Research on THRβ gene mutation in a patient with thyroid hormone resistance syndrome using whole-exome sequencing / 中华内分泌代谢杂志

Objective:To assess clinical and genetic features in a patient with thyroid hormone resistance syndrome(RTH) and explore the pathogenic mechanism.Methods:The clinical data of the proband was collected. The genomic DNA was extracted from peripheral blood samples of the patients. The pathogenic variant was identified using whole-exome sequencing and confirmed by Sanger sequencing. Then the function of the mutation sites was detected by bioinformatics.Results:The patient presented with chest distress, palpitation, and persistent atrial fibrillation, along with elevated levels of serum free triiodothyronine(FT 3), free thyroxine(FT 4), and thyroid stimulating hormone(TSH), which suggested RTH clinically. The genetic analysis identified a heterozygous mutant of THRβ(c.1313G>A) gene at exon 8, which was a missense mutation causing the substitution of arginine to histidine at 438 position of the protein(p.R438H). Its inheritance pattern was unknown. This mutation was considered as a new one that had not been reported. Conclusion:A novel pathogenic THRβ gene mutation was found in the patient with RTH, which might be the cause of this disease. This variant c. 1313G>A is located in the ligand binding domain of THRβ, which might result in low protein activity.

Study on genetic etiology of two cases with hypophosphatasia(HPP) using whole-exome sequencing / 中华内分泌代谢杂志

Objective:Clinical and genetic analysis were conducted in 2 patients with hypophosphatasia(HPP) and their families to explore the pathogenic mechanism of HPP.Methods:The genomic DNA was extracted from peripheral blood of two patients with HPP and their family members. Sanger sequencing and pedigree verification were performed on the pathogenic variants identified using whole-exome sequencing. Then the function of the mutation sites was analyzed with bioinformatics software.Results:Proband 1 presented with developmental retardation, pectus funnel and premature loss of deciduous tooth, of which the serum alkaline phosphatase level was slightly lower than the bound of the normal range. Two complex heterozygous missense variants c. 1120G>A and c. 1334C>G of ALPL gene were detected in the proband 1 which were inherited from his parents respectively, showing an autosomal recessive inheritance. Both the variants were predicted to inflict deleterious effects on ALPL gene function by multiple bioinformatics program, and were classified as likely pathogenetic variants according to American College of Medical Genetics and Genomics(ACMG) guidelines. Proband 2 showed three missing permanent teeth and the significantly lower level of serum alkaline phosphatase than normal range. A heterozygous variant c. 1190-3C>G of ALPL gene was detected in proband 2 whose pattern of inheritance was unknown. The clinical significance of this variant was unknown according to ACMG standards and guidelines. All of these variants were considered as novel since none of them has been reported. Along with the above combined results, proband 1 and 2 were diagnosed as childhood HPP and Odontohypophosphatasia, respectively.Conclusion:This study reinforced the relationship between HPP and variants in ALPL gene. Two variants, c. 1120G>A and c. 1334C>G, were located in the homodimer interface and crown domain of tissue-nonspecific alkaline phosphatase(TNSALP), respectively, while c. 1190-3C>G were located in the splice sites, which might result in low TNSALP activity.

Molecular diagnosis and clinical features of 206 patients with 46, XY disorders of sexual development / 中华内分泌代谢杂志

Objective:To investigate methods of molecular diagnosis and clinical features of 46, XY disorders of sexual development(DSD).Methods:A total of 206 cases of 46, XY DSD patients, who visited the Shanghai Ninth People′s Hospital, Shanghai Jiaotong University School of Medicine, from July 2009 to June 2021, underwent AA chip based on multiplex PCR and probe-capture-targeted next-generation sequencing. Clinical features of patients with genetic diagnosis were analyzed.Results:Among 206 patients, the diagnostic rate of patients with micropenis, hypospadias and cryptorchidism was the highest, up to 75.28%. Almost all patients had different degrees of undermasculinized external genitalia. The most frequent phenotype was micropenis with hypospadias(87.25%). Only one gene variant was detected in 81 patients(39.32%), multiple genetic variants were detected in 104 patients(50.49%), and no gene variant was identified in 21 patients(10.19%). 107 patients had definite genetic diagnosis, with a diagnostic rate of 51.94% by adding the pathogenic and likely pathogenic ratios following the American College of Medical Genetics and Genomics(ACMG) guidelines, including 40 patients of steroid 5α-reductase type 2(SRD5A2) variants(37.38%), 36 patients of androgen receptor(AR) variants(33.64%), 13 patients of steroidogenic factor 1(NR5A1) variants(16.82%), 6 patients of 17β-hydroxysteroid dehydrogenases 3(HSD17B3) variants(5.61%), 2 patients of 17α-hydroxylase/17, 20-lyase enzyme(CYP17A1), Wilms′ tumor 1(WT1) and GATA binding protein 4(GATA4) variants(1.87%), and one patient of luteinizing hormone receptor(LHCGR) variant(0.93%). Gynecomastia was found in 29 of 81 postpubertal patients, of which 25(86.21%) had AR variants.Conclusions:46, XY DSD presents complex clinical manifestations and molecular etiologies. Targeted nextgeneration sequencing has the advantages of high throughput, high efficiency and low cost, which has a high value especially in etiological diagnosis of 46, XY DSD with large genetic heterogeneity.

