长链非编码RNA肺腺癌转移相关转录本1对巨噬细胞极化影响的研究

目的研究长链非编码RNA(lnc RNA)肺腺癌转移相关转录本1(MALAT1)对巨噬细胞极化的影响。方法采用佛波酯诱导人THP-1细胞分化为成熟的巨噬细胞,IFN-γ/LPS刺激诱导M1型极化,IL-4刺激诱导M2型极化,采用实时荧光定量PCR(RT-q PCR)检测各组细胞中MALAT1的表达。si RNA干扰巨噬细胞MALAT1表达,加入IL-4继续诱导M2型极化,RT-q PCR检测极化相关基因表达;酶联免疫吸附试验(ELISA)检测TNF-α、IL-12、CCL22、IL-10的水平。si RNA干扰M2型巨噬细胞MALAT1表达,研究其对M2极化表型的影响。结果 MALAT1在M2型巨噬细胞中表达显著升高,M1向M2极化改变MALAT1表达升高,而M2向M1极化改变MALAT1表达降低。MALAT1干扰后,IL-4诱导的M2型巨噬细胞TNF-α、IL-12分泌升高,CCL22、IL-10分泌降低;CXCL10、CXCL11、HLA-DR表达升高,CCL17、CCL18、CD163表达降低。M2型巨噬细胞MALAT1干扰后,TNF-α、IL-12分泌升高,CCL22、IL-10分泌降低。结论 MALAT1参与调控巨噬细胞极化,干扰MALAT1表达有效抑制M2型巨噬细胞极化,促进M1型巨噬细胞极化。     

IL-22 对骨髓来源巨噬细胞相关炎症因子表达的影响

目的:探讨IL-22对骨髓来源巨噬细胞相关炎症因子表达的影响。方法:从小鼠的股骨分离骨髓细胞,经过分化培养获得骨髓来源巨噬细胞,再用脂多糖(LPS)刺激诱导为M1型巨噬细胞。IL-22干预M1型巨噬细胞后,采用流式细胞术检测巨噬细胞标志物;用ELISA分别检测IL-1β和TNF-α的分泌水平;用RT-PCR分别检测炎症因子i NOS、IL-1β和TNF-α的mRNA表达水平。结果:经过分化培养,骨髓来源巨噬细胞的纯率为(99. 3±0. 4)%,IL-22干预M1型巨噬细胞后,与LPS组相比,LPS+IL-22组的M1型巨噬细胞比例下降(P0. 05),细胞炎症因子i NOS、IL-1β和TNF-αmRNA表达下降(P0. 05),IL-1β和TNF-α分泌水平降低(P0. 05)。结论:IL-22可抑制LPS诱导的骨髓来源巨噬细胞炎症因子的表达,减轻其炎性反应。     

miR-34a调控的M2型巨噬细胞极化对食管癌细胞迁移和凋亡的影响

目的探究miR-34a在调控M2型巨噬细胞极化中的作用及其对食管癌细胞迁移和凋亡的影响。方法实时荧光定量PCR检测不同分化状态下的巨噬细胞miR-34a的表达水平;在M2型巨噬细胞中转染对照microRNA（miR-NC）和miR-34a mimics,使用荧光定量PCR检测CD163、CD206、CCL18和CCL22 m RNA的表达水平,使用酶联免疫吸附实验检测培养基中IL-10和TGF-β的表达水平;细胞划痕实验检测EC109细胞迁移能力;流式细胞术检测5-氟尿嘧啶（5-FU）诱导的EC109细胞凋亡水平;MTT法检测5-FU的半数抑制计量(IC₅₀)。结果与THP-1和M0型巨噬细胞相比,M2型巨噬细胞极化过程中miR-34a表达水平显著下降;与对照组细胞相比,转染miR-34a mimics的M2巨噬细胞CD163、CD206、CCL18和CCL22 mRNA表达水平显著下降,分泌到培养基中的IL-10和TGF-β含量显著下降。与对照组相比,使用过表达miR-34a的M2巨噬细胞条件培养基培养的食管癌EC109细胞迁移水平显著下降,5-FU诱导的细胞凋...     

