IL—2,IL—2R,IL—6mRNA在自身免疫性甲状腺病甲状腺组织中的原位表达

本实验分别用Ｄｉｇ标记的ＩＬ－２、ＩＬ－２Ｒ、ＩＬ－６ｃＲＮＡ探针在３７例自身免疫性甲状腺疾病患者甲状腺石蜡切片上行原位杂交，结果显示患者甲状腺浸润细胞中有ＩＬ－２、ＩＬ－２Ｒ、ＩＬ－６ｍＲＮＡ的异常表达，其分布与病理损伤部位一致，这种异常表达是引起甲状腺内免疫功能紊乱的重要因素之一。     

Graves病T,B细胞功能异常及甲巯咪唑体外调节作用的研究

目的：探讨Ｇｒａｖｅｓ病（ＧＤ）患者Ｔ、Ｂ细胞异常及甲巯咪唑体外对它们的影响。方法：将２５例ＧＤ病人 外周血单个核细胞（ＰＢＭＣ）在不同浓度的甲巯咪唑条件下进行体外细胞培养６天后,同时检测Ｔ细胞亚群及ｓＩＬ－ ２Ｒ和ＩｇＧ的含量。结果：ＧＤ病人ＰＢＭＣ中的ＣＤ_８~＋Ｔ细胞亚群百分率明显低于对照组,ＣＤ~＋_４／ＣＤ_８~＋比值显著升高 （ｐ＜０．０１）。 ｓＩＬ－２Ｒ及ＩｇＧ水平较对照组显著升高（ｐ＜０．０１）;Ｘ１０~（－５）ｍｏｌ／Ｌ的甲巯咪唑可明显升高ＧＤ患者ＰＢ－ ＭＣ培养后ＣＤ_８~＋ Ｔ细胞的百分率（Ｐ＜０．０１）,降低ＣＤ_４~８ＣＤ_８~＋比值及ｓＩＬ－２Ｒ和ＩｇＧ水平（Ｐ＜０．０１）。提示：ＧＤ病 人Ｔ细胞亚群对免疫应答发生调节失衡,甲巯咪唑具有调节ＧＤ患者Ｔ、Ｂ细胞功能异常的作用。     

维生素E对创伤小鼠淋巴细胞功能的调节作用及机制

研究了维生素Ｅ对创伤小鼠淋巴细胞功能的调节作用及机制。结果显示：ＶＥ体内应用（１００ｍｇ／ｋｇ·ｄ－１，ｉ·ｍ×４ｄ）对创伤小鼠淋巴细胞白介素２（ＩＬ－２）ｍＲＮＡ、ＩＬ－２受体（ＩＬ－２Ｒ）ｍＲＮＡ水平，ＩＬ－２的生成，ＩＬ－２Ｒ的表达以及Ｔ淋巴细胞转化具有明显的升高作用。可明显逆转创伤小鼠血清及淋巴组织中ＶＥ含量的降低，丙二醛（ＭＤＡ）含量的升高以及淋巴细胞膜流动性的降低。并可改善创伤小鼠淋巴组织的损害。结果表明：ＶＥ可通过降低创伤小鼠的脂质过氧化反应，保护淋巴细胞膜，进而纠正创伤后淋巴细胞功能的受抑状态。     

