哮喘患者外周血单个核细胞腺苷受体

目的通过检测哮喘患者外周血单个核细胞（PBMCs）腺苷受体mRNA表达,探讨腺苷受体在哮喘发病中的作用。方法用Ficoll液分离PBMCs,采用逆转录-多聚酶联反应法和图象分析半定量法检测PBMCs腺苷受体mRNA。结果正常人及哮喘患者PBMCs均表达A     

促甲状腺素受体抗体对Graves病患者治疗后促甲状腺激素水平长期抑制的影响

目的 探讨促甲状腺素受体抗体(TRAb)对Graves病患者抗甲状腺药物治疗后促甲状腺激素(TSH)水平长期抑制的影响。方法 90例Graves病患者根据TSH恢复正常水平的时间滞后于游离甲状腺素是否超过8周,分为长期组(>8周,28例)和短期组(≤8周,62例),比较两组一般资料及治疗前临床数据的差异。多因素logistic回归分析TSH水平抑制时间延长的影响因素。治疗6个月后根据TSH水平是否仍低于正常参考值下限0.27μIU/mL,将患者分为抑制组(<0.27μIU/mL,13例)和非抑制组(≥0.27μIU/mL,77例),比较两组治疗前、治疗6个月后TRAb水平及TRAb下降率差异。结果 短期组治疗前TRAb水平低于长期组(P<0.05),而两组治疗前TSH水平比较差异无统计学意义(P>0.05)。多因素logistic回归分析结果显示,治疗前TRAb高水平增加了TSH水平抑制时间延长的风险(P<0.05)。抑制组治疗前和治疗6个月后TRAb水平高于非抑制组(P<0.05),TRAb下降率低于非抑制组(P<0.05)。结论 抗甲状腺药...     

促甲状腺素受体抗体检测对Graves病患者的诊断价值分析

目的 研究促甲状腺素受体抗体对于Graves病的鉴别诊断和临床诊治的应用价值.方法 通过电化学发光法测定Graves病患者血清促甲状腺素受体抗体水平,将本院收治的189例Graves 病患者分为未接受治疗的初诊组、治疗效果不明显组和治疗后症状明显缓解组,同时与正常组作对照.结果 促甲状腺素受体抗体在初诊组、效果不明显组和治疗后缓解组患者中的血清阳性率均显著高于正常对照组;初诊组和效果不明显组的血清促甲状腺素受体抗体浓度较高,缓解组较低但高于正常对照组.结论 促甲状腺素受体抗体在血清中的浓度水平对于Graves病的鉴别诊断和临床诊治具有重要的意义.     

哮喘患者外周血单个核细胞腺苷受体mRNA的表达

目的：通过检测哮喘患者外周血单个核细胞（PBMCs)腺苷受体mRNA表达,探讨腺苷受体在哮喘发病中的作用。方法：用Ficoll液分离PBMCs,采用逆转录－多聚酶联反应法和图象分析半定量法检测PBMCs腺苷受体mRNA。结果：政党人及哮喘患者PBMCs均表达A1,A2,A2b,A3腺苷受体（A1AR,A2aAR,A2bAR,A3AR)mRNA,以A2bARmRNA所占百分比最高;哮喘患者PBMCsA1AR,A2bAR,A3AR mRNA表达较正常人明显增加,哮喘患者PBMCs A3AR mRNA表达与第一秒用力呼气容积实测值占预计值的百分率（FEV1％）呈负相关。结论：哮喘患者PBMCS表达A1AR,A2bAR,A3AR mRNA增加,以A2bAR mRNA所占百分比最高,A3AR mRNA表达与气道阻塞程度相关。     

