Objective This study was designed to determine the methylation profile of four CpGs and the genotypes of two CpG-SNPs located in promoter region of DIO2 in patients with Kashin-Beck disease (KBD). We also analyzed the interaction between the CpGs methylations and CpG-SNPs. Methods Whole blood specimens were collected from 16 KBD patients and 16 healthy subjects. Four CpGs and two CpG-SNPs in the promoter regions of DIO2 were detected using matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). The CpGs methylation levels were compared between samples from KBD patients and healthy subjects. The methylation levels were also analyzed in KBD patients with different CpG-SNP genotypes. Results The mRNA expression of DIO2 in whole blood of KBD patients was significnatly lower than in healthy controls (P <0.05). The methylation levels of DIO2-1_CpG_3 in KBD patients were significantly higher than those in healthy controls (P <0.05). The methylation levels of four CpGs were not significantly different between KBD patients and healthy controls. The methylation level of DIO2-1_CpG_3 in the promoter region of DIO2 in KBD patients with GA/AA genotype was significantly higher than that of KBD patients with GG genotype (P <0.05). Conclusion The methylation level of DIO2 increases in KBD patients. Similar trends exist in KBD carriers of variant genotypes of CpG-SNPs DIO2 rs955849187.
Case-Control Studies ; Humans ; Iodide Peroxidase/genetics* ; Kashin-Beck Disease/genetics* ; Methylation ; Polymorphism, Single Nucleotide ; Promoter Regions, Genetic

OBJECTIVE: To study whether polymorphisms in the iodothyronine deiodinase (DIO) gene region contribute to endurance exercise capacity and to validate whether TSHR gene can be used as genetic marks associated with aerobic endurance performance. METHODS: Three SNPs (C785T in DIO1 gene regions, Thr92Ala and Gly3Asp in DIO2 gene regions) were selected. The genotypes of the 123 elite long running athletes(EEA) and 127 college students from northern China(CG) were analyzed using the matrix assisted laser desorption ionisation-time of flight mass spectrometry method(MALDI-TOF). The athletes were divided into different groups according to the sports level and the items, which are international masters and masters (43 vs 80), 5/10 km and marathon (92 vs 31). RESULTS: There was no significant difference of the C785T loci in DIO1 gene and the Thr92Ala loci in DIO2 gene between the EEA and the CG(P＞0.05), while at Gly3Asp loci, the frequency distributions of the 3 genotypes were remarkably different in the groups of control and international masters of sports, as well as in the groups of control and marathon athletes(P＜0.05). The genotype TT only existed in EEA not in CG, however, the frequency distribution was very low. The Thr92Ala and Gly3Asp loci of DIO2 gene were in strong linkage disequilibrium. The frequency distributions of the haplotype CT were significantly different in the male CG and the female CG, the male CG and the male EEA(P＜0.05), the male CG and the male masters of sports, as well as in the male CG and the male marathon athletes(P＜0.05). The frequency distributions of the haplotype TC were remarkably higher in the groups of female international masters of sports and female 5 000 m/10 000 m than those in the female CG(P＜0.05). CONCLUSION The frequency distributions of the haplotype CT were different in male and female CG, and haplotype CT could be used as a genetic mark associated with aerobic endurance performance of the male EEA, especially for the long running athletes of masters of sports and marathon, while the haplotype CT was associated with the aerobic endurance performance of the female long running athletes of international masters of sports and 5 000 m/10 000 m.
Athletes ; China ; Female ; Genotype ; Haplotypes ; Humans ; Iodide Peroxidase ; genetics ; Male ; Physical Endurance ; genetics ; Polymorphism, Single Nucleotide ; Running

Congenital hypothyroidism (CH), which results from insufficient thyroid hormone biosynthesis, is one of the most common neonatal endocrine disorders. Thyroid dysgenesis and thyroid dyshormonogenesis are the two causes of CH and either one will lead to deficiencies of enzymes during thyroid hormone biosynthesis and insufficient thyroid hormone biosynthesis. Recently, researchers have performed extensive studies on genetics of CH. This paper reviews genes reported to be associated with CH in China.
Congenital Hypothyroidism ; genetics ; Humans ; Iodide Peroxidase ; genetics ; Membrane Proteins ; genetics ; PAX8 Transcription Factor ; genetics ; Receptors, Thyrotropin ; genetics ; Thyroglobulin ; genetics ; Transcription Factors ; genetics

OBJECTIVETo identify thyroid peroxidase (TPO) gene mutations in 35 patients with congenital hypothyroidism.

