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Objective:Exploring the role of thyrotropin receptor(TSHR) in lipotoxicity-induced thyroid function damage.Methods:Rat thyroid follicular epithelial cells(RTC) were stimulated with different doses of palmitic acid(PA), and the lipid content of the cells was observed through Oil Red O staining. The expression levels of TSH receptor(TSHR), Ttf1, and SSBP1 mRNA and protein in each group were detected using RT-PCR and Western blot. The TSHR protein level in the cell culture supernatant was measured using ELISA. Membrane TSHR was assessed through immunofluorescence and compared with the control group. We used PA to stimulate the TSHR over-expression(TSHR OE) and normal RTC, as PA+ TSHR OE group and PA group respectively, then testing Tg mRNA and protein, cAMP and Tg in cell supernatants levels, then comparing with the control.Results:RTC were stained into peau d′orange in PA groups. Compared with the control group, we found TTf1, SSBP1 and TSHR mRNA as well as protein levels in PA groups were decreased(all P<0.05), TSHR of the cell membrane and supernatants were reduced(all P<0.05), characterizing dose-dependent changes partly. Moreover, we found in PA group Tg mRNA level was downregulated( P<0.05), Tg protein levels were reduced in the supernatants and cells( P<0.05), cAMP level was decreased in cells( P<0.05); in TSHR OE group, Tg mRNA level was upregulated( P<0.05), Tg protein levels in cells and supernatants were increased(all P<0.05), cAMP level was similar. Compared with the PA group, we found in PA+ TSHR OE group Tg mRNA level was upregulated( P<0.05), Tg protein levels were increased in the supernatants and cells(all P<0.05), cAMP level was elevated in cells( P<0.05). Conclusion:PA induces lipid deposition in RTC, decreased synthesis and secretion of Tg. This effect is likely achieved through the downregulation of the TSHR/cAMP signaling pathway.
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Objective:To investigate the serum 25-hydroxyvitamin D [25(OH)D] level in patients with Graves' disease (GD) and its correlation with thyrotropin receptor antibody (TRAb) and bone metabolism markers.Methods:A total of 124 patients with newly diagnosed or relapsed GD were selected and divided into three groups according to serum 25(OH)D level, namely vitamin D deficiency group with 25(OH)D <12 μg/L, vitamin D insufficiency group with 25(OH)D of 12 to 20 μg/L, and vitamin D sufficiency group with 25(OH)D ≥ 20 μg/L. The levels of serum 25(OH)D, TRAb, type I procollagen N-terminal pro-peptide (PINP), type I collagen cross-linked C-terminal peptide (S-CTX), parathyroid hormone (PTH), total triiodothyronine (TT 3), total thyroxine (TT 4) and thyroid-stimulating hormone (TSH) were measured in all patients, and the differences of these biochemical indices were compared across groups. Oneway analysis of variance or Kruskal-Wallis test was used for comparison between groups, and Pearson or Spearman correlation analysis was applied for correlation test. Results:The levels of serum TT 3, TT 4, PINP, and S-CTX significantly increased ( P < 0.01) and the level of phosphorus (P) decreased ( P < 0.01) with the decreased vitamin D levels. The levels of PTH and calcium (Ca) were significantly lower in the vitamin D sufficiency group compared with the vitamin D insufficiency group and vitamin D deficiency group ( P < 0.01). Correlation analysis showed that serum 25(OH)D level was negatively correlated with the levels of TT 3, TT 4, PINP, S-CTX, PTH and Ca ( P < 0.01), and positively correlated with the levels of P and TSH ( P < 0.01). Conclusions:Decreased serum 25(OH)D level is closely related with increased bone turnover, PTH, and thyroid hormone levels in patients with GD, but not related with TRAb. Thyroid hormone levels have a certain predictive value regarding vitamin D deficiency in GD patients. It is necessary to monitor the vitamin D levels in patients with GD and provide vitamin D supplementation to reduce the incidence of osteoporosis, improve the effectiveness of antithyroid treatment and reduce the recurrence of GD.
