Correlation between low expression of protein disulfide isomerase A3 and lymph node metastasis in papillary thyroid carcinoma and poor prognosis: a clinicopathological study of 1,139 cases with long-term follow-up.

The incidence of papillary thyroid carcinoma (PTC) is increasing worldwide. The biomarkers to identify aggressive types of PTC are limited, illustrating the need to establish reliable novel biomarkers. Protein disulfide isomerase A3 (PDIA3) is a chaperone protein that modulates the folding of newly synthesized glycoproteins and stress-responsive proteins in the endoplasmic reticulum. Although the role of PDIA3 in various cancers such as breast, uterine cervix, head and neck, and gastrointestinal tract has been examined, its expression in thyroid cancer has not been reported. We retrospectively reviewed accumulated data with long-term follow-up of 1,139 PTC patients, and investigated the correlation between immunohistochemical expression of PDIA3 in PTC patients and clinicopathological features and prognosis. PDIA3 expression was significantly lower in PTCs compared to normal thyroid tissues (NTT; n = 80, p = 0.002). In PTCs, correlation between low PDIA3 expression and lymph node metastasis (p = 0.018) and the number of positive nodes (p = 0.004) was observed. Patients with low PDIA3 expression exhibited worse cause-specific survival compared to those with high PDIA3 expression (p = 0.013). Our findings indicate that low PDIA3 expression is related to poor clinical outcome in PTC patients, and that PDIA3 may potentially be a novel ancillary biomarker. Further clarification of the biological role of PDIA3 in PTC is warranted for the future clinical application.

Multifocality and Progression of Papillary Thyroid Microcarcinoma During Active Surveillance.

BACKGROUND: Prospective trials of active surveillance (AS) have shown low rates of progression in low-risk papillary thyroid microcarcinoma (PTMC; T1aN0M0). However, the significance of multifocality as a prognostic factor remains controversial. METHODS: Data from 571 patients (mean age, 53.1 years; 495 females) who underwent AS were reviewed. PTMC was unifocal in 457 patients (80.0%) and multifocal in 114 patients (20.0%), with 2-5 lesions each (261 tumors in total). Tumor progression was defined as tumor size enlargement ≥ 3 mm and/or development of clinically evident lymph node metastasis (LNM). RESULTS: After a mean duration of AS of 7.6 years, 53 patients (9.3%) showed tumor enlargement and 8 patients (1.4%) developed LNM. The 10-year progression rate was 13.1%. Age, sex, and calcification pattern did not differ significantly between uni- and multifocal diseases. However, anti-thyroglobulin antibody and/or anti-thyroid peroxidase antibody was more frequently positive with multifocal PTMCs (46.7%) than with unifocal disease (34.4%, p = 0.024). Patients with uni- and multifocal disease showed no significant differences in 10-year rate of tumor enlargement (11.4% vs. 14.8%), LNM development (1.1% vs. 2.4%), or progression (12.4% vs 15.9%). Multivariate analysis of predictors for progression showed multifocality was not a significant risk factor (odds ratio, 1.45; 95% confidence interval, 0.79-2.54; p = 0.22). Eventually, 9 patients (7.9%) with multifocal PTMCs underwent surgery and 7 needed total thyroidectomy, although 7 still showed T1N0M0 low-risk cancer. CONCLUSIONS: Even patients with multiple PTMCs (T1amN0M0) are good candidates for AS. Many patients can avoid total thyroidectomy and subsequent surgical complications.

Ultrasonographic Detective Flow Imaging for Evaluating Parathyroid Adenoma in Patients with Primary Hyperparathyroidism.