骨髓单个核细胞和外周血单个核细胞移植治疗外周动脉病的疗效分析

目的 观察自体骨髓单个核细胞(BMMNCs)移植、粒细胞集落刺激因子(G-CSF)动员的自体外周血单个核细胞(PBMNCs)移植治疗外周动脉病的临床疗效.方法 将69例外周动脉病患者随机分成3组:G-CSF动员的自体PBMNC移植组(PBMNC组)30例,给予G-CSF 600mg/d,皮下注射,连用4d,第5d采集PBMNCs移植至患肢;自体BMMNC移植组(BMMNC组)15例,采集骨髓液500～800 ml,分离BMMNCs移植至患肢;常规内科治疗组(对照组)24例,仅给予常规药物治疗.术后3月随访所有患者,观察评价各项指标.结果 PBMNC组CD34 细胞数[(3.31±1.39)×106 /kg]显著多于BMMNC组[(1.42±0.42)×106 /kg,P＜0.01)].移植后3月时复查,PBMNC组和BMMNC组的静息痛改善率分别为76.67%和73.33%均显著高于对照组16.67%(P＜0.01);PBMNC组的溃疡改善率为40.0%,BMMNC组为42.86%,与对照组比较差异均无统计学意义(P＞0.05);两个移植组之间静息痛改善率和溃疡改善率差异均无统计学意义(P＞0.05).两个移植组术后3月时,最大无痛步行距离(MWD)、踝臂压指数(ABI),经皮氧分压(TcO2)均明显高于治疗前(P＜0.01),对照组治疗前后无差异(P＞0.05).△MWD、△ABI、△TcO2分别指治疗前和治疗后3月时MWD、ABI、TcO2的差值,比较两个移植组[(108.75±66.70)m vs(113.75±63.86)m,(0.082±0.05)vs(0.077±0.051),(10.54±5.90)mm Hg vs(9.63±5.26)mm Hg)]差异均无统计学意义(P＞0.05).PBMNC组和BMMNC组的DSA显效率分别为66.67%和62.50%均显著高于对照组0%(P＜0.01,P＜0.05).两个移植组之间无差异(P＞0.05).结论 自体PBMNC移植和自体BMMNC移植均为治疗外周动脉病安全有效的新方法,两者疗效相当.     

栀子清肝汤对Graves病血清促甲状腺素受体抗体的影响

目的:观察栀子清肝汤对Graves病患者血清促甲状腺素受体抗体(TRAb)的影响。方法:将62例Graves病患者随机分为单纯规范化治疗组(GD单组)和栀子清肝汤加甲巯咪唑组(GD中组),通过长疗程治疗,观察TRAb的阳性率。结果 :治疗后GD中组TRAb阳性7例,阳性率22.58%;GD单组阳性15例,阳性率48.39%,两组有显著性差异(P<0.05)。结论:栀子清肝汤有利于降低Graves病的复发。     

促甲状腺素受体mRNA在甲状腺癌诊断中的作用

目的采用逆转录聚合酶链反应（RT-PCR）检测方法对甲状腺结节患者外周血中促甲状腺素受体mRNA（TSHR-mRNA）表达水平进行检测,分析其在甲状腺良恶性结节中的表达差异,并研究其在甲状腺癌诊断中的作用。方法选取130例外科住院甲状腺结节患者,通过细针穿刺（FNA）和（或）甲状腺手术获得组织标本并确诊,分为甲状腺良性疾病组（80例）和甲状腺癌组（50例）。对两组患者采用RT-PCR检测方法,比较两组患者的TSHR-mRNA的表达水平以及临床表现,分析TSHR-mRNA在甲状腺癌中的诊断意义。结果甲状腺癌组患者的TSHR-mRNA阳性率明显高于甲状腺良性疾病组,差异有统计学意义（P〈0.05）。结论促TSHR-mRNA表达水平的检测有助于甲状腺癌术前定性诊断。     

格雷夫斯病患者糖代谢特点及其影响因素分析

目的　了解格雷夫斯病(Graves′disease ,GD)患者胰岛素、C肽变化与糖耐量的关系。方法　未经治疗的格雷夫斯病患者6 5例,行口服葡萄糖耐量 胰岛素释放试验后,用化学发光法测血清胰岛素、C肽值并与正常对照比较。结果　格雷夫斯病组除空腹外其余各点血糖值明显增高(P <0 .0 0 1) ,基础免疫活性胰岛素正常,葡萄糖刺激后明显增高(P <0 . 0 0 2 7) ,总血清胰岛素/总血糖(∑IRI/∑BG)在弥漫性甲状腺肿伴格雷夫斯病组明显增加,游离三碘甲状腺原氨酸(FT3 )与多个时点血清胰岛素、C肽呈显著负相关。结论　格雷夫斯病在治疗前存在高血糖与高胰岛素血症并存的胰岛素抵抗状态。     

卡维地洛对人外周血单个核细胞CD25表达的影响

目的研究新型β受体阻滞剂卡维地洛(Car)对单个核细胞表达CD25的影响.方法分离外周血单个核细胞(PBMC),以PMA A23187作为刺激剂,分组体外培养.用MTT方法检测PBMC的活性;用流式细胞仪检测单个核细胞的凋亡率和CD25的表达水平.结果 (1)20 μmol以内的Car对PBMC没有毒性;(2)Car能显著抑制PBMC表达CD25,24 h时,浓度3组为(8.23±2.67)%,与对照组(30.73±4.10)%比较,差异有显著性(P＜0.01).结论 Car可以抑制PBMC表达CD25.这可能是卡维地洛治疗慢性心衰的机制之一.     

