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In the present paper, we compared the efficiency of six transfer learning models to detect malignant cells in urine cytology. We also applied an ensemble learning with weighted soft voting to assess its importance in the diagnostic accuracy in urine cytology. The voided urine samples of total 104 cases of urothelial cell carcinoma (UCC) and 86 cases with no malignancy (benign) were selected. All the 104 cases of UCC were histopathology proven high grade urothelial cell carcinoma (UCC). Urine with negative cytology reports were followed up clinically. There were 446 images of benign samples and 1369 images from malignant samples on 100 x objective. We applied six transfer learning models (DenseNet121, inception_v3, ResNet50, MobileNetV2, VGG16 and Xception) to detect malignant cells in urine. To compare the performance of different models, dynamic training optimization was done and each model was auto stopped after their maximum performance. In addition, an ensemble learning with soft voting was used including the top three models to enhance the diagnostic accuracy. Xception transfer model showed the highest sensitivity (88.57%), accuracy (86.55%), precision (80.52%) and FI score (84.35%). It was the best performing model. The other two top performing models were InceptionV3 and ResNet50. The area under curve of receiver operating characteristic (AUCROC) was ≤ 90 in all the transfer learning models. The accuracy, sensitivity, specificity, precision, F1-Score and AUCROC of the ensemble transfer learning model were as follows: 92.10%, 95.41%, 85.51%, 91.23%, 93.24% and 0.977 respectively. First time, we evaluated a large number of transfer learning models in urine cytology to detect malignant cells. All the models showed high sensitivity, specificity and accuracy. In addition, the ensemble learning technique with soft voting showed remarkable superior performance than individual top three transfer learning models. The techniques transfer learning and ensemble models have high potential to use in routine screening of urine.
Scientific writing (SW) is an essential component of academic development in cytopathology; however, many professionals face difficulties when attempting to initiate the writing process. This series arises from a commonly shared challenge: not knowing where or how to begin. To introduce a practical educational framework for SW in cytopathology, developed in response to the need for structured guidance among both early-career and experienced cytopathologists. This article aims to encourage structured scientific publication, strengthen mentorship, and promote academic continuity within the specialty. A narrative review of the literature was performed using major biomedical databases and leading cytopathology journals to identify publications addressing SW, medical education, mentorship, research training, and publication practices relevant to cytopathology. Educational initiatives from scientific societies and current resources supporting academic writing, including digital technologies and artificial intelligence, were also reviewed. Based on the available literature and the authors' educational experience, a practical framework was developed to describe the scientific writing pathway from idea generation to manuscript submission. The literature specifically addressing scientific writing in cytopathology was found to be limited, and existing educational resources are fragmented. This article introduces a practical and accessible framework for both junior and senior cytopathologists, cytotechnologists, and other biomedical professionals working in cytopathology, addressing key aspects such as how to begin writing, identifying publishable ideas, the fundamental elements of successful publication, the role of mentorship, responsible use of artificial intelligence and digital tools, and the importance of disseminating both positive and negative research findings. The framework is intended to guide readers from their first research idea through manuscript preparation and publication. SW is a learnable competency that should be incorporated into cytopathology education throughout professional life. Developing structured educational pathways, strengthening mentorship, and promoting the responsible use of modern digital resources may facilitate greater academic participation. Encouraging a culture of SW is essential to develop the next generation of authors, improve international collaboration, and ensure the long-term academic growth of cytopathology.
