【作者】 李恂；

【导师】 姜又红；

【作者基本信息】 中国医科大学 ， 流行病与卫生统计学， 2003， 硕士

【摘要】 前言 恶性肿瘤及肺结核均是目前严重危胁人类健康的疾病，两者的发生均与细胞免疫功能低下有关。大量研究表明T淋巴细胞在机体抗肿瘤、抗结核免疫应答过程中，处于关键地位。T淋巴细胞是胸腺依赖性淋巴细胞(thymus dependent lymphocyte)的简称。在机体的细胞免疫和体液免疫诱导中均有重要作用。 抗肿瘤免疫效应一般以细胞免疫为主，当机体免疫功能下降或瘤细胞的发生超出免疫监视和免疫清除时，就可能形成肿瘤；另一方面，肿瘤病人血清中出现的抑制性细胞因子使T淋巴细胞核中核糖体脱氧核糖核酸rDNA的转录活性下降，进而抑制了T淋巴细胞蛋白质的合成，即抑制了机体的免疫功能。 结核病主要是细胞免疫，即T淋巴细胞在抗原作用下产生的一种特异性免疫反应。结核菌属于细胞内寄生菌，当结核杆菌进入机体后产生细胞介导的免疫反应。对于结核菌感染个体的归宿，机体的免疫功能起着不可忽视的作用。 选择科学、有效的检测肿瘤和结核患者免疫功能状态的方法和指标，一直是广大学者十分关注的问题。通过查阅大量的文献和对比分析，我们拟采用KL型免疫分析系统对肿瘤和结核患者的免疫功能状态进行分析。因为，该系统是利用先进的图像分析技术定量分析T淋巴细胞内核仁形成区rDNA转录活性的变化，它集现代肿瘤学、免疫学、分子生物学和计算机图像分析技术于一体，从基因转录水平上定量分析肿瘤和结核病人外周血T淋巴细胞核仁形成区嗜银蛋白含量(Ag-NORs Argyrophilic nucleolar organizer region proteins)，以银染面积与细胞核面积比值(I.S％)作为Ag-NORs检测指标，反映T淋巴细胞核内rDNA的转录活性，从而间接反映肿瘤和结核患者的免疫功能状态。 为此，本研究利用该系统，通过对比分析正常人J瘤和肺结核患者外周血中T淋巴细胞核仁形成区嗜银蛋白（As－NORs）的含量是否有差异，以评价Ag－NORS检测对于鉴别正常人J瘤患者、肺结核患者的诊断及鉴别诊断是否有意义。以期为该系统在对肿瘤和结核患者的辅助诊断及预后的评估方面提供科学的依据。 材料与方 法 一、研究对象 1．肿瘤患者：来源于中国医科大学附属第一、第h临床医院，经组织病理学确诊的尚未接受放疗、化疗的恶性肿瘤患者，27例，乳腺癌：6例，肺癌：7例，胃癌：7例，其它肿瘤：7例；男性18例，女性9例；年龄范围：22－75岁，平均年龄50．7岁。 2．肺结核患者：来源于沈阳市胸科医院，根据临床表现、胸部X线片、痰结核分支杆菌检查确诊的无合并症初治肺结核患者，27例；痰菌阳性6例，阴性21例，男性17例，女性10例10；年龄范围：17－80岁，平均年龄 49．8岁。 3．正常人：来源于沈阳市疾病预防控制中心，健康体检者，健康无任何疾病，27例，男。四7例，女性10例；年龄范围：18－78岁，平均年龄 49．6岁。 二、实验方法 采用KL型免疫分析系统，检测肿瘤和结核病人外周血T淋巴细胞核仁形成区嗜银蛋白含量（Ag－NORSX以银染面积与细胞核面积比值门．S％）作为Ag－NORS检测指标。具体步骤包括： 1．细胞培养 2．细胞分离 ·二· 3．制片 4．银染 5．上机进行读片分析 三、统计学分析 采用 SPSS 11．5统计软件包，对各种数据迸行统计学描述，方差分析、两两比较 q检验及 X＼检验。 结 果 二．肿瘤患者和肺结核患者的1．S％值均明显低于正常人瞩＜0．01）。 2．肿瘤与肺结核患者相比1．S％值差异有统计学意义，肿瘤患者的1．3％值显著低于肺结核患者汀＜O．05人 3．不同类型的肿瘤患者之间1．6呢值无明显差异（P＞O．05八 4．不同类型的肺结核患者之间1．S％值亦无显著性差异（P＞0．05）。 讨 论 免疫系统是机体防御肿瘤的主要防线，在致突变因素作用下，机体每天约有＊＼‘细胞发生基因突变，部分细胞可转化为恶性表型的瘤细胞，正常的免疫监视功能可即时识别并清除这些细胞，而不致发生肿瘤，恶性肿瘤多发生在机体兔功能低下或相对低下时。T淋巴细胞是机体参与抗肿瘤疫免应答的主要免疫活性细胞，因此可通过监测病人T淋巴细胞功能，来反映肿瘤的存在和发展状态。 结核感染常伴细胞免疫功能的低下或缺陷，结核免疫学研究表明T淋巴细胞在结核病的免疫中起着重要作用，认为结核病的 ·3·发生与T淋巴细胞的功能状态呈负相关。 细胞核仁组成区相关蛋白是机体主要细胞形成区的相关蛋白瞩ORSX是一种酸性非组蛋白，它是调控rDNA转录活性的重要指标。用核仁组成区银染技术可清晰显示NORS的数量、大小和强度，它反映了核糖体蛋白基因卜DNA）转录活性及蛋白合成状态，也反映了该细胞的增殖活性。因此测定T淋巴细胞核仁区Ag－NORS的表达活性可知 T淋巴细胞的免疫活性，从而间接了解整个机体的免疫功能，即在理更多还原

