Background: Thyroid hormone (triiodothyronine [T3]) is essential for the development throughout vertebrates. Anuran metamorphosis mimics mammalian postembryonic development, a period around birth when plasma T3 level peaks and many organs/tissues mature into their adult forms. Compared with the uterus-enclosed mammalian embryos, tadpoles can be easily manipulated to study the roles of T3 and T3 receptors (TRs) in tissue remodeling and adult organ development. We and others have previously knocked out TR alpha or TR beta in the diploid anuran Xenopus tropicalis and reported distinct effects of the two receptor knockouts on metamorphosis. However, animals lacking either TR alpha or TR beta can complete metamorphosis and develop into reproductive adults. Methods: We have generated TR alpha and TR beta double knockout animals and carried out molecular and morphological analyses to determine if TR is required for Xenopus development. Results: We found that the TR double knockout tadpoles do not respond to T3, supporting the view that there are no other TR genes in X. tropicalis and that TR is essential for mediating the effects of T3 in vivo. Surprisingly, the double knockout tadpoles are able to initiate metamorphosis and accomplish many metamorphic changes, such as limb development. However, all double knockout tadpoles stall and eventually die at stage 61, the climax of metamorphosis, before tail resorption takes place. Analyses of the knockout tadpoles at stage 61 revealed various developmental abnormalities, including precocious ossification and extra vertebrae. Conclusions: Our data indicate that TRs are not required for the initiation of metamorphosis but is essential for the completion of metamorphosis. Furthermore, the differential effects of TR knockout on different organs/tissues suggest tissue-specific roles for TR to control temporal coordination and progression of metamorphosis in various organs.
Background: The correlation between the injured recurrent laryngeal nerve (RLN) with incomplete loss of signal (LOS) and the outcomes of vocal cord function is still not well understood. This large cohort validation study was aimed to evaluate the effectiveness of the warning criterion, the reduction of the R2p/R2d ratio, in neuromonitoring during thyroidectomy. Methods: A total of 1108 consecutive patients (1764 nerves) with normal vocal cord function undergoing monitored thyroidectomy were included. Standardized intraoperative neuromonitoring procedures were strictly followed, after complete dissection of RLN, the exposed RLN was routinely stimulated at the lowest proximal end (R2p signal) and the most distal end near the laryngeal entry point (R2d signal). If the reduction of the R2p/R2d ratio (([R2d - R2p]/R2d) x 100%) reached >20%, the whole exposed RLN would be checked to pinpoint the injured area of the nerve. Results: Visual anatomical integrity of the RLN was ensured in all 1764 nerves. Eighteen nerves had complete LOS, and the other 97 nerves had incomplete LOS where the reduction of the R2p/R2d ratio ranged from 21% to 84%. Postoperative temporary vocal cord paralysis (VCP) was noted in 11 (61.1%) RLNs with complete LOS and 16 (16.5%) RLNs with incomplete LOS, where the reduction of the R2p/R2d ratio ranged from 63% to 84%. The positive predictive value of a R2p/R2d ratio >63% for postoperative VCP was 79.4%. Conclusions: Testing and comparing the R2p and R2d signals were useful to detect RLN neurophysiologic injury, elucidating the mechanism of nerve injury and predicting vocal cord function. Determining R2p-R2d was found to be essential and can be applied in routine neuromonitoring thyroidectomy.
Mutations in the cell membrane thyroid hormone (TH) transporter monocarboxylate transporter (MCT) 8 produce severe neuropsychomotor defects and characteristic thyroid function test (TFT) abnormalities. Two children with mild neurological phenotypes and normal TFTs were found to harbor MCT8 gene variants of unknown significance (VUS), MCT8-R388Q that occurred de novo, and MCT8-Q212E. Normal TH transport and action in fibroblasts of MCT8-R388Q was demonstrated in a novel nonradioactive functional assay measuring the intracellular TH availability after L-T3 treatment. No genotype-phenotype correlation was found in additional family members carrying MCT8-Q212E. For the field of MCT8 deficiency, it is important to assess the significance of MCT8 gene VUS.
