-loop 3 is compensated by second-site mutations within the Gag protein of human immunodeficiency virus type 1

Encapsidation of human immunodeficiency virus type 1 (HIV-1) RNA involves specific interactions between viral Gag proteins and viral RNA elements located at the 5' untranslated region (UTR). These RNA elements are termed packaging (psi) or encapsidation (E) signals and mainly comprise the stem-loop 1 (SL1) and SL3 RNA structures. We have previously shown that deletion of the SL1 sequences is compensated by second-site mutations within Gag. Similar studies are now extended to SL3 and the results demonstrate that deletion of this RNA structure is rescued by two point mutations, i.e., A11V in p2 and I12V in nucleocapsid (NC). These two compensatory mutations are different from those associated with the rescue of SL1 deletion, suggesting that SL1 and SL3 may bind to different residues of Gag during viral RNA packaging. Analysis of virion-derived RNA in native agarose gels shows that deletion of SL3 leads to decreases in both viral RNA packaging and dimerization. These defects are corrected by the compensatory mutations A11V and I12V. Yet, defects in viral RNA dimerization at an early stage that were caused by the SL3 deletion in the context of a viral protease-negative mutation cannot be overcome by these two suppressor mutations. Therefore, the positive effects of A11V and I12V on dimerization of the SL3-deleted RNA must have taken place at the maturation stage.