肺癌患者肺泡巨噬细胞一氧化氮活性研究

用镀铜镉还原－Ｇｒｉｅｓｓ法检测肺癌患者ＢＡＬＦ及肺泡巨噬细胞（ＡＭ）培养上清液中ＮＯ水平；ＲＴ－ＰＣＲ检测ＡＭｉＮＯＳｍＲＮＡ表达。结果显示，肺癌组ＢＡＬＦ及ＡＭ培养上清液中ＮＯ水平均明显低于对照组。两组ＡＭｉＮＯＳｍＲＮＡ表达阳性率分别为６９％和９１％（Ｐ＞０．０５），但表达强度肺癌组明显弱于对照组（Ｐ＜０．０１）。经ＧＭ－ＣＳＦ刺激后，ＡＭｉＮＯＳ表达强度及培养上清液中ＮＯ水平均显著提高。提示肺癌局部ＡＭ抗肿瘤功能可能存在某些缺陷；ＧＭ－ＣＳＦ可刺激ＡＭ使其功能增强     

肺癌患者肺泡巨噬细胞抗肿瘤免疫功能研究

用镀铜镉还原法、放射免疫法、比色法、酶标双抗体夹心法及逆转录－聚合酶链式反应（ＲＴ－ＰＣＲ）研究肺癌患者肺泡巨噬细胞（ＡＭ）抗肿瘤免疫功能状态。结果显示肺癌患者荷瘤侧肺ＢＡＬＦ中ＮＯ，ＡＭ培养上清中ＮＯ和ＴＮＦα活性低于非荷瘤侧肺与对照组，ｓＩＬ－２Ｒ活性高于对照组；经ＧＭ－ＣＳＦ刺激后ＡＭ培养上清中ＮＯ，ＴＮＦα及ＳＯＤ活性增强，ｓＩＬ－２Ｒ水平降低；两组ＡＭ ｉＮＯＳ ｍＲＮＡ表达强度明显弱于     

肺癌患者肺泡巨噬细胞精氨酸酶活性研究

检测肺泡巨噬细胞（ＡＭ）培养液中精氨酸酶活性。结果表明，肺癌患者荷瘤侧肺ＡＭ较非荷瘤侧肺和非癌性肺病患者ＡＭ产生精氨酸酶活性低；ＡＭ经卡介菌（ＢＣＧ），干扰素－α（ＩＦＮ－α）刺激后产生精氨酸酶活性明显增强。提示肺癌患者肿瘤局部ＡＭ抗肿瘤活性存在缺陷；ＢＣＧ，ＩＦＮ－α诱导ＡＭ产生精氨酸酶可能为其抗肿瘤、抗感染的机制之一。     

肺癌患者肺泡巨噬细胞抗肿瘤免疫功能研究

用镀铜镉还原法、放射免疫法、比色法、酶标双抗体夹心法及逆转录－聚合酶链式反应（ＲＴ－ＰＣＲ）研究肺癌患者肺泡巨噬细胞（ＡＭ）抗肿瘤免疫功能状态。结果显示肺癌患者荷瘤侧肺ＢＡＬＦ中ＮＯ，ＡＭ培养上清中ＮＯ和ＴＮＦα活性低于非荷瘤侧肺与对照组，ｓＩＬ－２Ｒ活性高于对照组；经ＧＭ－ＣＳＦ刺激后ＡＭ培养上清中ＮＯ，ＴＮＦα及ＳＯＤ活性增强，ｓＩＬ－２Ｒ水平降低；两组ＡＭｉＮＯＳｍＲＮＡ表达强度明显弱于对照组。表明肺癌患者机体免疫功能及肿瘤局部ＡＭ抗肿瘤功能均存在不均一性缺陷。     

肺癌患者肺泡巨噬细胞表现HLA-DR的表达

目的 了解肺癌患者肺泡巨噬细胞表面人白细胞抗原-DR(HLA-DR)的表达。方法 经支气管肺泡灌洗获肺泡巨噬细胞，贴壁分离及培养，免疫组化方法检测HLA-DR表达的阳性细胞百分率。结果 ① 肺癌及良性肺病患者，其肺泡巨噬细胞未经刺激，均已有部分表达HLA-DR，但前者明显高于后者；② 干扰素-α(IFN-α)或脂多糖(LPS)刺激后，HLA-DR的表达均增加，且两者联合刺激优于单独刺激；③ 无论刺激与否，肺癌组荷瘤侧肺与非荷瘤侧肺比较，肺泡巨噬细胞表达HLA-DR的阳性细胞率明显增高。结论 肺癌患者，尤其是肿瘤局部，肺泡巨噬细胞可能通过表达HLA-DR提高抗原递呈功能。IFN-α、LPS均能明显增强肺癌患者肺泡巨噬细胞的抗原提呈功能，且两者有协同作用。     