【Abstract】 PrefaceThe tumor and tuberculosis is the two kinds of disease that threats human health. Both of them happens relate with the immunity lowli-head. Many studies suggested that T lymph cell plays an important role in immune responsion. T lymph cell what we always call thymus dependent lymphocyte, plays an important role on body cellular and humoral immune induction.The main effect of anti - carcinoma is cellular immune. Many studies showed T lymph cell is the key during body immune reaction. Tumor may form if body immune function decrease or immune monitor and clearance cant find it out. On the other hand , the transcription activity of T lymph cells rDNA may decrease, caused by inhibiting cy-tokin in the serum of patients with tumor. The result is T lymph cell protein synthesis and body immune function inhibited.The major immune reaction to tuberculosis is cellular immune. It is a specific immune reaction of T lymph cell by the effect of antigen. Tuberculosis, a kind of intra - cellular parasite, may cause cell - induced immune reaction when it invade the body. Then, whether the infected body is sick depends a lot on this bodyfc immune function.We used KL - immune analysis system, which has advanced image analysis, to analyze the development of rDNA transcription activi-ty, at the area nucleus formed. This system synthesizes modern oncology, immunology, cytobiology and computer image analysis technique. Quantitive analyzed Ag - NORs in T lymph cell of the patients with malignant tumor and pulmonary tuberculosis, which reflect the rDNA transcription activity. The index is the ratio between the surface areas of silver - dyed and the nucleus.The purpose of this research is to find out whether the index has difference between the normal person and the patients with tumor and pulmonary tuberculosis, and its value in the assistant diagnosis.Objects and Methods1. Objects( 1) Patients with TumorThey were the patients of the 1st and 2nd affiliated hospital of China Medical University. All the 27 patients were definitely diagnosed of malignant tumor in histology and had not got radiation therapy or chemical therapy.(2) Patients with Pulmonary TuberculosisAll the 27 patients were chosen from Chest Hospital of Shenyang. They were definitely diagnosed by X - ray and sputum smear.(3) Healthy peopleThe 27 healthy people come from Center for Disease Control And Prevention of Shenyang. The patients and those in control group match with one other in age ( 5 years) , sex and race.2. Epidemiological InvestigationWe investigated those whose blood samples were gathered in the morning by filling up well - designed tables looking up medical recordand patient reviewing.3. Experiment Methods ( 1) cells culture(2) cells isolation ( 3 ) making slices(4) Ag - straining(5) Analysis by computer4. Statistical AnalysisStatistical soft we used is SPSS11.5. All the data were indicated by XS. Comparation of data among groups is made by q - test, x2- a~ nalysis.ResultsThe value of I. S% is much lower in patients with tumor and tuberculosis than in health people. And obvious differences were found between them (P < 0.001).There were obvious differences between I. S% of tumor patients and those of tuberculosis patients ( P <0.05).No differences were found in both different types of tumor ( P > 0.05 ) and different type of tuberculosis ( P > 0. 05 ) .DiscussionImmune system is mainly line of defence of anti - carcinoma in body. On the effect of cause mutation factor, There are about 107 ~ 8cells happened gene saltation everyday. Some cells can translate into Tumor cell. Normal immune monitor function can distinguish and clearthese cells. Tumor can’t happens. The malignancy often occurs in body immune function decrease or comparatively decrease. T lymph cell is the mainly cell in the body that participate anti - carcinoma immune responsory. So, though monitor the patients function of T lymph cell to reflect the positive and development statue o更多还原