灵芝多糖体外对小鼠脾细胞IL-2,IL-3 mRNA表达的影响

目的观察灵芝多糖体外对小鼠脾细胞ＩＬ－２、ＩＬ－３ｍＲＮＡ的表达水平是否有影响。方法逆转录多聚酶链式反应（ＲＴ－ＰＣＲ）检测ｍＲＮＡ表达，凝胶图象光密度分析系统进行相对定量。结果在原代小鼠脾细胞培养开始时加入不同浓度的灵芝多糖ＧＬＢ７（５，１０，２０，４０ｍｇ·Ｌ－１），培养１２ｈ及２１ｈ后观察ＧＬＢ７与ＩＬ－２及ＩＬ－３ｍＲＮＡ表达水平之间的量效关系。发现在一定范围内，随着ＧＬＢ７浓度的升高，ＩＬ－２及ＩＬ－３ｍＲＮＡ表达水平也相应提高，对ＩＬ－２ｍＲＮＡ的提高率分别为１０．４％，２１．９％，３７．８％，４６．６％；对ＩＬ－３ｍＲＮＡ的提高率分别为１５．４％，３５．２％，５２．４％，７４．５％。１９９８－０４－０８收稿，１９９８－０７－２６修回＊国家自然科学基金资助课题，Ｎｏ３９４００１６５作者简介：王庆彪，男，２６岁，硕士；雷林生，男，３８岁，医学博士，副教授培养１２ｈ和３０ｈ的上清液中ＩＬ－２样活性及ＩＬ－３样活性处理组与对照组相比亦有明显升高，且有一定的浓度依赖关系。结论灵芝多糖的免疫增强作用与其在转录水平上促进ＩＬ－２及ＩＬ－３ｍＲＮＡ的表达有关     

慢性肺心病患者血清可溶性白介素—2受体等含量的变化

观察６８例肺心病急性发作期与缓解期ｓＩＬ－２Ｒ、β２－ｍＧ、ＭＤＡ的变化。结果表明，肺心病发作期ｓＩＬ－２Ｒ、β２－ｍＧ、ＭＤＡ均明显升高，且缺氧、心衰、肾衰时升高更明显。随病情缓解，有８４．６％的患者ｓＩＬ－２Ｒ下降，８５．３％的β２－ｍＧ下降，９２．６％的ＭＤＡ下降。由此显示，肺心病发作期脂质过氧化过程加强而免疫功能减低。这３项指标的测定有助于判断病情和预后。     

1-(2,6-二甲基苯氧基)-2-(3,4-二甲氧基苯乙氨基)丙烷盐酸盐对内皮素促血管平滑肌细胞增殖及癌基因表达的影响

采用内皮素－１（ＥＴ－１０．１μｍｏｌ·Ｌ－１）建立培养的血管平滑肌细胞增殖模型，用［３Ｈ］胸腺嘧啶核苷（［３Ｈ］ＴｄＲ）参入法，流式细胞术，免疫细胞化学及Ｎｏｒｔｈｅｒｎｂｌｏｔ方法，观察了１－（２，６－二甲基苯氧基）－２－（３，４－二甲氧基苯乙氨基）丙烷盐酸盐（ＤＤＰＨ０．１μｍｏｌ·Ｌ－１）对血管平滑肌细胞增殖的作用及对原癌基因及抑癌基因的影响．结果发现：ＤＤＰＨ能逆转ＥＴ－１所致［３Ｈ］ＴｄＲ参入量增多，阻止血管平滑肌细胞由静止期（Ｇ０／Ｇ１期）进入ＤＮＡ合成期（Ｓ期）和有丝分裂期（Ｇ２／Ｍ期），并能逆转ＥＴ－１引起的ｃ－ｆｏｓ，ｃ－ｍｙｃ，ｃ－ｓｉｓ原癌基因相关抗原及ｍＲＮＡ表达增强，Ｐ５３抑癌基因相关抗原及ｍＲＮＡ表达减弱．提示ＤＤＰＨ能抑制血管平滑肌细胞增殖，与癌基因调控的分子生物学机理有关．     

膀胱癌多药耐药基因与p53基因表达的临床意义

目的 探讨膀胱癌中多样耐药基因（ＭＤＲ１）与ｐ５３基因表达的临床意义。方法 采用免疫组化（ＳＡＢＣ）法对３５例膀胱移行细胞癌中ＭＤＲ１表达产物Ｐ－ＧＰ与ｐ５３基因产物ｐ５３蛋白进行了检测。结果 ３５例膀胱移行细胞癌中Ｐ－ＧＰ、ｐ５３阳性率分别为４０％和４８．６％,随膀胱癌的病理分析和临床分期而增高。Ｔ２～Ｔ４期同Ｔｉｓ～Ｔ１期相比,ＭＤＲ１及ｐ５３的表达均有显著性差异,ＭＤＲ１与ｐ５３的表达无相关     