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Objective:To investigate the correlation between the level of thyrotropin receptor antibody(TRAb) and bone turnover markers(BTMs) in the patients with newly-diagnosed Graves′ disease(GD).Methods:The clinical data of GD patients who were newly-diagnosed in the First Affiliated Hospital of Zhengzhou University from October 2016 to June 2021 were collected, including free triiodothyronine(FT 3), free thyroxine(FT 4), thyroid stimulating hormone, thyroid related antibodies, N-terminal procollagen of type I collagen(PINP), N-terminal osteocalcin(N-MID), β-cross-linked C-telopeptide of type I(β-CTX), blood lipid and renal function, etc. Results:There were 618 GD patients with an average age of(43.7±13.2) years(male∶female=1∶1.99). The PINP and β-CTX level in male GD patients were significantly higher than those in female(all P<0.05). Spearman correlation analysis showed that PINP, N-MID and β-CTX were positively correlated with FT 3, FT 4, TRAb, serum calcium and serum phosphorus; and negatively correlated with body mass index and low density lipoprotein cholesterol(all P<0.05). Linear regression analysis showed that TRAb was positively correlated with lg-PINP, lg-N-MID and sqrt-β-CTX in the univariate model of total GD patients( β were 0.006, 0.005, and 0.006, respectively; all P<0.001); positive correlation remained after adjusting for thyroid function(all β=0.004, all P<0.001); and for multiple confounding factors(model 3 and 4, all P<0.05). Results of univariate and adjusted thyroid function models with GD in different genders were consistent with the total patients(all P<0.05). Conclusion:TRAb is a risk factor for accelerated bone turnover in GD patients which is independent of thyroid function.
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Objective:To observe the efficacy of methimazole (MMI) combined with 1α-hydroxyvitamin D3 (alfacalcidol, ALF) in patients with Graves disease of high-titer thyrotropin receptor antibodies (TRAb) and to explore new clinical strategies to reduce serum TRAb in Graves disease.Methods:120 patients with Graves disease initially diagnosed in Quanzhou First Hospital Affiliated to Fujian Medical University and the People′s Hospital Affiliated to Quanzhou Medical College from June 2017 to June 2019 were prospectively selected as the research objects. All patients received conventional dose of MMI for anti hyperthyroidism treatment. The patients were randomly divided into three groups: group A [ n=40, treated with MMI combined with high-dose ALF (0.5 μg/d)], group B [ n=37, treated with MMI combined with low-dose ALF (0.25 μg/d)] and group C ( n=43, treated with MMI only). The treatment lasted for 24 weeks. The serum free triiodothyronine (FT 3), free thyroxine (FT 4), thyroid stimulating hormone (TSH) and TRAb in patients before and after above treatments were detected. The blood routine, liver function, alkaline phosphatase (ALP), 25(OH)D, serum calcium (CA) and serum phosphorus were detected regularly. Results:After drug treatment: ⑴ the thyroid function of the three groups returned to normal. The average daily dosage of MMI in group A was significantly lower than that in group B and C ( P<0.05), and that in group B was also lower than that in group C ( P<0.05), with significant difference. After 24 weeks of treatment, the daily dosage of MMI in group A and B was significantly lower than that in group C ( P<0.05). ⑵ There was no significant difference in thyroid function among the three groups. The concentration of serum TRAb in group A was significantly lower than that in group B and C ( P<0.05), and that in group B was also lower than that in group C ( P<0.05). ⑶ During the 24 week follow-up, there was no significant difference in serum 25(OH)D, ALP, Ca and P among the three groups ( P>0.05); no leukopenia in peripheral blood and no abnormal liver function were found in the three groups. Conclusions:MMI combined with ALF can effectively treat Graves′ disease, reduce the dosage of MMI drugs, decline the level of TRAb in the serum of Graves′ patients, and improve the prognosis of Graves′ disease.
