血清TSH水平轻微增高的新生儿190例随访分析

对 1998 ,1999年出生时血清TSH在 5～ 2 0mU/L范围的新生儿于 2 0 0 2年 4月通知来院复查 ,实际召回复检小儿 190例。结果 1例血TSH为 8.8mU/L ,可列为亚临床甲状腺功能减退Ⅱ级。     

重组人TSHR片段蛋白建立人TRAb ELISA及临床应用

Objective Thyrotropin receptor antibodies(TRAb)include thyroid stimulating antibody (TSAb) and thyroid stimulating blocking antibody(TSBAb).It is very helpful to detect seFum TSAb and TSBAb levels in patients with thyroid diseases.The aim of this study was to establish two enzyme linked immunosorbent assay(EUSA)methods(N method and C method)that may determine serum TSAb and TSBAb levels respectively.The initial clinical application in thyroid diseases was also investigated.Methods Recombinant human thyroid stimulating hormone receptor(TSHR)-ecd N-terminal fragment was used in N method and C-terminal fragment was used in C method as antigens respectively.Two ELISA methods for detection of TSAb and TSBAb were established and evaluated.The absorbance value A405 was used as the quantitative parameter to reflect the level of TRAb.The serum TSAb and TSBAb levels were measured in 229 healthy controls,and 675 patients.They were 451 untreated Graves disease(GD),104 untreated chronic lymphocytic thyroiditis(Hashimoto disease,HT)complicating hypothyroidism,35 simple goiters,47 nodular goiters and 38 subacute thyroiditis.The ANOVA and X2-test were used for statistic data analysis.Results N method:(1)the serum TRAb levels measured with N method were positively correlated with commercial TSAb kit(r=0.7422.P＜0.05).(2)The intra-assay coefficient of variation(CV)and the inter-assay CV were 3.96%-4.67% and 7.32%-8.33%,respectively.(3)A405 (x-±s)value in healthy group was 0.44±0.15 with a cut-off value(x-+2s)of 0.74.(4)In untreated GD group,A405 was 0.93±0.28,which was significantly higher than that in healthy group(F=2.0341,P=0.001).The positive rate was 82.8%.(5)In untreated HT complicating hypothyroidism group,A405 was 0.64±O.23,which was significantly higher than that in healthy group(F=1.7822,P=0.003).The positive rate was 62.1%.C method:(1)intrs-assay CV and inter-assay CV were 3.24%-4.72% and 7.22%-8.55%,respectively.(2)The measured A405 value in healthy group was 0.56±0.11 with a cut-off valRe(x-+2s) of 0.78.(3)In untreated HT complicating hypothyroidism group,A405(x-±s) was 1.17±0.26,which was significantly higher than that in healthy group(F=3.0333,P=0.002).The positive rate was 93.4%.(4)In untreated GD group,A405 was 0.74±0.23,which was significantly higher than that in healthy group(F=1.8339,P=0.03).The positive rate was 66.3%.There were no significant difierences among simple goiter,nodular goiter,subacute thyroiditis and healthy groups(N method:F=1.6134,1.5772,1.6208,C method:F=1.5910,1.5369,1.6055,all P＞0.05)assayed with both N method and C methods.Conclusions The present novel N method may mainly measure TSAb level and C method mainly measure TSBAb levels.The two ELISA methods showed to be accurate,stable and convenient.It may be helpful for clinical evaluation in GD,HT and other thyroid diseases.     

全球主要甲状腺学会

目前全世界范围内主要有4个致力于甲状腺生物学及甲状腺疾病研究的学会.这些学会分布在亚洲/澳大利亚、欧洲、北美洲和南美洲.     

甲状腺过氧化物酶B细胞抗原决定簇研究进展

甲状腺过氧化物酶(TPO)作为一种重要的甲状腺自身抗原，在自身免疫性甲状腺疾病(AITD)的发病机制中占有重要地位。大多数TPO-B细胞抗原决定簇是高度构象性的，并集中于一个有限的区域——免疫优势区(IDR)中，而TPO表位“指纹”也具有一定的指导意义。IDR定位仍不清楚，可能主要由TPO中MPO样区与CCP样区相接的区域高度折叠而构成。TPO中也存在少量的非IDR B细胞抗原决定簇和线性决定簇。TPO—B细胞抗原决定簇及IDR构成、分布、定位等问题的解决很可能有助于阐明AITDs的发病机制，并为开辟特异性分子治疗途径提供理论基础。     

人甲状腺过氧化物酶膜外区基因的克隆及其杆状病毒表达载体的构建

摘要 目的：尝试克隆人甲状腺过氧化物酶（hTPO）膜外区基因并构建其杆状病毒表达载体,为其在昆虫细胞中的表达打好基础。方法：PCR扩增hTPO膜外区基因,并将其先后重组入pGEM3zf(+)质粒和pFastBac1质粒,以hTPO-pFastBAC1质粒转染E.coli DH10Bac大肠杆菌,获得重组hTPO杆状病毒表达载体（hTPO-Bacmid）,分别以多种方法鉴定其正确性。结果：经PCR及酶切、基因测序等方法证实得到的重组hTPO-pGEM3zf(+)质粒、hTPO-pFastBAC1质粒和hTPO-Bacmid均与预期相符。结论：成功的克隆了hTPO膜外区基因,并将其正确重组入pGEM3zf(+)质粒和pFastBac1质粒,后者经位点特异性转座整合至Bacmid,成功的构建了重组hTPO杆状病毒表达载体,为进一步实现hTPO在昆虫细胞中的真核表达作好了准备。     

