Abnormalities of serum magnesium levels in dialysis patients undergoing parathyroidectomy.

In primary hyperparathyroidism, postoperative hypocalcemia can be exacerbated by magnesium deficiency. However, the significance of magnesium homeostasis in surgery for secondary hyperparathyroidism is unknown. In this study, 268 consecutive adult patients on renal replacement therapy who underwent parathyroidectomy for secondary hyperparathyroidism were included for analysis. We found that about one fifth presented with hypomagnesemia (5.6%) or hypermagnesemia (14.6%). Hypomagnesemia was associated with lower calcium levels and longer postoperative hospital stays. Hypermagnesemia was associated with higher calcium-phosphorus products and lower parathyroid hormone levels. In multivariate analysis, patient age, alkaline phosphatase, and osteocalcin were independent predictors of prolonged stay after parathyroidectomy. There was a positive correlation between serum magnesium levels and severity of itching in these patients. Calcium-phosphorus products and serum magnesium levels were independently associated with pruritus. In conclusion, magnesium abnormalities play a minor role in hungry bone syndrome after parathyroidectomy for secondary hyperparathyroidism. Patients with higher serum magnesium levels had greater severity of pruritus.

Sublethal thermal stress promotes migration and invasion of thyroid cancer cells.

OBJECTIVE: Radiofrequency ablation is a viable option in the treatment of benign thyroid nodules. Some reports suggest that thermal ablation may also be safe for the management of low-risk thyroid cancer. In this study, we applied transient heat treatment to thyroid cancer cells to mimic clinical scenarios in which insufficient ablation leads to incomplete eradication of thyroid cancer. METHODS: Differentiated thyroid cancer cell lines B-CPAP, TPC-1, and FTC-133 were subjected to heat treatment at different temperatures for 10 min. Effects on cell growth, clonogenicity, wound healing assay, and Transwell invasion were determined. RESULTS: Heat treatment at 45°C or higher reduced cell growth, whereas viability of thyroid cancer cells was not changed after heat treatment at 37, 40, or 42°C. Heat treatment at 40°C increased the number of colony formations by 16% to 39%. Additionally, transient heat treatment at 40°C resulted in a 1.75-fold to 2.56-fold higher migratory activity than treatment at 37°C. Invasive capacity was increased after heat treatment, ranging from 115% to 126%. Expression of several epithelial-mesenchymal transition markers, including ZEB1, N-cadherin, and MMP2, was upregulated following heat treatment at 40°C. CONCLUSION: We for the first time demonstrate that sublethal thermal stress may increase clonogenicity, migration, and invasion of thyroid cancer cells.

SOX4 is a pivotal regulator of tumorigenesis in differentiated thyroid cancer.

The SOX family consists of about 20 transcription factors involved in embryonic development, reprogramming, and cell fate determination. In this study, we demonstrated that SOX4 was significantly upregulated in differentiated thyroid cancer. Immunohistochemical analysis revealed that high SOX4 expression was associated with papillary histology, extrathyroidal extension, lymph node metastasis, and advanced disease stage. Patients whose tumors exhibited high SOX4 expression had a shorter recurrence-free survival, though significance was lost in multivariate Cox regression analysis. SOX4 silencing in thyroid cancer cells slowed cell growth, attenuated clonogenicity, and suppressed anoikis resistance. Additionally, SOX4 knockdown impeded xenograft tumor growth in nude mice. Knockdown of SOX4 expression was accompanied by reduced phosphorylation of AKT and ERK. Furthermore, CRABP2 expression correlated with SOX4 expression, and SOX4 silencing decreased CRABP2 expression and its downstream effectors such as integrin ß1 and ß4. These results indicate that SOX4 has both prognostic and therapeutic implications in differentiated thyroid cancer, and targeting SOX4 may modulate tumorigenic processes in the thyroid.

Downregulation of cellular retinoic acid binding protein 1 fosters epithelial-mesenchymal transition in thyroid cancer.