免疫性血小板减少症Th1和Th2趋化因子及其受体表达的研究

目的免疫性血小板减少症(immune thrombocytopenia,ITP)是一种获得性自我免疫紊乱导致的出血性疾病,本文旨在研究ITP患者体内T-helper 1(Th1)趋化因子CCL5、CXCL11及其受体CCR5、CXCR3和Th2趋化因子CCL11及其受体CCR3的表达变化,以探讨趋化因子及其受体在ITP免疫异常中的作用。方法选择28名活动性ITP患者为研究对象,随机选取21名与ITP患者相匹配的健康人作对照组。以酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)检测ITP患者和对照组血浆中CCL5、CXCL11和CCL11的含量,采用实时聚合酶链式反应(real-time polymerase chain reaction,RT-PCR)检测外周血单个核细胞(PBMC)CCL5、CXCL11、CCL11、CCR5、CXCR3以及CCR3 mRNA的表达。结果 Th1相关趋化因子CCL5和Th2相关趋化因子CCL11在活动性ITP患者血浆中含量均低于正常对照组(P0.05),而Th1趋化因子CXCL11升高(P0.05);ITP患者PBMC的CXCL11 mRNA表达高于对照组,CCL11 mRNA表达低于对照组(P0.05),而CCL5 mRNA在ITP组与对照组差异无显著性(P0.05)。ITP患者血浆中Th1相关趋化因子受体CCR5和CXCR3表达量高于对照组(P0.05),Th2相关趋化因子受体CCR3降低(P0.05)。经过糖皮质激素等治疗有效的22例患者,Th1趋化因子CXCL11、趋化因子受体CCR5和CXCR3均下降(P0.05),血浆CCL5含量回升,但是仍低于对照组(P0.05);而Th2相关趋化因子CCL11及其受体CCR3均回升(P0.05)。结论 Th1/Th2相关趋化因子及其受体的异常表达可能参与了ITP的免疫紊乱,是ITP发病因素之一;阻断Th1相关趋化因子与其受体的作用途径,可望成为ITP的生物调控靶点。     

Gpnmb在小鼠M1和M2型骨髓源性巨噬细胞中表达的差异

目的了解小鼠M1和M2型骨髓源性巨噬细胞（ BMMφs）中非转移性黑色素瘤糖蛋白b（ Gpnmb）表达的差异。方法原代培养小鼠BMMφs,免疫荧光染色F4/80和流式细胞仪检测CD11b鉴定巨噬细胞;用IFN-γ和LPS诱导BMMφs向M1型分化,用IL-4诱导向M2型分化。实时荧光定量PCR检测M1型巨噬细胞标志物（ TNF-α、iNOS）、M2型巨噬细胞标志物（ MMR、Arg-1）和Gpn-mb的mRNA表达;免疫荧光双染色、Western印迹、流式细胞仪检测Gpnmb与MMR的蛋白表达。结果（1）免疫荧光染色结果示BMMφs中F4/80高表达;流式细胞仪检测结果示BMMφs中有（92．7±6．1）％细胞表达CD11b,提示BMMφs培养成功;（2）相对于未分化的M0型BMMφs,TNF-α、iNOS mRNA在M1型BMMφs中高表达（P均＜0．01）,而MMR、Arg-1 mRNA在M2型BMMφs中高表达（P均＜0．01）,提示原代M1、M2型BMMφs分化成功;（3）M2型BMMφs 的Gpnmb mRNA和蛋白表达均较M0型和M1型BMMφs显著增高（P均＜0．01）;免疫荧光双染色及流式细胞仪结果显示,BMMφs中Gpnmb与MMR共表达,在M2型BMMφs中MMR阳性BMMφs有（83．2±9．7）％表达Gpnmb。结论 M2型BMMφs的Gpnmb表达较M1型BMMφs显著增高,提示Gpnmb可能作为鉴别M1、M2型巨噬细胞的标志物,在巨噬细胞的表型分化中起作用。     