Background: Papillary thyroid microcarcinoma (PTMC) typically carries an excellent prognosis regardless of the treatment option pursued. Radiofrequency ablation (RFA) for thyroid disease has been utilized for benign lesions and in patients who are poor surgical candidates for thyroid cancers with compression symptoms, but the efficacy and limitations of RFA as first-line therapy for PTMC has not been described in adequate detail. The purpose of our study was to investigate RFA versus thyroidectomy in terms of efficacy, oncologic outcomes, quality of life, complications, and costs over a 5-year period of follow-up. Methods: From January 2013 to November 2013, 174 consecutive patients with an isolated, solitary intrathyroidal PTMC were identified in a Chinese teaching hospital. Those with more aggressive or advanced PTMCs were not included. Ninety-four patients elected RFA and 80 patients elected surgery for treatment of these PTMC. Nodules were confirmed to be PTMC without an aggressive histological type by core needle biopsy in the RFA group and by final surgical pathology in the surgery group. The extent of surgery was decided based on patient preferences in consultation with the surgeons. Of all the patients, 58 (72.5%) underwent lobectomy, 22 (27.5%) underwent total thyroidectomy, and 53 (66.3%) underwent lymph node dissection. Pre- and post-treatment variables were compared between the two groups, including demographics, tumor characteristics, treatment, local tumor progression, lymph node metastasis, distant metastasis, local recurrence, complications, and quality-of-life findings. Results: Patient-specific variables were similar between groups, as were oncologic outcomes after 5 years of follow-up. When compared with RFA, surgery took longer, had a longer hospitalization time, and was costlier (all p < 0.001). The surgery group had three complications, with 2 (2.5%) permanent recurrent laryngeal nerve injuries and 1 (1.3%) case of permanent hypoparathyroidism, while none was reported in the RFA group (p = 0.095). The surgery group had a lower post-treatment thyroid-related quality of life. Conclusions: For carefully selected low-risk intrathyroidal PTMC, RFA was not oncologically inferior to open surgery, and it was associated with a higher quality of life and lower overall costs.
Background: Iodine deficiency (ID) is a global problem in individuals living in an iodine-deficient environment, specifically in mountainous regions. However, data regarding the iodine nutritional status of Tibetan people in the plateau are limited. Methods: A population-based survey was conducted from July 2016 to July 2017 in Lhasa, Tibet, including 12 communities in Lhasa city and 10 surrounding rural areas. The iodine nutritional status of Tibetan people was evaluated using the traditional iodine nutrition indexes: urinary iodine concentration (UIC), thyroid size, serum thyroxine, thyrotropin, thyroglobulin antibody and thyroid peroxidase antibody (TPOAb). Results: A total of 2295 healthy participants were screened, and 2160 participants who had completed all the required examinations were enrolled in this study (response rate, 94.1%). Urinary iodine showed a skewed distribution, with a median (upper and lower quartiles) of 154 (99-229) mu g/L. The percentages of low iodine (UIC <100 mu g/L), adequate iodine (UIC, 100-199 mu g/L), and high iodine (UIC >= 200 mu g/L) were 25.6%, 42.0%, and 32.4%, respectively. The urinary iodine level in the urban region was higher than that in the rural region (p < 0.05). Urinary iodine levels were lower with increasing age (p < 0.05). The prevalence of hyperthyroidism, hypothyroidism, goiter, TPOAb positivity, and thyroglobulin antibody positivity was 1.0%, 21.8%, 4.7%, 6.6%, and 10.4%, respectively. Logistic regression analysis found that urinary iodine was an independent risk factor for TPOAb positivity (odds ratio = 0.997 [95% confidence interval, 0.995-0.999]; p < 0.001). Conclusions: Compared with individuals living in the plains of China, Tibetan adults have a higher rate of ID. UIC was an independent risk factor for TPOAb positivity. This public health issue should be further investigated.