肺癌患者肺泡巨噬细胞表现HLA—DR的表达

目的 了解肺癌患肺泡巨噬细胞表面人白细胞原－ＤＲ（ＨＬＡ－ＤＲ）的表达。方法 经支气管肺泡灌洗获肺泡巨噬细胞,贴壁分离及培养,免疫组化方法检测ＨＬＡ－ＤＲ表达的阳性细胞百分率。结果 ①肺癌及良性肺病患,其肺泡巨噬细胞未经刺激,均忆有部分表达ＨＬＡ－ＤＲ,但前明显高于后;②干扰素α（ＩＦＮ－α）或脂多糖（ＬＰＳ）刺激后,ＨＬＡ－ＤＲ的表达均增加,且两联合刺激优于单独刺激;③无论刺激与否,肺     

肺癌患者肺泡巨噬细胞精氨酸酶活性研究

检测肺泡巨噬细胞培养液中中精氨酸活性。结果表明，肺癌患者荷瘤侧肺ＡＭ较非荷瘤侧肺和非癌性肺癌患者ＡＭ产生精氨酸活性低；ＡＭ经卡介菌，干扰素－α刺激后产生精氨酸活性明显增强。     

肺癌患者肺泡巨噬细胞上HLA—Ⅱ类抗原的表达

通过检测肺癌患者肺泡巨噬细胞上人主要组织相容抗原Ⅱ类原的表达来抗究肺癌者局部的免疫情况。经支气管肺泡灌洗获肺癌患者病变累及处及对照组的ＡＭ。     

温石棉对兔肺泡巨噬细胞合成一氧化氮及抗氧化酶活性的影响

为探讨一氧化氮及抗氧化酶在石棉致病中的作用,用改良Ｍｙｒｖｉｋ法收集兔肺泡巨噬细胞（ＡＭ）进行体外培养,采用硝酸还原酶法测定ＵＩＣＣ温石棉处理后兔ＡＭ培养液中亚硝酸根（ＮＯ＾－２）的含量（ＮＯ的静态氧化形式）,同时利用黄嘌呤及黄嘌呤氧化酶反应系统测定ＡＭ的细胞裂解液中ＳＯＤ的活性,和酶－底物反应形成有色物质来测定ＡＭ的细胞裂解液中ＮＯＳ和ＧＳＨ－Ｐｘ的酶活性。     

硝酸甘油、地塞米松对哮喘肺泡巨噬细胞源性一氧化氮、内皮素的影响

研究哮喘患者肺泡巨噬细胞(AM)源性一氧化氮(NO)、内皮素(ET)的变化及硝酸甘油(NTC)、地塞米松(DXM)对两者的影响及机制。对15例轻、中度过敏性支气管哮喘发作期患者的AM(分为未干预组、DXM干预组、NTG干预组),7名健康自愿受试者的AM(未干预组)培养48h,用镀铜镉还原法、放射免疫法和原位杂交法分别测定AM培养上清液中NO,ET水平和iNOS-mRNA,ET-mRNA的表达。结果发现,哮喘AM iNOS-mRNA,ET-mRNA表达增强,分别导致NO,ET水平升高;NTG以直接作用的方式促进AM源性NO的产生,反馈抑制iNOS-mRNA表达并明显抑制ET mRNA的表达,降低ET的水平;DXM降低哮喘AM源性NO,ET水平及iNOS-mRNA,ET-mRNA的表达,尤以抑制iNOS-mRNA表达和降低NO水平为甚,使NO,ET处于低水平的异常状态。     

Immunological status of alveolar macrophages in patients with lung cancer and benign pulmonary diseases]

