穿心莲内酯对S180小鼠体内抗肿瘤作用的研究

观察穿心莲内酯对小鼠移植性肿瘤S180生长的抑制作用,并探讨其可能的机制.采用MTT法检测穿心莲内酯对体外培养的S180细胞增殖的抑制作用;建立小鼠移植瘤模型,采用荷瘤小鼠抑瘤率检测穿心莲内酯对小鼠肉瘤S180的体内抗肿瘤作用.以对荷瘤小鼠免疫器官质量,巨噬细胞对中性红的吞噬率,巨噬细胞产生TNF-α的影响评价穿心莲内酯对小鼠免疫功能的作用.结果穿心莲内酯能够剂量依赖型的抑制体外培养的S180细胞的增殖;25、50 mg/kg的穿心莲内酯对移植性动物肿瘤S180的抑瘤率分别为45.27%和56.87%,对荷瘤小鼠脾脏和胸腺质量无明显的影响.经穿心莲内酯刺激后,小鼠巨噬细胞对中性红的吞噬功能明显地增强,并能显著刺激巨噬细胞分泌TNF-α.说明穿心莲内酯具有明显的体内抗肿瘤作用,其抗肿瘤活性是通过抑制肿瘤细胞的生长,激活小鼠巨噬细胞两方面共同产生的.     

草鱼肾脏吞噬细胞吞噬活性的研究

分离草鱼肾脏细胞,并检测其中吞噬细胞对福尔马林灭活的柱状嗜纤维菌和福建爱德华氏菌的吞噬活性与饲养水温,测试温度的关系,及其血清的调理作用。结果表明,草鱼肾脏细胞中有吞噬的主要是嗜中性白细胞和巨噬细胞,饲养低温虽然能抑制血清中凝集抗体的产生,但对肾脏细胞的吞活性影响不大。     

绵羊肺泡巨噬细胞的分离培养和形态学观察

从绵羊肺组织中分离肺泡巨噬细胞进行原代培养,通过台盼蓝拒染试验和鸡红细胞吞噬试验检测肺泡巨噬细胞活力。利用电子显微镜对原代培养的巨噬细胞进行形态学观察。结果表明,本试验采用的方法适合分离培养绵羊肺泡巨噬细胞,活细胞数能达到90％以上。能为开展细胞病理学和细胞内寄生菌存活机理研究提供很好的试验材料。     

接骨木果实油对小鼠腹腔巨噬细胞吞噬活性的影响

作者以小鼠腹腔巨噬细胞吞噬活性及巨噬细胞酸性磷酸活性指标，检测了接骨木骨实油对小鼠胜利腔巨噬细胞功能的影响。     

家蝇蛹凝集素的纯化及对巨噬细胞免疫活性的影响

采用缓冲液浸提、亲和层析、超滤和离子交换4种方法,从家蝇蛹中分离纯化出一种D–半乳糖特异性结合的凝集素(MPL-1),该组分具有明显的凝血活性和免疫调节活性.聚丙烯酰胺凝胶电泳和高效液相色谱分析结果表明,MPL-1为单一成分的蛋白,其分子质量为80 ku.中性红吞噬实验结果显示,此种凝集素能提高巨噬细胞的吞噬活性,其作用效果具有时间和浓度依赖性.扫描电镜(SEM)观察结果表明,MPL-1作用后的巨噬细胞呈现明显的活化状态,证明MPL-1对巨噬细胞具有明显的免疫调节作用.     