METHODGenomic DNA was isolated from peripheral blood samples of 35 patients with congenital hypothyroidism. All of the 17 exons and flanking introns of TPO gene were amplified by PCR, then the PCR products were sequenced bi-directionally and were analyzed by restriction endonucleases.

RESULTOne patient had compound heterozygous mutations c.961A>G/c.2422delT, one was c.2268insT/c.1477G>A, and three was homozygous mutation c.2268insT. The TPO gene mutation c.961A>G [p. Thr321Ala] was one novel mutation.

CONCLUSIONHigh frequency mutation in TPO gene was detected in patients with congenital hypothyroidism.

Autoantigens ; genetics ; Case-Control Studies ; Child ; Child, Preschool ; Congenital Hypothyroidism ; genetics ; DNA Mutational Analysis ; Exons ; Female ; Humans ; Infant ; Iodide Peroxidase ; genetics ; Iron-Binding Proteins ; genetics ; Male ; Mutation

BACKGROUND AND OBJECTIVELung cancer harms people's health or even lives severely. Especially, the therapy of non-small cell lung cancer (NSCLC) has not been obviously improved for many years. The aim of this study is to transfer the human sodium/iodide symporter (hNIS) and the human thyroperoxidase (hTPO) genes into H460 lung cancer cell line, and to study the uptake ability of iodide after co-transfected hTPO and hNIS gene in cell lines.

METHODSThrough cloning, recombination, packaging and amplifying, the recombinant adenosine virus (AdTPO) was constructed. Then the protein expression of AdTPO was tested by Western blot. After transfected hNIS gene into human lung cancer cell line H460 through liposome, stably expressing hNIS gene cell lines (hNIS-H460) selected by G418 antibiotics was determined as hNIS-H460 group. Using AdTPO, hTPO gene was transducted into hNIS-H460, as AdTPO-hNIS-H460 group. H460 cell without hNIS gene was applied as control group (H460). Then, we investigated the 125I uptake assay of the above cells.

RESULTSWe were successful in co-transfecting hNIS and hTPO gene into human lung cell lines H460, and were obtained hNIS and hTPO gene lung cancer cell lines (hNIS-H460 and AdTPO-hNIS-H460). In AdTPO-hNIS-H460, hNIS-H460 and H460, the uptake ability of 125I was (59 637.67 +/- 1 281.13), (48 622.17 +/- 2 242.28) and (1 440.17 +/- 372.86) counts x min(-1). The uptake ability of 125I was 41 fold higher in AdTPO-hNIS-H460 than in blank control H460 (P < 0.01), and 34 fold higher in hNIS-460 than in blank control H460 (P < 0.01), and 1.2 fold higher in AdTPO-hNIS-H460 than in hNIS-H460 (P < 0.01).

CONCLUSIONThe uptake ability of 125I could increase by co-transfected hNIS and hTPO genes into human lung cancer cell lines H460.

Adenoviridae ; genetics ; Cell Line, Tumor ; Genetic Therapy ; Humans ; Iodide Peroxidase ; genetics ; Iodine Radioisotopes ; pharmacokinetics ; therapeutic use ; Lung Neoplasms ; metabolism ; therapy ; Symporters ; genetics ; Transfection

BACKGROUNDType 1 deiodinase (D1) plays an important role in the metabolism of thyroid hormone and has close relationship with thyroid function. In this study we explore the effects of iodine intake on D1 activity and its mRNA expression and its possible mechanism.

METHODSForty-eight Wistar rats were randomly divided into six groups with 8 in each: low iodine (LI), normal iodine (NI), five-fold iodine (HI(5)), ten-fold iodine (HI(10)), fifty-fold iodine (HI(50)), one hundred-fold iodine (HI(100)) group. Three months, six months and twelve months after admistration of potassium iodate, they were sacrificed and thyroids were excised. The expression of D1 mRNA in the thyroid tissue was determined by RT-PCR and D1 activity was analyzed by (125)I-rT3 as substrate. The thyroid hormone was measured with radioimmunoassay method.

RESULTSCompared with NI group, D1 mRNA expression in LI groups slightly decreased, and D1 activity greatly increased. Both T(3) and T(4) in thyroid tissue significantly decreased, but the T(3)/T(4) ratio increased. D1 mRNA expression decreased in all HI groups, and D1 activity was significantly lower in HI groups. There was a tendency of decrease in D1 activity with increased doses of iodine intakes. There was no significant difference in T(4) in thyroid tissue between HI groups and NI group, but a tendency of decrease in T(3) level was found in all HI groups.