格雷夫斯眼病患者TGAb TMAb TRAb的检测及临床意义

目的探讨甲状腺球蛋白抗体(TGAb)、甲状腺微粒体抗体(TMAb)、甲状腺受体抗体(TRAb)与格雷夫斯眼病(GO)的关系,了解各指标在GO患者发病中的作用。方法采用放射免疫分析法检测64例格雷夫斯病(GD)患者及121例GO患者血清TGAb、TMAb、TRAb值,并对两组各指标的检测值及TRAb阳性检出率进行比较。结果两组患者血清TGAb、TMAb活性差异无统计学意义(P>0.05);GO组及GD组TRAb活性分别为(28±6)U/L和(11±10)U/L,GO组明显高于GD组(P<0.05);GO组及GD组阳性检出率分别为72.7%和51.6%,GO组显著高于GD组(P<0.05)。结果TRAb在GO的诊断中具有重要意义;推测GD患者血清TRAb显著增高是合并或随病程延长可能合并GO的重要标志。     

环瓜氨酸肽对类风湿关节炎外周血T淋巴细胞增殖的影响

目的观察环瓜氨酸多肽（CCP）对类风湿关节炎（RA）患者外周血T细胞的激活作用。方法用CCK-8法检测CCP对24例RA患者外周血T细胞增殖的影响,并与16名健康志愿者和24例系统性红斑狼疮（SLE）患者做对照。分9个组别观察,分别是RA空白对照组、RA植物血凝素（PHA）对照组、RA CCP组、健康空白对照组、健康PHA对照组、健康CCP组、SLE空白对照组、SLE PHA对照组及SLE CCP组。结果体外培养72 h后,RA PHA对照组、RA CCP组与RA空白对照组相比,淋巴细胞增殖水平（0.618±0.068、0.699±0.077）均较（0.494±0.057）有明显升高,差异有统计学意义（P〈0.01,P〈0.01）;SLEPHA对照组、SLECCP组与SLE空白对照组相比,淋巴细胞增殖水平（0.496±0.079、0.451±0.073）均较（0.364±0.061）有明显升高,差异有统计学意义（P〈0.01,P〈0.01）。RA CCP组增殖幅度（0.205±0.037）较SLE CCP组增殖幅度（0.087±0.018）明显升高,差异有统计学意义（P〈0.01）。结论CCP可特异性刺激RA患者T细胞增殖,瓜氨酸化抗原是诱导RA发病并参与RA致病过程的重要抗原成分,进一步干预此过程将有助于更好的治疗RA。     

冠心病患者外周血单个核细胞TLR5的测定及临床意义

目的:检测Toll样受体5(Toll-like receptor 5,TLRS)在冠心病(CHD)患者外周血单个核细胞(PBMCs)的表达水平,并探讨其与冠状动脉粥样硬化发生发展的关系.方法:用逆转录聚合酶链扩增法(RT-PCR)的方法检测50例CHD患者及40例健康对照者PBMCs TLR5 mRNA的表达情况.结果:CHD患者外周血单个核细胞Toll样受体5 mRNA表达水平高于健康对照组,分别为(0.793±0.284)和(0.211±0.319)(P＜0.01).结论:TLR5及其介导的天然免疫应答与动脉粥样硬化的发生发展存在一定的相关性.     