汉丹肝乐对肝纤维化的免疫调节作用

目的 从免疫学方法探讨汉丹肝乐抗肝纤维化的作用。方法 慢性肝炎、早期肝硬化病人４１例服且汉丹肝乐１．６ｇ,每日３次,三个月为一个疗程,收集治疗前后静脉血分离血清检测ＴＮＦ－α（肿瘤坏死因子）、ｓＩＬ－２Ｒ（可溶性白细胞介素－２受体）、ＳＯＤ（超氧化物歧化酶）、ＳＦ（血清铁蛋白）、Ｉｇ（免疫球蛋白）、Ｇ３水平。结果 慢性肝炎、早期肝硬化患者血清ＴＮＦ－ａ、ＳＩＬ－２Ｒ、ＩｇＭ、ＩｇＧ、ＩｇＡ均升高,     

甲状腺疾病患者血清抗三碘甲腺原氨酸抗体的研究及其临床意义

采用放免法检测１６５例Ｇｒａｖｅｓ病（ＧＤ）患者，１０５例桥本氏甲状腺炎（ＨＴ）患者及１０７例正常人血清抗三碘甲状腺原氨酸抗体（Ｔ３Ａｂ）。结果显示，ＧＤ未治组、缓解组和ＨＴ组Ｔ３Ａｂ的阳性率分别为０．０３６、０．０６９和０．０１９，其中ＧＤ缓解组中２例Ｔ３Ａｂ阳性者，其临床表现与血清Ｔ３和／或ＦＴ３不相符合。正常人Ｔ３Ａｂ皆为阳性。所有Ｔ３Ａｂ阳性的病人血中均可检出甲状腺球蛋白抗体（ＴＧＡｂ）。     

急性白血病患者多药耐药基因表达的检测及意义

为研究急性白血病患者多药耐药基因（ＭＤＲ１）民临床耐药、预后判断等关系，应用逆转录酶／多聚酶链反应（ＲＴ－ＰＣＲ）检测了３６例急性白血病及１０例正常人骨ＭＤＲ１基因的表达，并以ＭＤＲ１／β２ＭＧ≥０．２定为ＭＤＲ１ｍＲＮＡ阳性。结果显示，１０便正常人ＭＤＲ１ｍＲＮＡ表达均阴性。初治患者中ＭＤＲ１ｍＲＮＡ阳性率为２５％，而复发和未缓解患者则达８０％。化疗缓解率在ＭＤＲ１ｍＲＮＡ阴性及阳性的急性白血病     

甲巯咪唑与甲状腺素钠对自身免疫性甲状腺疾病患者血清细胞因子的影响

目的:观察甲巯咪唑与甲状腺素钠对自身免疫性甲状腺疾病患者血清细胞因子的影响。方法:将40例自身免疫性甲状腺疾病患者分为Graves病治疗组(n=15)和桥本甲状腺炎治疗组(n=25),另选20人为健康对照组。Graves病治疗组采用甲巯咪唑治疗;桥本甲状腺炎治疗组采用甲状腺素钠治疗。采集各组患者治疗前、后血清,分别用酶标记免疫吸附测定法检测血浆白介素(IL)-17、IL-4、干扰素(INF)-γ、转化生长因子(TGF)-β1水平。结果:在Graves病治疗组中,治疗前IL-17、IL-4、IFN-γ均较健康对照组明显升高(P<0.05),治疗后该组患者INF-γ水平有所下降,IL-4水平进一步上升,而IL-17水平并无明显下降(P>0.05)。桥本甲状腺炎治疗组治疗前上述4种细胞因子均明显升高(P<0.05),治疗后各种细胞因子水平均无明显变化(P>0.05)。结论:甲巯咪唑治疗可使Graves病患者血清细胞因子由Th1类细胞因子向Th2类细胞因子漂移,而对IL-17、TGF-β1无明显作用。甲状腺素钠则对桥本甲状腺炎中细胞因子无明显影响。     

