Effect of Dihydrotestosterone on CostimulatoryMolecules in a Mouse Model of Graves' Disease / 中国医学科学杂志(英文版)

Objective Graves' disease is the most common autoimmune thyroid disease and its prevalence and clinical manifestations are disparate between females and males. Costimulatory molecules play an essential role in regulating autoimmune responses. The objective of this study was to determine if expression of inhibitory molecules was correlated with treatment by dihydrotestosterone (DHT) in an BALB/c mouse model of experimental autoimmune Graves' disease.Methods Female BALB/c mice were immunized three times with thyroid stimulating hormone receptor A-subunit encoded by adenovirus to establish a Graves' disease model. Three different doses of DHT or a matching placebo were administered by implantation of slow-release pellets a week before the first immunization. Four weeks after the third immunization, the mice were euthanatized, and then the spleen and thymus were removed. Total thyroxine and free thyroxine levels in serum of mice were detected using a radioimmunoassay kit. Real-time polymerase chain reaction was performed to estimate the expression of costimulatory molecules in lymphocytes from the spleen and thymus. Flow cytometry was used to analyze the percentage of CD4 T cells in splenic lymphocytes. Quantitative data were compared with unpaired -tests. Correlation between two variables was analyzed using Analysis of Variance.Results Treatment with DHT can dramatically reduce total thyroxine and free thyroxine levels. Higher expression of programmed death-1 was found in the spleen of Graves' disease mice receiving 5 mg of DHT treatment (0.635±0.296 . 0.327±0.212; =2.714, =0.014), similarly, T-cell immunoglobulin domain and mucin domain 3 (TIM-3) in both the spleen (1.004±0.338 . 0.646±0.314; =2.205, =0.022) and the thymus (0.263±0.127 . 0.120±0.076; =3.221, =0.004) also increased after 5 mg of DHT treatment compared with the parallel placebo model mice. Moreover, the percentage of CD4 T cells declined in the splenic lymphocytes of Graves' disease mice treated with 5 mg of DHT (19.90%±3.985% . 24.05%±2.587%; =2.804, =0.012). A significant negative association was observed between expression of TIM-3 in the spleen and serum levels of total thyroxine (=-0.7106, =0.014) as well as free thyroxine (=-0.6542, =0.029).Conclusion This study demonstrates that DHT can ameliorate experimental autoimmune Graves' disease, which may occur by up-regulating expression of programmed death-1 and TIM-3 and inhibiting development of CD4 T cells.

Glycosylation of recombinant human thyroid peroxidase ectodomain of insect cell origin has little effect on recognition by serum thyroid peroxidase antibody / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Thyroid peroxidase (TPO) is an important autoantigen in Hashimoto's thyroiditis (HT), and almost all epitopes are located in TPO ectodomain. The glycosylation of TPO might contribute to breaking self-tolerance, therefore, purified glycosylated recombinant TPO ectodomain is prerequisite of elucidating its role in the pathogenesis of HT. The aim of our study was to investigate whether the glycosylation has influence on the antigenic determinants of recombinant TPO.</p><p><b>METHODS</b>Bac-to-Bac baculovirus expression system was used to generate recombinant human TPO ectodomain. The antigenicity was analyzed by antigen specific enzyme-linked immunosorbant assays (ELISAs). The glycosylation of recombinant human TPO ectodomain of High Five insect cell origin was detected by lectin-ELISAs.</p><p><b>RESULTS</b>TPO ectodomain was recovered from the culture media as a soluble protein, and it was fused with a hexahistidine tag which allowed purification by nickel-affinity chromatography. The recombinant TPO ectodomain could be recognized by all the 54 HT patients and three TPO monoclonal antibodies. Fucose, sialic acid and galactose were all detected on the recombinant TPO ectodomain. Sera TPOAb binding decreased slightly after non-specific deglycosylation of TPO by periodic acid.</p><p><b>CONCLUSIONS</b>High Five insect cells derived recombinant human TPO ectodomain had N-glycosylation sites, which might have little effect on recognition by serum TPOAb.</p>

Surgical treatment of hyperthyroidism / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>In order to explore the management of peri-operation and the therapeutic effect in the surgical treatment of hyperthyroidism.</p><p><b>METHODS</b>Fifty five cases of hyperthyroidism were undergone near-total thyroidectomy, during the operation recurrent laryngeal nerve was exposed, and the parathyroid was found with microscope when necessary. The third rank of inferior thyroid arteries were ligated to guarantee the blood supply for parathyroid.</p><p><b>RESULTS</b>All cases underwent near-total thyroidectomy. There was no mortality, and no permanent recurrent laryngeal nerve palsy occurred, and no permanent hypoparathyroidism, and no recurrent hyperthyroidism. Follow-up was carried out 16 months to approximately 5 years after near-total thyroidectomy patients, Hypothyroidism occurred in 15 cases (57.7%), serum calcium levels were 2.15-2.45 mmol/L.</p><p><b>CONCLUSIONS</b>Special attention should be given to the management of peri-operation, the above the method can prevent operative complication in the surgical treatment of hyperthyroidism, with excellent result.</p>

Study on TR beta gene mutation in a thyroid hormone resistance syndrome family / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To detect the gene mutation of thyroid hormone receptor beta (TR beta) in a family with thyroid hormone resistance syndrome.</p><p><b>METHODS</b>The genomic DNA was extracted from peripheral blood leukocytes of the patient, 14 family members and 7 healthy subjects. The exons 7-10 of TR beta gene were amplified by PCR. The products of PCR were purified and sequenced directly to detect the gene mutation.</p><p><b>RESULTS</b>Five members of this family were confirmed to have the C to G transition mutation at nucleotide 1642 site within exon 10 of TR beta gene, which was a missense mutation causing the substitution of Proline to Alanine (P453A); and also to have the C to T transition mutation at nucleotide 1020 within exon 7 of TR beta gene, which was a synonymous mutation that didn't cause the change of amino acid at this position (F245F). The two mutations were heterozygote. No mutation in the exons 7-10 of TR beta gene were identified in other family members.</p><p><b>CONCLUSION</b>A family with thyroid hormone resistance syndrome caused by TR beta gene mutation is first founded in Chinese people.</p>

Variation of CD4~+ CD25~+ regulatory T cells in patients with Graves' disease / 中华内分泌代谢杂志

The level of CD4~+ CD25~+ regulatory T cells was assayed in the peripheral blood from the patients with Graves' disease (GD) by floweytometry.CD4~+ CD25~+ regulatory T cells as a kind of non-specific immune cells appear to have no relationship with thyroid function,however,their lowered number in early GD patients suggests that they seem to play an essential role in the pathogenesis of GD.