脂肪间充质干细胞促进M1型巨噬细胞向M2型巨噬细胞转化

目的探索脂肪间充质干细胞(ADSC)对M1/M2型巨噬细胞的影响以及ADSC能否促使M1型巨噬细胞向M2型巨噬细胞转化。方法利用脂多糖(LPS)、γ干扰素(IFN-γ)刺激J774.1巨噬细胞24 h诱导为M1型巨噬细胞,利用白细胞介素4(IL-4)刺激J774.1巨噬细胞24 h诱导为M2型巨噬细胞;将M1/M2型巨噬细胞分别与ADSC共培养24 h后,收集巨噬细胞和上清液,用实时定量PCR和ELISA检测巨噬细胞IL-6、肿瘤坏死因子α(TNF-α)、诱导型一氧化氮合酶(i NOS)、CC趋化因子配体2(CCL2)、CD86、精氨酸酶1(Arg1)、甘露糖受体/CD206(MR/CD206)、IL-10、炎症区分子1(found in inflammatory zone 1,FIZZ1)、几丁质酶3样分子3(chitinase 3-like 3,即Ym-1)的变化。结果 ADSC使M1型巨噬细胞分泌的IL-6、TNF-α、i NOS、CCL2、CD86明显减少,Arg1、CD206、IL-10明显增加,且上清液中IL-6和TNF-α含量明显减少,CD206明显增加;使M2型巨噬细胞分泌的IL-6、TNF-α、i NOS、CD86明显减少,Arg1、CD206、FIZZ-1、Ym-1、IL-10明显增加,且上清液中IL-6和TNF-α含量明显减少,CD206明显增加。结论 ADSC抑制M1型巨噬细胞的特异性基因的表达,促进M2型巨噬细胞特异性基因的表达,并使M1型巨噬细胞向M2型巨噬细胞转化。     

活动性结核病人PBMCs中转录因子PLZF的表达降低与iNKT细胞的减少相关

目的:比较活动性结核病人和健康人外周血单个核细胞(PBMCs)中转录因子PLZF(Promyelocytic leukemiazinc finger)的表达和iNKT细胞含量的差异,研究活动性结核病人PBMCs中PLZF的表达与iNKT细胞含量的相关性。方法:采集活动性结核病人和健康人新鲜的抗凝血,淋巴细胞分离液分离PBMCs。每个样本的PBMCs分两份,一份细胞用Trizol法提取总RNA,荧光定量PCR检测PLZF mRNA的相对表达量;另一份细胞用特异性抗体标记iNKT细胞,流式细胞仪检测iNKT细胞的含量。统计学软件分析两组样本PLZF的表达和iNKT细胞含量的差异,以及PLZF的表达与iNKT细胞含量的相关性。结果:活动性结核病人PMBCs中PLZF的表达和iNKT细胞含量均显著低于健康对照(P值分别为0.010 2和0.001 2),PLZF的表达与iNKT细胞的含量成正相关(r=0.480 4,P=0.027 5)。结论:活动性结核病人PLZF的表达降低与iNKT细胞百分含量的减少相关。     