Background: Thyroid autoimmunity (TAI) is the most frequent autoimmune disease among reproductive-aged women. It has been related to premature ovarian insufficiency, but the mechanisms remain elusive, and its association with ovarian reserve in euthyroid women is debatable. Moreover, the impact of TAI on assisted reproduction is controversial: especially for women with diminished ovarian reserve (DOR), few studies are available. Therefore, the present study was aimed to look for an association between TAI and DOR, and to evaluate the effect of TAI on pregnancy outcomes and offspring health following assisted reproductive technology stratified by ovarian reserve. Methods: A total of 6213 euthyroid women from the Reproductive Hospital Affiliated to Shandong University between 2012 and 2017 were retrospectively included. The prevalence of DOR in women with negative or positive TAI was calculated, and pregnancy and neonatal outcomes after in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) cycles were compared between the TAI-positive and TAI-negative groups both in women with DOR and in those with normal ovarian reserve (NOR). Longitudinal growth parameters and temperament type of the offspring were also observed in the TAI-positive and TAI-negative groups. Results: The prevalence of DOR in women with positive TAI and those with negative TAI was not significantly different (4.09% vs. 2.96%, p = 0.053), even after stratifying patients by age. In women with DOR, the live birth rate, pregnancy loss rate, neonatal complication rate, and offspring outcomes between the TAI-positive and TAI-negative groups were comparable (p > 0.05). In women with NOR, a higher rate of live births (44.94% vs. 40.34%, p = 0.027) and a higher prevalence of congenital anomalies (4.68% vs. 2.14%, p = 0.005) were observed in the TAI-positive group. Conclusions: TAI had no impact on ovarian reserve in euthyroid women and had no association with IVF/ICSI outcomes in women with DOR. Although an increased incidence of congenital anomalies in the TAI-positive group was observed in women with NOR, an association between neonatal anomalies and TAI cannot be demonstrated. Large cohort studies to evaluate the effects of TAI on offspring health are warranted, and further experimental studies are required to explore the underlying mechanisms.
Background: It has been advocated to apply individualized strategies to evaluate thyroid nodules due to the growing awareness that the pathogenesis of thyroid cancer is not uniform. Molecular markers in fine needle biopsies (FNBs) may be helpful for the diagnosis and management decisions. Unlike the detection of BRAF mutations, the clinical utility of rat sarcoma viral oncogene homolog (RAS) mutations has not been fully elucidated. This study aimed at presenting a real-world performance of RAS mutations in identifying thyroid malignancies, at investigating the nature of thyroid tumors carrying RAS mutations, and at providing an additional reference for interpreting how to utilize the presence of RAS mutations in the decision-making process of thyroid nodule management. Methods: Between February 2015 and December 2017, 1400 sequential thyroid biopsies were performed at Boston Medical Center. Of these, 546 FNBs were evaluated for RAS mutations by using a ThyroSeq next-generation sequencing panel. Nodules carrying RAS mutations were prospectively followed, and medical records were collected. Results: ThyroSeq successfully provided molecular information in 504 nodules; 173 with molecular alteration(s); and 80 positive for mutations in the Kirsten-, Neuroblastoma-, or Harvey-RAS genes. RAS gene mutations constituted up to 46.2% of the total molecular alterations found in the study. Fifty-six of the 80 RAS-positive nodules underwent surgery, 33 (58.9%) were confirmed to be benign, 7 (12.5%) were noninvasive follicular thyroid neoplasms with papillary-like nuclear features (NIFTP), and 16 (28.6%) were thyroid carcinomas. The positive predictive value, negative predictive value, and accuracy of RAS mutations for identifying malignancies among cytologically indeterminate nodules were 25.5%, 89.7%, and 54.0% when NIFTP was not counted as cancer. A combination of RAS and other mutations increased the risk of malignancy. Twelve histopathologically proved RAS-only-positive malignant nodules all showed low-risk features and favorable prognosis. RAS isoforms added little assistance for predicting a malignancy and the response to therapy in our series. Conclusions: RAS mutations represent the most frequently detected genetic alterations in our series. RAS mutations, when occurring alone, are not helpful markers to identify malignancy among Bethesda III/IV cytologies, but may predict favorable behavior, and hence should be considered to guide initial management.