17β-雌二醇对大鼠腹腔巨噬细胞活性的影响

①目的:研究17β-雌二醇(Estradiol,E)对大鼠腹腔巨噬细胞的影响。②方法:以大鼠腹腔巨噬细胞为对象,采用吞噬中性红法,观察不同剂量、不同时间雌激素处理后腹腔巨噬细胞的吞噬功能的变化。并HE染色进行形态学观察。③结果:剂量为10-4、10-6可引起腹腔巨噬细胞吞噬功能抑制,该作用在E处理后的2h、6h、16h均明显,随时间延长,作用增加。剂量10-8可增强吞噬功能。并有形态学变化。④结论:17β-雌二醇具免疫增强和免疫抑制双重功效,并随时间、剂量大小有变化。     

绞股蓝多糖的抗肿瘤作用及其机制研究

通过建立小鼠移植瘤模型,采用荷瘤小鼠抑瘤率检测绞股蓝多糖对小鼠肉瘤S180的体内抗肿瘤作用;采用MTT法检测绞股蓝多糖对体外培养的肿瘤细胞增殖的抑制作用.此外,采用比色法测定了绞股蓝多糖对巨噬细胞吞噬率和产生细胞因子NO的影响;采用ELISA法测定了绞股蓝多糖对巨噬细胞产生IL-1β和TNF-α的影响. 绞股蓝多糖可明显抑制移植性动物肿瘤S180的生长,并呈明显的剂量依赖关系;经绞股蓝多糖刺激后,小鼠巨噬细胞对中性红的吞噬功能明显的增强,并能显著刺激巨噬细胞分泌NO、TNF-α和IL-1β.为绞股蓝多糖的进一步开发提供了实验数据.     

CHO细胞中性红试验参数研究

为探讨中性红摄入法测定中国仓鼠卵巢细胞(CHO细胞)活性的有效性,为中性红比色法用于卷烟烟气毒理学评价提供参考.采用中性红吸入法,当中性红质量浓度为50～5000μg/mL,培育时间3h,溶解液pH值为3.0～4.0范围内,吸光值(A540nm)与CHO细胞的活细胞数呈正相关,是较为合适的实验参数.结果表明中性红比色法灵敏准确,结果稳定,操作简便,可以用于卷烟烟气细胞毒性的评价.     

“枸杞花粉口服液”对巨噬细胞功能的影响

作者观察了“枸杞花粉口服液”对小鼠腹腔巨噬细胞形态、酸性磷酸酶变化及对酶母细胞的吞噬活性，结果表明：“枸札花粉口服液”可以激活巨噬细胞，吴活化状态ＡＣＰａｓｅ活性增高，对酵母细胞吞噬活性增强，说明“枸杞花粉口服液”可提高机体免疫功能。     

白桦树皮提取物的抗腹水型肿瘤作用及对腹腔巨噬细胞功能的影响

利用体内荷腹水瘤模型 ,采用中性红吞噬法、染料释放法、Griess法和放射性释放法研究白桦树皮提取物 ( Extraction of Betula Platyphylla,EBP)在体内抗腹水型 S1 80和艾氏腹水癌以及对腹腔巨噬细胞功能的影响 .结果表明 ,EBP可明显延长荷腹水型 S1 80和艾氏腹水癌小鼠的生存期 ;巨噬细胞吞噬功能及分泌肿瘤坏死因子 ( TNF)和一氧化氮的功能明显增强 ,其细胞毒活性亦明显增加 ( p<0 .0 5)     

Uptake of Pneumocystis carinii mediated by the macrophage mannose receptor

Human exposure to Pneumocystis carinii is common but, in the absence of acquired or genetic dysfunction of either cellular or humoral immunity, exposure rarely leads to illness. Although alveolar macrophages can degrade P. carinii, macrophage receptors involved in P. carinii recognition have not been clearly defined. Characterization of a predominant surface glycoprotein of the high mannose type led us to investigate the role of the macrophage mannose receptor in this process. We report here that binding and uptake of cultured rat P. carinii by human and rat alveolar macrophages is reduced by 90% in the presence of competitive inhibitors of mannose receptor activity and by adherence of alveolar macrophages to mannan-coated surfaces. Further, only those COS cells transfected with the human macrophage mannose receptor complementary DNA that express surface mannose receptors bind and ingest P. carinii. These studies establish that the macrophage mannose receptor is sufficient for uptake of P. carinii and emphasize the role of the alveolar macrophage in first-line host defence against P. carinii.     