微小RNA-155在小鼠骨髓来源M1和M2巨噬细胞中表达的差异**

背景：目前对微小RNA-155在巨噬细胞中的表达和功能的研究还集中在总体单核巨噬细胞水平。 目的：了解小鼠骨髓来源M1和M2巨噬细胞中微小RNA-155表达的差异。 方法：用100 U/mL干扰素γ和5 μg/L脂多糖诱导骨髓细胞来源的巨噬细胞向M1分化,用10 μg/L白细胞介素4诱导出M2巨噬细胞。 结果与结论：流式细胞检测显示实验诱导的M1和M2巨噬细胞纯度分别达91%和95%。RT-PCR检测显示诱导型一氧化氮合酶mRNA在M1巨噬细胞中高表达,在M2中则基本不表达;而Ⅰ型精氨酸酶、发现于炎症区域1 mRNA在M2中高表达,在M1中则表达较低;炎症因子肿瘤坏死因子α mRNA在M1中的表达明显高于M2,相反白细胞介素10 mRNA在M2中的表达高于M1(P < 0.05)。实时荧光定量PCR检测显示M1巨噬细胞中微小RNA-155的表达量明显高于M2(P < 0.05)。提示微小RNA-155可作为鉴别M1,M2巨噬细胞的标志。     

慢性支气管炎患者肺泡巨噬细胞上CD11c、CD14及胞内TGF-β1 mRNA表达的研究

目的　研究慢性支气管炎患者肺泡巨噬细胞上CD11c和CD14及其胞内TGF β1mRNA的表达情况。方法　对慢性支气管炎缓解期患者和正常对照者各 10例进行局部支气管肺泡灌洗。将支气管肺泡灌洗液中的肺泡巨噬细胞分离、纯化后 ,与藻红蛋白 (phycoerythrin ,PE)和异硫氰酸荧光素 (fluoresceinisothiocyanate,FITC)标记的CD11c和CD14单克隆抗体共同孵育 ,以流式细胞仪分析其CD11c和CD14表达的百分率 ;应用转化生长因子β1(transforminggrowthfactor beta 1,TGF β1)的cDNA探针原位杂交 ,观察肺泡巨噬细胞内TGF β1的表达。结果　 (1)慢性支气管炎组肺泡巨噬细胞上CD11c和CD14表达的百分率 (39.17%± 5 .5 6 % ,35 .73 %± 8.0 5 % )均明显高于正常对照组 (16 .2 9%±3 .78% ,15 .2 6 %± 5 .96 % ) (均P <0 .0 0 1) ;(2 )慢性支气管炎患者肺泡巨噬细胞内TGF β1mRNA的表达量明显高于正常对照者 (P <0 .0 1)。结论　慢性支气管炎缓解期患者肺泡巨噬细胞上CD11c和CD14及其胞内TGF β1mRNA的表达增加 ,上述分子可能共同参与气道内的慢性炎症的病理过程     

整合素β3 mRNA表达与胃癌微血管密度、进展及预后的关系

目的　探讨整合素 β3mRNA在胃癌中的表达及其与肿瘤微血管密度 (MVD)、生长方式、浸润转移和预后的关系。方法　应用原位杂交和免疫组织化学技术 ,检测 10 5例胃癌组织中整合素 β3mRNA和CD3 4的表达。结果　整合素 β3mRNA在非肿瘤胃黏膜中的阳性表达率为 3 0 % (6/ 2 0 ) ,显著低于胃癌组 (61 0 % ,64/ 10 5,χ2 =8 85,P =0 0 3 7) ;在浸润性生长、T3 ～T4 期、有淋巴结转移和远处转移的病例中 ,阳性率分别为 70 2 % (40 / 57)、72 1% (44/ 61)、68 6% (48/ 70 )和 85 7% (3 6/ 42 ) ,均显著高于膨胀性生长 (χ2 =14 97,P =0 0 0 2 )、T1～T2 期 (χ2 =15 2 1,P =0 0 15)、无淋巴结转移 (χ2 =17 89,P =0 0 2 5)及无远处转移的病例 (χ2 =2 0 2 2 ,P =0 0 0 5;χ2 =2 1 3 5,P =0 0 3 5) ;同样 ,它们的平均MVD均显著高于膨胀性生长 (t =10 10 5,P =0 0 0 1)、T1～T2 期 (t=5 961,P =0 0 0 1)、无淋巴结转移 (t= 3 819,P =0 0 1)和无远处转移的病例 (t =10 578,P =0 0 0 1;t =7 882 ,P =0 0 0 1) ;阳性表达组的平均血管数 (41 0 2± 8 55)个 / 0 72mm2 明显高于阴性表达组的平均微血管数 (2 5 2 6± 11 2 5)个 / 0 72mm2 (t =11 2 5,P =0 0 2 5) ,两者之间呈显著正相关 (rs=0 3     