Purpose: To systemically investigate the prognostic value of pre-ablation stimulated thyroglobulin (s-Tg) in children and adolescents with differentiated thyroid cancer. Methods: Clinical records from 118 children and adolescents were retrospectively reviewed. Results: The median age was 16 years, and the majority were female (79.7%). All children and adolescents underwent total thyroidectomy and received radioactive iodine therapy. After a median follow-up of 5.3 years, 68 (57.6%) patients were disease free, while 50 patients (42.4%) had persistent/recurrent disease. In multivariate analysis, pre-ablation s-Tg and M1 were the independent predictive factors for persistent/recurrent disease. According to the receiver operating curve analysis, the best pre-ablation s-Tg cutoff to predict disease-free status was 17.8 ng/mL with a negative predictive value of 96.8%. Integration of pre-ablation s-Tg into American Thyroid Association pediatric risk categories indicated that the presence of pre-ablation s-Tg <= 17.8 ng/mL was associated with a decreased chance of having persistent/recurrent disease in intermediate- and high-risk patients (22.6% to 2.6% in intermediate-risk patients, and 64.4% to 5.6% in high-risk patients). Conclusions: Our findings suggest that pre-ablation s-Tg has the capability of predicting the clinical outcomes in children and adolescents with thyroid cancer.
Background: A population-based reference interval (RI) of thyroid hormones in pregnancy using a standardized methodology is crucial for clinicians to make accurate diagnoses and important for the comparison of test results obtained from different analytic platforms. Methods: We enrolled 600 healthy Chinese women to obtain longitudinal serum samples across gestation, after exclusion of subjects with antibodies to thyroid peroxidase, thyroglobulin or thyrotropin receptor. Gestational age-specific RIs were constructed by using polynomial regression equations with MLwiN. Results: Free thyroxine (fT4) levels rose to a peak at the 7th-8th gestational weeks and then declined gradually till 28th week, while thyrotropin (TSH) level decreased from early pregnancy to a nadir at the 9th week. The data support the recent notion by the American Thyroid Association to raise the TSH upper RI to 4.0 mIU/L. We also demonstrate that thyroid hormone reference ranges are not affected in a mildly iodine-deficient population and by including women with the presence of antibodies against thyroid peroxidase and thyroglobulin who are otherwise healthy. Conclusions: The study highlights a methodology in constructing gestational age-specific thyroid function test RIs on different analytic platforms to provide a better interpretation and comparison of results obtained across different platforms.
Background: Remarkable achievements have been made for over two decades by implementing a universal salt iodization policy in China. However, changes in the standards and manufacturing of iodized salt have made it necessary to regularly monitor iodine intake and thyroid function in the population. Therefore, we aimed to evaluate iodine nutritional status using thyroid function and urinary iodine levels in pregnant women via a national survey. Methods: Participants included pregnant women enrolled in the 2015 Chinese Adults Chronic Diseases and Nutrition Surveillance (CACDNS). Urinary iodine concentration (UIC) was measured using arsenic and cerium catalysis spectrophotometry. Plasma levels of thyrotropin (TSH), fT4 (free thyroxine), thyroid peroxidase antibodies (TPO-Ab), and thyroglobulin antibodies (TG-Ab) were determined using an automated chemiluminescence immunoassay analyzer. Results: UIC was analyzed in a total of 6173 urinary samples collected from pregnant women residing in 31 provinces of China. The median urinary iodine concentration was 146 mu g/L. Pregnant women with UIC >250 mu g/L had higher TSH levels than those with UIC 120-149 and 150-249 mu g/L. After excluding women with positive TPO-Ab and TG-Ab, a total 2097 plasma samples collected from pregnant women during three periods were analyzed for TSH and fT4. The frequency of normal thyroid function in this study was >85%. The most common type of abnormal thyroid function, subclinical hypothyroidism, was similar to the 2010-2012 China Nutrition and Health Surveillance (CNHS), but its prevalence decreased to 6.8% compared with that in the 2010-2012 CNHS. Compared with pregnant women who had UIC <250 mu g/L, the prevalence of subclinical hypothyroidism was significantly increased in those with UIC >250 mu g/L. Even with UIC levels 120-149 mu g/L, the iodine status of Chinese pregnant women in the 2015 CACDNS is acceptable. Conclusions: The iodine status of pregnant women is generally adequate, but urinary iodine levels are close to the cutoff for suitable iodine status (150 mu g/L). With noniodized salt widely available, continued improvement in monitoring of iodine nutritional status and thyroid function is important in this population.
