Detection Of Papillary Thyroid Carcinoma Research Articles

The value of Korean, American, and Chinese ultrasound risk stratification systems combined with BRAF(V600E) mutation for detecting papillary thyroid carcinoma in cytologically indeterminate thyroid nodules.

To investigate the value of Korean, American, and Chinese ultrasound risk stratification systems combined with BRAF(V600E) mutation in the detection of papillary thyroid carcinoma (PTC)within cytologically indeterminate thyroid nodules (CITNs). A single-center retrospective study encompassed 511 CITNs selected from 509 patients between January 2020 and July 2023.Each nodule underwent surgical treatment and was classified according to three distinct systems. Receiver operating characteristic (ROC) curves were plotted using histopathological diagnosis as the reference standard, and diagnostic performance was compared. The three ultrasound stratification systems showed an elevated malignant risk with increasing grades (all P for trend2 < 0.001). The cut-off values for Korean, American, and Chinese systems were 5, 5, and 4c, and their respective area under the curves (AUCs) were 0.735, 0.778, and 0.783.The combination of BRAF (V600E) mutation significantly enhanced the diagnostic efficacy for the Korean(0.773vs0.735, P < 0.001), American (0.809vs0.778, P < 0.001) and Chinese (0.815vs0.783, P < 0.001) stratification systems in distinguishing CITNs without compromising specificity. When the three stratification systems were applied individually or combined with BRAF (V600E) mutation, the AUCs of the American and Chinese systems were similar (all P > 0.05), both of which were higher than the AUC of the Korean system (all P < 0.05). The American system exhibited higher specificity compared to the Chinese and Korean systems (all P < 0.001), whereas the Chinese system demonstrated higher sensitivity and accuracy when compared to the American and Korean systems (all P < 0.001). Korean, American and Chinese stratification systems present potential in the differential diagnosis of CITNs. BRAF (V600E) mutation can significantly improve the detection rate of malignant nodules within CTNs, particularly PTC. Notably, the American and Chinese systems demonstrate superior overall diagnostic performance among these systems.

The Role of TSHR, PTEN and RASSF1A Promoters' Methylation Status for Non-Invasive Detection of Papillary Thyroid Carcinoma.

Aim: We investigated whether a difference exists between TSHR, PTEN and RASSF1A methylation status in plasma of subjects with papillary thyroid cancer (PTC). Methods: Peripheral blood samples were collected from 68 patients with PTC and 86 healthy controls (HC). Thyroid cancer tissue and corresponding adjacent normal tissue methylation levels were analyzed. DNA methylation level changes in TSHR, PTEN and RASSF1A genes were analyzed by quantitative methylation-sensitive polymerase chain reaction. Results: We observed that the methylation level of TSHR was significantly higher in the thyroid cancer tissue compared to adjacent normal tissue (p = 0.040). TSHR methylation levels in the PTC group plasma samples were significantly higher compared to HC (p = 0.022). After surgery, PTC plasma samples showed lower TSHR and PTEN methylation levels compared to the levels before surgery (p = 0.003, p = 0.031, respectively). The TSHR methylation level was significantly higher in PTC with larger tumor size (>2 cm) (p < 0.001), and lymph node metastases (p = 0.01), lymphovascular invasion (p = 0.02) and multifocality (p = 0.013) 0ROC analysis revealed that the TSHR methylation level provides high accuracy in distinguishing PTC from HC (p = 0.022, AUC of 0.616). Conclusion: TSHR methylation in peripheral blood samples is expected to be a sensitive and specific minimally invasive tool for the diagnosis of PTC, especially in combination with other diagnostic means.

