IL-18基因修饰增强肿瘤抗原多肽致敏的树突状细胞体内诱导的抗肿瘤免疫反应

目的 :研究肿瘤抗原多肽致敏的白细胞介素 18(IL 18)基因修饰的树突状细胞体内诱导的抗肿瘤免疫反应。方法 :①以Lewis 3LL肺癌细胞特异性抗原肽mut1冲击致敏IL 18基因修饰的骨髓来源的树突状细胞 (DC IL 18 mut1) ,每次用其 1× 10 5 只皮下免疫小鼠 2次 ,然后测定脾细胞的NK活性及CTL杀伤活性 ;②以DC IL 18 mut1每次 2× 10 5 只皮下免疫 1次 ,然后再以 5× 10 53LL细胞攻击 ,在诱导及效应阶段分别以单抗阻断不同免疫成份 ,观察肿瘤的生长。结果 :以DC IL 18 mut1皮下免疫后可诱导出比DC mut1等免疫组更高水平的 3LL肺癌细胞特异性CTL ,并使NK活性明显增加 ;单抗体内阻断实验提示在DC IL 18 mut1免疫诱导阶段 ,CD4 + T细胞和抗原共刺激分子、IFN γ均起到重要作用 ,而效应阶段CD8+ T、IFN γ、NK起作用 ,而CD4 + T则是非必需的。结论 :DC IL 18 mut1皮下免疫后可诱导高水平的抗肿瘤免疫活性 ,其机理与抗原有效提呈、特异性CTL诱导、NK活性增加以及CD4 + 、CD8+ T、NK细胞、IFN γ参与密切相关。     

探讨BCG初次免疫结核杆菌DNA疫苗加强免疫方案的效应

目的:探讨BCG初次免疫(BCG-prime),结核杆菌共表达DNA疫苗加强免疫(DNA疫苗-boost)的策略对小鼠的免疫效果。方法:将BCG及结核杆菌重组DNA疫苗依次免疫小鼠,通过检测CTL和NK细胞的杀伤活性和特异性淋巴细胞增殖,以及小鼠血清抗体及细胞因子的水平,观测BCG-prime、共表达结核杆菌Ag85A/GM-CSFDNA疫苗boost策略对小鼠的免疫效果。结果:采用prime-boost免疫策略组的小鼠CTL的杀伤活性明显增强、特异性淋巴细胞明显增殖、IFN-γ的水平明显增高,NK细胞杀伤活性与对照组相比也有一定提高,但未超过BCG单独免疫效果。免疫小鼠血清特异性抗体的滴度超过单独DNA疫苗免疫组。结论:在采用BCG-prime-结核杆菌DNA疫苗boost免疫策略后,能增强对小鼠的免疫效应,尤其是Th1型细胞免疫反应增强明显,为进一步在动物体内进行保护性效应试验的研究提供了实验依据。     

低氧和高二氧化碳作用下肺动脉高压形成过程中骨桥蛋白表达与分布的实验研究

目的： 观察慢性低O2高CO2大鼠肺动脉骨桥蛋白（OPN）及其基因的表达与分布情况，探讨OPN在肺动脉高压发病机制中的作用。方法： 48只雄性SD大鼠随机分为4组：即正常对照组（NC组），低O2高CO2 1周、2周和4周组（1HH、2HH和4HH）。采用RT-PCR法测定不同低O2高CO2 组大鼠离体肺动脉和肺组织骨桥蛋白（OPN）mRNA，免疫组织化学染色法测定肺细小动脉骨桥蛋白分布表达情况，ELISA 法定量测定肺组织匀浆OPN水平，Western blotting 法测定离体肺动脉OPN表达水平。结果： ①低O2高CO2各组大鼠mPAP和RV/LV+S均明显高于NC组（P<0.01），4组大鼠间mCAP比较无显著差异（P>0.05）；②低O2高CO2 各组大鼠肺组织和离体肺主动脉OPN mRNA表达水平显著高于对照组（均P<0.01）；③免疫组化法显示NC组大鼠肺组织OPN表达主要见于支气管、肺泡上皮细胞内，1HH组大鼠肺细小动脉内皮细胞、平滑肌细胞以及周围巨噬细胞均见有OPN的表达，其中以中膜平滑肌细胞最为明显，而2HH和4HH组大鼠OPN表达更加明显, 低O2高CO2 各组肺细小动脉OPN相对含量与对照组比较有显著差异（均P<0.01），且随缺氧时间的延长，骨桥蛋白表达也呈增高趋势；④ 低O2高CO2 各组大鼠肺组织匀浆OPN含量分别为正常对照组的1.69倍、2.28倍和2.87倍（均P<0.01）；⑤ Western blotting 分析1HH、2HH和4HH各组大鼠离体肺动脉OPN表达明显增强，与NC组比较有显著差异（均P<0.01）。结论： 慢性低O2高CO2能够刺激大鼠肺组织和肺动脉OPN及其基因表达增强，OPN可能在慢性低O2高CO2性肺动脉高压发病的病理和病理生理过程中起着重要的作用。     

CFTR 5T variant has a low penetrance in females that is partially attributable to its haplotype.