Hydroxyl radicals induced by quartz particles in lung alveolar macrophages： the role of surface iron

Previous studies have shown that hydroxyl radical generation is a key step in the mechanism of pathogenic process caused by airborne particles to the lung. However, there is no direct evidence for dose-response relationship between airborne particles and hydroxyl radical generation. In this study, hydroxyl radicals generated in lung alveolar macrophages exposed to quartz particles were measured using a highly sensitive capillary electrophoresis-fluorescence detection method. The results demonstrated that quartz particles induced the generation of hydroxyl radical in a dose-dependent manner, and the amount of the hydroxyl radicals was 10^-10 mol/10⁶ cells. The viability of alveolar macrophages exposed to quartz particles decreased with the increase of quartz concentration, showing a clear dose-response relationship. Hydroxyl radical scavenger mannitol could increase the viability of quartz-treated cells, suggesting that hydroxyl radical contributed directly to cell death. In this study this contribution accounted for about 5%?20% of cell death. The hydroxyl radical generating potential was found to be related to surface iron content of the quartz particles.     

全自动免疫组化筛查ALK基因融合非小细胞肺癌及其病理特征

目的探讨全自动免疫组化筛查间变性淋巴瘤激酶(ALK)基因融合非小细胞肺癌的临床特点及病理特征.方法选取经病理检查确诊的554例非小细胞肺癌组织,采用Ventana抗ALK试剂和全自动免疫组化(IHC)染色检测ALK状态,分析ALK基因融合非小细胞肺癌的临床特点和病理特征.结果本次研究的554例非小细胞肺癌患者组织中,共筛选出34例ALK阳性,占6.14%;年龄60岁的非小细胞肺癌患者ALK阳性率8.69%,明显高于年龄≥60岁的3.62%,差异具有显著统计学意义(P0.01);男性患者ALK阳性率6.71%高于女性5.21%,但差异无统计学意义(P0.05).组织形态学方面,34例ALK阳性非小细胞肺癌中28例为肺腺癌,6例为非肺腺癌.16例实体型为主腺癌合并黏液产生,7例腺泡型为主腺癌,1例为乳头型为主腺癌,4例为浸润性黏液腺癌,4例为鳞状细胞癌.EGFR基因突变检测显示:仅有1例合并该基因突变,其余均为野生型.9例IHC阳性样本,9例ALK基因融合非小细胞肺癌,9例IHC阴性样本经荧光原位杂交技术检测和RT-PCR检测均为阴性结果,6例IHC染色可能为阳性,经荧光原位杂交技术检测均显示为ALK融合阴性.结论 ALK基因融合肺癌是非小细胞肺癌一新的分子亚型,具有独特的临床表现和病理形态;Ventana抗ALK试剂和IHC染色是检测ALK阳性非小细胞肺癌首选方法,对提高该类型肺癌的检出率及个体化治疗具有重要意义.     

微量元素锰与肺癌的关系

本文对锰含量的测定表明:肺癌患者发锰含量显著低于对照者(P<0.01)。在动物诱癌实验中观察到,随着肿瘤的发生和发展,肝、肺中锰含量降低,而肿瘤部位锰含量则升高,提示锰含量低可能是肺癌发病的结果。     

Recombinant human TNF induces production of granulocyte-monocyte colony-stimulating factor

Tumor necrosis factor (TNF) is synthesized by macrophages exposed to endotoxin. It produces haemorrhagic necrosis of a variety of tumours in mice and is cytostatic or cytocidal against various transformed cell lines in vitro, but viability of normal human or rodent cells is unaffected. The role of TNF is unlikely to be restricted to the rejection of tumours. Colony-stimulating factors (CSFs) are required for survival, proliferation and differentiation of haematopoietic progenitor cells. The haematopoietic growth factor known as granulocyte-monocyte colony-stimulating factor (GM-CSF) has the ability to stimulate proliferation and differentiation of normal granulocyte-monocyte and eosinophil stem cells and enhance the proliferation of pluripotent, megakaryocyte and erythroid stem cells. In addition, GM-CSF stimulates a variety of functional activities in mature granulocytes and macrophages, for example inhibition of migration, phagocytosis of microbes, oxidative metabolism, and antibody-dependent cytotoxic killing of tumour cells. We show here that TNF markedly stimulates production of GM-CSF messenger RNA and protein in normal human lung fibroblasts and vascular endothelial cells, and in cells of several malignant tissues.     