Graves'病~(131)I治疗对IL-2、IL-6和TNF-α细胞因子的影响

目的通过对131I治疗前后Graves’病患者血清IL-2、IL-6和TNF-α细胞因子水平的比较,探讨细胞因子与Graves病发生、发展的内在联系。方法放射免疫分析法检测45例接受131I治疗的GD患者治疗前后60d外周血清IL-2、IL-6和TNF-α浓度变化,分析其与血清甲状腺激素水平的相关性分析。GD患者治疗前血清IL-6,TNF-α水平显著高于对照组(P<0.01),131I治疗后IL-6,TNF-α水平明显降低。而IL-2治疗前显著降低,治疗后逐渐回升(P<0.01);血清IL-6、TNF-α水平与FT3、FT4呈明显正相关(P<0.05),而IL-2与FT3、FT4明显负相关。结论131I治疗Graves’病不仅可以安全有效地控制甲亢症状且可改善自身免疫紊乱,多种细胞因子的变化和131I治疗的效果有关。     

Increased proportions of Tc17 cells and NK cells may be risk factors for disease progression in Hashimoto's thyroiditis

Both cellular and humoral responses play important roles in the pathogenesis of Hashimoto's thyroiditis (HT). However, the immunological differences between euthyroid (mild HT) and hypothyroid (severe HT) patients are unknown. This study aimed to investigate the distribution of lymphocyte subsets and cytokine profiles in HT patients with differences in thyroid function.Peripheral blood was drawn from 18 healthy controls and 54 HT patients (33 patients with mild HT, 21 patients with severe HT). The percentages of B cell subsets, T cell subsets and NK cells were analyzed by flow cytometry. The levels of IL-21 and Bcl-6 mRNA were assessed using real-time PCR. The levels of serum IFN-γ, TNF-α, TGF-β, IL-4, IL-6, IL-10 and a proliferation-inducing ligand (APRIL) were measured by ELISA or cytometric bead array.The percentages of double-negative memory B cells, plasma cells, Tfh cells and Tc17 cells were higher in HT patients than in the healthy controls. The percentages of Tc17 cells and NK cells were higher in the patients with severe HT than in the patients with mild HT. The levels of serum APRIL, IL-6 and IL-10 were higher in the patients with severe HT than in those with mild HT. The percentage of NK cells was positively correlated with TSH levels in the HT patients.Our data indicate that the distribution of lymphocyte subsets and cytokine profiles is aberrant in HT patients, and the increased percentages of Tc17 cells and NK cells and increased cytokine levels might be involved in the progression of HT.     

Diosgenin relieves goiter via the inhibition of thyrocyte proliferation in a mouse model of Graves＇ disease

Aim： To investigate the effects of diosgenin （Dio）, a naturally occurring steroid saponin, on goiter formation in a mouse model of Graves’ disease （GD） and the underlying mechanisms.
Methods： Female BALB/c mice were injected with adenovirus expressing the A subunit of thyrotropin receptor to induce GD. The mice were treated with Dio （20, 100 mg·kg-1·d-1, ip） for 12 or 24 d. The serum levels of TT4 and TRAb were examined using radioimmunoassay and electrochemiluminescence. The size and morphology of thyroid glands were examined. Thyrocyte proliferation was determined using BrdU incorporation assay. The expression of proliferation-associated proteins IGF-1, NF-κB, cyclin D1, and PCNA in thyroids was analyzed using immunohistochemistry and real-time PCR.

Results： The GD mice showed significantly high serum levels of TRAb and TT4 compared to the normal mice. Treatment of the GD mice with Dio for 24 d dose-dependently reduced the TT4 level and thyroid size, but did not affect the abnormal level of TRAb. Furthermore, Dio treatment dose-dependently reversed the morphological changes and reduced excessive thyrocyte proliferation in thyroids of the GD mice. Dio treatment also dose-dependently reduced the mRNA and protein levels of IGF-1, NF-κB, cyclin D1, and PCNA in thyroids of the GD mice.