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Objective: To investigate the expression and expression efficiency of recombinant adeno-associated viral containing human thyrotropin receptor A subunit (rAAV2/9-hTSHR289-IRES-ZS Green) in mouse embryonic fibroblasts (NIH3T3) or in mice. Methods: NIH3T3 was infected by rAAV2/9-hTSHR289-IRES-ZsGreen and its TSHR289 protein expression was detected by Western blotting. Female BALB/c mice were intramuscularly injected with different doses of rAAV2/9-hTSHR289-IRES-ZsGreen; the expressions of the target protein in different organs were determined by immunofluorescence while the serum TSHR antibody (TRAb) titer was determined by radioimmunoassay. Results: NIH3T3 cells transfected with rAAV2/9-hTSHR289-IRES-ZsGreen expressed hTSHR289 protein both at 48 h and 72 h. The expression of target protein at 48 h or 72 h was significantly higher than that at 24 h (P<0.05). There were different levels of TSHR289 expression in leg skeletal muscle, heart, liver and spleen under fluorescence microscope. The results of radioimmunoassay showed that the higher dose injection produced a higher titer of TSHR antibody, but only the TRAb level in high dose and control injection exhibited a statistical difference (P<0.05) at week 4, week 8 and week 12. Furthermore, strong antibody response was observed in the mice injected with high dose and medium dose of rAAV2/9-hTSHR289-IRES-ZsGreen at week 4 and gradually weakened between 4 and 8 weeks. In addition, the antibody lasted for 12 weeks. Results: rAAV2/9-hTSHR289-IRES-ZsGreen can highly express hTSHR289 protein in vivo and in vitro, and the hTSHR289 protein displays strong immunogenicity. It indicates that rAAV2/9-hTSHR289-IRES-ZsGreen might become a more effective means of preparing Graves' disease model.
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Objective Higher expression of B-cell activating factor (BAFF) in patients with Graves' disease can activate B cells and increase proportion of plasma cells. However, the mechanism is still unclear. This study aims to investigate the effects of T3 on the BAFF level and plasma cell ratio in bone marrow, spleen and peripheral blood of mice, and to explore the mechanism of T3 in affecting the mature and differentiation of B cells. Methods 80 C57BL/6J mice were randomly divided into control group and T3 group, and were given isotonic saline or T3 5/10μg once a day for 6 weeks, respectively. The levels of T3 in peripheral blood of each group were measured with ELISA. Flow cytometry was used to detect the proportion of B220+CD138+ plasma cells and IgM, IgG and IgD expression of B cells in the spleen, bone marrow and peripheral blood. Immunohistochemistry, Western blot and PCR were performed to determine the expression of BAFF in spleen and thyroid. ELISA was used to determine the expression of BAFF in peripheral blood. Results Compared with control group, the levels of T3 in peripheral blood, diet and drinking water in the T3 group were significantly increased after 6 weeks T3 intervention. The mRNA and protein expression of BAFF in spleen mononuclear cells of T3 group (2.03±0.52, 0.50±0.03) were higher than those in control group (1.06±0.19, 0.05±0.01) (P<0.01). HE staining showed that the white medulla in the spleen of the T3 group increased and merged. Flow cytometry indicated that the proportion of spleen plasma cells and antibody expression of B cells in T3 group [(3.92±1.55)%, (75.76±8.88)%] increased compared with control group [(2.43±1.18)%, (65.26±8.38)%] (P<0.05); Proportion of bone marrow plasma cells [(8.48±3.62)%] and antibody expression [(40.63±18.96)%] in T3 group were significantly increased compared with control group [(4.96±3.11)%, (22.89±7.32)%](P<0.05); Peripheral plasma cell ratio [(8.56±4.27)%] and antibody expression [(76.15±9.44)%] were lower than those in control group [(14.70±4.76)%, (84.20±3.98)%](P<0.05); Compared with control group [(5.98±0.78) pg/mL], the BAFF level in peripheral blood increased [(7.61±1.72) pg/mL] (P<0.01). Immunohistochemistry showed that the expression of BAFF increased in mononuclear cells of thyroid of the T3 group. Conclusion T3 could activate BAFF expression in bone marrow, spleen, peripheral blood and thyroid mononuclear cells, and induce differentiation of bone marrow and spleen B cells, thus causing pathological changes in thyroid tissue. Such mechanisms might play an important role in the pathogenesis of thyroid autoimmune diseases.