This is a report of the study on the immunological status of alveolar macrophages (aM phi) in patients with lung cancer (LC, n = 27) and benign pulmonary diseases (BD, n = 26). Patients were undergone bronchoalveolar lavage by fiberoptic bronchoscopy. aM phi in the lavage fluid isolated by adherence on plastic surface were examined in vitro for their cytostatic and cytolytic activities against tumor target cells, secretion of interleukin 1 (IL-1) and tumor necrosis factor (TNF), intracellular IL-1 activity and mRNA expression of IL-1 beta and TNF-alpha. aM phi, both non-activated and activated, were shown to be highly cytostatic against P815 cells by 3H-TdR post-labelling assay. There was no statistical difference between the LC and BD group. As shown by isotope release assay, regardless of being activated or not, aM phi were not cytolytic against P815 and NS-1 cells in both groups of patients. TNF activity could be demonstrated in the culture supernatants of aM stimulated with LPS. Statistically, the TNF activity was not different in the two groups of patients. Spontaneous release of TNF activity was occasionally detected in unstimulated aM phi. While both intracellular and extracellular IL-1 activity of unstimulated aM phi was demonstrated in the two patient groups, the former activity was 1 to 5 times as high as the latter. When stimulated with LPS, there was some increase in extracellular but not intracellular IL-1 activity. mRNA expression of IL-1 beta and TNF-alpha by dot blot hybridization was demonstrable in aM phi from both patient groups irrespective of activation. These results indicate that the immune status of aM phi in lung cancer patients examined does not differ from that in patients with benign pulmonary diseases.     

硝酸甘油、地塞米松对哮喘肺泡巨噬细胞源性一氧化氮、内皮素的影响

研究哮喘患者肺泡巨噬细胞 (AM)源性一氧化氮 (NO)、内皮素 (ET)的变化及硝酸甘油 (NTG)、地塞米松(DXM)对两者的影响及机制。对 15例轻、中度过敏性支气管哮喘发作期患者的AM(分为未干预组、DXM干预组、NTG干预组 ) ,7名健康自愿受试者的AM(未干预组 )培养 48h ,用镀铜镉还原法、放射免疫法和原位杂交法分别测定AM培养上清液中NO ,ET水平和iNOS mRNA ,ET mRNA的表达。结果发现 ,哮喘AMiNOS mRNA ,ET mRNA表达增强 ,分别导致NO ,ET水平升高 ;NTG以直接作用的方式促进AM源性NO的产生 ,反馈抑制iNOS mRNA表达并明显抑制ETmRNA的表达 ,降低ET的水平 ;DXM降低哮喘AM源性NO ,ET水平及iNOS mRNA ,ET mRNA的表达 ,尤以抑制iNOS mRNA表达和降低NO水平为甚 ,使NO ,ET处于低水平的异常状态。     

肺癌患者血浆游离性DNA的研究

目的：研究（1）肺癌患者血浆游离DNA的肿瘤特异性；（2）肺癌患者血浆游离DNA3p部位微卫星不稳定（microsat-ellite instability,MSI）和杂合性缺失（loss of heteroxygosity,LOH）在肺癌诊断、监测中的意义。方法：本研究以37例肺癌术后新鲜标本、94例肺癌患者的全血标本作为研究对象；15例健康正常人和15例结核病人的全血标本作为对照。通过PCR－银染法检测血浆游离DNA3p部位3个微卫星位点的LOH、MSI情况；并和组织标本结果进行对比。结果：（1）大多数肺癌患者的血浆游离DNA的浓度在微克水平以上，而15例正常对照和15例结核病人多在微克水平以下；（2）联合3个位点进行LOH、MSI检测时，组织和血浆的阳性率均在70％以上，一致性在80％以上；而对照组未检测到有LOH、MSI；（3）不同分期、不同病理类型的LOH、MSI之间差异无显著性意义；（4）有淋巴结转移与无淋巴结转移之间差异有显著性意义。结论：血浆游离DNA的未可微克含量水平具有一定的诊断意义，其3p部位的微卫星检测在肺癌的诊断中具有较高的敏感性和特异性，并和组织标本有较高的一致性变化。肺癌患者血浆游离DNA可以作为基因检测的新途径，具有广阔的应用前景。     

健择对非小细胞肺癌患者肺泡巨噬细胞免疫功能影响的研究

目的探讨健择对非小细胞肺癌患者肺泡巨噬细胞(AM)凋亡及其分泌细胞毒效应分子功能的影响。方法ELISA法、酶法分别检测35例非小细胞肺癌患者支气管肺泡灌洗液中及其AM培养上清夜中TNF-α和NO水平,并分别用IFN-α、健择和两药联合干预AM培养,检测其培养上清液中TNF-α、NO变化水平。流式细胞仪检测健择对AM凋亡的影响。结果所有非小细胞肺癌患者其肺泡巨噬细胞均可自发地产生一定量的TNF-α和NO。非小细胞肺癌患者荷瘤侧肺NO和TNF-α活性无论是整体状态,还是细胞水平均明显低于非荷瘤侧肺。健择单独干预组肺泡巨噬细胞培养上清液中TNF-α和NO的浓度无明显变化(P>0.05);经IFN-α刺激肺泡巨噬细胞培养上清液中TNF-α和NO的浓度明显升高,与对照组比较差异呈显著性(P<0.01);健择和IFN-α联合干预下肺泡巨噬细胞培养上清液中TNF-α和NO水平较对照组、IFN-α干预组明显升高,差异呈显著性(P<0.05)。不同时间不同浓度健择对肺泡AM凋亡率无明显差异(P>0.05)。结论健择单独应用对肺泡巨噬细胞免疫功能无明显影响,联合干扰素时能提高非小细胞肺癌患者肺泡巨噬细胞抗肿瘤免疫功能。其机制有待进一步研究。     