Approximately 5% of all cases of papillary thyroid cancer (PTC) are inherited. However, the susceptibility gene(s) for nonsyndromic familial PTC (FPTC) remain unclear. We performed whole genome sequencing of peripheral blood DNA samples from two affected family members with PTC. CHEK2 transcript expression and the protein levels of CHK2 and p53 were evaluated in the thyroid tissues from two affected members of the kindred and sporadic PTC cases. The entire CHEK2 coding sequence was examined by Sanger sequencing in blood DNA samples from 242 sporadic PTC patients. We identified a novel heterozygous germline mutation in CHEK2 (c.417C -> A) that was detected in all available affected members of a kindred with FPTC. This variant was found in only 1 out of 264,200 persons in the Genome Aggregation Database and the NHLBI Trans-Omics for Precision Medicine program. The CHEK2 c.417C -> A variant introduces a premature termination codon (Y139X). We found reduced CHK2 protein expression in tumor samples from the two patients who carried the variant as compared with sporadic cases without the variant. The Y139X loss-of-function variant led to reduced p53 phosphorylation and decreased p53 protein level. In addition, two rare missense variants (R180C and H371Y) in CHEK2 were identified in 5 (2%) of 242 patients with sporadic PTC. Our findings suggest that the CHEK2 Y139X variant may be associated with FPTC.
Background: Accurate preoperative prediction of cervical lymph node (LN) metastasis in patients with papillary thyroid carcinoma (PTC) provides a basis for surgical decision-making and the extent of tumor resection. This study aimed to develop and validate an ultrasound radiomics nomogram for the preoperative assessment of LN status. Methods: Data from 147 PTC patients at the Wuhan Tongji Hospital and 90 cases at the Hunan Provincial Tumor Hospital between January 2017 and September 2019 were included in our study. They were grouped as the training and external validation set. Radiomics features were extracted from shear-wave elastography (SWE) images and corresponding B-mode ultrasound (BMUS) images. Then, the minimum redundancy maximum relevance algorithm and the least absolute shrinkage and selection operator regression were used to select LN status-related features and construct the SWE and BMUS radiomics score (Rad-score). Multivariate logistic regression was performed using the two radiomics scores together with clinical data, and a nomogram was subsequently developed. The performance of the nomogram was assessed with respect to discrimination, calibration, and clinical usefulness in the training and external validation set. Results: Both the SWE and BMUS Rad-scores were significantly higher in patients with cervical LN metastasis. Multivariate analysis indicated that the SWE Rad-scores, multifocality, and ultrasound (US)-reported LN status were independent risk factors associated with LN status. The radiomics nomogram, which incorporated the three variables, showed good calibration and discrimination in the training set (area under the receiver operator characteristic curve [AUC] 0.851 [CI 0.791-0.912]) and the validation set (AUC 0.832 [CI 0.749-0.916]). The significantly improved net reclassification improvement and index-integrated discrimination improvement demonstrated that SWE radiomics signature was a very useful marker to predict the LN metastasis in PTC. Decision curve analysis indicated that the SWE radiomics nomogram was clinically useful. Furthermore, the nomogram also showed favorable discriminatory efficacy in the US-reported LN-negative (cN0) subgroup (AUC 0.812 [CI 0.745-0.860]). Conclusions: The presented radiomics nomogram, which is based on the SWE radiomics signature, shows a favorable predictive value for LN staging in patients with PTC.