Elevated serum levels of CCL17 correlate with increased peripheral blood platelet count in patients with active tuberculosis in China

The serum levels of Th2 markers, including CCL17 (thymus and activation-regulated chemokine [TARC]), CCL22 (macrophage-derived chemokine [MDC]), and soluble CD30, were measured in 101 HIV-negative tuberculosis patients, 103 healthy community controls, and 18 tuberculosis patients in recovery. The levels of CCL17/TARC (249.8 ± 19.91 versus 143.9 ± 10.54, P < 0.0001) and sCD30 (7.78 ± 0.44 versus 4.93 ± 0.23, P < 0.0001) were significantly higher in patients with active tuberculosis than in controls; however, the CCL22/MDC serum level had no statistical difference between the groups (579.9 ± 16.42 versus 556.5 ± 15.29, P = 0.298). The counts of platelet and eosinophil in the peripheral blood of patients with active tuberculosis are significantly increased as well (289.4 ± 8.14 versus 248.3 ± 5.34 [P < 0.0001] and 165.1 ± 14.33 versus 102.5 ± 10.72 [P = 0.0005], respectively), and the platelet counts were positively correlated with serum TARC levels (Pearson r = 0.456, P < 0.0001), which indicates a new source of Th2 bias showing in active TB patients.     

Progression of clinical tuberculosis is associated with a Th2 immune response signature in combination with elevated levels of SOCS3

In this study, we explored the local cytokine/chemokine profiles in patients with active pulmonary or pleural tuberculosis (TB) using multiplex protein analysis of bronchoalveolar lavage and pleural fluid samples. Despite increased pro-inflammation compared to the uninfected controls; there was no up-regulation of IFN-γ or the T cell chemoattractant CCL5 in the lung of patients with pulmonary TB. Instead, elevated levels of IL-4 and CCL4 were associated with high mycobacteria-specific IgG titres as well as SOCS3 (suppressors of cytokine signaling) mRNA and progression of moderate-to-severe disease. Contrary, IL-4, CCL4 and SOCS3 remained low in patients with extrapulmonary pleural TB, while IFN-γ, CCL5 and SOCS1 were up-regulated. Both SOCS molecules were induced in human macrophages infected with Mycobacterium tuberculosis in vitro. The Th2 immune response signature found in patients with progressive pulmonary TB could result from inappropriate cytokine/chemokine responses and excessive SOCS3 expression that may represent potential targets for clinical TB management.     