肾炎患者外周血单个核细胞尿激酶型纤溶酶原激活剂受体及相关细胞因子的表达

目的　探讨肾炎患者外周血单个核细胞尿激酶受体及相关细胞因子的表达。方法　采用流式细胞术测定肾炎患者外周血单个核细胞尿激酶型纤溶酶原激活剂受体、IL - 2R和整合蛋白 β2 水平 ,双抗体夹心ELISA法测定血清IL - 2 ,IL - 4和IL - 10水平。结果　肾炎患者外周血单个核细胞尿激酶型纤溶酶原激活剂受体、IL - 2R和整合蛋白 β2 表达水平分别为 (15 .94± 3.2 1) %、(10 .93± 4.2 4) %和 (12 .87± 4.79) % ,均显著高于正常组的 (4.2 1± 1.0 2 ) %、(1.49± 0 .47) %和 (2 .0 8± 1.0 1) % (P <0 .0 1) ;血清IL - 2和IL - 4水平分别为 (2 1.14± 8.2 4) μg/L和 (43.5 6± 10 .2 4) μg/L ,亦均显著高于正常组的 (8.73± 4.0 2 ) μg/L和 (2 3.0 4± 4.78) μg/L(P <0 .0 1) ;IL - 10水平 (8.34± 2 .0 1) μg/L则显著低于正常组的 (12 .0 1± 2 .34 ) μg/L(P <0 .0 1) ;89例肾炎患者外周血单个核细胞尿激酶型纤溶酶原激活剂受体表达水平随病情好转而逐渐恢复正常 ,13例患者激素治疗 2周病情无缓解 ,但外周血单个核细胞尿激酶型纤溶酶原激活剂受体表达水平也逐渐恢复。结论　肾炎患者外周血单个核细胞尿激酶型纤溶酶原激活剂受体表达水平与多种细胞因子有关 ,参与肾小球肾炎的发病 ,在肾小球基底膜?     

Effects of silymarin on the proliferation and glutathione levels of peripheral blood mononuclear cells from beta-thalassemia major patients

Iron toxicity in beta-thalassemia major is the main cause of oxidative stress and cell mediated immune deficiencies. Despite indicative signs of severe oxidative deficiencies associated with beta-thalassemia major, such as decreased level of plasma antioxidants and depletion of erythrocyte glutathione, little is known about intracellular redox status of immune cells. Since glutathione is a primary intracellular antioxidant and plays an essential role in several functions in T cells, in this study intracellular glutathione (GSH) levels as well as proliferation of PHA-activated peripheral blood mononuclear cells (PBMC) were investigated in 28 beta-thalassemia major patients and 28 healthy age-matched individuals. Considering the potential benefits of flavonoids in the therapy of oxidative stress, the effects of silymarin on the GSH levels and proliferation of PBMC from normal and thalassemia individuals were further examined. Quantitative determination of intracellular GSH and proliferative response of PBMC to PHA were performed before and after 72 h incubation of PBMC with various concentrations of silymarin (0, 5, 10, or 20 mug/ml). Results demonstrated a significant reduction of GSH and proliferation in beta-thalassemia major cells; however treatment with silymarin led to restoration of both GSH levels and PBMC proliferation in thalassemia patients. Considerably low levels of GSH and depressed proliferative response of PBMC in beta-thalassemia major may be responsible for the cell mediated immune abnormalities in iron overload conditions. Moreover, the GSH restoration and improvement of PBMC growth by silymarin is a possible explanation for its recently reported antioxidant and immunostimulatory activities. These data suggest the benefit of using flavonoids to normalize immune dysfunction in beta-thalassemia major. The immunomodulatory effects of silymarin in beta-thalassemia major are currently under further investigation in a double blind clinical trial.     

Mibefradil-induced inhibition of proliferation of human peripheral blood mononuclear cells

To evaluate the role of intracellular calcium and particularly Ca2+-uptake in the initiation of lymphocyte mitogenesis, the effect of mibefradil, which blocks both L- and T-type calcium channels with a more selective blockade of T-type channels, on the proliferation of human peripheral blood mononuclear cells (PBMCs) is compared with the effect of nifedipine, which blocks only the L-type calcium channel. The rate of [3H]thymidine incorporation into control and concanavalin A-stimulated PBMCs in the presence or absence of the calcium channel blockers mibefradil or nifedipine (1, 10, or 50 microM), and of the intracellular calcium antagonist 3,4,5-trimethoxybenzoic acid 8-(diethylamino) octyl ester (TMB-8; 1, 10, 25, or 50 microM) was assayed in the cells cultured for 3 days. The cellular cytotoxicity and the cell number in growing cultures also was determined in mibefradil- or nifedipine-treated control or stimulated cells. Restoration of the proliferative response in mibefradil- or nifedipine-treated cells was investigated by addition of exogenous interleukin-2. Interleukin-2-receptor expression in the cells was monitored by using anti-activated T-cell antigen (Tac) antibody, and the interleukin-2 production in the cell supernatants of the cultures was determined by an enzyme-amplified sensitive immunoassay. Mibefradil and nifedipine concentration-dependently reduced the cell number and the [3H]thymidine incorporation or the de novo DNA synthesis in control and concanavalin A-stimulated human PBMCs. Mibefradil exhibited a more pronounced inhibition of the proliferation of human PBMCs than did nifedipine. The inhibitory effect of mibefradil or nifedipine on DNA synthesis was dependent on the timing of treatment with the drugs. The inhibitory effect of mibefradil or nifedipine on the lymphoproliferative response was nearly abolished if the drugs were added 20 h after cell stimulation. A markedly reduced inhibitory effect was found when mibefradil or nifedipine was added 1-7 h after cell stimulation. However, regardless of time of addition, TMB-8 caused a persistent inhibition of the proliferation of human PBMCs. The inhibitory effect of mibefradil or nifedipine on the proliferation of human PBMCs is nearly abolished by addition of the calcium channel activator Bay K 8644. The proliferative response of mibefradil- or nifedipine-treated cells is restored by addition of exogenous interleukin-2. The normal expression of interleukin-2 receptors was preserved, whereas the interleukin-2 production was blocked in the presence of mibefradil or nifedipine. Our data show that mibefradil has a more pronounced inhibitory effect on the proliferation of human PBMCs than nifedipine and that this inhibitory effect on DNA synthesis is dependent on the timing of treatment with both drugs.     