Cellular retinoic acid binding protein 1 (CRABP1) participates in the regulation of retinoid signaling. Previous studies showed conflicting results regarding the role of CRABP1 in tumor biology, including protumorigenic and tumor-suppressive effects in different types of cancer. Our bioinformatics analyses suggested that CRABP1 expression was downregulated in thyroid cancer. Ectopic expression of CRABP1 in thyroid cancer cells suppressed migratory and invasive activity without affecting cell growth or cell cycle distribution. In transformed normal thyroid follicular epithelial cells, silencing of CRABP1 expression increased invasiveness. Additionally, CRABP1 overexpression was associated with downregulation of the mesenchymal phenotype. Kinase phosphorylation profiling indicated that CRABP1 overexpression was accompanied by a decrease in phosphorylation of epidermal growth factor (EGF) receptor and downstream phosphorylation of Akt, STAT3, and FAK, which were reversed by exogenous EGF treatment. Immunohistochemical analysis of our tissue microarrays revealed an inverse association between CRABP1 expression and disease stage of differentiated thyroid cancer. Taken together, our results suggest that CRABP1 expression is aberrantly lost in thyroid cancer, and this downregulation promotes the epithelial-mesenchymal transition at least partly through modulating EGF receptor signaling.

Impact of completion thyroidectomy on postoperative recovery in patients with differentiated thyroid cancer.

While an increasing number of patients now undergo lobectomy for low-risk differentiated thyroid cancer, a subset of patients require completion thyroidectomy to facilitate radioactive iodine therapy. Completion thyroidectomy is generally as safe as the initial operation, but a previous study showed that a longer hospitalization is required for completion thyroidectomy. In this study, we reviewed 61 consecutive patients who had been treated with an initial lobectomy and subsequent completion thyroidectomy at our institution from 2012 to 2021. We detected a changepoint in 2016 for the proportion of patients who were treated with a thyroid lobectomy (Pettitt's test P = 0.049). The rate of completion thyroidectomy remained stable throughout the study period. There was no difference in operating time, intraoperative blood loss, perioperative drop in calcium levels, and postoperative hospital stay between surgeries. The patients reported higher pain scores on the day of operation (P = 0.007) and the postoperative day 1 (P = 0.022). Occult papillary microcarcinomas were identified in the contralateral thyroid lobe in 13 (21%) patients. Multifocality was the only predictor for residual malignancy in multivariate regression. In conclusion, patients with differentiated thyroid cancer experienced more pain in the immediate postoperative period following completion thyroidectomy. Hospital stays do not change with appropriate opioid-free pain control.

Aberrant Expression of Thymosin Beta-4 Correlates With Advanced Disease and BRAF V600E Mutation in Thyroid Cancer.

Thymosin beta-4 (TMSB4X) was recently identified as a differentially expressed gene between malignant and non-malignant thyroid cells via single-cell RNA sequencing. In the present study, we aimed to study the immunostaining pattern of TMSB4X in benign and malignant thyroid neoplasms. Immunohistochemical analysis revealed that normal thyroid tissue or benign thyroid disorders exhibited undetectable immunoreactivity against TMSB4X except for positive staining of inflammatory infiltrates and stromal cells associated with autoimmune thyroid disease. By contrast, overexpression of TMSB4X was observed in a variety of thyroid malignancies, including papillary, follicular, poorly differentiated, and undifferentiated thyroid cancer. Among 141 patients with differentiated thyroid cancer, higher TMSB4X expression was associated with papillary tumor type, extrathyroidal extension, lymph node metastasis, and BRAF V600E mutation. The results were consistent with those from the public transcriptomic datasets. In summary, TMSB4X expression was aberrantly increased in various types of thyroid cancer, and higher TMSB4X expression was correlated with advanced disease characteristics. Thymosin beta-4 may be a novel downstream effector of the BRAF V600E mutation.

CRABP2 Is Associated With Thyroid Cancer Recurrence and Promotes Invasion via the Integrin/FAK/AKT Pathway.

Cellular retinoic acid-binding protein 2 (CRABP2) participates in retinoid partitioning between different nuclear receptors. Recently, we identified that CRABP2 is one of the progression-associated genes in thyroid cancer. To explore the prognostic and functional significance of CRABP2, immunohistochemical analysis was performed in thyroid tissues and neoplasms. Overexpression of CRABP2 was observed in malignant thyroid neoplasms but not in benign thyroid lesions. CRABP2 expression was an independent predictive factor for recurrence-free survival in patients with differentiated thyroid cancer. Knockdown of CRABP2 reduced the sensitivity of thyroid cancer cells to retinoic acid. Importantly, CRABP2 expression in thyroid cancer cells was associated with epithelial-mesenchymal transition properties, including anoikis resistance, migration, and invasion capacity. Furthermore, invasion promoted by CRABP2 was mediated at least partly by the integrin/focal adhesion kinase/AKT pathway. In summary, CRABP2 expression is upregulated in thyroid cancer with adverse prognostic implications. The invasion-stimulating effects appear independent of canonical retinoic acid signaling and may serve as a potential therapeutic target.