DLL3蛋白在人小细胞肺癌细胞中的功能及其机制

为探讨DLL3(human notch ligand delta-like 3)过表达对人小细胞肺癌细胞的影响及可能的作用机制,通过PCR方法扩增人DLL3基因全长序列,并克隆至慢病毒表达载体Lenti-EFS-FLAG-puro而构建人DLL3基因过表达慢病毒表达质粒,酶切及测序鉴定质粒正确。通过慢病毒包装及感染,构建DLL3稳定过表达的人小细胞肺癌细胞株,并通过Western blot验证DLL3蛋白的表达。采用CCK-8法检测DLL3过表达对人小细胞肺癌细胞的增殖的影响。采用平板克隆实验检测DLL3过表达对人小细胞肺癌细胞的克隆形成的影响。Western blot检测DLL3过表达对细胞周期相关蛋白Cyclin D1,Cyclin D3的表达水平的影响。结果表明:人DLL3过表达慢病毒表达质粒构建成功; CCK-8实验显示DLL3过表达促进人小细胞肺癌细胞的增殖。平板克隆实验显示DLL3过表达提高人小细胞肺癌细胞的克隆形成能力。Western blot结果表明DLL3过表达增加细胞周期蛋白Cyclin D1,Cyclin D3的表达水平。可见DLL3过表达对人小细胞肺癌细胞的增殖具有促进作用。     

Functional genomic analysis of phagocytosis and identification of a Drosophila receptor for E. coli

The recognition and phagocytosis of microbes by macrophages is a principal aspect of innate immunity that is conserved from insects to humans. Drosophila melanogaster has circulating macrophages that phagocytose microbes similarly to mammalian macrophages, suggesting that insect macrophages can be used as a model to study cell-mediated innate immunity. We devised a double-stranded RNA interference-based screen in macrophage-like Drosophila S2 cells, and have defined 34 gene products involved in phagocytosis. These include proteins that participate in haemocyte development, vesicle transport, actin cytoskeleton regulation and a cell surface receptor. This receptor, Peptidoglycan recognition protein LC (PGRP-LC), is involved in phagocytosis of Gram-negative but not Gram-positive bacteria. Drosophila humoral immunity also distinguishes between Gram-negative and Gram-positive bacteria through the Imd and Toll pathways, respectively; however, a receptor for the Imd pathway has not been identified. Here we show that PGRP-LC is important for antibacterial peptide synthesis induced by Escherichia coli both in vitro and in vivo. Furthermore, totem mutants, which fail to express PGRP-LC, are susceptible to Gram-negative (E. coli), but not Gram-positive, bacterial infection. Our results demonstrate that PGRP-LC is an essential component for recognition and signalling of Gram-negative bacteria. Furthermore, this functional genomic approach is likely to have applications beyond phagocytosis.     

人类逆转录病毒的长末段重复序列（VRTVLTR）的表达与肺癌转移的关系

肺癌是最常见的恶性肿瘤之一。肿瘤癌很容易发生转移。实验以17例肺癌（5例肺腺癌、1例腺鳞癌、7例肺鳞癌及4例小细胞末分化癌）的原发灶、癌旁正常组织及转移淋巴结为材料,采用PCR及非放射性标记的RNA斑点杂交分析LTR在基因组的存在及其表达情况。研究结果发现：LTR序列在17例肺癌病人的正常组织及肿瘤组织的基因组中普遍存在,LTR致肺癌转移与其插入基因组中无关;LTR致肺癌转移与其表达增高有关,而且其高表达与小细胞肺癌的转移无关,而与非小细胞肺癌的转移密切相关。提示LTR参与了非小细胞肺癌的转移过程。实验结果为从细胞系获得的结论提供了进一步的证据。