PURPOSE: The study's purpose was to understand the molecular basis for different clinical phenotypes of the 5T variant, a tract of 5 thymidines in intron 8 of the cystic fibrosis transmembrane conductance regulator (CFTR) gene, which disrupts processing of CFTR mRNA and reduces synthesis from the corresponding CFTR alleles. METHOD: We analyzed the polymorphic TG dinucleotide repeat adjacent to the 5T variant in intron 8 and the codon 470 in exon 10. Patients selected for this study were positive for both the 5T variant and the major cystic fibrosis mutation, Delta F508. Almost all Delta F508 mutation alleles occur in a 10TG-9T-470M haplotype. Therefore, it is possible to determine the haplotype of the 5T variant in trans. RESULTS: Of the 74 samples analyzed, 41 (55%) were 11TG-5T-470M, 31 (42%) were 12TG-5T-470V, and 2 (3%) were 13TG-5T-470M. Of the 49 cases for which we had clinical information, 17.6% of females (6/34) and 66.7% of males (10/15) showed symptoms resembling atypical cystic fibrosis. The haplotype with the highest penetrance in females (42% or 5/12) and more than 80% (5/6) in males is 12TG-5T-470V. We also evaluated 12 males affected with congenital bilateral absence of vas deferens and positive for the 5T variant; 10 of 12 had the 12TG-5T-470V haplotype. CONCLUSION: Overall, the 5T variant has a milder clinical consequence than previously estimated in females. The clinical presentations of the 5T variant are associated with the 5T-12TG-470M haplotype.     

Aberrations of growth factor control in metastatic follicular thyroid cancerin vitro

The aggressiveness of follicular thyroid cancer (FTC) varies widely, and metastasis is the primary cause of death. Uncontrolled proliferation of cancer cells may be associated with loss of growth factor control. We investigated the effects of stimulating (epidermal growth factor [EGF]; thyreotropin [TSH] in low concentrations) and inhibiting growth factors (transforming growth factor beta 1 [TGF beta 1]; TSH in high concentrations) on invasion and growth of FTC cell lines from the thyroid tumor (FTC133) and from the lymph node (FTC236) and lung (FTC238) metastases of the same patient. Invasion—penetration through an 8m pore membrane, covered by Matrigel (basement membrane)—and growth were measured using the MTT-method. EGF (10 ng/ml) and TSH in low concentrations (1 mU/ml) stimulated invasion and growth of all FTC cell lines, but the amplitude of stimulation differed significantly. The parental cell line FTC133 was considerably more responsive to growth factor stimulation than the metastatic clones. Invasion of FTC133 was enhanced by 42% (EGF;p<0.02) and 21% (TSH;p<0.01), invasion of FTC236 by 8% (EGF;p<0.02) and 8% (TSH;p<0.01), and invasion of FTC238 by 9% (EGF;p<0.02) and 8% (TSH;p<0.01). Conversely, invasion and growth of FTC133 were significantly more inhibited by TGF beta 1 (10 ng/ml) and supraphysiologic concentrations of TSH (100 mU/ml) than the cell lines from the lymph node and lung metastases. At day 7, invasion of FTC133 was inhibited by 32% (TGF beta 1;p<0.02) and 21% (TSH;p<0.01), invasion of FTC236 by 18% (TGF beta 1;p<0.02) and 11% (TSH;p<0.01), and invasion of FTC238 by 16% (TGF beta 1;p<0.02) and 12% (TSH;p<0.01). Moreover, we analyzed growth factor independence in minimally supplemented or unsupplemented medium. Growth, but no invasion was evident when cells were cultured completely unsupplemented over 7 days. These results suggest that metastatic FTCs may have developed by escaping from the normal control of TSH and other growth factors.     

幽门螺杆菌疫苗免疫机制的研究进展

幽门螺杆菌(H.pylori)是定植于人体胃粘膜的重要致病菌,而粘膜疫苗以其有效的保护性免疫成为防治H.pyiori的热点,但对相应的免疫机制的认识目前尚不明确.故本文对H.pyiori疫苗的保护性免疫机制中的抗原递呈机制,抗体的作用以及TH细胞反应的作用等研究进展做一概要介绍.     