Propensity Score-Matched Analysis to Identify Pathways Associated with Loss of Sodium Iodide Symporter in Papillary Thyroid Cancer.

Sodium iodide symporter (NIS) expression in thyroid follicular cells plays an important role in normal physiology and radioactive iodine therapy for thyroid cancer. Loss of NIS expression is often seen in thyroid cancers and may lead to radioiodine refractoriness. To explore novel mechanisms of NIS repression beyond oncogenic drivers, clinical and RNA-seq data from the thyroid cancer dataset of The Cancer Genome Atlas were analyzed. Propensity score matching was used to control for various genetic background factors. We found that tumoral NIS expression was negatively correlated with tumor size. Additionally, low NIS expression was the only factor associated with recurrence-free survival in a Cox multivariate regression analysis. After matching for clinicopathologic profiles and driver mutations, the principal component analysis revealed distinct gene expressions between the high and low NIS groups. Gene set enrichment analysis suggested the downregulation of hedgehog signaling, immune networks, and cell adhesions. Positively enriched pathways included DNA replication, nucleotide excision repair, MYC, and Wnt/ß-catenin pathways. In summary, we identified several potential targets which could be exploited to rescue the loss of NIS expression and develop redifferentiation strategies to facilitate radioactive iodine therapy for thyroid cancer.

ACLY inhibitors induce apoptosis and potentiate cytotoxic effects of sorafenib in thyroid cancer cells.

PURPOSE: ATP-citrate lyase (ACLY) is a critical enzyme at the intersection of glucose and lipid metabolism. ACLY is often upregulated or activated in cancer cells to accelerate lipid synthesis and promote tumor progression. In this study, we aimed to explore the possibility of utilizing ACLY inhibition as a new strategy in the treatment of thyroid cancer. METHODS: Bioinformatics analysis of the public datasets was performed. Thyroid cancer cells were treated with two different ACLY inhibitors, SB-204990 and NDI-091143. RESULTS: Bioinformatics analysis revealed that ACLY expression was increased in anaplastic thyroid cancer. In thyroid cancer cell lines FTC-133 and 8505C, ACLY inhibitors suppressed monolayer cell growth and clonogenic ability in a dose-dependent and time-dependent manner. Flow cytometry analysis showed that ACLY inhibitors increased the proportion of sub-G1 cells in the cell cycle and the number of annexin V-positive cells. Immunoblotting confirmed caspase-3 activation and PARP1 cleavage following treatment with ACLY inhibitors. Compromised cell viability could be partially rescued by co-treatment with the pan-caspase inhibitor Z-VAD-FMK. Additionally, we showed that ACLY inhibitors impeded three-dimensional growth and cell invasion in thyroid cancer cells. Isobolograms and combination index analysis indicated that ACLY inhibitors synergistically potentiated the cytotoxicity rendered by sorafenib. CONCLUSIONS: Targeting ACLY holds the potential for being a novel therapeutic strategy for thyroid cancer.

Multicenter Study of Benign Thyroid Nodules with Radiofrequency Ablation: Results of 762 Cases over 4 Years in Taiwan.

BACKGROUND: To evaluate the effectiveness and safety of radiofrequency ablation (RFA) treatment for benign thyroid nodules (BTNs) in five medical centers in Taiwan. METHODS: From April 2016 to July 2020, 762 patients underwent ultrasound guided RFA treatment of 826 benign thyroid nodules at five medical centers in Taiwan. The RFA procedure was performed by radiologists, otolaryngologists, or surgeons. Patients were grouped into three subgroups according to the initial volume of BTNs. The volume reduction ratio (VRR) of each nodule, and complications were analyzed at 1, 3, and 6 months after RFA treatment and the three groups compared. RESULTS: The large nodular group showed greater VRR compared to the other two groups at first 1-month follow-up. At 6-months follow-up, there was no significant difference of VRR among the three groups. Goiters with difference in size can attain a successful VRR (>50%) although different specialists demonstrated variable VRR after 6-months follow-up. A total of 40 (4.8%) complications were reported. All patients recovered spontaneously without surgery intervention. CONCLUSIONS: The reliability and safety of RFA for benign thyroid nodules had been established. RFA has gradually become an alternative to surgery in the treatment of benign thyroid nodules in Taiwan.