CONCLUSIONSIn the case of iodine deficiency, D1 activity increased greatly in order to convert more T(4) to T(3). Excess iodine can inhibit both D1 mRNA expression and its activity to protect organism from being injured by excessive T(3).

Animals ; Iodide Peroxidase ; genetics ; metabolism ; Iodine ; administration & dosage ; RNA, Messenger ; analysis ; Rats ; Rats, Wistar ; Thyroid Gland ; enzymology ; Thyroxine ; blood ; Triiodothyronine ; blood

OBJECTIVETo investigate the effects of ammonium perchlorate (AP) on thyroid functions and mRNA expression levels of thyroglobulin (Tg) and thyroperoxidase (TPO) genes of rats.

METHODSThirty SD male rats were randomly divided into six groups: control group, iodine-deficient group, low dose AP group (130 mg/kg), moderate dose AP group (260 mg/kg), high dose AP group (520 mg/kg) and high iodine-combined group. After the rats were exposed orally for 90 days, serum free-thyroxine (FT(4)), free-triiodothyronine (FT(3)) and thyroid stimulating hormone (TSH) were measured using radioimmunoassays. mRNA expression levels of thyroglobulin (Tg) and thyroperoxidase (TPO) genes were detected by real-time quantitative PCR.

RESULTSSerum FT(4) levels in moderate dose AP group and high dose AP group were [(9.540 ± 1.327) fmol/ml] and [(6.509 ± 1.949) fmol/ml] respectively, which were significantly lower than that [(13.505 ± 1.276) fmol /ml] in control group (P < 0.05 or P < 0.01). Serum TSH level in high dose AP group was [(1.227 ± 0.295) mIU/L], which was significantly higher than that [(0.545 ± 0.282) mIU/L] in control group (P < 0.05). The mRNA expression levels of thyroglobulin (Tg) gene in all groups exposed to AP were significantly lower than that in control group (P < 0.01). The mRNA expression level of thyroperoxidase (TPO) gene in high dose AP group was significantly higher than that in control group (P < 0.05).

CONCLUSIONAP can reduce the serum FT(3) and FT(4) levels of rats, increase the serum TSH level of rats and decrease obviously the mRNA expression levels of Tg and TPO genes. In addition, high iodine can reduce the toxic effects of AP on thyroid gland of rats to some extent.

Animals ; Iodide Peroxidase ; genetics ; metabolism ; Iodine ; administration & dosage ; Male ; Perchlorates ; toxicity ; Quaternary Ammonium Compounds ; toxicity ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Thyroglobulin ; genetics ; metabolism ; Thyroid Gland ; drug effects ; metabolism ; Thyrotropin ; blood ; Thyroxine ; blood ; Triiodothyronine ; blood

OBJECTIVETo investigate the effect of selenium supplementation on the selenium status and selenoenzyme, especially the activity and mRNA expression of type 1 deiodinase (D1) in mice with excessive iodine (EI) intake and to explore the mechanism of selenium intervention on iodine-induced abnormities.

METHODSWeanling female BALB/c mice were given tap water or 3 mg/L of iodine or supplemented with 0.5 mg/L or 1.0 mg/L of selenium in the presence of excessive iodine for 5 months. Selenium status, thyroid hormone level, hepatic and renal D1 activity and mRNA expression were examined.

RESULTSExcessive iodine intake significantly decreased the selenium concentration in urine and liver, and the activity of glutathione peroxidase (GSH-Px) in liver. Meanwhile, serum total T4 (TT4) increased while serum total T3 (TT3) decreased. Hepatic D1 enzyme activity and mRNA expression were reduced by 33% and 86%, respectively. Renal D1 enzyme activity and mRNA were reduced by 30% and 55%, respectively. Selenium supplementation obviously increased selenium concentration, activity of GSH-Px and Dl as well as mRNA expression of D1. However, increasing the supplementation of Se from 0.5 to 1.0 mg/L did not further increase selenoenzyme activity and expression.

CONCLUSIONRelative selenium deficiency caused by excessive iodine plays an essential role in the mechanism of iodine-induced abnormalities. An appropriate dose of selenium supplementation exercises a beneficial intervention.