The role of ACTB status for early non-invasive detection of papillary thyroid carcinoma

Searchable abstracts of presentations at key conferences in endocrinology ISSN 1470-3947 (print) | ISSN 1479-6848 (online)

Combined Application of Fine-Needle Aspiration Cytology with BRAFV600E Mutation Screening in the Detection of Different Grades of Papillary Thyroid Carcinoma

The present study aimed to evaluate the application of fine-needle aspiration cytology (FNAC) of the thyroid coupled with the screening of serine/threonine-protein kinase B-Raf (BRAF) gene mutation in the detection of papillary thyroid carcinoma (PTC). From October 2016 to June 2019, 1244, liquid-based specimens originated from the patients diagnosed with PTC and benign lesions were collected from the Pathology Department of First Central Hospital of Baoding. BRAFV600E mutation was screened in all the liquid-based specimens using quantitative fluorescence polymerase chain reaction. Combined with the pathological results after surgery, the results of FNAC alone and FNAC combined with BRAFV600E screening results in PTC of different grades were compared and analyzed. Of the 1244 cases, there were 818 cases with definite cytological diagnosis and 426 cases with uncertain diagnoses. Combined with BRAFV600E screening, 36 out of 90 cases with atypical cells and 150 out of 223 suspected PTC were diagnosed as PTC. In conclusion, coupled with BRAFV600E mutation screening, the pre-operative detection rate of PTC can be increased in patients with atypical cells and suspected PTC which cannot be confirmed by pre-operative FNAC.

Relationship between malignancies and autoimmunity. A literature review and analysis of a clinical case

There is a complex bidirectional relationship between rheumatic diseases and cancer. Certain rheumatic diseases, in particular dermatomyositis, polymyositis, rheumatoid arthritis, systemic lupus erythematosus, Sjogren’s syndrome, and systemic sclerosis, on the one hand, are associated with an increased risk of malignant neoplasms against the background of the disease, contributing to cancer due to immunological stimulation. On the other hand, it can manifest as a result of autoimmune reactions caused by primary cancer in the form of paraneoplastic syndrome. Paraneoplastic syndrome is a symptom or set of symptoms that are secondary to the primary cancer process. These symptoms can result from substances secreted by the tumor (hormones and other biologically active substances) or due to the immune reaction to tumor cells (autoimmune reactions, the formation of immune complexes, suppression of the immune system), involving various organs and systems distant from the primary focus or metastases. This paper presents a literature review about the relationship between autoimmune diseases, to a greater extent systemic lupus erythematosus, and malignant neoplasms, and a clinical case in which the manifestation of lupus nephritis coincided with the detection of papillary thyroid carcinoma.

Інтраопераційна цитологія як додатковий метод виявлення папілярної карциноми щитоподібної залози

Актуальність. Папілярну карциному (ПК) щитоподібної залози (ЩЗ), як і будь-яку іншу, важливо своєчасно ідентифікувати, оскільки від цього залежить алгоритм лікування пацієнта. Основним та ефективним методом її передопераційної діагностики є тонкоголкова аспіраційна пункційна біопсія (ТАПБ). Між тим, чітко виявити ПК на даному етапі вдається не завжди. Вирішенню цієї проблеми часто допомагають інтраопераційні експрес-гістологічні дослідження (ЕГД), однак не в усіх випадках вони ефективні. Тому актуальним залишається пошук додаткових методів виявлення ПК ЩЗ, а як одного з кандидатів можна розглядати інтраопераційну цитологію (ІЦ). Мета дослідження: оцінка результативності ІЦ як додаткового методу виявлення ПК ЩЗ. Матеріали та методи. ІЦ виконана в 213 випадках вузлових утворень ЩЗ, а її результати порівняні з ЕГД з урахуванням висновків ТАПБ, які були класифіковані відповідно до Бетездівської системи висновків тиреоїдної цитопатології. Результати. Серед проаналізованих випадків було 127 ПК, в яких чітко ідентифікувати карциному за допомогою ТАПБ не уявлялося можливим. Порівняння результатів ІЦ та ЕГД показало, що у випадках неінкапсульованих ПК ЩЗ результативність ЕГІ становила 100 %, а ІЦ їх лише дублювали. Однак при інкапсульованих ПК ефективність ІЦ та ЕГД була практично однаковою: 18 із 81 (22,2 %) та 20 із 81 (24,7 %) відповідно. При цьому виконання ІЦ дозволило виявити додатково 7 ПК: в категорії ТАПБ «підозра на карциному» — 5 із 34 (14,7 %), в категорії «фолікулярна неоплазія» — 1 із 30 (3,3 %), в категоріях «атипія/доброякісний процес» — 1 із 17 (5,9 %). Висновки. У випадках інкапсульованих утворень ЩЗ комплексне виконання ЕГД та ІЦ дозволяє підвищити ефективність інтраопераційної діагностики ПК ЩЗ.