肺癌患者与吸烟及非吸烟者支气管肺泡灌洗液T淋巴细胞...

The measurements of T-lymphocyte subsets (as expressed by, CD3 CD4 and CD8) in the peripheral blood and bronchial alveolar lavage fluid (BALF) on 30 cases of lung cancer, 26 cases of smoker and 25 cases of nonsmoker have been done. The results showed that there existed no differences in the peripheral blood of the above said subsets of lymphocyte. In BALF the percentage of CD3 in all the lymphocytes did not show any significant difference among the three tested groups, either (P > 0.05). But the percentage of CD4 and the ratio of CD4/CD8 manifested the following peculiarity: patients with lung cancers < smokers < nonsmokers (P < 0.01). On the contrary, the percentage of CD8 showed just the opposite: patients with lung cancer > smokers nonsmokers (P < 0.01). It was suggested that the cellular immunity as shown in the peripheral blood did not correspond with the findings in the BALF. The cellular immunity of the lungs was decreased both in patients with lung cancer and smokers, but more severely in the group of lung cancer patients. Smokers who have had changes of above said subsets of lymphocytes in the lungs may develop lung cancer. As the lung cancer advances, it may suppress body immunity and in turn enhance the cancer growth. Both the cause and the effect are interrelated. Therefore, the provocative agents for cellular immunity may act as one of the adjunctive therapies for patients with lung cancer. Giving up smoking should play an important role in recovering or promoting the immunity of the lungs and decreasing the incidence as well as improving the prognosis in patients with lung cancer.     

肺癌患者血清及癌组织中NO和NOS变化及意义

目的：测定肺癌患血清及癌组织中的一氧化氮（NO）,一氧化氮合酶（NOS）,了解肺癌患与其体内NO及NOS之间做出评价。方法：对57例肺癌患及40例健康献血,清晨空腹抽取静脉血,对46例开胸手术的肺癌患切取肿瘤组织及正常肺组织,采用硝酸原原酶法测定NO,NOS含量并进行比较,结果：1,肺癌患血清NO含量较正常对照组为低（P＜0．05）,NOS较对照组明显偏低（P＜0．01）,2,不同病理类型肺癌患的肿瘤组织中NO,NOS均较对照肺组织为高（P＜0．05）,3不同分化程度的非小细胞肺癌患中,其高,低分化肿瘤组织中的NO及高,迥,低分化肿瘤组织中的NOS,较其对照肺组织为高（P＜0．05）,4．不同临床分期的肺癌患,其,I,Ⅲ期肺癌肿瘤组织中NO及Ⅰ,Ⅱ,Ⅲ期肿瘤组织中的NOS,较对照肺组织明显增加（P＜0．05）,结论：肺癌患血清中NO,NOS含量较正常人为低,而较多存在于肺癌患肿瘤组织中,提示NO,NOS在肺癌的代谢及病理过程中,可能起重要作用。     

一氧化氮调节脂多糖致炎大鼠肺泡巨噬细胞糖皮质激素受体功能的研究

目的研究肺泡巨噬细胞在脂多糖(LPS)作用条件下一氧化氮供体(SNP)对糖皮质激素受体(GR)功能的调节作用.方法将GR荧光表达质粒pGFP-GR转染大鼠肺泡巨噬细胞,经LPS和SNP作用后,荧光显微镜观察质粒表达产物GFP-GR核移位情况;相对荧光素酶法检测GR转录激活活性;EMSA检测检测细胞NF-κB活性.结果 500 μmol/L SNP作用2 h出现GFP-GR核移位,同时GR转录激活活性显著增强,NF-κB活性被显著抑制.运用GR特异性拮抗剂RU486后,NF-κB活性抑制现象消失.结论肺泡巨噬细胞在致炎因子作用条件下,一氧化氮供体(500 μmol/L SNP )通过活化GR发挥抗炎活性.