Computer-Analyzed Ultrasound Predictors of the Treatment Efficacy of Radiofrequency Ablation for Benign Thyroid Nodules.

AIM: Radiofrequency ablation (RFA) is a relatively safe and efficient alternative to surgery for patients with benign thyroid nodules. We investigated predictive factors associated with volume reduction using digital imaging analysis. METHODS: In this retrospective study, a prospectively maintained database containing the data of patients who received treatment from April 2019 to March 2020 was analyzed. Computerized analysis for quantitative measurement of echogenicity, heterogeneity, and the proportion of cystic components was performed on ultrasonographic images. The volume reduction rate (VRR) was calculated during follow-up. Treatment efficacy was defined as a volume reduction greater than 50% of baseline volume. RESULTS: The median volume of 58 benign thyroid nodules before RFA was 22.7 mL. Of 53 nodules with sufficient follow-up, the median VRR was 46.4%, 61.5%, 63.4%, and 67.4% at 1, 3, 6, and 12 months, respectively. Overall, at one-year follow-up, treatment efficacy was achieved in 39 (74%) nodules. In a multivariate regression analysis, the proportion of cystic components and RFA treatment time were independently associated with treatment efficacy. A subgroup analysis focusing on solid nodules indicated a negative correlation between echogenicity and VRR. CONCLUSIONS: The proportion of cystic components in thyroid nodules is the main predictor of RFA treatment efficacy. In solid nodules, higher echogenicity is associated with a lower volume reduction.

SREBP1 promotes invasive phenotypes by upregulating CYR61/CTGF via the Hippo-YAP pathway.

Aberrant lipid metabolism provides bioenergetic, biosynthetic, and redox supplies to cancer cells. Previous studies have reported differential lipid profiling in thyroid malignancies. Sterol regulatory element-binding protein 1 (SREBP1), encoded by the SREBF1 gene, is a master regulator of cellular lipid homeostasis. The clinical and functional significance of SREBP1 in thyroid cancer is not well understood. Here, we showed that SREBP1 expression is significantly upregulated in invasive thyroid cancer than in normal thyroid tissue or benign thyroid nodules. High tumoral SREBP1 expression was associated with extrathyroidal extension, advanced disease stage, and shorter disease-specific survival in patients with differentiated thyroid cancer. SREBP1 overexpression significantly increased the oxygen consumption rate, filopodia formation, and migratory and invasive capacities of thyroid cancer cells. Knockdown of SREBF1 or treatment with an SREBP1 activation inhibitor fatostatin had the opposite effect. RNA-Seq analysis showed that modulation of SREBP1 expression was accompanied by corresponding changes in the expression of epithelial-mesenchymal transition markers and CYR61/CTGF. SREBP1-facilitated cell invasion could be abrogated by treatment with a YAP inhibitor such as verteporfin or genetic silencing of CYR61 or CTGF. In summary, SREBP1 upregulation can be used as a prognostic indicator for thyroid cancer and SREBP1 overexpression is involved in cancer invasiveness, at least partly, through upregulation of CYR61/CTGF via the Hippo-YAP pathway.

Learning curve analysis of radiofrequency ablation for benign thyroid nodules.

BACKGROUND: Radiofrequency ablation (RFA) has been increasingly accepted as an alternative to surgery in the treatment of symptomatic benign thyroid nodules. However, the learning curve of thyroid RFA has yet to be defined. We hypothesized a temporal relationship between proficiency of the skill and midterm volume reduction. METHODS: Consecutive patients who underwent RFA and had at least a six-month follow-up were identified from an institutional database. The cumulative sum (CUSUM) analysis was applied to visualize the learning curve on the adjusted volume reduction rate (VRR). RESULTS: A total of 102 nodules in 93 patients were included in the analysis. Linear regression revealed that nodule composition was the main predictor of the VRR. The composition-adjusted VRR increased with the chronological treatment order. The series was divided into three phases based on inflection points of the CUSUM analysis: the initial learning phase (case 1-20), consolidation phase (case 21-65), and proficiency phase (case 66-102). In the later phase, more solid nodules were treated, power output used by the operator was higher, and RFA treatment time was longer. CONCLUSION: The treatment efficiency of thyroid RFA was associated with technical proficiency, suggesting the presence of a learning curve effect.