Single-cell transcriptome analysis of T cells in thyroid and peripheral blood of patients with Hashimoto′s thyroiditis / 中华内分泌代谢杂志

Objective:To construct single-cell transcription landscape of T cell in peripheral blood mononuclear cells(PBMCs) and thyroid tissue of patients with Hashimoto ′s thyroiditis(HT), and to analyze the changes in the proportion and functionality of T cell clusters in HT disease state.Methods:Single cell RNA sequencing was performed on PBMCs and thyroid tissue from 5 HT patients. Single cell RNA sequencing data of PBMCs from 5 healthy individuals were retrieved from public databases. After preliminary clustering, the clusters expressing CD3E were extracted and clustering again, and the names of each cluster were determined according to the known cell markers. The proportion of each cell subtype was compared, and the differentially expressed genes in different samples were analyzed.Results:After quality control, the 71 533 T cells were classified into 19 cell clusters. Among them, the proportion and function of C1_CD4 + Naive T cell clusters, C3_CD4 + Treg cell clusters, C7_CD8 + Naive T cell clusters, C8_GNLY -CD8 + T cell clusters, C10_RORC + CD8 + T cell clusters, C11_ GZMK + CD8 + T cell clusters, C12_CCL4 + CD8 + T cell clusters, and C18_PTGDS + NK cell clusters in thyroid tissue of HT patients were significantly different from those in PBMCs of healthy controls and HT patients. Conclusion:The proportion of multiple T cells in thyroid tissue of HT patients were significantly different from those in PBMCs. Among them, the proportion of three of CD8 + T cell subsets with high expression of cell killing-related genes in thyroid tissue T cells of HT patients is higher than that in PBMCs T cells, and it is statistically significant. In addition, the functionality of various T cells in the thyroid tissue of HT patients are also significantly different from those in PBMCs. A cluster of GZMK + CD8 + T cells showes significantly lower expression of genes related to PD1 pathway in thyroid tissues of HT patients compared with cells in PBMCs of HT patients, also a cluster of CCL4 + CD8 + T cells showes significantly lower expression of genes related to IL-12 pathway.

Clinical features and treatment outcome of four patients with Kallmann syndrome caused by fibroblast growth factor receptor-1 gene mutation / 中华内分泌代谢杂志

Objective:To investigate the clinical features and treatment outcome of Kallmann syndrome(KS) caused by fibroblast growth factor receptor-1(FGFR1) gene mutation in 4 patients.Methods:Targeted next-generation sequencing(NGS) was performed on thirty KS and normosmic idiopathic hypogonadotropic hypogonadism(nIHH) patients. FGFR1 mutation was identified in four KS patients. The clinical data, laboratory and imaging examinations, and treatment outcome were retrospectively analyzed.Results:Four male patients, aging from 11 to 22 years old, presented as micropenis, and with olfactory dysfunction. Among them, two had history of cryptorchidism, three had history of cleft lip and palate repair surgery. The most severe patient presented with short stature, left microtia and dental agenesis. FGFR1 heterozygous mutation was identified in all four patients, two were point mutation(p.Y374X; p. E670K), and the other was frameshift mutation(p.S346Yfs*61; p.S723*fs*1). One patient, who started treatment of the pulsatile GnRH pump during his youth, succeeded in having two babies.Conclusion:Patients with Kallmann syndrome caused by FGFR1 mutation presents complex clinical manifestations. Besides dysosmia, micropenis, microrchidia, and delayed pubertal development are the main clinical manifestations in male patients. Symptoms such as cleft lip and palate are helpful for early recognition. Genotyping analysis is crucial to confirm the diagnosis. The pulsatile GnRH pump can produce satisfactory therapeutic effect, but the age of initiating therapy should be carefully considered.