BACKGROUND: Detective flow imaging (DFI) is a new imaging technology that displays low-velocity blood flow, which is difficult to visualize on conventional color Doppler ultrasonography (CDU). In this study, we compared the usefulness of DFI with that of CDU and methoxy-isobutyl-isonitrile (MIBI) scintigraphy for detecting parathyroid adenoma (PA) in patients with primary hyperparathyroidism (PHPT). METHODS: From March 2021 to March 2023, 87 PHPT patients underwent surgery, and 66 had a single PA. We performed preoperative conventional ultrasonography with CDU, MIBI scintigraphy, and DFI for 42 patients (5 males and 37 females; mean age: 61.6 ± 15.4 years). RESULTS: MIBI scintigraphy detected PA in 85.7% (36/42) patients, and both CDU and DFI detected PA in all patients. The rates of vascularity in PA detected by CDU and DFI were 71.4% (30/42) and 85.7% (36/42), respectively. Vascularity was detected by DFI in 6 patients who were negative for vascularity on MIBI scintigraphy. Furthermore, DFI detected blood supply in 6 of the 12 patients with undetectable blood supply on CDU. Fisher's exact test revealed that high or low blood flow, as determined by DFI, was significantly associated with detection of feeding vessels in PA by CDU (P < 0.001). CONCLUSIONS: DFI was useful for preoperative detection of PA blood flow.

Use of Ultrasonographic Shear Wave Measurements to Diagnose Thyroid Metastasis from Breast Carcinoma.

A metastatic thyroid tumor (MTT) arising from breast carcinoma (BC) is rare and sometimes difficult to diagnose. We present a case of MTT from BC; we suspected anaplastic thyroid carcinoma at initial presentation. The patient was a 58-year-old female with a hard nodule in the right anterior neck and a history of breast cancer. Computed tomography indicated tumors on both thyroid lobes, and ultrasonography (US) with shear wave measurement (SWM) showed malignant features. We performed fine needle aspiration cytology (FNAC), the results of which led us to strongly suspect MTT from BC. The surgically resected specimen was evaluated histopathologically, including by immunohistochemistry (IHC), and the diagnosis was confirmed. In addition to FNAC and IHC, SWM is useful to diagnose MTT from BC.

Associations of AminoIndex Cancer Screening (Breast) Grade with Clinical and Laboratory Variables.

BACKGROUND: Altered metabolism in the blood of cancer patients is closely related to changes in amino acids. Amino acids play an important physiological role as essential metabolites and regulators of metabolism. AminoIndex Cancer Screening (AICS) uses multivariate analysis of plasma-free amino acid profiles to screen for seven cancer types, including breast cancer. METHODS: To determine the clinical utility of AICS (breast), we retrospectively analyzed associations of AICS (breast) score with clinical and laboratory variables in 390 patients who underwent AICS (breast) testing. The mean age of participants was 50.7 years (range: 26-87 years) and all were female. RESULTS: The AICS (breast) grade was A, B, and C for 250 (64.1%), 90 (23.1%), and 50 (12.8%) participants, respectively. AICS (breast) was significantly correlated with AICS (gastric) (r = 0.487, p < 0.0001) and AICS (lung) (r = 0.523, p < 0.0001). Multivariate linear regression analysis showed no significant difference of AICS (breast) grade with age, body mass index, estimated glomerular filtration rate, dyslipidemia, or blood pressure. However, neutrophil-to-lymphocyte ratio significantly differed in relation to AICS (breast) grade (cut-off value, 1.7; p = 0.030), although only data from 72 patients were analyzed. CONCLUSION: To our knowledge, this is the first study to report associations of AICS (breast) grade with clinical variables.

VsN, a Reliability-index of Shear-wave Measurement in Sonoelastography, Is Useful for the Diagnosis of Thyroid Tumor Malignancy.

BACKGROUND/AIM: Shear wave measurement (SWM) is a new elastography modality that quantifies the shear wave velocity (Vs) and the percentage of the net effective shear wave velocity (VsN). This study examined whether these parameters could be used to differentiate between malignant and benign thyroid tumors. PATIENTS AND METHODS: The study of SWM enrolled 111 patients (133 nodules) who underwent thyroid surgery. Overall, 61 nodules were diagnosed as benign and 72 as malignant, of which 68 nodules were diagnosed as papillary thyroid carcinoma (PTC) and 4 as follicular thyroid carcinoma (FTC). RESULTS: A preoperative SWM revealed that Vs was significantly higher and VsN significantly lower in the PTC compared to the benign nodules. The VsN of FTC was significantly lower than that of follicular adenoma. Multivariate analysis revealed that VsN significantly correlated with nodule malignancy. CONCLUSION: SWM parameters, especially VsN, can potentially differentiate between benign and malignant thyroid nodules non-invasively.

RNA recovery from specimens of duct-washing cytology performed contemporaneously with mammary ductoscopy.