Differences in Clinicopathological Characteristics of Papillary Thyroid Carcinoma between Symptomatic and Asymptomatic Patients with Primary Hyperparathyroidism.

Background Popularization of cervical ultrasound led to higher detection of papillary thyroid carcinoma (PTC) and primary hyperparathyroidism (PHPT), as well as increasing percentage of asymptomatic PHPT in China. Although the coexistence of PTC and PHPT has been reported, it is unknown whether the clinicopathological features of PTC differ between asymptomatic and symptomatic PHPT patients. Methods We retrospectively reviewed the medical records of 304 PHPT patients treated in our hospital between January 2009 and July 2020, including 217 females and 87 males with the average age of 53.27 ± 13.54 years. Of the 304 patients, 181 were symptomatic PHPT patients and 123 were asymptomatic PHPT patients. We analyzed the laboratory results, postoperative pathology, and the TNM stage of PTC between asymptomatic and symptomatic PHPT patients. Results Concurrent thyroid nodules were found in 61.51% of PHPT patients, and the prevalence of PTC in thyroidectomized PHPT patients was 29.89% in our cohort. Lower serum parathyroid hormone (PTH) (p < 0.05) and calcium (p < 0.05) were found in PHPT patients with PTC compared to patients with benign thyroid lesion. Compared with the symptomatic PHPT patients, the asymptomatic PHPT patients showed lower serum calcium (p < 0.05), serum chlorine (p < 0.05), alkaline phosphatase (p < 0.05), PTH (p < 0.05), and bone turnover markers (p < 0.05) but higher prevalence of thyroid nodules (70.73% versus 55.24%, p < 0.05) and PTC (15.44% versus 3.87%, p < 0.05). All the PTC in symptomatic PHPT patients were papillary microcarcinoma limited to the thyroid, while 68.42% and 15.78% of the PTC in asymptomatic PHPT patients showed microscopic extrathyroidal extension and lymph node metastases, respectively. Moreover, 36.84% (7/19) of the PTC patients in asymptomatic group showed multifocality, which was much higher than 14.29% (1/7) in the symptomatic group; however, no statistical significance was found (p=0.24). Conclusions The concomitant PTC in asymptomatic PHPT patients showed a higher rate of microscopic extrathyroidal invasion when compared to symptomatic PHPT patients. So the FNA is essential to the asymptomatic PHPT patients with suspicious thyroid nodules and once the PTC is confirmed, concurrent parathyroidectomy should be performed with thyroidectomy in asymptomatic PHPT patients.

Development of Flow Cytometric Assay for Detecting Papillary Thyroid Carcinoma Related hsa-miR-146b-5p through Toehold-Mediated Strand Displacement Reaction on Magnetic Beads

In this work, a simple enzyme-free flow cytometric assay (termed as TSDR-based flow cytometric assay) has been developed for the detection of papillary thyroid carcinoma (PTC)-related microRNA (miRNA), hsa-miR-146b-5p with high performance through the toehold-mediated strand displacement reaction (TSDR) on magnetic beads (MBs). The complementary single-stranded DNA (ssDNA) probe of hsa-miR-146b-5p was first immobilized on the surface of MB, which can partly hybridize with the carboxy-fluorescein (FAM)-modified ssDNA, resulting in strong fluorescence emission. In the presence of hsa-miR-146b-5p, the TSDR is trigged, and the FAM-modified ssDNA is released form the MB surface due to the formation of DNA/RNA heteroduplexes on the MB surface. The fluorescence emission change of MBs can be easily read by flow cytometry and is strongly dependent on the concentration of hsa-miR-146b-5p. Under optimal conditions, the TSDR-based flow cytometric assay exhibits good specificity, a wide linear range from 5 to 5000 pM and a relatively low detection limit (LOD, 3σ) of 4.21 pM. Moreover, the practicability of the assay was demonstrated by the analysis of hsa-miR-146b-5p amounts in different PTC cells and clinical PTC tissues.