The establishment of zebrafish Dio3b gene knockout model / 中华内分泌代谢杂志

Objective To set up the model of deiodinase ( Dio) 3b-/- zebrafish and to observe the effect of which on embryo development. Methods The zebrafish model of Dio3b-/- was set up by CRISPR-Cas9 gene editing technology, PCR and sequencing was used to confirm the efficiency of deletion. The heart rate of embryos at 48 hours post fertilization was counted. The locomotor activity of 5-7 days post fertilization larve was detected using behavior tracking system. Results The model of Dio3b knockout zebrafish was set up successfully. The heart rate of embryos Dio3b-/- increased ( P＜0. 001) and the locomotor activity of 5-7 days post fertilization larves lacking Dio3b gene increased (P＜0.05) significantly compared with that of wild type control respectively. Conclusion The deletion of zebrafish Dio3b gene results in the phenotype of hyperthyroidism and the model of Dio3b-/- is proper for studying the effect of partial excess thyroid hormone on embryo development.

DUOX2 mutations are frequently associated with congenital hypothyroidism in a Chinese Patients / 中华内分泌代谢杂志

Objective To investigate the prevalence of DUOX2 mutations in Chinese patients with congenital hypothyroidism (CH) and to discuss the inheritance pattern of DUOX2 gene.Methods Blood samples were collected from 91 CH children and their genomic DNA was extracted from peripheral blood leukocytes.All exons and exon-intron boundaries of DUOX2 were analyzed by target next-generation sequencing and family trios was established to study the inheritance pattern of DUOX2 gene.Results Fifty-four out of 91 children with CH carried DUOX2 mutation, with a prevalence of 59.34%.Of the 54 CH children, 36 carried DUOX2 biallelic mutations.In all 12 family trios with probands carrying biallelic DUOX2 mutations, the parents carried heterozygous DUOX2 mutations while still showing normal thyroid function, suggesting that CH caused by DUOX2 mutations is inherited in an autosomal recessive manner.Conclusion DUOX2 gene is one of the most frequently mutated genes in Chinese CH patients and its inheritance pattern is an autosomal recessive one.

Correlation of FOXE1 rs925489 gene polymorphism and high normal TSH level in Chinese Han population / 中华内分泌代谢杂志

Objective To explore the correlation of the gene polymorphism of the two single nucleotide polymorphisms(SNPs)rs1443434andrs925489onforkheadboxEl(FOXE1)withthehighnormalthyroidstimulating hormone ( TSH) level in Chinese Han population. Methods 1 400 subjects with normal serum TSH and thyroid peroxidase antibody(TPOAb) levels were included. According to TSH or TPOAb levels, the subjects were divided into high normal TSH group(H-TSH group,n=195) and normal TSH control group(TSH control group,n=1 205) or high normal TPOAb group ( H-TPOAb group, n=711 ) and low normal TPOAb group ( L-TPOAb group, n=689 ) , respectively. The genotypes on the two SNPs of all the subjects were performed by whole-genome genotyping chips. Results There were significant differences in rs925489 genotypic distributions and allele frequencies between H-TSH group and TSH control group(both P<0. 05). The genotype TT and allele T in H-TSH group were significantly higher than those in TSH control group(89. 75% vs 83. 15%, 94. 62% vs 91. 29%). The normal TSH levels were positively associated with rs925489 genotypic distributions after adjustment for sex, age, and high density lipoprotein cholesterol(P<0. 01). There were no significant differences in rs1443434 genotypic distributions and allele frequencies between two TSH groups or two TPOAb groups. Conclusion FOXE1 rs925489 gene polymorphism may be correlated with the high normal TSH level in Chinese Han population.