Preoperative Factors Associated with Extrathyroidal Extension in Papillary Thyroid Cancer.

OBJECTIVE: Extrathyroidal extension may not be accurately recognized during thyroidectomy and can increase the risk of positive margins and even recurrence. This study aimed to investigate the preoperative factors associated with extrathyroidal extension. METHODS: We analyzed 887 patients with papillary thyroid cancer (PTC) who underwent surgery in the period of 2005-2017. Binary logistic regression analyses and generalized additive models were used to identify associations. RESULTS: Minimal extrathyroidal extension was present in 233 (26%) patients and advanced extrathyroidal extension was found in 60 (7%) patients. Age, BMI, and tumor size were independent predictors of all or advanced extrathyroidal extension. Among the 493 patients whose BRAF mutation status was available, age (OR = 1.025), BMI (OR = 1.091), tumor size (OR = 1.544), and BRAF V600E mutation (OR = 2.311) were independently associated with extrathyroidal extension. CONCLUSIONS: Older age, a greater BMI, a larger tumor size, and presence of the BRAF mutation were predictive of extrathyroidal extension. These factors should be taken into consideration in decision-making before surgery is performed.

Lymphovascular invasion of papillary thyroid carcinoma revisited in the era of active surveillance.

INTRODUCTION: Lymphovascular invasion (LVI) is associated with disease recurrence and compromised survival in patients with thyroid cancer. Nonetheless, LVI is not identifiable on preoperative ultrasound or cytologic assessment. We aimed to explore the clinicopathological features associated with LVI. PATIENTS AND METHODS: We conducted a retrospective review of our prospectively maintained database from 2009 to 2018. Multivariate analyses were performed to determine the associations between clinicopathological parameters and LVI. Generalized additive models were used to examine the nonlinear relationship between continuous variables and LVI. RESULTS: A total of 795 patients were included in the analysis, and 174 (22%) had LVI. Patients' age (odds ratio [OR] = 0.982), tumor size (OR = 1.466), clinical lymphadenopathy (OR = 6.975), and advanced extrathyroidal extension (OR = 2.938) were independently associated with LVI. In the subset analysis of 198 patients with available genetic information, tumor size (OR = 1.599), clinical lymph node metastasis (OR = 3.657), and TERT promoter mutation (OR = 4.726) were predictive of LVI. Among 573 patients who had no clinical lymphadenopathy or advanced extrathyroidal extension, tumor size was the only predictor of LVI. Tumor size >1.5 cm had an increased risk of LVI based on the generalized additive model plot and receiver operating characteristic curve analysis. CONCLUSION: Tumor size is positively associated with the risk of LVI in papillary thyroid cancer. To avoid delayed treatment in patients with LVI, a tumor size of 1.5 cm may be considered as the safe upper limit for active surveillance.

Effects of Preoperative Iodine Administration on Thyroidectomy for Hyperthyroidism: A Systematic Review and Meta-analysis.

OBJECTIVE: The current guidelines recommend that potassium iodide be given in the immediate preoperative period for patients with Graves' disease who are undergoing thyroidectomy. Nonetheless, the evidence behind this recommendation is tenuous. The purpose of this study is to clarify the benefits of preoperative iodine administration from published comparative studies. DATA SOURCES: We searched PubMed, Embase, Cochrane, and CINAHL from 1980 to June 2018. REVIEW METHODS: Studies were included that compared preoperative iodine administration and no premedication before thyroidectomy. For the meta-analysis, studies were pooled with the random-effects model. RESULTS: A total of 510 patients were divided into the iodine (n = 223) and control (n = 287) groups from 9 selected studies. Preoperative iodine administration was significantly associated with decreased thyroid vascularity and intraoperative blood loss. Significant heterogeneity was present among studies. We found no significant difference in thyroid volume or operative time. Furthermore, the meta-analysis showed no difference in the risk of postoperative complications, including vocal cord palsy, hypoparathyroidism/hypocalcemia, and hemorrhage or hematoma after thyroidectomy. CONCLUSION: Preoperative iodine administration decreases thyroid vascularity and intraoperative blood loss. Nonetheless, it does not translate to more clinically meaningful differences in terms of operative time and postoperative complications.