A Panel of Plasma Exosomal miRNAs as Potential Biomarkers for Differential Diagnosis of Thyroid Nodules

Background: A liquid biopsy using circulating exosomal genetic materials provides new insights for thyroid cancer diagnosis. This study aimed to identify plasma-derived exosomal biomarkers that could be used for early detection of papillary thyroid carcinoma (PTC).Method: Exosomal miRNAs in plasma were isolated from patients with benign thyroid nodules and patients with PTC. Profiling of exosomal miRNA was performed using RNA sequencing (RNA-seq) to identify miRNA candidates and differentiate the benign from malignant. The validation cohort consisted of 30 patients with benign thyroid nodules, 35 PTC patients, and 31 healthy individuals. Real-time PCR was used to quantify the expression of miRNA candidates. The diagnostic potential of the candidates was evaluated by receiver operating characteristic (ROC) curves.Results: After RNA-seq, eight plasma exosomal miRNAs were selected as candidates. Further validation indicated that the levels of exosomal miR-16-2-3p, miR-223-5p, miR-34c-5p, miR-182-5p, miR-223-3p, and miR-146b-5p were significantly lower in nodules compared to healthy controls (p < 0.0001), whereas miR-16-2-3p and miR-223-5p were significantly higher in the PTC cases than in those with benign nodules (p < 0.05). ROC analyses revealed that the above six miRNAs were potent indicators for detection of thyroid nodules. Meanwhile, miR-16-2-3p and miR-223-5p can be utilized for detecting PTC from benign nodules. Additionally, combined miRNA panels showed increased diagnostic sensitivities and specificities compared to single miRNA markers.Conclusion: Six aberrantly expressed plasma exosomal miRNAs may be used as diagnostic biomarkers to differentiate thyroid nodules from healthy individuals. The panel consisting of miR-16-2-3p, miR-223-5p, miR-101-3p, and miR-34c-5p are eligible for discriminating benign from malignant thyroid nodules.

Rapid detection of papillary thyroid carcinoma by fluorescence imaging using a \u03b3-glutamyltranspeptidase-specific probe: a pilot study

BackgroundNodular lesions of the thyroid gland, including papillary thyroid carcinoma (PTC), may be difficult to diagnose by imaging, such as in ultrasonic echo testing, or by needle biopsy. Definitive diagnosis is made by pathological examination but takes several days. A more rapid and simple method to clarify whether thyroid nodular lesions are benign or malignant is needed. Fluorescence imaging with γ-glutamyl hydroxymethyl rhodamine green (gGlu-HMRG) uses γ-glutamyltranspeptidase (GGT), a cell-surface enzyme, to hydrolyze the γ-glutamyl peptide and transfer the γ-glutamyl group. GGT is overexpressed in several cancers, such as breast, lung, and liver cancers. This imaging method is rapid and useful for detecting such cancers. In this study, we tried to develop a rapid fluorescence detection method for clinical samples of thyroid cancer, especially papillary carcinoma.MethodsFluorescence imaging with gGlu-HMRG was performed to detect PTC using 23 surgically resected clinical samples. A portable imaging device conveniently captured white-light images and fluorescence images with blue excitation light. Hematoxylin-eosin (HE) staining was used to evaluate which fluorescent regions coincided with cancer, and immunohistochemical examination was used to detect GGT expression.ResultsAll 16 PTC samples exhibited fluorescence after topical application of gGlu-HMRG, whereas the normal sections of each sample showed no fluorescence. HE staining revealed that each fluorescent region corresponded to a region with carcinoma. The PTC samples also exhibited GGT expression, as confirmed by immunohistochemistry.ConclusionsAll PTC samples were detected by fluorescence imaging with gGlu-HMRG. Thus, fluorescence imaging with gGlu-HMRG is a rapid, simple, and powerful detection tool for PTC.

DNA microarray-based resonance light scattering assay for multiplexed detection of DNA mutation in papillary thyroid cancer.