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Objective: To construct an adeno-associated viral vector serotype2/9 containing human thyrotropin receptor (rAAV2-9-hTSHR289-IRES-ZsGreen) so as to provide a better means for establishing an ideal animal model and the gene prevention against Graves' disease. Methods: AAV skeleton plasmid pAAV-IRES-ZsGreen and PDC315 plasmid containing hTSHR289 were digested by EcoR+BamH. The digestion products were connected. And pAAV-hTSHR289-IRES-ZsGreen vector was generated after transformation of JM109 by ligation products. The recombinant plasmid was identified by gel electrophoresis and sequencing. Three plasmids (pAAV-hTSHR289-IRES-ZsGreen, pHelper, and pAAV-2/9) were transfected into 293AAV cell line by calcium phosphate method and a large amount of rAAV2/9-hTSHR289-IRES-ZsGreen was obtained. After purification, the packaging efficiency of recombinant adeno-associated virus was observed by using fluorescence microscope, and its titer was determined by rQ-PCR. HEK293 cells were transfected with rAAV2/9-hTSHR289-IRES-ZsGreen; then the expression level of hTSHR289 was analyzed by ELISA at different time points. Results: The success of hTSHR289 gene inserted into the AAV skeleton vector was confirmed by double digestion and sequencing. After 72 h of transfection of 293AAV cells, the packaging efficiency of the virus was 92%-94% under fluorescence microscope. The titer of the recombinant adeno-associated virus was 1×1013vg/mL. ELISA assay showed that the expression level of hTSHR289 protein mediated by rAAV2/9 was significantly higher at 96 h than that at 72 h and 48 h. Conclusion: The rAAV2/9-hTSHR289-IRES-ZsGreen was successfully constructed. Furthermore, it could be transfected and expressed in cells effectively.
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Most cases of congenital hyperthyroidism are autoimmune forms caused by maternal thyroid stimulating antibodies. Nonautoimmune forms of congenital hyperthyroidism caused by activating mutations of the thyrotropin receptor (TSHR) gene are rare. A woman gave birth to a boy during an emergency cesarean section at 33 weeks of gestation due to fetal tachycardia. On the 24th day of life, thyroid function tests were performed due to persistent tachycardia, and hyperthyroidism was confirmed. Auto-antibodies to TSHR, thyroid peroxidase, and thyroglobulin were not found. The patient was treated with propylthiouracil and propranolol, but hyperthyroidism was not well controlled. At 3 months of age, the patient had craniosynostosis and hydrocephalus, and underwent a ventriculoperitoneal shunt operation. Direct sequencing of the TSHR gene showed a heterozygous mutation of c.1899C>A (p.Asp633Glu) in exon 10. No mutations were discovered in any of the parents in a familial genetic study. We have reported a case of sporadic nonautoimmune congenital hyperthyroidism, by a missense mutation of the TSHR gene, for the first time in South Korea.