Conclusion： Dio relieves goiter in a mouse model of GD through the inhibition of thyrocyte proliferation. The mechanisms involve the suppression of IGF-1, NF-κB, cyclin D1, and PCNA expression.     

甲状旁腺腺瘤大小与甲状旁腺功能及甲状腺功能的相关性研究

目的　分析甲状旁腺腺瘤（PTA）患者合并甲状腺疾病的情况,探讨PTA的大小与甲状旁腺功能及甲状腺功能的关系。方法　选取华北理工大学附属医院收治的100例PTA患者,依据是否合并甲状腺疾病将患者分为合并甲状腺疾病组（n=55）和非合并甲状腺疾病组（n=45）。收集患者临床资料,分析年龄、病程以及术前碱性磷酸酶（ALP）、校正血钙、血磷、肌酐、总三碘甲状腺原氨酸（TT3）、总甲状腺激素（TT4）、游离三碘甲状腺原氨酸（FT3）、游离甲状腺素（FT4）、促甲状腺激素（TSH）、甲状旁腺激素（PTH）水平与PTA大小的相关性。采用Logistic回归分析偏大腺瘤的危险因素,绘制受检者工作特征（ROC）曲线分析术前指标预测偏大腺瘤的临床价值。结果　合并甲状腺疾病组患者较非合并甲状腺疾病组患者腺瘤的病程短（P＜0.05）。PTA大小与校正血钙、PTH、血肌酐及病程呈正相关,与FT4呈负相关（P＜0.01）;术前PTH与校正血钙、ALP、肌酐水平及病程呈正相关,与血磷、TT4和FT4呈负相关（P＜0.01）;术前TT4水平与校正血钙、ALP、肌酐水平及病程呈负相关,与血磷、FT4水平呈正相关（P＜0.05）;术前FT4水平与校正血钙、FT3水平及病程呈负相关（P＜0.01）。Logistic回归分析显示术前PTH高及病程长为预测偏大腺瘤的危险因素。ROC曲线提示术前PTH及病程联合检测可进一步提高预测偏大腺瘤的敏感度。结论　PTA大小与术前PTH水平呈正相关性,术前PTH水平及病程的联合考虑可成为偏大PTA的预测因子,推测甲状旁腺功能亢进对甲状腺的功能起到抑制性的作用。     

老年2型糖尿病患者甲状腺功能状况及其与心率变异性的关系

目的：探讨老年2型糖尿病（DM）患者甲状腺功能状况及其与心率变异性（HRV）的关系。方法：分别测定20例老年健康对照组和老年2型DM患者的甲状腺功能及DM患者的HRV。结果：DM组的血FT3、TT3水平降低，与对照组相比具有显著性差异，DM组的FT3、FT4与超低频段功率/高频段功率之比值密切正相关。结论：老年2型DM患者在病程较长，病情控制不佳时可出现低T3现象，其游离甲状腺激素水平可能是影响DM患者HRV的原因之一。     

不孕症病人血清白细胞介素-6和甲状腺功能的相关性研究

目的 了解不孕症病人血清白细胞介素-6(IL-6)的水平及其与甲状腺功能的相关性.方法 选取66例不孕症女性为不孕组,根据其血清甲状腺激素水平将其分为不孕合并甲状腺功能减退组(32例)和不孕未合并甲状腺功能减退组(34例),另外选取健康孕龄期妇女29例为对照组,分别测定三组血清中IL-6、甲状腺激素及甲状腺自身抗体的水平.结果 (1)与对照组相比,不孕组病人血清IL-6水平升高(P<0.01);(2)在不孕症病人中,不孕合并甲减病人较不孕未合并甲减病人血清IL-6水平升高(P<0.01);(3)不孕症女性中,血清抗甲状腺球蛋白抗体(ATG)、抗甲状腺过氧化物酶抗体(TPO-Ab)阳性病人较阴性病人血清IL-6水平升高(P<0.01);(4)不孕症病人血清IL-6水平与游离三碘甲状腺原氨酸(FT3)、游离甲状腺素(FT4)、促甲状腺激素(TSH)、ATG、TPO-Ab均呈线性相关(P<0.01).多元线性回归分析调整混杂因素后,IL-6与FT3、FT4的相关性更为显著(P<0.05).结论 不孕症病人血清IL-6水平升高,尤其在合并甲减时更为明显,提示IL-6与甲状腺激素水平有一定的相关性.同时,甲状腺自身抗体阳性的不孕症病人也可通过影响IL-6的水平参与不孕症的发生发展.     