OBJECTIVE: Conventional cytological diagnosis including duct-washing cytology (DWC) is sometimes performed using ductal epithelial cells collected during mammary ductoscopy; it is useful for detection of early-stage breast cancer such as ductal carcinoma in situ (DCIS). However, conventional cytological diagnosis focuses exclusively on cellular morphology; false negatives and false positives may be caused by inadequate specimen preparation (triggering cell degeneration) or poor examiner diagnostic skills. Molecular diagnosis using RNA biomarkers is expected to compensate for the weaknesses of cytological diagnosis. We previously employed microarray analysis to identify highly expressed genes in DCIS, suggesting that they may be useful for DCIS diagnosis. Here, we explored whether DWC samples yielded RNA of sufficient quantity and quality for RNA biomarker-based diagnosis. RESULTS: We extracted RNAs from 37 DWC samples. RNA from 12 samples exhibited RNA integrities of ≥ 6, indicative of moderate-to-high quality. We then showed that cocaine and amphetamine regulated transcript prepropeptide (CARTPT) and breast cancer-associated transcript 54 (BRCAT54) mRNA-previously shown by microarray analysis to be highly expressed in DCIS-were detectable in these samples. Therefore, DWC samples may be useful for molecular diagnosis involving RNA biomarkers.

Usefulness of ultrasonographic detective flow imaging to diagnose parathyroid tumors: two case reports.

Parathyroid tumors (PTs) are sometimes difficult to diagnose because they are small and have a low-velocity blood flow, which can be missed by current imaging modalities. PTs consist of parathyroid adenoma (PA), parathyroid cyst, and parathyroid carcinoma (PC). Detective flow imaging (DFI) is a new imaging technology that displays a low-velocity blood flow. Herein, we report two cases in which DFI was useful for the diagnosis of PTs. One case consisted of a PA and a parathyroid cyst in close proximity, and the other was a PC. To the best of our knowledge, this is the first report to demonstrate the usefulness of DFI in the diagnosis of PTs.

Learning Curve for Endoscopic Thyroidectomy Using Video-Assisted Neck Surgery: Retrospective Analysis of a Surgeon's Experience with 100 Patients.

BACKGROUND: Endoscopic thyroidectomy offers excellent cosmetic outcomes but requires some time for surgeons to become proficient. We examined the learning curve for the first 100 patients treated by a single surgeon using a subclavian approach for video-assisted neck surgery (VANS). METHODS: We retrospectively studied the records of 100 patients (99 women, 1 man; mean age, 36.2 years) with either benign or malignant thyroid disease treated between 2016 and 2020. RESULTS: Preoperative diagnosis was papillary thyroid carcinoma (PTC) in 36 cases and other (non-PTC) in 64 cases. All patients underwent lobectomy, with additional unilateral central node dissection for patients with PTC. Mean operative time was 125 min for non-PTC cases and 129 min for PTC cases (p = 0.43); blood loss was 33.8 mL and 7.6 mL, respectively (p = 0.01). Recurrent laryngeal nerve paralysis (RNP) was observed in 12 patients (12%) and hemorrhage in 2 patients (2%). In a comparison of the first 30 cases and subsequent 70 cases, no significant differences in operative time or blood loss were evident, although tumor size was significantly greater among later non-PTC cases (32.4 mm vs. 39.5 mm, p = 0.039). RNP was significantly lower in later cases (26.7% vs. 5.7%, p = 0.003). Multivariate analysis revealed that tumor size was a significant risk factor for increased blood loss, and increased experience significantly correlated with a decrease in RNP. CONCLUSIONS: In VANS, satisfactory surgical proficiency was reached after treating 30 patients.

Clinical Utility of Fine-Needle Aspiration Cytology for Adenoid Cystic Carcinoma of the Trachea with Thyroid Invasion: A Case Report.

