A total of 187 consecutive cases were clinically and cytologically suspected as FTs (category III or IV of the Bethesda System) before surgery at Kuma Hospital in Kobe between August 2018 and July 2019. Of these, 183 cases were available in this study. The remaining four cases were excluded from the study because the subjects declined to join in this study. All cytology samples were obtained from surgically resected thyroid glands by FNAC, and sent to the Department of Tumor and Diagnostic Pathology, Nagasaki University. Final diagnosis of all cases was histologically confirmed at Kuma Hospital. LBC specimens were prepared for conventional Papanicolaou staining to confirm the presence of FT cells from FNA samples, and used to quantitatively analyze 53BP1 expression in FT cells by IF at Nagasaki University. Exclusion criteria were, (i) technically inadequate samples, such as either too many cells showing severe nuclear overlapping (unable to identify the number of NF) or too few cells (less than 10 clusters of follicular cells) to evaluate type of 53BP1 expression, (ii) cases finally diagnosed as papillary carcinoma, well differentiated tumor-uncertain malignant potency (UMP), poorly differentiated carcinoma, and other nodular lesions by histopathology. The 53BP1 expression profile in each case was determined before obtaining their clinicopathological information from Kuma Hospital. A schematic representation of these analytic procedures is shown in Fig. 2.

Schematic representation of analytic procedures in this study. 53BP1, p53-binding protein 1; LBC, liquid-based cytology; AG, adenomatous goiter; FA, follicular adenoma; FC, follicular carcinoma; FT-UMP, follicular tumor-uncertain malignant potential; PDC, poorly differentiated carcinoma; WDT-UMP, well differentiated tumor-UMP.

This study was conducted in accordance with the tenets of the Declaration of Helsinki and approved by the institutional ethical committee for medical research at both Nagasaki University (approval date: June 20, 2019; #15062617-3) and Kuma Hospital (approval date: July 6, 2017; #170706-4). Profiles of all patients were anonymized by coding, and collectively summarized with the obtained data as final datasets.

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