小鼠甲状腺球蛋白的提取、纯化和鉴定

目的 进行小鼠甲状腺球蛋白的纯化和鉴定。方法 小鼠甲状腺组织匀浆后进行盐析和Sphadex G-200凝胶层析2次纯化,纯化后的蛋白质产物经十二烷基磺酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和Western免疫印迹鉴定。结果 经纯化的蛋白产物为高纯度的甲状腺球蛋白。结论 Sphadex G-200凝胶层析和Western免疫印迹是进行甲状腺球蛋白纯化和鉴定的一种较好的方法。     

Premenarchal and postmenarchal girls with insulin-dependent diabetes mellitus: ovarian and other organ-specific autoantibodies, menstrual cycle

STUDY OBJECTIVE: To estimate various organ-specific autoantibodies and detect other endocrine autoimmune disorders and menstrual cycle characteristics in girls with Type 1 insulin dependent diabetes mellitus (IDDM). DESIGN: Prospective cohort study from 1993 to 1998, duration 4.5 years. SETTING: Diabetes & Endocrine Clinic of the University Hospital, Motol, Prague. PATIENTS: 53 IDDM girls (group A--43 postmenarchal, group B--10 premenarchal), 15.5 +/- 2.5 (8-19) years old, 6.2 +/- 4.3 years after IDDM onset. MAIN OUTCOME MEASURES: Ovarian autoantibodies directed to ooplasm, zona pellucida, membrana granulosa, theca folliculi interna, and lutein cells, insulin autoantibodies, thyroid peroxidase and thyroglobulin autoantibodies. Menstrual cycle character, endocrine glands disturbance. Diabetes control, body mass index, duration of IDDM. RESULTS: Ovarian autoantibodies in at least one of the followed structures were found in 67.9% of the IDDM girls. In the control group of 21 healthy girls of corresponding age, the positive findings in lutein cells were found in only 4.8% of the girls (P < 0.01 versus IDDM girls). The lutein cells commonly associated with theca folliculi interna cells were the most frequent immunopositive structures in diabetic girls (P < 0.05 versus another positive ovarian autoimmune structure). Autoantibodies directed to ovarian steroid producing cells were frequent in IDDM patients with both irregular and normal menstrual cycles. Irregular menstrual cycles were diagnosed in 27.9% of IDDM girls, polymenorrhea in half of them, and oligomenorrhea in the remainder. Diabetes control in our patients (glycosylated hemoglobin HbA1c in postmenarchal girls 10.1 +/- 2.0%) did not differ between those with regular and those with irregular menstrual cycles. Over a follow-up period one-third of the girls with oligomenorrhea and a long-term noncompliance (HbA1c 13.5%) developed secondary amenorrhea. Insulin autoantibodies were found in 67.8%, thyroid peroxidase autoantibodies in 12.5%, and thyroglobulin autoantibodies in 10.4% of the IDDM girls. Autoimmune thyroiditis was diagnosed in 5 IDDM patients (9.4%); hypothyroidism developed in 3 of them. Menstrual cycle was irregular in 4 of the 5 girls with autoimmune thyroiditis (polymenorrhea in 1, oligomenorrhea in another 3 girls). CONCLUSIONS: An increased incidence of various circulating autoantibodies may be markedly demonstrated in IDDM girls. Their reproductive function might have an important relationship to an evidence of ovarian autoantibodies. Menstrual cycle disturbances could be linked to the poor diabetes control, to the presence of ovarian and other autoantibodies, and also to other autoimmune disease.     

Serum thyroglobulin as a marker of thyroid neoplasms after childhood cancer

AIM: To evaluate serum thyroglobulin (Tg) level as a marker of the development of thyroid disease when following individuals who received neck irradiation therapy in childhood. METHODS: In a non-randomized cross-sectional study Tg was assessed in 172 survivors of childhood cancer 10.8 y (1.9-24) median (range) after diagnosis and 7.9 y (0.9-24.3) median (range) after the end of treatment. The patients were divided into two groups: group 1 included 47 patients who had received irradiation to the neck and group 2 included 125 patients who did not receive irradiation to the neck. RESULTS: Patients who had received irradiation to the neck had significantly higher Tg levels compared with those who did not receive neck irradiation: median 14.0 (1.0-189.0) microg/L vs median 8.8, (0.7-112.2) microg/L (p < 0.001). Six out of seven patients with elevated Tg levels (>70 microg/L) had received neck irradiation. Among these six patients, two patients developed secondary differentiated thyroid cancer and two patients developed benign thyroid neoplasms. None of the patients who had normal levels of Tg developed thyroid cancer. CONCLUSION: A high Tg level should be a cause for further investigation in the follow-up of individuals who have received irradiation therapy in childhood.     