BACKGROUND: Adenoid cystic carcinoma of the trachea (ACCT) is a rare cancer; ACCT with thyroid invasion is particularly rare. We first suspected anaplastic thyroid carcinoma (ATC) but diagnosed ACC after performing fine-needle aspiration cytology (FNAC). Tracheal origin was confirmed postoperatively. CASE DESCRIPTION: A 77-year-old woman presented to our hospital with acute inspiratory dyspnea requiring emergency tracheotomy. Physical examination revealed swelling of the right anterior neck and a hard, immobile mass. Computed tomography (CT) and ultrasonography (US) showed tumor extension to the right thyroid lobe and between the first and third tracheal rings, which caused severe stenosis of the lumen. We performed FNAC. Clinical findings were highly suggestive of ACCT with thyroid invasion. She underwent total laryngectomy, cervical esophagectomy, and thyroidectomy with bilateral selective neck dissection at another hospital. The tumor was located in the right posterior wall of the trachea and extended into the right thyroid gland. Pathological examination showed infiltrative carcinomatous proliferation with tubular and cribriform patterns. The tumor was classified as pT4N1. A definite diagnosis was made after histopathological analysis of the surgical specimen confirmed ACCT. The tumor was positive for FABP7, a putative prognostic marker of ACC, and metastasized to the lungs 3 years after surgery. CONCLUSIONS: ACCT with thyroid invasion is an extremely rare malignant neoplasm. FNAC was useful for differentiating ACCT from other diagnoses and enabled appropriate surgical treatment.

Microarray analysis of ductal carcinoma in situ samples obtained by puncture from surgical resection specimens.

OBJECTIVE: The incidence of ductal carcinoma in situ (DCIS) is increasing due to more widespread mammographic screening. DCIS, the earliest form of breast cancer, is non-invasive at the time of detection. If DCIS tissues are left undetected or untreated, it can spread to the surrounding breast tissue. Thus, surgical resection is the standard treatment. Understanding the mechanism underlying the non-invasive property of DCIS could lead to more appropriate medical treatments, including nonsurgical options. DATA DESCRIPTION: We conducted a microarray-based genome-wide transcriptome analysis using DCIS specimens obtained by puncture from surgical specimens immediately after surgery.

Congenital hemophilia A diagnosed with postoperative hemorrhage after thyroidectomy for papillary thyroid carcinoma: a case report.

BACKGROUND: Postoperative bleeding in thyroid surgery is a serious complication with fatal outcomes. Risk factors for postoperative hemorrhage have been reported as old age, male sex, Graves' disease, use of anticoagulants, and hematological disorders. Among the hematological diseases, congenital hemophilia is an inherited bleeding disorder characterized by absence or reduced levels of clotting factors VIII or IX. Most patients with hemophilia display bleeding symptoms during infancy or childhood, but diagnosis could be delayed in mild cases. We report a case of congenital hemophilia A that was diagnosed after three episodes of postoperative bleeding after thyroid surgery. CASE PRESENTATION: A 46-year-old man developed repeated postoperative hemorrhage after thyroid surgery for thyroid cancer. In this case, several irregularities were seen in the postoperative course, such as a relatively long interval between surgery and bleeding, the lack of an obvious bleeding point, fresh red blood dripping from the drain insertion site on the second postoperative day, and repeated bleeding three times. We therefore considered that the cause of postoperative hemorrhage might be other than the surgical operations. After a thorough examination, hemophilia A was diagnosed. CONCLUSIONS: Hemophilia is a risk factor for postoperative bleeding in thyroid surgery. However, mild hemophilia shows normal prothrombin time and activated partial thromboplastin time. We encountered a case of papillary thyroid carcinoma associated with congenital hemophilia A, which was diagnosed after repeated bleeding.

Microarray analysis of formalin-fixed, paraffin-embedded follicular thyroid carcinoma samples from patients who developed postoperative distant metastasis.

OBJECTIVE: Although follicular thyroid carcinoma (FTC) generally has a good prognosis, it occasionally metastasises, leading to poor prognosis. Unfortunately, minimally invasive FTC (mi-FTC) and encapsulated angioinvasive FTC (ea-FTC) cannot be distinguished cytopathologically from thyroid follicular adenoma (FTA), a benign tumour with a good prognosis. Therefore, a molecular diagnosis to distinguish mi- or ea-FTC from FTA is needed for clinical treatment. Several transcriptomics/proteomics studies have searched for FTC biomarkers. However, the results of these studies were not consistent, which could be partly explained by inaccurate diagnosis of the specimens analysed. DATA DESCRIPTION: We conducted a microarray-based genome-wide transcriptome analysis using formalin-fixed paraffin-embedded mi- or ea-FTC specimens from patients who developed distant metastasis up to 10 years postoperatively, which ensured the accuracy of diagnosis.