男性抗精子抗体对体外受精一胚胎移植的影响

目的;评价男性抗精子抗体对体外受精-胚胎移植(IVF-ET)的影响。方法:以ELISA方法测定施行IVF-ET的150对不育夫妇中男方精浆抗精子抗体(AsAb)并分组:AsAb阳性组22例,AsAb阴性组128例。对比分析抗精子抗体及抗体类型对体外受精、卵裂和早期胚胎发育的影响,同时对移植后的妊娠率和流产率进行分析。结果:AsAb阳性组体外受精数明显低于AsAb阴性组(P<0.01),两组的卵裂数则无明显差异(P>0.05),而在阳性组中,不同抗体类型间体外受精、卵裂及胚胎质量无明显差异(P>0.05)。在移植日,AsAb阳性组的I级胚胎较阴性组低,而ⅢⅣ级胚胎和2原核受精细胞(2PN)明显增加(P<0.01)。两组移植后临床妊娠流产率无显著差异(P>0.05)。结论:AsAb能明显降低精子体外受精能力,对早期胚胎发育有损害。精子体处理不能完全清除AsAb对IVF影响。     

精神分裂症与KCNN3基因1137～1140缺失移码突变传递不平衡研究

目的 探讨KCNN3基因1137～1140的4个碱基缺失所致移码突变与精神分裂症的关系。方法 95个核心家系共289名家庭成员，包含107例精神分裂症患者纳入本研究。精神分裂症采用CCMD-Ⅱ-R诊断标准。KCNN3基因1137～1140的4个碱基缺失基因型检测使用PCR技术和限制性内切酶DdeI消化方法。精神分裂症与KCNN3基因1137～1140缺失4个碱基的关联分析采用基于单倍型的单倍型相对风险率和传递／不平衡检验。结果 患者组与正常父母组比较，KCNN3基因1137～1140的4个碱基缺失的基因型频数和等位基因频率分布差异无统计学意义（χ^2=0．253，P〉0．05和χ^2＝0．010，P〉0．05）。基于单倍型的单倍型相对风险分析发现父母传递与不传递给患者的KCNN3基因等位基因之间差异无统计学意义（χ^2=0．042，P〉0．05）。传递不平衡检验结果发现KCNN3基因等位基因传递不存在连锁不平衡（χ^2=3．000，P＝0．0833）。结论 本组研究对象中，KCNN3基因第1外显子1137～1140的4个碱基缺失的发生率少；分析结果提示KCNN3基因第1外显子1137～1140的4个碱基缺失的等位基因与精神分裂症无关联。     

Comparison of nasal pressure transducer and thermistor for detection of respiratory events during polysomnography in children

STUDY OBJECTIVES: The results of small studies have suggested that a nasal-cannula pressure transducer has a higher sensitivity than a thermistor in detecting hypopneas and diagnosing sleep-disordered breathing in both adults and children. We compared a thermistor alone, and in conjunction with a pressure transducer, for detection of sleep-disordered breathing in children during in-home polysomnography. DESIGN: Retrospective analysis of a subsample of a prospective cohort study. SETTING: Students attending elementary school in the Tucson Unified School District. PARTICIPANTS: A subsample of the Tucson Children's Assessment of Sleep Apnea study population. MEASUREMENTS AND RESULTS: Polysomnographic recordings of 40 children (24 girls and 16 boys, mean age 9.2 +/- 1.7 years; range 6-11 years) were analyzed to compare the detection of sleep-disordered breathing events by 2 different methods of measuring airflow: thermistor alone and thermistor with nasal-cannula pressure transducer (transducer) used simultaneously. The transducer detected all the respiratory events detected by the thermistor, but the thermistor detected only 84% of the transducer-defined events. Consequently, the transducer-derived mean respiratory disturbance index was higher than that detected by the thermistor (7.0 +/- 3.8 vs 5.9 +/- 3.4, P < .001). The bias error between transducer respiratory disturbance index and thermistor respiratory disturbance index on a Bland-Altman plot was 1.08 (95% confidence interval, 0.8 - 1.4). There was good agreement between the thermistor and the transducer for making the diagnosis of sleep apnea using a cutoff of a respiratory disturbance index greater than 5 (kappa = 0.69). The quality of the tracings with the transducer was comparable to that of the thermistor, but the transducer dislodged more frequently. CONCLUSION: The use of a nasal transducer in conjunction with a thermistor was more sensitive than the thermistor alone in detecting sleep-disordered breathing in children during unattended polysomnography.