Sujets)
Femelle , Humains , Mâle , Grossesse , Césarienne , Craniosynostoses , Urgences , Exons , Mutation germinale , Hydrocéphalie , Hyperthyroïdie , Immunoglobulines thyréostimulantes , Iodide peroxidase , Corée , Mutation faux-sens , Parents , Parturition , Propranolol , Propylthiouracile , Récepteur TSH , Tachycardie , Thyroglobuline , Tests de la fonction thyroïdienne , Dérivation ventriculopéritonéaleRésumé
Objective To analysis the expression of thyroid peroxidase antibody (TPOAb) ,thyroglobulin antibody (TGAb) and thyrotropin receptor antibody (TRAb) in vitiligo patients with different stages .Methods A total of 161 cases of vitiligo patients were divided into advanced group (n=84) and stable group (n=77) according to their condition ,and 60 healthy subjects were cho-sen as the control group .The serum levels of TPOAb ,TGAb and TRAb were detected in 3 groups ,and the difference of the expres-sion level and positive rate between the 3 groups were compared .Results The difference of TPOAb ,TGAb and TRAb levels a-mong the 3 groups was statistically significant (H=14 .371 ,6 .335 ,8 .284 ,P<0 .05) .Multiple comparison showed that :TPOAb , TGAb ,TRAb levels of the advanced group were higher than those of the stable group and the control group (Uvs .stable group =9 .380 , 7 .923 ,8 .381 ,P<0 .05 ;Uvs .control group =23 .244 ,19 .026 ,25 .873 ,P<0 .05);TPOAb ,TGAb ,TRAb levels of the stable group were higher than those of the control group(U=11 .356 ,12 .450 ,16 .351 ,P<0 .05) .The levels of TPOAb ,TGAb and TRAb in the 3 groups were as follows :the advanced group> the stable group> the control group .The positive rates of TPOAb ,TGAb and TRAb in the 3 groups were statistically significant(χ2 =18 .676 ,23 .618 ,23 .857 ,P<0 .05) .Multiple comparison showed that :the positive rates of TPOAb ,TGAb ,TRAb of the advanced group were higher than those of the stable group and the control group (χ2vs .stable group=5 .273 ,6 .484 ,6 .305 ,P< 0 .017;χ2vs .control group = 14 .997 ,18 .352 ,17 .829 ,P< 0 .017);the positive rates of TPOAb , TGAb ,TRAb of the stable group were higher than those of the control group(χ2 =5 .233 ,5 .036 ,6 .719 ,P<0 .017) .The positive rates of TPOAb ,TGAb and TRAb in the 3 groups were as follows :the advanced group> the stable group> the control group .Con-clusion The expression of thyroid autoantibodies (TPOAb ,TGAb ,TRAb) is abnormal in vitiligo patients ,and the progression of the disease is also related to the expression level of such antibodies .
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Objective To analysis the expression of thyroid peroxidase antibody (TPOAb) ,thyroglobulin antibody (TGAb) and thyrotropin receptor antibody (TRAb) in vitiligo patients with different stages .Methods A total of 161 cases of vitiligo patients were divided into advanced group (n=84) and stable group (n=77) according to their condition ,and 60 healthy subjects were cho-sen as the control group .The serum levels of TPOAb ,TGAb and TRAb were detected in 3 groups ,and the difference of the expres-sion level and positive rate between the 3 groups were compared .Results The difference of TPOAb ,TGAb and TRAb levels a-mong the 3 groups was statistically significant (H=14 .371 ,6 .335 ,8 .284 ,P<0 .05) .Multiple comparison showed that :TPOAb , TGAb ,TRAb levels of the advanced group were higher than those of the stable group and the control group (Uvs .stable group =9 .380 , 7 .923 ,8 .381 ,P<0 .05 ;Uvs .control group =23 .244 ,19 .026 ,25 .873 ,P<0 .05);TPOAb ,TGAb ,TRAb levels of the stable group were higher than those of the control group(U=11 .356 ,12 .450 ,16 .351 ,P<0 .05) .The levels of TPOAb ,TGAb and TRAb in the 3 groups were as follows :the advanced group> the stable group> the control group .The positive rates of TPOAb ,TGAb and TRAb in the 3 groups were statistically significant(χ2 =18 .676 ,23 .618 ,23 .857 ,P<0 .05) .Multiple comparison showed that :the positive rates of TPOAb ,TGAb ,TRAb of the advanced group were higher than those of the stable group and the control group (χ2vs .stable group=5 .273 ,6 .484 ,6 .305 ,P< 0 .017;χ2vs .control group = 14 .997 ,18 .352 ,17 .829 ,P< 0 .017);the positive rates of TPOAb , TGAb ,TRAb of the stable group were higher than those of the control group(χ2 =5 .233 ,5 .036 ,6 .719 ,P<0 .017) .The positive rates of TPOAb ,TGAb and TRAb in the 3 groups were as follows :the advanced group> the stable group> the control group .Con-clusion The expression of thyroid autoantibodies (TPOAb ,TGAb ,TRAb) is abnormal in vitiligo patients ,and the progression of the disease is also related to the expression level of such antibodies .