Relations between cyclin D1 and chromosome 11 in thyroid carcinoma: analysis by dual stain using FISH and immunostaining method.

Cyclin D1 is an important cell-cycle regulator that drives the cell cycle from the G1 phase to the S phase. Elevated nuclear cyclin D1 expression has been found in human tumors, including thyroid carcinoma. Protein production is known to require DNA amplification in each cell, but reports of such amplification have not been published. This study aimed to analyze the relationship between cyclin D1 protein production and chromosome 11 in cultured cells by means of dual staining with fluorescence in situ hybridaization (FISH) and immunostaining. In addition, we immunostained anaplastic thyroid carcinoma tissue. The results indicate that cyclin D1 is not related to chromosome 11 in cultured cells. Furthermore, tissue study showed that cyclin D1 is produced in the cytoplasm and in nuclei in various ratios.

Proteinase K treatment improves RNA recovery from thyroid cells fixed with liquid-based cytology solution.

OBJECTIVE: Fine-needle aspiration biopsy (FNAB), an important diagnostic tool given its simplicity, safety, and cost-effectiveness, is fast becoming a popular procedure in the diagnosis of thyroid diseases. Generally, cells isolated from biopsies are transferred directly to microscope slides to prepare smears for cytopathological examination; however, the technical difficulties of this procedure often cause poor reproducibility, which limits the accuracy of diagnostic results. Liquid-based cytology (LBC), in which isolated cells are collected in a fixative solution, is advantageous in that it facilitates the preparation of homogenous cytological specimens. However, LBC has not been applied to molecular diagnoses, such as RNA expression-based diagnosis, mainly because of difficulties in cell recovery and RNA isolation. This study was aimed to improve RNA extraction from papillary cancer-derived K1 cells and thyroid FNAB specimens suspended in LBC solutions. RESULTS: K1 cells suspended in CytoRich-Red and CytoRich-Blue, fixatives for LBC, were efficiently recovered by trapping to glass-fiber filters. Importantly, subsequent Proteinase K treatment was essential for efficient RNA extraction from the fixed cells. This finding was also applicable to RNA extraction from CytoRich-Red-fixed thyroid FNAB specimens processed in the same way. Consistently, U6 small nuclear RNA was detected in these RNA samples by reverse transcription-polymerase chain reaction.

The Reality of Multiple Endocrine Neoplasia Type 2B Diagnosis: Awareness of Unique Physical Appearance Is Important.

BACKGROUND: Multiple endocrine neoplasia type 2B (MEN2B) is an extremely rare syndrome mainly caused by RET918 germline mutations. MEN2B typically causes medullary thyroid carcinoma (MTC), pheochromocytoma, and unique physical characteristics including mucosal neuroma, distinctive facial appearance, and Marfanoid habitus. Most patients have abdominal symptoms such as bloating, intermittent constipation, and diarrhea. MTC is the most important determinant of mortality in patients with MEN2B. Establishing the diagnosis of MEN2B at a curative stage of MTC is crucial. CASE PRESENTATION: We have encountered four patients with MEN2B. Two were hereditary cases from the same family, and two were considered de novo cases with phenotypically normal parents. Mean age at diagnosis was 25.5 years (range, 13-39 years). Although all patients had shown mucosal neuroma on the lips and tongue, in addition to gastrointestinal symptoms from infancy, diagnoses were made from symptomatic MTC even for the hereditary patients (our index case was a 14-year-old girl, whose mother was subsequently diagnosed with advanced MTC). Genetic tests for RET mutations revealed the M918T mutation in all patients. Two patients developed pheochromocytoma, two died from distant metastases of MTC, and two received treatment for multiple metastases of MTC (one with vandetanib). CONCLUSIONS: In our patients with MEN2B, prophylactic or early thyroidectomy could not be performed. The characteristic phenotype associated with MEN2B is almost always seen prior to detection of MTC or pheochromocytoma. Knowledge about the non-endocrine manifestations of MEN2B needs to be shared among pediatricians and gastroenterologists.

Characteristic ultrasonographic features of the encapsulated variant of papillary carcinoma of the thyroid.