Effects of free fatty acids on catalytic activity of P450c17 in cultured NCI-H295R cells of adrenal gland / 中华内分泌代谢杂志

[Summary] The activities of 17α-hydroxylase and 17,20 lyase of cytochrome P450c17 were evaluated by ELISA in NCI-H295R cells after treatment with palmitate and oleate. The results showed that 0. 75 mmol/L plamitate did not influence the activity of 17α-hydroxylase, but increased the activity of 17, 20 lyase by 74. 3% ( P<0. 05). Oleate at the same concentration did not change the activity of 17,20 lyase. There were no significant changes in the protein expressions of P450c17, P450 oxidoreductase, and cytochrome b5 after treatment with palmitate and oleate. However,reactive oxygen species in cells were elevated by palmitate. The results suggest that exposure to palmitate may increase androgen production by inducing 17, 20 lyase activity of P450c17 in NCI-H295R cells, which is related with oxidative stress-mediated post-translational regulation of the enzyme.

Correlation of a high normal serum TSH with blood pressure, fasting plasma glucose, and blood lipids in Chinese Han population / 中华内分泌代谢杂志

[Summary] A total of 1 510 subjects undergoing physical examination in the Central Hospital of Xuzhou were included in this study. According to the level of TSH, the subjects were divided into low TSH group(0. 30-0. 99 mIU/L,n=351), moderate TSH group(1. 00-1. 89 mIU/L, n=703), and high TSH group(1. 90-4. 80 mIU/L, n=456). Analysis of variance and linear regression were used for data analysis. The results showed that systolic blood pressure ( SBP) , diastolic blood pressure ( DBP) , triglyceride ( TG) , and high-density lipoprotein cholesterol( HDL-C) revealed significant differences among 3 group(P<0. 05 or P<0. 01). In the univariate linear regression model, serumTSHwithinthereferencerangewasnegativelyassociatedwithSBPandDBP(P<0.05orP<0.01),and positively associated with TG and HDL-C (P<0. 05 or P<0. 01). However, the correlations disappeared after adjustment for gender, age, body mass index, fasting plasma glucose ( FPG ) , and TG. In the multiple linear regression model, a significant negative correlation of TSH with SBP and FPG was found in males(P<0. 05).

Association of the rs6832151 within chromosomal band 4p14 with Graves′disease / 中华内分泌代谢杂志

[Summary] The genotypes of rs6832151 in the 4p14 were genotyped by Taqman probe technique on FluidigmEPl platform in 617 patients with Graves′disease( GD) and 4 915 health control subjects. The result showed thatRs6832151 Gin4p14wasstronglyassociatedwithGD(OR=1.39,P0. 05). The result suggests that rs6832151 G in 4p14 is the susceptibility genes of Graves′ disease in Bengbu population, and is related to the high risk of GD.

SCGB3A2: A validated susceptibility gene to Graves' disease in different ethnic populations / 中华内分泌代谢杂志

Graves' disease is a common autoimmune disease triggered by the susceptibility genes and environmental factors.Among the 9 risk genes related to Graves' disease,SCGB3A2 is the first Graves' disease-predisposing gene identified by our group using tagSNPs,strategy of candidate genes,and positional clones.The association between SCGB3A2 and Graves' disease has been confirmed by two independent cohorts from UK and Russia.So far,the geneticists on Graves' disease regard SCGB3A2 as a validated susceptibility gene of that disease.

Association between pulse wave velocity and impaired fasting glucose / 中国医师杂志

ObjectiveTo examine the relation between pulse wave velocity (PWV) and impaired fasting glucose (IFG),then evaluate the modification effects of age,BMI,hypertension and lipids in Chinese adults.Methods5099 cases from a community-based health examination survey in Xuzhou,Jiangsu prownce,China,were enrolled in this study.Blood pressure,weight,height,waist circumference,neck circumference,body fat ratio and determination of fasting glucose,lipidsand pulse wave velocity were measured in all cases.IFG was defined as 6.1 mmol/L≤FBG ＜7.0 mmol/L.ResultsThe odds ratios (OR,95％ CI ) of IFG across increasing variable of cf-PWV were 1.00,1.07(0.83 - 1.39),1.20( 1.08 -1.34),1.13(1.04 - 1.23),1.14(1.05 - 1.25) ( Pfor trend ＜0.01).Age and neck circumference levels significantly interacted with cf-PWV in relation to IFG risk ( P ＜0.01 ).ConclusionsThe present data indicate serum cf-PWV concentration was associated with the risk of IFG,and the association was modified by age and neck circumference levels.