Animals ; Antioxidants ; pharmacology ; Creatinine ; metabolism ; urine ; Dietary Supplements ; Female ; Iodide Peroxidase ; genetics ; metabolism ; Iodine ; toxicity ; urine ; Kidney ; metabolism ; Liver ; metabolism ; Mice ; Mice, Inbred BALB C ; RNA, Messenger ; metabolism ; Selenium ; pharmacology ; urine ; Thyroxine ; blood ; Triiodothyronine ; blood

Radioiodine is a routine therapy for differentiated thyroid cancers. Non-thyroid cancers can intake radioiodine after transfection of the human sodium iodide symporter (hNIS) gene. The human telomerase reverse transcriptase (hTERT) promoter, an excellent tumor-specific promoter, has potential value for targeted gene therapy of glioma. We used the hTERT promoter to drive the expression of the hNIS and human thyroid peroxidase (hTPO) gene as a primary step for testing the effects of radioiodine therapy on malignant glioma. The U87 and U251 cells were co-transfected with two adenoviral vectors, in which the hNIS gene had been coupled to the hTERT promoter and the hTPO gene had been coupled to the CMV promoter, respectively. Then, we performed Western blot, 125I intake and efflux assays, and clonogenic assay with cancer cells. We also did 99mTc tumor imaging of nude mice models. After co-transfection with Ad-hTERT-hNIS and Ad-CMV-hTPO, glioma cells showed the 125I intake almost 1.5 times higher than cells transfected with Ad-hTERT-hNIS alone. Western blots revealed bands of approximately 70 kDa and 110 kDa, consistent with the hNIS and hTPO proteins. In clonogenic assay, approximately 90% of co-transfected cells were killed, compared to 50% of control cells after incubated with 37 MBq of 131I. These results demonstrated that radioiodine therapy was effective in treating malignant glioma cell lines following induction of tumor-specific iodide intake by the hTERT promoter-directed hNIS expression in vitro. Co-transfected hNIS and hTPO genes can result in increased intake and longer retention of radioiodine. Nude mice harboring xenografts transfected with Ad-hTERT-NIS can take 99mTc scans.
Adenoviridae ; genetics ; Animals ; Autoantigens ; genetics ; metabolism ; Cell Line, Tumor ; Cell Survival ; Cytomegalovirus ; genetics ; Genetic Vectors ; Glioma ; diagnostic imaging ; genetics ; metabolism ; pathology ; Half-Life ; Humans ; Iodide Peroxidase ; genetics ; metabolism ; Iodine Radioisotopes ; metabolism ; Iron-Binding Proteins ; genetics ; metabolism ; Mice ; Mice, Nude ; Promoter Regions, Genetic ; Recombinant Proteins ; genetics ; metabolism ; Symporters ; genetics ; metabolism ; Technetium ; Telomerase ; genetics ; Tomography, Emission-Computed, Single-Photon ; Transfection

OBJECTIVETo investigate the mRNA expressions of RASSF1A, Galectin-3 and TPO in papillary thyroid carcinoma and some other thyroid benign lesions, and evaluate their diagnostic significance.

METHODSReverse transcription polymerase chain reaction (RT-PCR) was used to detect the mRNA expression of RASSF1A, galectin-3 and TPO in the samples from 73 cases, including 23 cases with papillary thyroid cancer, 16 with nodular goiter, 29 with thyroid adenoma and 5 with Hashimoto's disease.

RESULTSA statistically significant difference in the mRNA expression of RASSF1A, Galectin-3 and TPO was observed between papillary thyroid carcinoma and follicular benign lesions (P<0.05). However, there was no significant difference among various kinds of benign lesions (P>0.05). A negative correlation of the expression of RASSF1A and Galectin-3 mRNA was found between thyroid benign lesions and malignant ones (P = 0.000). While the mRNA expression of RASSF1A and TPO was positively correlated between benign and malignant lesions (P = 0.028).

CONCLUSIONLoss of expression of RASSF1A and TPO mRNA but high expression of Galectin-3 mRNA in papillary thyroid carcinoma are common. Therefore, the products of these three genes may be closely related to the development of thyroid papillary carcinoma, and may be used as useful markers in differential diagnosis of papillary thyroid carcinoma from the benign lesions. The results are more reliable if this detection method is used in combination with other techniques.

Adolescent ; Adult ; Aged ; Autoantigens ; genetics ; metabolism ; Biomarkers, Tumor ; metabolism ; Carcinoma, Papillary ; genetics ; metabolism ; pathology ; Diagnosis, Differential ; Female ; Galectin 3 ; genetics ; metabolism ; Goiter, Nodular ; genetics ; metabolism ; pathology ; Hashimoto Disease ; genetics ; metabolism ; pathology ; Humans ; Iodide Peroxidase ; genetics ; metabolism ; Iron-Binding Proteins ; genetics ; metabolism ; Male ; Middle Aged ; RNA, Messenger ; metabolism ; Thyroid Neoplasms ; genetics ; metabolism ; pathology ; Tumor Suppressor Proteins ; genetics ; metabolism ; Young Adult