PURPOSE: The purpose of this study was to evaluate the ultrasonographic and clinical features of the encapsulated variant of papillary carcinoma of the thyroid. METHODS: Of a total of 2020 patients whose papillary carcinoma of the thyroid was treated surgically between 1998 and 2002, 56 patients had the encapsulated variant. Ultrasonographic features of the encapsulated variant of papillary thyroid carcinoma were classified into the following two types based on the shape and character of the border: papillary carcinoma (PC) and follicular tumor (FT). Ultrasonographic features of the PC type are a jagged border and an irregular tumor shape. Those of the FT type are a smooth border and a regular shape (spherical to oval). RESULTS: The PC type was seen in 36 patients and the FT type was found in 20 patients. Both types of encapsulated variant of papillary carcinoma showed cervical lymph node metastases. CONCLUSION: Ultrasonographic findings of the FT type have characteristic features that are similar to those of benign follicular tumors of the thyroid. We should remember that some patients do not show typical and classical features of papillary carcinoma on ultrasonography.

Hyalinizing trabecular tumor of the thyroid gland: characteristic features on ultrasonography.

We report a case of hyalinizing trabecular tumor of the thyroid gland and describe the characteristic ultrasonographic features of this tumor. This was a rare tumor of follicular cell origin with a trabecular pattern of growth and marked intratrabecular hyalinization. The tumor had an irregular shape, a delicately jagged border, and hypoechoic and heterogeneous internal echoes on B-mode ultrasonography. Very rich intratumoral blood flow, the so-called "tumor inferno" was evident on power Doppler ultrasonography. In the clinical management of thyroid nodules, clinicians should be aware of this peculiar type of thyroid tumor and its characteristic ultrasonographic findings.

Delta Np73 expression in thyroid neoplasms originating from follicular cells.

AIMS: p73, a homologue of p53, is known as a negative regulator of tumour progression. However, delta Np73, an isoform of p73 lacking the NH2-terminal transactivation domain plays an oncogenic role by interfering with the activity of p53 and TA (full-length transactivating isoforms) p73. In this study, we investigated the expression of delta Np73 in human thyroid neoplasms originating from follicular cells. METHODS: We immunohistochemically investigated delta Np73 expression in 223 thyroid neoplasms. Delta Np73 expression level was evaluated as the sum of positivity score and intensity score. RESULTS: Normal follicular cells did not express delta Np73, but 27.3% of follicular adenoma, 85.4% of follicular carcinoma, 99.2% of papillary carcinoma, and 95.7% of anaplastic carcinoma were positive for the transcript. Delta Np73 expression level did not differ between widely invasive and minimally invasive follicular carcinomas. In papillary carcinoma, the level was inversely linked to tumour size, extrathyroid extension, and clinically apparent metastasis. Furthermore, in anaplastic carcinoma, delta Np73 expression level was significantly lower than that in papillary carcinoma. CONCLUSIONS: Our findings indicate that delta Np73 plays a role predominantly in the early phase of papillary carcinoma progression.

An extended family with familial medullary thyroid carcinoma and Hirschsprung's disease.

Familial medullary thyroid carcinoma (FMTC) is an autosomal dominant inherited disease that has highly characteristic clinical features, including medullary thyroid carcinoma (MTC). Mutation of the RET proto-oncogene is known to be responsible for development of FMTC and for multiple endocrine neoplasia types 2A and 2B. Hirschsprung's disease is the most common form of structural intestinal obstructive disease in human newborns. Hirschsprung's disease is defined by the absence of neural crest-derived enteric ganglia along a variable length of the bowel that invariably involves the rectoanal junction. Co-segregation of FMTC and Hirschsprung's disease is uncommon; nevertheless, in 3 generations of 1 family, we observed 5 patients with FMTC, 2 patients with Hirschsprung's disease, and 1 patient with characteristics of both FMTC and Hirschsprung's disease. Moreover, a Cys620Ser mutation in RET was identified in 4 of the 8 patients. This mutation had both activating and inactivating effects on the RET (REarranged during Transfection) protein. There were individual differences in the penetrance of Hirschsprung's disease due to the RET mutation, but the penetrance of MTC was uniform and high. Genetic testing is important for making decisions about treatment and follow-up in families of this kind.