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Objective To investigate the detection value of thyrotropin receptor antibody(TRAb)in children with autoimmune thyroid disease.Methods During the January 2015 and October 2016,a total of 72 cases of children with autoimmune thyroid dis-ease admitted to our hospital were selected,including 36 cases of Graves disease as the Graves disease group,and 36 cases of Hashi-moto's thyroiditis as Hashimoto's thyroiditis group;36 cases of healthy children in this hospital at the same period were selected as the control group.Children in the Graves disease group and Hashimoto's thyroiditis group were taken routine treament,ane the changes of the serum TRAb level before the treatment and one year after the treatment between the two groups.The positive value range of TRAb level is more than 1.75 U/L,and the positive rate of serum TRAb level before the treatment and one year after the treatment was recorded in the two groups.Results Before treatment,TRAb level of the Graves disease group[(12.79 ± 6.38)U/L]was higher than those in Hashimoto's thyroiditis group[(2.03 ± 0.29)U/L]and the control group[(0.85 ± 0.23)U/L],and the difference was statistically significant(P<0.05);TRAb level of the Hashimoto's thyroiditis group[(2.03 ± 0.29)U/L]was significantly higher than that of the control group[(0.85 ± 0.23)U/L],(P<0.05).After treatment:TRAb level of the Graves disease group[(9.36 ± 12.14)U/L]was significantly higher than those in Hashimoto's thyroiditis group[(1.78 ± 0.38)U/L]and the control group[(0.84 ± 0.29)IU/L],and the difference was statistically significant(P<0.05);TRAb level of the Hashimoto's thyroiditis group was significantly higher than the control group,and the difference was statistically significant(P<0.05).TRAb levels in patients with graves'disease and Hashimoto thyroiditis after treatment were significantly lower than those before treat-ment,and the difference was statistically significant(P<0.05).The positive rates of TRAb level in the Graves disease group before and after treatment were 100%,and the positive rate of TRAb level in Hashimoto's thyroiditis group after treatment(11.11%)de-creased in comparison with the one before treatment(22.22%),and the difference was statistically significant(P<0.05).Conclusion There is a significant change of TRAb level in children with autoimmune thyroid disease.Especially,the change of TRAb level in Graves disease before and after treatment may provide a reference for clinical diagnosis and prognosis evaluation of Graves disease.
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The establishment animal model of Graves’ disease contributes to the study of etiology, pathogenesis and therapeutic modalities. After decades of studies and making improvements, the method of building mice model of Graves’disease has achieved a great development. Although there were many reports of animal model building in Graves’disease, as a mature technology A-subunit of thyrotropin receptor( TSHR)-expressing adenovirus was used to establish Graves’disease mice model, which has been accepted widely because of its high efficacy.
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AlM:To construct recombination eukaryotic expression plasmid of human thyrotropin receptor extracellular domain encapsulated with cationic liposomes. METHODS:We amplified the target gene of shuttle vector PHMCMVTSHR289, conjugated the target gene and eukaryotic expression plasmid pcDNA3. 1 +, and accredited whether pcDNA3. 1+/TSHR289 was connected or not by enzymatic digestion and sequencing. Cationic liposomes encapsulated the recombination plasmid pcDNA3. 1+/TSHR289. RESULTS: Recombination plasmid pcDNA3. 1+/TSHR289 digested with enzyme Hindlll and the fragment through 0. 8% gel electrophoresis showed 512bp strip. Recombination plasmid pcDNA3. 1+/TSHR289 were found synonymous mutation through forward ( AAC to AAT ) and reverse sequencing ( GCG to GCT) . The volume ratio of cationic liposomes and recombinant plasmid was 3:1. CONCLUSlON: lt is successful to construct the recombination plasmid pcDNA3. 1+/TSHR289 by accredit it through enzymatic digestion and sequencing.