ICAM-1与内分泌和代谢病关系的研究进展

细胞问粘附分子-1(ICAM-1)有着十分重要的生理、病理作用,它的变化与很多疾病都密切相关,本文综述了近年来国外学者对于ICAM-1与自身免疫性甲状腺疾病、高脂血症以及糖尿病并发症的研究,发现:ICAM-1/淋巴细胞功能相关抗原-1(LFA-1)途径介导了Graves病和桥本甲状腺炎的甲状腺组织损伤,Graves眼病的球后炎症反应,高脂血症、动脉粥样硬化及糖尿病所致的血管内皮损伤。     

糖尿病大鼠甲状腺组织TG,TPO,TSHR与RAGE mRNA表达的实验研究

目的 探讨糖尿病大鼠甲状腺组织甲状腺球蛋白(TG)、甲状腺过氧化物酶(TPO)、促甲状腺激素受体(TSHR)与晚期糖基化终末产物受体(RAGF)mRNA表达的变化.方法 Wistar大鼠112只,随机分为糖尿病组(DM 组)27只、糖尿病氨基胍治疗组(AG组)28只、糖尿病胰岛素治疗组(INS组)32只和对照组(N组)25只,采用逆转录-聚合酶链反应检测造模后12和20周各组大鼠甲状腺组织TG、TPO、TSHR和RAGE mRNA的表达.结果 TG mRNA表达:12周各组之间无统计学差异(P>0.05).20周DM组低于AG组(P<0.01)、INS组(P<0.01)和N组(P<0.01).后3组间无统计学差异(P>0.05).TPO mRNA表达:12周DM组低于AG组(P<0.05)、INS组(P<0.001)和N组(P<0.001),AG组低于INS组(P<0.001)和N组(P<0.001),INS组低于N组(P<0.01).20周DM组低于INS组(P<0.01)和N组(P<0.001),与AG组无统计学差异,AG组低于INS组(P<0.05)和N组(P=0.001),INS组与N组无统计学差异.TSHR mRNA表达:12周DM组高于AG组(P<0.05)、INS组(P<0.05)和N组(P<0.01),后3组间无统计学差异(P>0.05).20周各组之间差别无统计学意义(P>0.05).RAGE mRNA表达:12周和20周均表现为DM组高于AG组(P<0.001)、INS组(P<0.01)和N组(P<0.001),INS组高于AG组(P<0.05)和N组(P<0.01),AG组与N组无统计学差异.结论 糖尿病大鼠甲状腺组织TG mRNA、TPO mRNA表达降低,TSHR mRNA表达增高,可能与高糖毒性所致甲状腺组织RAGE mRNA表达增高有关.     

siRNA体内导入策略

以small interfering RNA (siRNA)为基础的RNA 干扰(RNA interfering,RNAi),是指当与内源性mRNA编码区某段序列同源的双链RNA(dsRNA)导入细胞后,该mRNA发生特异性降解,而导致该基因表达的沉寂.是一种dsRNA分子在mRNA水平上关闭相应序列基因的表达或使其沉默的过程,也就是序列特异性的转录后基因沉默。     

糖尿病大鼠甲状腺组织NISmRNA表达的变化

目的:探讨糖尿病(DM)大鼠甲状腺组织钠,碘转运体(NIS)mRNA表达的变化及胰岛素、氨基胍对其的干预作用.方法:高脂饲料喂养联合链脲佐菌素制备DM大鼠49只,随机分为DM组17只、胰岛素十预组(DI)16只、氨基胍干预组(DA)16只,设正常对照组(NC)9只,成模12周末处死,取甲状腺组织,以半定量RT-PCR方法测定NISmRNA的表达.结果:DM组大鼠甲状腺组织NISmRNA与GAPDH mRNA表达水平比为0.17±0.09显著低于NC组的0.47±0.16(P<0.05),经胰岛索或氨基胍干预后DI组与DA组大鼠甲状腺组织NIS mRNA与GAPDH mRNA表达水平比分别为0.45±0.20、0.40±0.17显著高于DM组(P<0.05).结论:高糖毒性可能通过非酶糖基化途径导致DM大鼠甲状腺组织NIS mRNA表达下降.     

Effect of atorvastatin and pravnstatin on insulin synthesis from islet β cell in rat and its mechanisms

Objective To evaluate the effect of atorvastatin and pravastatin on insulin synthesis from islet β cell in rat and its mechanisms.Methods In the experiment,it was divided into control group,atorvastatin group and pravastatin group.Pancreatic islets were isolated by the collagenase method.Cultured with 100 μmol/L atorvastatin or pravastatin for 24 hours respectively,the alteration of insulin content was measnred by RIA,and the expression of insulin mRNA from rats'islet β cells was accessed by quantitative PCR.The human insulin promoter-luciferase vector was constructed and transfected to MIN6 cells by using Lipofectamine 2000.Cultured with 100 μmol/L atorvastatin or pravastatin for 24 and 48 hours respectively,the activity of lnciferase was measured by the dual-luciferase reporter assay system to evaluate the activity of insulin promoter.Difference between the three groups was determined by one-way analysis of variance.Results Cultured with 100 μmol/L atorvastatin for 24 hours,the insulin content was significantly decreased to 76.3%of the control group(P<0.05),the mRNA expression levels[(0.125 232 ±0.014 827)vs.(0.264 896±0.029 541),P<0.05]and the activity of luciferase[(0.763 616±0.253 780)vs.(1.290 601±0.386 566),P<0.05]were all significantly inhibited compared with the control group,but the inhibitory effects were not shown in the pravastatin group.Conclusion In a higher concentration,atorvastatin may reduce the synthesis of insulin by inhibiting the expression of insulin mRNA of islet β cells.This effect is relative to its inhibition on the activity of insulin promoter,and the degree of inhibition is relative to its lipophilicity.