乙型肝炎病毒感染者外周血CD4+T细胞CD25、CD127不同亚群表达的检测及临床意义

目的 研究乙型肝炎病毒(HBV)感染者外周血CD4+T细胞表面CD25、CD127不同亚群的表达情况及临床意义。方法 用荧光抗体CD127-FITC、CD4-PECY5、CD25-PE标记T细胞。用流式细胞仪分别测定53例慢性乙型肝炎患者和53例HBV携带者CD4+T细胞表面CD25、CD127不同亚群的表达情况。对20例HBV-DNA阳性乙型肝炎病毒感染者干扰素治疗进行随访。结果与健康对照组[7.26％(6.15％,8.50％)]比较,慢性乙型肝炎患者[11.23％(9.10％,14.86％)]、HBV携带者[13.34％( 10.73％,18.90％)]CD25-CD 127-均显著升高,差异均有统计学意义(Q=4.559,P＜0.05;Q=6.230,尸＜0.05)。慢性乙型肝炎患者CD25hiCD127low/-[8.78％ (7.62％,10.44％)]显著高于健康对照[6.76％(5.73％,8.23％)]和HBV携带者[6.99％(5.77％,9.34％)],差异均有统计学意义(Q=3.497,P＜0.05;Q=3.103,P＜0.05)。HBV-DNA阳性组CD25-CD127-显著低于阴性组,两者差异有统计学意义[(12.92±5.20)％比(15.78±6.91)％,t=2.290,P=0.024],而CD25+/-CD127+显著高于阴性组,两者差异有统计学意义[(79.27±5.20)％比(76.02±7.04)％,t=2.194,P=0.030]。与治疗前比较,干扰素治疗12周CD25hiCD127low/-显著升高[(9.29±2.51)％比(11.08±2.38)％,t=2.820,P=0.011],而CD4+CD25-CD127-显著降低,两者差异有统计学意义[(13.86±5.72)％比( 10.86±3.60)％,t=2.469,P=0.024]。结论 HBV感染者外周血CD4+T细胞中CD25-CD127-亚群的表达与病毒的感染和清除有关;CD25hiCD 127low/-亚群的表达升高与发病有关。外源性干扰素可升高CD25hiCD127low/-的表达,降低CD25-CD127-的表达,从而抑制免疫反应。     

Abnormal immune responses in persons with previous extrapulmonary tuberculosis in an in vitro model that simulates in vivo infection with Mycobacterium tuberculosis

Persons with previous extrapulmonary tuberculosis have reduced peripheral blood mononuclear cell cytokine production and CD4(+) lymphocytes compared to persons with previous pulmonary tuberculosis or latent tuberculosis infection, but specific defects related to Mycobacterium tuberculosis infection of macrophages have not been characterized. The objective of this study was to further characterize the in vitro immune responses to M. tuberculosis infection in HIV-seronegative persons with previous extrapulmonary tuberculosis. Peripheral blood mononuclear cells were isolated from HIV-seronegative persons with previous extrapulmonary tuberculosis (n = 11), previous pulmonary tuberculosis (n = 21), latent M. tuberculosis infection (n = 19), and uninfected tuberculosis contacts (n = 20). Experimental conditions included M. tuberculosis-infected macrophages cultured with and without monocyte-depleted peripheral blood mononuclear cells. Concentrations of interleukin 1β (IL-1β), IL-4, IL-6, CXCL8 (IL-8), IL-10, IL-12p70, IL-17, CCL2 (monocyte chemoattractant protein 1), tumor necrosis factor alpha (TNF-α), and gamma interferon (IFN-γ) were measured by multiplex cytokine array. When M. tuberculosis-infected macrophages were cocultured with monocyte-depleted peripheral blood mononuclear cells, IFN-γ (P = 0.01), TNF-α (P = 0.04), IL-10 (P < 0.001), and IL-6 (P = 0.03) exhibited similar continua of responses, with uninfected persons producing the lowest levels, followed by extrapulmonary tuberculosis cases, pulmonary tuberculosis controls, and persons with latent M. tuberculosis infection. A similar pattern was observed with CXCL8 (P = 0.04), IL-10 (P = 0.02), and CCL2 (P = 0.03) when monocyte-depleted peripheral blood mononuclear cells from the four groups were cultured alone. Persons with previous extrapulmonary tuberculosis had decreased production of several cytokines, both at rest and after stimulation with M. tuberculosis. Our results suggest that persons who develop extrapulmonary tuberculosis have a subtle global immune defect that affects their response to M. tuberculosis infection.     