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It has been 50 years since the discovery of thyrotropin receptor autoantibody (TRAb). Advances in the knowledge of thyrotropin receptor ( TSHR) structure and function, combined with the elucidation of TSHR signaling and TSHR-autoantibody interaction have greatly facilitated our understanding of TRAb and their clinical applications. Measurement of TRAb activity plays an important role in the diagnosis of Graves' disease ( GD) and Graves' opthalmopathy. It has also been well recognized that TRAb is an effective predictor of GD relapse or remission after antithyroid drug and radioactive iodine treatment. TRAb test is of particular help in pregnant women and lactating mothers with recent iodine load, where radioactive iodine or technetium tests are contraindicated. In addition, it is useful in the diagnosis and differential diagnosis of fetal and neonatal hyperthyroidism as well as some rare forms of thyrotoxicosis in clinical practice. Accumulating evidence also indicates the possible correlation between thyroid cancer occurring in GD patients with positive TRAb and adverse outcomes. However, further innovation and standardization of TRAb tests are required to help pave the way for clinical applications.
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Objective To explore the relationship of thyrotropin receptor antibody (TRAb) and soluble intercellular adhesion molecule-1 ( sICAM-1 ) in patients with thyroid-associated ophthalmopathy ( TAO), and the role played by TRAb and sICAM-1 in the pathogenesis of TAO. Method Twenty-three TAO patients were assigned to groups according to the clinical activity score and EUGOGO. All patients were treated with intravenous methylprednisolone pulse. The levels of serum TRAb and sICAM-1 were evaluated by a competitive radioimmunoassay and enzyme-linked immunosorbent assay respectively before treatment and by the end of each methylprednisolone pulse. Results The differernce in serum TRAb levels was associated with activity scores of TAO (P=0. 020). The change in serum sICAM-1 was associated with durations of TAO ( P = 0.015). During methylprednisolone treatment in active TAO patients, the levels of TRAb kept on decreasing gradually and markedly declined after the third methylprednisolone pulse in active TAO patients (P＜0.05). The trends of changes in serum TRAb and sICAM-1 levels were both different in active and inactive TAO patients by tendency analysis. Conclusion TRAb level was related to the activity of TAO and might serve as a significant predictor of response to methylprednisolone therapy. The negative correlation between sICAM-1 levels and duration of TAO corroborates the role played by ICAM-1 during the early stage of TAO. Higher sICAM-1 levels are not expected to be specific to TAO and may not predict a response to methylprednisolone therapy.
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Objective To construct an animal model of Graves' disease (GD) by immunizing BALB/c mice with hM12 cells co-expressing major histocompatibility complex (MHC) class Ⅱ molecules and human thyrotropin receptor (TSHR) molecules. Methods BALB/c mice in experimental group (H-2d) were immunized with hM12 cells Intraper-itoncally every 2 weeks for six times, while mice in control group were immunized with M12 cells. Five weeks later, the thyroids were histologically examined, and serum samples were tested for thyroid-stimulating antibodies (TSAb) and thyroid hormone levels. Results One BALB/c mouse in experimental group developed Graves'-like disease. Total T4 and T3 levels in this mouse were above the upper limit of normal, TSAb activity was displayed in its serum. The thyroid histologically showed the features of thyroid hyperactivity including thyrocyte hypercellularity and colloid absorption.None of control mice developed Graves'-like disease. Conclusion An animal model with some characteristics of human Graves' disease was successfully induced and the model will facilitate studies aimed directly at understanding the patho-genesis of autoimmunity in Graves' disease.