Background: Sporadic medullary thyroid carcinoma (MTC) is a relatively uncommon neuroendocrine malignancy and the molecular tumorigenesis of its sporadic type (sMTC) is only partially understood. In this study, we performed a study focusing on the genomic and transcriptomic characterization of sMTC. Methods: Twenty-nine sMTC patients were included. Whole-exome sequencing (WES) was carried out in 18 patients, including both tumor samples and matched noncancerous tissues. Whole transcriptome sequencing (RNA-Seq) was performed in all 29 tumors. WES, RNA-Seq, and copy number alteration (CNA) data were analyzed. A Cell Counting Kit-8 (CCK-8) assay was used to evaluate cell proliferation. Results: Among the somatic mutations, RET was the only recurrently cancer-related mutated gene (5/18, 27.8%). In the germline, FAT1 and FAT4, two members of the FAT gene family, were identified as the two most common mutated genes. CNA analysis found that FAT1 and FAT4, both located on chromosome 4q, were also two of the genes most commonly affected by somatic chromosomal deletions (4/18, 22.2%). Using TT and MZ-CRC-1 cell lines, the CCK-8 assay showed that FAT1 and FAT4 knockdown could promote MTC cell proliferation. Based on the gene expression profile, patients were clustered into two molecular subtypes: the mesenchymal-like subtype is characterized by epithelial-mesenchymal transition, while the proliferative-like subtype is associated with enrichment of cell cycle pathways. Most events of structural recurrence (80%) occurred in the proliferative-like subtype. Conclusion: In addition to RET, these findings demonstrate that FAT1/FAT4 genomic alterations appear to be frequent in sMTC. Two molecular subtypes of sMTC with distinct biological behavior could be identified. However, these results need to be validated by larger samples and more comprehensive experiments in the future, especially for the frequency and function of FAT1/FAT4 germline variants.
Background: Mandatory universal salt iodization (USI) has been implemented in China for 20 years. Although iodine deficiency disorders are effectively controlled, the risk of excess iodine have been debated. Methods: A nationally representative cross-sectional study with 78,470 enrolled participants, aged 18 years or older, from all 31 provincial regions of mainland China was performed. The participants were given a questionnaire and underwent B-mode ultrasonography of the thyroid. Serum concentrations of thyroid hormones, thyroid antibodies, and urine iodine concentration (UIC) were measured. Results: The median UIC of the adult population was 177.89 mu g/L. The weighted prevalence of thyroid disorders in adults were as follows: 0.78% of overt hyperthyroidism, 0.44% of subclinical hyperthyroidism, 0.53% of Graves' disease, 1.02% of overt hypothyroidism, 12.93% of subclinical hypothyroidism, 14.19% of positive thyroid antibodies, 10.19% of positive thyroid peroxidase antibodies, 9.70% of positive thyroglobulin antibodies, 1.17% of goiter, and 20.43% of thyroid nodules. Iodine excess was only associated with higher odds of overt hyperthyroidism and subclinical hypothyroidism, while iodine deficiency was significantly associated with higher odds of most thyroid disorders. In addition, increased iodine intake was significantly associated with elevated serum thyrotropin levels but was inversely associated with thyroid antibodies and thyroid nodules. Conclusions: The long-term mandatory USI program with timely adjustments is successful in preventing iodine deficiency disorders, and it appears to be safe. The benefits outweigh the risks in a population with a stable median iodine intake level of up to 300 mu g/L.