SNPs in the SCGB3A2 promoter are associated with susceptibility to Graves' disease / 中华内分泌代谢杂志

Objective To investigate the association of single nucleotide polymorphisms (SNPs) in the SCGB3A2(secretoglobin family 3A member 2) gene promoter with susceptibility of Graves' disease.Methods One-hundred and seventy-nine SNPs within a 3.0 Mb region surrounding marker D5s2090 were scanned in a case-control study.The size of the region(s) associated with GD was then narrowed.Results Total 179 SNPs within a 3.0 Mb region surrounding marker D5s2090 were analyzed.The most significant association signal was found at SNP rs1368408 (P =3.69 × 10-5).Subsequent association analysis was then performed and the results suggested that the SNP76 (P =4.11 × 10-8) and SNP75 (P =1.37 × 10-8) in the promoter of SCGB3A2 gene may be the causal variants of GD.Logistic regression analysis suggested these 2 SNPs in this region may contribute to GD susceptibility.Conclusion A significant association seems to exist between GD with the SCGB3A2 gene.

Progress of genetic studies on serum TSH levels / 中华内分泌代谢杂志

Thyroid hormones play major roles in the regulation of a wide range of metabolie and physiologic processes.Serum thyroid-stimulating hormone( TSH ) concentration,a sensitive barometer of thyroid function,shows significant individual difference in which genetic variation is a major factor.After using traditional genetic linkage studies and candidate gene association studies to explore the suseeptibility genes of serum TSH,many progresses have been made and new susceptibility genes have been identified by genome-wide association study (GWAS).In this review,we focus on the susceptibility genes of serum TSH levels and also the future prospect that may be obtained from these studies.

TRβ gene mutation in a family with thyroid hormone resistance syndrome / 中华内分泌代谢杂志

Objective To detect the gene mutation of thyroid hormone receptor β ( TRβ ) in a family with thyroid hormone resistance syndrome.Methods The genomic DNA was extracted from peripheral blood leukocytes of the patient and his 5 family members.The exons 1-10 ofTRβ gene were amplified by PCR.The products of PCR were sequenced directly to detect the gene mutation.Results Two members of this family were confirmed to have the C y A transition mutation at nucleotide 1642 site within exon 10 of TRβ gene,which was a missense mutation causing the substitution of Proline to Threonine (P453T).The mutation was Heterozygous.Conclusions It was confirmed that the patient has TRβ gene mutation P453T in exon 10.The mutation may lead to the occurrence of thyroid hormone resistance syndrome.

The single nucleotide polymorphisms in the intron 1 of TSHR gene were associated with Graves' disease / 中华内分泌代谢杂志

Objective To investigate the association between single nucleotide polymorphisms in the intron 1 of thyroid stimnulating hormone receptor gene (TSHR) and Graves' disease (GD) in the Chinese Han population from Linyi city,Shandong Province.Methods A total of 1759 GD patients and 1740 control subjects were recruited for genotyping in TSHR intron 1 with genome-wide association study (GWAS) and Taqman probe technique.At the same time,serum thyroid hormone and TSH receptor antibody (TRAb) levels of patients were determined.Results Five SNPs were selected for further replication.The rs12101261 _T was significantly associated with GD risk ( OR=1.257,95％CI 1.137-1.390,P =8.23 × 10-6 ). Logistic regression identified that rs12101261 was an independent susceptibility locus of GD ( P=1.61 × 10-3 ).Furthermore,rs12101261 _T was strongly associated with GD ( OR =1.317,95％ CI 1.171-1.481,P=4.14× 10-4 ) in TRAb positive patients,but no association in TRAb negative patients ( OR=1.056,95％ CI 0.892-1.251,P=0.524 ).Serum TRAb concentration showed remarkable difference among three genotype groups of rs12101261.Conclusions Five SNPs in TSHR intron 1 are associated with GD.rs12101261 contributes to increased GD risk independently and is associated with serum TRAb level.