Elevated serum CCL2 concomitant with a reduced mycobacterium-induced response leads to disease dissemination in leprosy

Mycobacterium leprae and Mycobacterium tuberculosis are successful intracellular pathogens which down regulate host immune responses. T-cell interferon-gamma (IFNgamma) and macrophage tumour necrosis factor-alpha (TNFalpha) activate chemokines such as, C-C chemokine ligand-2 (CCL2) and CCL5, which play a role in granuloma formation. Lepromatous leprosy is characterized by defective granulomas with lowered T-cell- and macrophage-mediated responses. Tuberculosis (TB) can be localized to the lung, whereby discreet granulomas are formed. The role of chemokines in leprosy infections is as yet unclear. We compared chemokine responses in lepromatous leprosy and pulmonary tuberculosis patients. Circulating serum CCL2 was raised while CCL5 was lowered in leprosy, as compared with TB patients and healthy controls. However, both Mycobacterium bovis BCG- (P=0.08) and M. leprae-induced (P=0.05) CCL2 secretion was reduced in leprosy. In leprosy, BCG induced greater CCL2 (P=0.01), TNFalpha (P=0.02) and somewhat greater CCL5 (P=0.08) than M. leprae, while CXCL8 induction was comparable. Overall levels of Mycobacterium-induced CCL2, TNFalpha and CXCL8 were two to threefold lower, and CCL5 was 10-fold lower in leprosy as compared with TB. Reduced inducible CCL2 combined with a lowered TNFalpha response in lepromatous leprosy may contribute to the unrestricted growth and dissemination of mycobacteria found in the disease.     

脉冲振荡法测定慢性阻塞性肺疾病患者呼吸阻抗的影响因素分析

目的 探讨连接管道、体位和通气对脉冲振荡法(IOS)测定慢性阻塞性肺疾病(COPD)患者呼吸阻抗的影响,以评价IOS的临床意义及其在重症监护中的应用.方法 分别在坐位无管、坐位有管、卧位无管、卧位有管4种状态下采用IOS对COPD患者组和健康组(各40例)进行呼吸阻抗测定;30例机械通气COPD患者在通气和断开通气两种情况下进行呼吸阻抗测定.结果 健康组中,外加连接管后黏性阻力稍增大,共振频率左移[(9.849+1.811)Hz比(7.850+1.043)Hz,P＜0.05],弹性阻力减少,总阻抗无明显变化;由坐位转为平卧位后,黏性阻力明显升高,高频不如低频明显,共振频率右移[(9.849±1.811)Hz比(13.604±1.702)Hz,P＜0.05],弹性阻力增大,总阻抗增加[(0.328±0.080)kPa·L-1·s-1比(0.443±0.120)kPa·L-1·s-1,P＜0.05].COPD患者呼吸阻抗的变化规律与健康组类似.4种状态下区别COPD患者与健康者最敏感的指标均为共振频率(F=159.5、114.3、98.6、97.0,均P＜0.05).与断开通气比较,COPD患者机械通气时部分阻力值下降,以高频时明显,但主要阻力参数的变化差异无统计学意义.结论 外加管道或体位改变后,IOS测定值变化有规律,且与呼吸机连接后不相互影响.IOS测定可广泛应用于平卧位、病情严重或机械通气的患者.     