Highly accurate analysis of single-nucleotide polymorphisms (SNPs) plays an important role in both disease diagnostics and personalized medicine development. In this work, a DNA microarray-based resonance light scattering (RLS) assay has been developed for multiplexed detection of papillary thyroid carcinoma (PTC) related mutation points including BRAFV600E (t1m), NRAS codon 61 (t2m), TERT promoter g.1295228 (t31m) and TERT promoter g.1295250 (t32m) with high sensitivity and selectivity by the attachment of polyvalent ssDNA modified 13 nm gold nanoparticles (ssDNAs@GNPs) followed by silver deposition for signal enhancement. The microarray-based RLS assay provides a detection limit (S/N = 3) at the sub-nanomolar level for the target ssDNAs and determines allele frequencies as low as 0.2% for t1m, 0.2% for t2m, 0.5% for t31m, and 0.5% for t32m in the cocktail of target ssDNAs, respectively. The practicability of the DNA microarray-based RLS assay is demonstrated by profiling of t2m in 50 clinical thyroid tissue samples of PTC patients, and satisfactory results are obtained.

The Stature of Human Thyroid Cancer Related to Over Expressed mir-221 and mir-146b

Purpose: The ability of miRNAs for biomarker tools was further built by recent research showing that cancerspecific miRNAs are traceable in cell-free blood fluids such as plasma and serum. In this study, the levels of miR-221 and miR146b in human’s peripheral blood as tumor markers for early detection of Papillary Thyroid Carcinoma [PTC] were investigated. Methods: A stem-loop quantitative reverse transcription-polymerase chain reaction [qRT-PCR] method was performed to determine the serum levels of miR-221 and miR-146b from 25 healthy controls, 25 PTC patients the day before thyroidectomy, 25 patients with benign nodules and 25 patients after thyroidectomy during the scheduled medical examinations. Results: There was statistical difference in the serum levels for miR-221 and miR-146b between the controls and the PTC patients. It also showed no statistical difference for miR-221 and miR-146b between controls and thyroidectomy patients. Conclusion: The overexpression of miR-221 and miR-146b could be potentially served as serum indicator biomarkers in the approach for Thyroid Papillary Carcinoma detection from benign nodules.

Noninvasive surface detection of papillary thyroid carcinoma by Fourier transform infrared spectroscopy

Previously Fourier transform infrared(FTIR) spectroscopy has been applied to detecting thyroid cancer during operations and to discriminating cervical metastatic ones from non-metastatic lymph nodes. This study explored the possibility of establishing a sensitive, accurate and noninvasive screen or diagnosis by preoperative FTIR spectroscopy. 111 patients undergone a thyroid operation and 50 healthy volunteers were enrolled in the study. The FTIR spectra were obtained by two mid-infrared optical fibers with an attenuated total reflectance(ATR) probe closely contacting the subjects’ skin on the thyroid nodules. The FTIR spectra obtained from normal thyroid, nodular goiter(NG) and papillary thyroid carcinoma(PTC) patients were compared. A Fisher’s discriminant analysis was created based on these data. There were 41 PTC patients and 70 NG patients according to their histopathological examinations. A total of 23(of 39) parameters were statistically different among the three groups(P<0.05). The F1300 and F1080 parameters were significantly different between the three groups. In total, 9 out of 39 FTIR parameters were selected as independent factors by the Wilks’ lambda stepwise discriminant analysis. The discrimination accuracy of papillary thyroid carcinoma in the three groups was 88.8%. Surface detection of PTC by FTIR spectroscopy is feasible. FTIR spectroscopy can be used for rapid and noninvasive PTC screen and auxiliary diagnosis.

Incidental papillary thyroid carcinoma: diagnostic findings in a series of 287 carcinomas.