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Objective To explore the mechanism of persistent thyrotropin suppression in euthyroid patients with Graves′ disease after antithyroid drugs (ATD) treatment. Methods A prospective clinical study was performed in 122 patients with newly diagnosed Graves′ disease. All the patients were treated with 30 mg methimazole or 300 mg propylthiouracil daily, to whom L-T4was added, aiming at normalizing FT3 and FT4 but avoiding elevated TSH level. When the patients were clinically and biochemically euthyroid for at least 3 months, their blood levels of thyroid hormones, TSH, TSH receptor antibody(TRAb) and thyroid peroxidase antibody(TPOAb) were detected again and the cases were divided into two groups according to negative or positive TRAb. Results After treatment as long as (7.1±1.1) months, stable euthyroid status was restored for 3 months. When the patients reached the euthyroid state, 64 of them still had detectable TRAb levels, and 58 became negative TRAb. The two groups had similar levels of FT3 and FT4, but patients with positive TRAb had lower TSH level than patients with negative TRAb[0.044 mIU/L(0.001-4.163 mIU/L) vs 1.749 mIU/L(0.079-4.646 mIU/L),P＜0.01]. In addition, the TSH level was negatively correlated with TRAb level (r=-0.539, P＜0.01), and not with FT3, FT4 levels or other factors. Conclusion The present study showed that elevated TRAb level is associated with persistent suppression of TSH in patients with Graves′ disease after being rendered euthyroid. This finding may be due to the binding of TRAb to pituitary TSH receptor.
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One hundred and fifty-seven patients with Graves' disease treated with 131I from July 2001 to March 2003 were studied retrospectively. According to the thyroid function one year after 131I therapy, the patients were divided into 3 groups: remission group with normal FT3, FT4, partial remission group with above normal levels of FT3, FT4, and hypothyroid group with lowered levels of FT3, FT4. Prior to 131I treatment the thyroid-stimulating hormone receptor antibody (TRAb) was positive in 88.5% of the patients. After the therapy, TRAb levels rose transiently and reached the peak within 3-6 months and then dropped gradually thereafter. The weight of pretreatment thyroid, high levels of TRAb (≥405 U/L) and FT3, FT4 were predictors for the outcome of treatment.
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Objective:To establish an animal model of Graves’disease by immunizing syngeneic BALB/c mice with hTSHR-ac-tivated splenocytes. Methods:The female BALB/c mice(6～8 weeks) were randomly divided into 2 groups(group a and group b), which were immunized with recombinant plasmid pcDNA3.1/hTSHR and blank plasmid pcDNA3.1(+) respectively for 3 times at 3-week intervals.The animals were sacrificed at 18th week and their splenocytes were isolated.Other two groups of female syn- geneic BALB/c mice(6~8 weeks) were divided randomly into experimental and control groups, which received splenocytes from group a and group b respectively. Four weeks later, the thyroid glands were removed for histological examination and the sera were determined for Total T4,TSH and TRAb measurements. Results:The thyroid tissues under light microscope displayed dif- fuse hypercellularity with the follicular epithelia being tall cuboidal cells and reduced colloid in 44% of experimental mice with irregular apical poles protruding into the follicular space; there was minimal immune cell in the interstitium with hyper- aemia and edema. Electron microscope examination revealed that the follicular epithelia contained numerous mitochondria and rough endoplasmic reticula,and many long microvilli existed in the luminal surface.The serum level of TT4 in experimental mice was higher than that in control(7.13?1.02 vs 6.43?0.57 ?g/dL) (P
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Thyrotropin receptor(TSHR) mutations must be considered when congenital hyperthyroidism has persisted, but there has been no evidence for autoimmunity. TSHR mutations leading to constitutive activation of the thyroid gland were identified as the molecular cause of autosomal dominant nonautoimmune hyperthyroidism and sporadic congenital hyperthyroidism. We report two cases of hyperthyroidism caused by germline TSHR mutation who presented with exessive sweating and no evidence of autoimmune thyroid disease. They were brothers and their mother had undergone thyroidectomy because of hyperthyroidism. Direct sequencing of the polymerase chain reaction-amplified exon 10 of the TSHR genomic DNA revealed a transition of GCT to GTT, resulting in an exchange of alanine 627 to valine in the patients and their mother. This might be a novel mutation or polymorphism, but we did not perform any functional gene study. But considering the clinical profiles, we can conclude that hyperthyroidism of these two brothers might come from the point mutation described above.