Influences of multiple gene interactions on bone mineral density and osteoporotic fractures in postmenopausal women / 中华内分泌代谢杂志

Objective To investigate the effects of the genetic polymorphisms in osteoporosis-related genes and the gene-gene interaction on bone mineral density (BMD) and osteoporotic fractures.Methods Thirty-nine single nucleotide polymorphism (SNP) sites in 23 genes that related to bone mineral density ( BMD ) and osteoporotic fractures were scanned in 683 Shanghai Han postmenopausal women.TaqMan SNP Genotyping Assay or Sequenom Mass ARRAY System were applied for genotyping analysis.The relation of these SNP sites with BMD and osteoporotic fractures were analyzed.Results Altogether,12 SNPs in 9 candidate genes ( rs7524102 and rs6696981 in ZBTB40 gene,rs9479055 in ESR1 gene,rs6993813,rs6469804,and rs11995824 in OPG gene,rs3736228 in LRP5 gene,rs1107748 in SOST gene,rs87938 in CTNNB1 gene,rs1366594 in MEF2C gene,rs7117858 in SOX6 gene,and rs10048146 in FOXL1 gene) were associated with BMD at lumbar spine(L1-L4) or total hip.In addition,rs11898505 in SPTBN1 gene was related to osteoporotic fractures ( OR 0.522,95％ CI 0.326-0.838,P =0.007 ).Gene-gene interaction involving rs1038304 in ESR1 gene,rs1366594 in MEF2C gene,and rs10048146 in FOXL1 gene was associated with osteoporotic fractures ( P =0.010 7 ).Conclusions ( 1 ) SNPs in gene ZBTB40,ESR1,OPG,LRP5,SOST,CTNNB1,MEF2C,SOX6,FOXL1,and SPTBN1 are associated with BMD of lumbar spine or total hip,as well as osteoporotic fractures.(2) Gene-gene interaction involving rs1038304,rs1366594,and rs10048146may contribute to the risk of osteoporotic fractures.

TSHR gene intron 1 region polymorphism and its relationship with Graves' disease / 中华内分泌代谢杂志

Objective To investigate the association between the six single nucleotide polymorphisms ( SNP),named as rs179247,rsl2101261,rs2284722,rs4903964,rs2300525,rsl7111394 in the intron 1 of thyroid stimulating hormone receptor gene (TSHR) and Graves' disease (GD).MethodsThe genotypes of the six SNPs were genotyped by Taqman probe technique on Fluidigm EP1 platform in 618 GD patients and 646 control subjects.Meanwhile,TSH receptor antibodies (TRAb) of the patients were determined.ResultsAmong the six SNPs,five S NPs were strongly associated with GD,with the most signals at rs179247_G,rs12101261_C,rs4903964 _G (P=2.85×10-10,OR=1.73,95％CI1.46-2.05;P=1.74×10-10,OR=1.73,95％CI 1.46-2.05;P=2.24×10-10,OR=1.69,95％ CI 1.44-1.99 ).The results of logistic regression analysis indicated that rs12101261 and rs4903964 were main susceptibility loci of GD in the intron 1 of TSHR.rs179247_G,rs1210126 1_C,and rs4903964_G were associated with subset of the GD patients with positive TRAb (P=4.24× 10-13,p=5.48× 10-13,P =3.89×10-12 ).Conclusionrs179247,rs12101261,and rs4903964 in TSHR intron 1 were significantly associated with GD in the Chinese Han population from Bengbu city.rs12101261 and rs4903964 were the major susceptibility SNPs associated with GD.TSHR gene may play a main role of susceptibility gene in the subset of GD patients with persistent positive TRAb.

Recent progress in identification of candidate genes of Graves' disease / 中华内分泌代谢杂志

Intensive but quite chaotic and decentralized candidate gene studies on susceptibility to Graves' disease ( GD ) carried out in small size population in the past half century have provided a quantity of inconsistent data,which,however,resulted in finding of a proven association of GD with the MHC class Ⅱ region that exerts a major effect on the genetics of GD.Using low-resolution microsatellite-based human genome-wide linkage analysis,several regions of linkage harboring putative susceptibility variants but no gene susceptibility to GD were identified.Further,high throughput genotyping of large population cohorts with help of high dense panels of single nucleotide polymorphisms (SNPs) and application of advanced tools for analysis of extended blocks of linkage disequilibrium within a candidate gene ( tagging SNP,etc.) have found several genes susceptible to GD,including immune-related genes such as MHC,CTLA4,SCGB3A2/UGRP1,FCRL3,and thyroid specific genes ( such as TSHR,etc.).Less consistent results have been obtained in cases of PTPN22 and thyroglobulin.In the nearest future,implementation of even more robust technology such as genome-whole associated analysis (GWAS) and whole-genome re-sequencing are expected to catch more genes susceptibilities to GD.