Allergic rhinitis induced by intranasal sensitization and challenge with trimellitic anhydride but not with dinitrochlorobenzene or oxazolone in A/J mice.

Allergic airway diseases induced by low molecular weight (LMW) chemicals, including trimellitic anhydride (TMA), are characterized by airway mucus hypersecretion and an infiltration of eosinophils and lymphocytes. Many experimental models have linked LMW chemical-induced allergic airway disease to Th2 cytokines. Most murine models, however, use dermal exposure to sensitize mice. The present study was designed to test the hypothesis that intranasal sensitization and challenge with the known chemical respiratory allergen TMA, but not the nonrespiratory sensitizers dinitrochlorobenzene (DNCB) and oxazolone (OXA), will induce characteristic features of LMW chemical-induced allergic airway disease in the nasal and pulmonary airways. A/J mice were intranasally sensitized and challenged with TMA, DNCB, or OXA. Only mice that were intranasally sensitized and challenged with TMA had a marked allergic rhinitis with an influx of eosinophils, lymphocytes, and plasma cells, increased intraepithelial mucusubstances, and a regenerative hyperplasia. Cytokine mRNA levels in the nasal airway of TMA treated mice also revealed an increase in the mRNA levels of the Th2 cytokines IL-4, IL-5, and IL-13, but no change in the level of the Th1 cytokine IFN-gamma. No lesions were found in the nasal airways of mice exposed to DNCB or OXA. TMA increased lung-derived IL-5 mRNA while DNCB and OXA caused no change in lung-derived cytokine mRNA levels. Both TMA and DNCB caused increases in total serum IgE, unlike OXA-exposed mice. However, no adverse alterations were found microscopically in the lungs of mice treated with TMA, DNCB, or OXA. This study is the first to demonstrate that intranasal administration of a known chemical respiratory allergen is an effective method of sensitization resulting in the hallmark features of allergic rhinitis after challenge with a concomitant increase in nasal airway-derived Th2 cytokine mRNA, lung-derived IL-5 mRNA, and total serum IgE. In contrast, DNCB and OXA failed to elicit the pathologic changes in the nasal airways and cytokine changes in the lung. This model may be useful for identifying other chemical respiratory allergens.     

重组人甲状腺球蛋白检测TgAb间接ELISA法的建立及其初步应用

目的:探讨用重组人甲状腺球蛋白(hTg)建立检测甲状腺球蛋白抗体(TgAb)的间接ELISA法及其临床意义。方法:用该法测定300例健康体检者(正常对照组)和各种甲状腺疾病患者484例;其中桥本甲状腺炎(HT)219例,Graves’病(GD)212例(初诊91例、复诊121例),非自身免疫性甲状腺疾病(AITD)患者53例。结果:确立本法正常界限值为0.74(A405值),血清TgAb水平HT组与正常对照组、非AITD组、GD组相比差异均有统计学意义(均P<0.01);GD组与正常对照组、非AITD组相比差异均有统计学意义(P<0.05或P<0.01);非AITD组与正常对照组相比差异无统计学意义(P>0.05)。HT、GD初诊、GD复诊患者的阳性率分别为70.8%、38.8%和28.6%,3类非AITD患者阳性率均不超过5.3%。结论:以重组hTg为抗原建立检测TgAb的ELISA法有助于各种甲状腺疾病的临床鉴别。