The recent increase in the detection of papillary thyroid carcinoma (PTC) has been influenced by the finding of incidental tumours. To this group, carcinomas measuring less than 1 cm (the so-called microcarcinomas) as well as those above 1 cm belong. Analyzing a case series from our own experience, this paper focuses on the current pre-operative diagnostic challenges that can lead to PTC incidental discovery. For this retrospective study, 287 patients with a PTC diagnosis were selected. For each, the following variables were analysed: sex, age, ultrasound (US) appearance, number of thyroid nodules, PTC size, PTC variants and presence of other associated pathology. Pre-operative fine needle aspiration (FNA) results were classified according to the five-tiered SIAPEC system. For 281 patients, the US-guided FNA results were available. Cytohistological correlation was evaluated in terms of FNA sensitivity and false negative rate. An incidental PTC was found in 45.2 % of patients. The majority of these were due to unsuccessful US detection of malignant nodules (103 cases); incorrect cytological diagnosis was responsible for the other 24 cases. The most powerful clinical confounding factors were: multinodular background versus single nodule presentations (p < 0.001) and histotype (follicular vs conventional variant, p < 0.05). Of course, tumour size remains a strongly influential feature on pre-operative diagnosis, with greater difficulties arising for carcinomas <5 mm. Moreover, FNA sensitivity was lower also in large PTCs (>2 cm) due to tumour heterogeneity. Although with limitations related to the tumour's intrinsic features and the thyroid background, US-guided FNA, especially if performed by a dedicated multidisciplinary team, is a powerful diagnostic tool for detecting malignant thyroid nodules. To the state of the art, we propose a practical clinical-pathological cut-off for this procedure, setting it at 5 mm.

Correlation between maximum intensity and microvessel density for differentiation of malignant from benign thyroid nodules on contrast-enhanced sonography.

The purpose of this study was to retrospectively evaluate contrast-enhanced sonography for differentiation of benign and malignant thyroid nodules by analyzing the correlation between maximum intensity and microvessel density. From February 2010 to May 2012, 122 patients (85 female and 37 male; mean age ± SD, 45 ± 9.1 years) with thyroid nodules (62 papillary thyroid carcinomas, 30 nodular goiters, and 30 adenomas) that underwent routine thyroid sonography and were diagnosed by surgery were included in this study. Contrast-enhanced sonography was performed, and enhancement patterns were classified into 3 groups: high, equal, and low enhancement. As a time-intensity curve parameter, the correlation of maximum intensity with CD31 and CD34 microvessel density counts was analyzed. On contrast-enhanced sonography, most patients with papillary thyroid carcinomas showed a heterogeneous low enhancement pattern, whereas most patients with nodular goiters showed an equal enhancement pattern, and patients with adenomas showed a high enhancement pattern. The detection of papillary thyroid carcinomas with low enhancement had sensitivity of 96.8%, specificity of 95.0%, and accuracy of 95.9%. Compared with the papillary thyroid group, the mean microvessel density counts were significantly higher in the nodular goiter and adenoma groups (P< .05). We also found that the maximum intensity was significantly associated with CD31 and CD34 counts (CD31, r = 0.963; P < .01; CD34, r = 0.968; P < .01). Maximum intensity has a significant relationship with microvessel density. Contrast-enhanced sonography is a practical and convenient means for differentiating benign from malignant thyroid nodules.

Thyroglobulin Measurement in Fine Needle Aspirates (Tg-FNA) of Cystic Neck Masses for Detection of Papillary Thyroid Carcinoma (PTC)

Introduction: Nuclear pseudoinclusions are one of the most specific pathologic features in diagnosing papillary thyroid carcinoma (PTC). However, the biologic nature of these pseudo-inclusions is largely unknown. Beta-catenin is an oncoprotein involved in Wnt signaling pathway which is activated in several malignancies including PTC. Using immunohistochemistry, we studied the expression of beta-catenin in PTC and hyalinizing trabecular adenoma (HTA), with particular focus on its expression in pseudoinclusions, both on histologic sections and on cytologic specimens. Materials and Methods: A cohort of 13 cases, including 8 cases of PTC and 5 cases of HTA, was applied in this study. Beta-catenin immunohistochemitry was performed on histology block recuts and cytology slides of each case. The beta-catenin labeling index of pseudoinclusions was counted in 15 high power fields and calculated in comparison with those counted on immediate next H&E stained histologic slides. Cytology slides were digitally scanned before destaining and restaining with beta-catenin. The number of beta-catenin-labeled pseudo-inclusions was compared with those digitally scanned slides. Results: On histologic sections, neoplastic cells from both PTC and HTA show much stronger membranous staining pattern of beta-catenin, compared to adjacent normal follicles or colloid nodules. However, labeling index of beta-catenin for nuclear pseudo-inclusions is significantly different between PTC and HTA. Strong intranuclear staining of beta-catenin was seen in 93.5% (range of 72-100%) of the pseudoinclusions in PTC, while only 6.4% (range of 0-13%) of pseudo-inclusions in HTA showed positive staining pattern (p<0.001). For cytologic slides, although it demonstrated similarly strong membranous staining pattern for beta-catenin in PTC cells, none of the nuclear pseudo-inclusions in both PTC and HTA was labeled with beta-catenin antibody on cytologic smears. Conclusions: Overexpression of the beta-catenin oncoprotein was seen in majority of the nuclear pseudo-inclusions in PTC, but only rarely seen in HTA. This differential expression suggests that activation of the betacatenin is likely involved in the formation of “malignant peudoinclusions” in PTC, but not in “benign peudoinclusions” in HTA. The failure of betacatenin labeling of pseudoinclusions on cytologic slides may be in part attributed to inability of the antibody to penetrate into nucleus through the intact cell membranes in cytologic preparations.