The usefulness of detecting thyroglobulin in fine-needle aspirates from patients with neck lesions using a sensitive thyroglobulin assay

The aim of this study was to assess the diagnostic utility of thyroglobulin (Tg) in fine needle aspirates (Tg-FNAB) of nonthyroidal neck masses using a sensitive in-house method for detecting Tg in washout specimens. A total of 256 samples from 145 patients were evaluated for Tg in washout specimen from FNAB and compared to corresponding cytological smear and histology of 46 surgical specimens. Tg was measured by a sensitive in-house time-resolved immunofluorometric assay. The sensitivity for Tg-FNAB alone or in combination with cytological findings was found to be 100% in both the follow-up group and before primary surgery. In the follow-up group the specificity of Tg-FNAB was 100%. Fifty-nine of 60 follow-up specimens with malignant cytology were Tg-FNAB positive (n = 195). Histological examination of one lymph node with malignant cytology and negative Tg-FNAB showed metastasis from carcinoma of the salivary gland. Tg-FNAB was positive in 25 specimens with suspicious or cystic cytology. Tg-FNAB values were high (median 4557 microg/l, range 122-37200 microg/l) in washout specimen from cystic metastasis from which cytology did not confirm malignancy. Of the 20 lymph nodes with histology confirming metastasis from differentiated thyroid carcinoma (DTC), the Tg-FNAB was positive in 19 and intermediate in one. However, before primary surgery, two Tg-FNABs were false positive compared to the histology of the lymph nodes. TgAb in serum did not interfere with FNAB-Tg measurements. Tg-FNAB measurement is accurate with high sensitivity (100%) and of great importance in detecting cystic metastasis when cytology is not conclusive. Even metastases to small neck lymph nodes may be detected by using sensitive Tg-assay. Serum thyroglobulin antibodies appear to have ignorable effect on the clinical performance of Tg-FNAB.     

The effect of vitamin D on thyroid autoimmunity in euthyroid men with autoimmune thyroiditis and testosterone deficiency

BackgroundAutoimmune (Hashimoto’s thyroiditis) is characterized by a strong female preponderance, which may suggest that sex hormones have an impact on thyroid autoimmunity. The aim of this study was to investigate whether testosterone determines vitamin D action on thyroid antibody titers and thyroid function tests in men with autoimmune thyroiditis and low testosterone levels.MethodsThe study included 36 men with testosterone deficiency, 17 of whom had been treated for at least 26 weeks with oral testosterone undecanoate (120 mg daily). Because of coexistent euthyroid Hashimoto’s thyroiditis, all participants were then treated with vitamin D (100 μg daily). Serum titers of thyroid peroxidase and thyroglobulin antibodies, serum levels of thyrotropin, free thyroid hormones, testosterone and 25-hydroxyvitamin D, as well as Jostel’s thyrotropin index, SPINA-GT and SPINA-GD were assessed before vitamin D treatment and 26 weeks later.ResultsWith the exception of testosterone levels, there were no significant differences between both study groups in serum hormone levels, antibody titers and thyroid function tests. All participants completed the study. In addition to increasing 25-hydroxyvitamin D levels, vitamin D increased SPINA-GT and reduced thyroid peroxidase and thyroglobulin antibody titers. In testosterone-treated men, vitamin D increased testosterone levels. Vitamin D did not affect serum levels of thyrotropin, free thyroid hormones, Jostel’s thyrotropin index and SPINA-GD. Treatment-induced changes in thyroid antibody titers and SPINA-GT were more pronounced in testosterone-treated than testosterone-naïve men.ConclusionsThe obtained results suggest that the beneficial effect on thyroid autoimmunity and thyroid secretory function is stronger in men receiving testosterone therapy.     