川芎嗪对类风湿性关节炎患者PBMC免疫球蛋白合成的影响

目的：探讨川芎嗪对培养的类风湿性关节炎(RA)活动期、静止期外周血单个核细胞(PBMC)产生免疫球蛋白(Ig)的影响。方法：应用细胞培养技术，采用ELISA检测方法，检测细胞培养上清液IgG、IgM的含量。结果：川芎嗪明显抑制类风湿性关节炎活动期及静止期IgG、IgM的合成。结论：川芎嗪可以抑制RA患者中IgG、IgM的合成，可减轻RA的发展。     

Effects of ozone on isolated peripheral blood mononuclear cells.

We have investigated the release of cytokines from isolated peripheral human blood mononuclear cells (PBMC) exposed to various ozone concentrations for 10 min and the release of both proinflammatory and immunosuppressive cytokine after 24, 48 and 76 h incubation. Ozonation was performed by exposing for 10 min equal cell numbers and volumes of cell suspension to equal volumes of a gas mixture (1:1 ratio) composed of oxygen-ozone with precise ozone concentrations ranging from 1.0 up to 80 microg/ml (0.02 up to 1.68 mM). Markers of oxidative stress showed a significant relationship between ozone doses and both lipid peroxidation and protein thiol groups content. With the exception of the lowest ozone concentration, the cytokine production of PBMC was depressed particularly at concentrations from 40 mug/ml upwards. There was no significant effect on IL-6 production between exposed or unexposed cells, up to 72 h of incubation. IL-4 production was markedly affected by ozone exposure, showing a marked decrease even at the lowest ozone concentration (2.5 microg/ml) already after 24 h incubation. On the other hand, production of IFN-gamma and TNF-alpha was slightly stimulated by the lowest ozone dose either at all times or only after 72 h incubation, respectively. Analysis of the proliferation index (PI) is consistent with these results showing that, while the lowest concentration stimulates it, progressively increasing O(3) concentrations inhibit the PI. These data show that there is a significant relationship between cytokine production and ozone concentrations and that PBMC are very sensitive to oxidation particularly in presence of serum with low antioxidant capacity.     

甲氨蝶呤对类风湿关节炎环瓜氨酸肽抗原特异性T细胞体外增殖的影响

目的观察甲氨蝶呤（MTX）对类风湿关节炎（Rheumatoid Arthritis,RA）患者环瓜氨酸肽抗原特异性T细胞（Circum-citrul-linated peptide antigen special Tcell,CCP/AST）体外增殖水平的影响。方法体外分离、培养RA患者外周血淋巴细胞,分别设：空白组：不加任何干预药物;CCP组：仅加CCP干预（CCP终浓度为20 mg·L^-1）;甲氨喋呤（MTX）组：同时加CCP与MTX干预（CCP与MTX终浓度均为20 mg.L-1）;体外培养72 h后,采用CCK-8法检测细胞增殖（OD）水平。结果CCP可以显著诱导RA患者环瓜氨酸肽抗原特异性T细胞（CCP/AST）体外增殖（P〈0.01）,在此基础上加入MTX后,CCP/AST增殖状态被显著抑制（P〈0.01）。结论MTX对RA患者CCP/AST的增殖水平有显著的抑制作用,推断MTX做为RA临床治疗的经典用药,抑制RA患者CCP/AST的活化、增殖,可能是其治疗RA的有效途径之一。