Development of a MicroRNA-Based Molecular Assay for the Detection of Papillary Thyroid Carcinoma in Aspiration Biopsy Samples

Although thyroid nodules are common and diagnosed in over 5% of the adult population, only 5% harbor malignancy. Patients with clinically suspicious thyroid nodules need to undergo fine-needle aspiration biopsy (FNAB). The main limitation of FNAB remains indeterminate cytopathology. Only 20%-30% of the indeterminate nodules harbor malignancy, and therefore up to 80% of patients undergo unnecessary thyroidectomy. The aim of this study was to identify and validate a panel of microRNAs (miRNAs) that could serve as a platform for an FNAB-based diagnostic for thyroid neoplasms. The study population included 27 consecutive patients undergoing total thyroidectomy for FNAB-based papillary thyroid cancer (n = 20) and benign disorders (n = 7). Aspiration biopsy was performed from the index lesion and from the opposite lobe normal tissue in all study patients at the time of operation. RNA was extracted from all aspiration biopsy samples. Quantitative polymerase chain reaction on a panel of previously selected miRNAs was performed. Polymerase chain reaction results were compared with final histopathology. miRNA from tumor tissues was amplified using the highest value of each miRNA expression in normal tissue as a threshold for malignancy detection. Diagnostic characteristics were most favorable for mir-221 in differentiating benign from malignant thyroid pathology. mir-221 was overexpressed in 19 patients (p < 0.0001) with a sensitive yield of 95%. Specificity, negative and positive predictive value, and accuracy of the miRNA panel were 100%, 96%, 100%, and 98%, respectively. miRNA quantification for differential diagnosis of thyroid neoplasms within aspiration biopsy samples is feasible and may improve the accuracy of FNAB cytology.

Detection of Papillary Thyroid Carcinoma by Analysis of BRAF and RET/PTC1 Mutations in Fine‐needle Aspiration Biopsies of Thyroid Nodules

Activating mutations of the oncogene BRAF or rearrangements of the tyrosine kinase receptor RET are observed in up to 80% of papillary thyroid carcinomas (PTCs). The predominant BRAF V600E mutation has not been detected in benign thyroid tissue so far, so consequently, this assumedly pathognomonic alteration is qualified to improve the preoperative diagnosis of PTC. Two hundred ninety preoperatively harvested fine-needle aspiration biopsies (FNABs) underwent routine cytologic assessment. BRAF V600E mutation analysis was performed by mutation-specific PCR using the same cell material; a hybrid-specific RT-PCR assay was used for detection of RET/PTC1 rearrangements. Detected genetic alterations were verified by direct sequencing. Definitive histopathology was obtained in 93/290 lesions following surgery of the respective thyroid nodule. While cytology alone diagnosed 13/30 malignancies (22 PTCs, 4 FTCs, 1 MTC, 1 UTC, 2 metastases), five additional malignancies were identified by supplementary mutation analysis. Cytology classified eight FNABs as benign, while postoperative histology demonstrated a thyroid malignancy (6 PTCs, 1 FTC, 1 metastasis). In four of these eight cases, the genetic analysis detected a BRAF V600E mutation or a RET/PTC1 rearrangement. Classifying both suspicious and malignant FNAB results as positive cytology results, supplementary genetic testing increased the overall sensitivity of FNAB from 70.4 to 85.7%, the positive predictive value (PPV) from 59.4 to 64.9%, and the negative predictive value (NPV) from 84.0 to 91.3%. Supplementary mutation analysis of RET and especially of the BRAF V600E mutation in FNABs is a fast and probably cost-effective assay in routine diagnostic setting. Mutation analyses of PTC-specific genetic alterations improve the preoperative identification and prognostic assessment of thyroid malignancies and therefore enable an optimized surgical strategy.