术后分化型甲状腺癌131Ⅰ清甲治疗疗效及影响因素分析

目的：探讨手术后分化型甲状腺癌131Ⅰ清甲治疗的临床疗效及影响因素.方法：对376例分化型甲状腺癌患者行131Ⅰ治疗,并进行2-10年随访,以全身131Ⅰ扫描,甲状腺摄131Ⅰ率、血清TG浓度来评判清甲治疗疗效.结果：376例患者中266例1次131Ⅰ消融成功,1次成功率70.74％.1次消融有效病例333例,有效率为88.56％.消融无效病例43例,无效率11.44％.不同病理类型间一次消融率无统计学差异,不同治疗时间一次消融成功率有统计学差异.结论：甲状腺外科手术＋ 131Ⅰ＋ TSH抑制治疗的综合治疗方法在分化型甲状腺癌的治疗中有重要作用,外科手术方式和手术后到131Ⅰ治疗时间是疗效的重要影响因素.     

Defective Thyroglobulin Endocytosis and Hydrolysis in Thyroid Cold Nodules1

Abstract. The catabolic phase of thyroid secretion has been studied by incubating thyroid slices with 0.1%¹²⁵I thyroglobulin (Tg) in the following cases: 11 follicular adenomas (FA), 8 hyerplastic nodules in multinodular glands (NM), 3 carcinomas (follicular or papillary, FC., PC) and normal (N) or micronodular tissue (M). Tg pinocytosis is severely impaired in all cold nodules (25 to 60 % reduction). In the follicular tumours (FA, FC), Tg hydrolysis is more affected than would be expected based only on reduced Tg endocytosis: the hydrolysis products remain in the phagolysosomes instead of being secreted. As far as the acid hydrolases are concerned these enzymatic activities are increased in nearly all nodules, in particular in follicular adenomas and carcinomas.     

3种血清标志物检测在桥本甲状腺炎中的诊断探讨

目的探讨甲状腺过氧化物酶抗体(anti-TPO)、甲状腺球蛋白抗体(anti-Tg)、促甲状腺激素受体抗体(TRAb)水平对桥本甲状腺的临床诊断意义。方法收集2014年1月至2015年12月在本院已确诊桥本甲状腺炎(HT)患者89例,根据其甲状腺功能分为甲状腺功能低下组(HT1)21例,甲状腺功能亢进组(HT2)9例,甲状腺功能正常组(HT3)59例;临床确认为Graves病(GD)35例;单纯性结节性甲状腺肿(SG)40例;80例来本院健康体检者作为对照组,其中男18例,女62例。用电化学发光免疫分析(CLIA)方法测定5组anti-TPO、anti-Tg、TRAb水平并统计分析。结果不同疾病患者血清中的anti-TPO、anti-Tg和TRAb水平及阳性率间差异均有统计学意义(P0.05);在HT患者中,HT1组anti-TPO水平高于HT2及HT3组,anti-Tg水平在HT1、HT2、HT3组之间差异无统计学意义(P0.05),anti-TPO和anti-Tg水平在HT组高于GD组,GD组高于SG组及健康对照组,且差异有统计学意义(P0.05),但SG组与健康对照组差异无统计学意义(P0.05)。TRAb阳性率在GD组最高,与其他4组阳性率比较,差异均有统计学意义(P0.05)。结论联合检测anti-TPO,anti-Tg是诊断HT的重要指标,TRAb对鉴别诊断HT与GD有重要意义。     

Prognostic value of postsurgical stimulated thyroglobulin levels after initial radioactive iodine therapy in well-differentiated thyroid carcinoma

BACKGROUND: In well-differentiated thyroid carcinoma, predictors of future positivity of stimulated thyroglobulin (>2 microg/L) after initial radioactive iodine treatment are not known. METHODS: In a retrospective study, we used logistic regression analysis to determine whether postoperative stimulated thyroglobulin measurements and pathologic stage independently predict future stimulated thyroglobulin positivity. RESULTS: We followed 141 patients with well-differentiated thyroid carcinoma for a median of 35 months; follow-up stimulated thyroglobulin measurements were positive in 20.6% (29/141). The natural logarithm of the postsurgical stimulated thyrogolobulin was independently associated with a positive stimulated thyroglobulin at long-term follow-up (odds ratio [OR], 4.44; 95% confidence interval [CI], 2.33-8.45; p < .001); there was a trend for a positive association of TNM stage with positive follow-up stimulated thyroglobulin (p = .054). Lymph node positivity predicted a positive stimulated thyroglobulin in papillary cancer. CONCLUSIONS: Stimulated thyroglobulin measurements prior to initial radioactive iodine treatment independently predict future stimulated thyroglobulin positivity in well-differentiated thyroid carcinoma.