Th17细胞在Graves’病中的变化及意义

目的:探讨Th17细胞在中国人Graves’病(GD)中的变化及意义。方法:应用电化学发光法测定30例初发GD患者及30例健康体检者血清中促甲状腺激素(TSH)、游离三碘甲腺原氨酸(FT3)、游离甲状腺激素(FT4)、甲状腺球蛋白抗体(TgAb)和甲状腺过氧化酶抗体(TPOAb)的水平;采用流式细胞术检测外周血单个核细胞(PBMC)中CD4+IL-17+T(Th17)细胞的数量;用ELISA法测定初发GD患者和健康对照者PBMC产生IL-17的水平;应用实时定量PCR(qRT-PCR)检测PBMC中ROR-γt、IFN-γ、IL-4 mRNA的表达量;采用免疫组化染色检测甲状腺组织中IL-17+细胞的分布和数量。结果:初发GD组外周血中Th17细胞占CD4+T细胞的比例[(20.59±4.63)%]、PBMC分泌的IL-17水平[(83.22±34.28)pg/ml]以及PBMC中ROR-γt mRNA表达(1.67±0.98)均明显高于正常对照组Th17[(11.77±4.18)%](P<0.01)、IL-17[(19.74±5.99)pg/ml](P<0.01)和ROR-γt mRNA(0.92±0.18)(P<0.05)的水平。与正常对照组比较,GD患者PBMC中IFN-γmRNA的表达(0.31±0.07)、IL-4 mRNA的表达(2.53±0.70)均明显降低(P<0.01)。初发GD组患者PBMC中Th17细胞百分率与ROR-γt mRNA表达呈正相关(r=0.5047,P=0.01);与IFN-γmRNA表达量呈负相关(r=-0.5085,P<0.01);与IL-4 mRNA表达量呈负相关(r=-0.5372,P<0.01)。正常对照组甲状腺组织中未发现IL-17+细胞,GD组甲状腺组织滤泡间质内可见散在的IL-17+细胞。结论:初发GD患者PBMC中Th17细胞数量、PBMC分泌IL-17的水平和ROR-γt mRNA表达明显增高。GD患者甲状腺组织中有IL-17+细胞浸润,提示Th17细胞可能参与了中国人GD的发生。     

APOPTOSIS OF HUMAN MONOCYTES/MACROPHAGES IN MYCOBACTERIUM TUBERCULOSIS INFECTION

Tuberculosis (TB) is still a major health problem, both as a single disease entity and as a cofactor in AIDS. The interaction between macrophage and Mycobacterium tuberculosis (MTB) is a critical step in the establishment of an early chronic infection. This study analyses the capacity of MTB to induce apoptosis in cells obtained by broncho-alveolar lavage (BAL) from patients with reactive pulmonary tuberculosis and from AIDS patients with disseminated pulmonary tuberculosis. Apoptosis was increased three-fold in BAL cells obtained from patients with pulmonary tuberculosis and even more markedly in alveolar macrophages of MTB-infected AIDS patients, compared with controls. Apoptosis was analysed and characterized by propidium iodide (PI) incorporation, terminal deoxy transferase (TDT)-mediated dUTP-biotin nick end labelling (TUNEL), and tissue transglutaminase (tTG) expression. The MTB–macrophage interaction was also investigated in vitro by infecting monocyte-derived macrophages (MDM) with MTB (virulent strain H37Rv). The induction of apoptosis by MTB required viable bacteria, was dose-dependent, and was restricted to H37Rv. Infection with either Mycobacterium avium complex (MAC) or HIV-1 and treatment with heat-killed MTB failed to induce apoptosis. © 1997 by John Wiley & Sons, Ltd.     

Diminished Adherence and/or Ingestion of Virulent Mycobacterium tuberculosis by Monocyte-Derived Macrophages from Patients with Tuberculosis

The interaction between the macrophage and Mycobacterium tuberculosis is mediated by a variety of macrophage membrane-associated proteins. Complement receptors have been implicated in the adherence of M. tuberculosis to macrophages. In the present work, the adherence and/or ingestion of M. tuberculosis H37Rv to human monocyte-derived macrophages (MDM) from patients with tuberculosis (TB) and healthy controls was measured by microscopical examination, [³H]uracil incorporation, and CFU. The adherence and/or ingestion was enhanced by fresh serum and inhibited by heat inactivation, EDTA treatment, and anti-CR1 and anti-CR3 antibodies. Comparison of MDM from TB patients and healthy controls showed that the former exhibited a significantly decreased capacity to adhere and/or ingest M. tuberculosis, as determined by the number of CFU and ³H incorporation. The expression of CR1 (CD35) and CR3 (CD11b/CD18) on MDM from TB patients and healthy controls, as determined by flow cytometry, did not show significant differences. These results suggest that the lower ingestion of M. tuberculosis by MDM from TB patients is not due to defects in complement receptors, and therefore, there might be other molecules involved in the adherence and/or ingestion process that render MDM from TB patients ingest less mycobacteria than those from healthy controls.