Cytological detection of papillary thyroid carcinomas by nuclear membrane decoration with emerin staining

Decoration of the nuclear membrane by emerin staining enhances detection of nuclear irregularities typical of papillary thyroid carcinoma (PTC). Our study aims to verify whether staining with emerin is a helpful diagnostic marker in fine-needle aspiration (FNA) cytology. We first designed a prospective study on smears, Thin Prep, and cell block specimens to prove the feasibility of the procedure; subsequently, we designed a retrospective study of 78 FNA cell blocks from cases that, after surgery, turned out to be either benign (34 cases) or malignant lesions (44, of which 31 PTC). From each sample, we obtained two slides, one stained with hematoxylin and eosin (H&E) and the other with emerin. In cases classified as Thy3, HBME-1 and galectin 3 (Gal3) stains had also been performed. Two blinded observers made the judgment concerning Thy categories (as proposed by the British Thyroid Association), first on H&E, then on emerin, HBME-1, and Gal3 stained slides. On cytological preparation, emerin staining represents an effective tool for the detection of nuclear irregularities, allowing for the identification of cases of PTC. In Thy3 cases, emerin staining's sensitivity and specificity (64% and 96%) proved higher than HBME-1's (60% and 88%) and Gal3's (61% and 68%). In conclusion, the immunohistochemical definition of the nuclear membrane, as determined by emerin stain, is a useful tool in the cytological diagnosis of thyroid lesions and can help to solve inconclusive cases by highlighting nuclear irregularities typical of PTC.

Assessment of Biomarkers for Clinical Diagnosis of Papillary Thyroid Carcinoma with Distant Metastasis

Early diagnosis and treatment of thyroid cancers are critical for better prognosis and better survival rates. The purpose of this study was to identify potential diagnostic markers for papillary thyroid carcinomas with distant metastasis. Fifty-eight papillary thyroid tumor specimens (27 papillary thyroid carcinomas with distant metastasis and 31 without metastasis) were examined, and protein expression of pituitary tumor-transforming gene (PTTG), E-cadherin, p27kip1, vascular endothelial growth factor (VEGF)-C, metalloproteinase (MMP) 2, MMP9, chemokine receptor CXCR4, and basic fibroblast growth factor (bFGF) in these tumors was assessed by immunohistochemistry. The clinicopathological variables with diagnostic significance were determined by multivariate analysis, and their diagnostic values were evaluated by ROC curve analysis. PTTG, VEGF-C, MMP2, MMP9, CXCR4, and bFGF were overexpressed in metastatic papillary thyroid carcinomas, whereas p27kip1 expression was elevated only in carcinomas lacking metastasis. Multiple-factor binary ordinal logistic regression analysis revealed that PTTG, VEGF-C, MMP2, and bFGF were independently related to biological metastatic behavior in thyroid tumors, suggesting their potential use as biomarkers. ROC curve analysis showed that among these four proteins, VEGF-C and bFGF were the best diagnostic biomarkers. A VEGF-C and bFGF cluster was the most useful factor for the differential diagnosis between metastatic and non-metastatic papillary thyroid cancers. Thus, the combined use of VEGF-C and bFGF as biomarkers may improve the diagnostic accuracy of papillary thyroid carcinoma and may be useful in multimodal screening programs for the clinical diagnosis of papillary thyroid carcinoma and early detection of papillary thyroid carcinoma with distant metastasis.