To understand the efficacy of pleural effusion cytology in the diagnosis of malignant mesothelioma, we conducted this retrospective study to review the pleural fluid cytologies of patients diagnosed on concurrent/recent pleural biopsy histology. The clinicopathologic features and common cytomorphology were analysed. Patients who underwent pleural biopsy/stripping and had concurrent/recent pleural fluid cytology between May 2004 and December 2023 at Rush University Medical Center in Chicago were included in our study. All pleural fluid specimens initially collected and preserved in CytoLyt Solution (Hologic, Marlborough, MA) were processed into ThinPrep slides using a standardised laboratory protocol. The cytology diagnoses were made based on a combination of Papanicolaou stained ThinPrep slides, H&E stained cell block and immunohistochemistry on cell block. A total of 26 patients with malignant pleural mesothelioma diagnosed on pleural biopsy/stripping who also had concurrent/recent pleural fluid cytology were identified. About 53.8% (14/26) of pleural fluid cytologies were negative. The most common initial interpretations for the negative findings were reactive mesothelial cells (8/14), mixed inflammatory cells (12/14) and macrophages (2/14). Indeterminate pleural fluid cytologies accounted for 19.2% (5/26) of specimens in our study, with 11.5% being atypical and 7.7% suspicious for malignancy. Malignant pleural fluid cytologies accounted for 26.9% (7/26) of our cases. Surgical pathology specimens included 22 pleural biopsies and 4 pleural strippings, all of which confirmed malignant pleural mesothelioma. The subtypes of malignant pleural mesothelioma included 18 epithelioid, 4 sarcomatoid and 4 biphasic mesothelioma subtypes. The sensitivity of pleural fluid cytology alone in our study was low, at 26.9%. However, when including the 'Atypical cells present' and 'Suspicious for malignancy' categories as abnormal cytology, the sensitivity increased to 46.2%. In conclusion, we report that a diagnosis of mesothelioma can be made from pleural fluid cytology samples in a subset of patients, albeit with a relatively low sensitivity (26.9%).
Cytopathologists communicate with clinicians via cytopathology reports. For decades Papanicolaou Classes (Pap Classes), descriptive reports or surgical pathology terminologies have been used. The Bethesda System for reporting cervical cytology has been the first organ-specific terminology system and since 2010, there has been a plethora of new reporting terminology systems. We aimed to map the implementation of reporting systems via a survey designed and led by the Scientific Committee of European Federation of Cytological Societies. The survey was available online between May 2022 and May 2023. For each classification, respondents were asked whether each was implemented, barriers to implementation and if there were data on national validation of the system. A total of 37 countries participated. Department-level implementation decision was the most reported (81.6%) with a strong involvement of national cytology societies. 65.8% of respondents indicated that Pap Classes were used prior to organ-specific terminologies and 60.5% of respondents used descriptive-only text prior to organ-specific terminologies. Overall half of the respondents used national or local systems (50.0%). The Bethesda System for reporting cervical cytology and the Bethesda System for reporting thyroid cytopathology have been implemented by most of the responding countries (92.1% and 94.7%, respectively). Overall, the most reported barriers to implementing terminologies were a lack of awareness of the details of the terminology (44.7%) and the difficulty in implementing the terminology (34.2%). Despite the variabilities in practices, regulations and legislations, cytopathology reporting systems are widely accepted and implemented in Europe.
Fine needle aspiration cytology (FNAC) is an affordable diagnostic method for assessing salivary gland lesions. However, interpreting cytology results of salivary gland FNACs can be difficult due to the wide variety of lesion morphologies and overlapping features among different tumour subtypes. To address this challenge, the "Milan System for Reporting Salivary Gland Cytopathology" (MSRSGC) was introduced to aid in diagnosis, management, and to determine the risk of malignancy (ROM) across different categories. Our study aims to assess the cytological spectrum of salivary gland lesions on fine needle aspiration cytology and evaluate the diagnostic utility of MSRSGC with histological correlation. Cytology slides of FNAC over an 11-year retrospective period were retrieved, reviewed and categorized based on the MSRSGC. Histological findings were correlated with cytological findings in cases where excision specimens were received for histopathological evaluation. A total of 111 cytology cases were categorized by MSRSGC. All cases were clinically followed up, and histopathological correlation was available for 34 cases. The overall specificity for diagnosis of neoplastic lesions on cytology was 71.40%, sensitivity was 96%, and accuracy was 90.63%. The specificity for diagnosis of malignant lesions on cytology was 95.83%, sensitivity was 100%, accuracy was 96.29%, positive predictive value was 75%, and negative predictive value was 100%. FNAC is an excellent, non-invasive and resource-efficient tool for diagnosing salivary gland lesions. The MSRSGC six-tier system aids in managing tumours with complex morphology and overlapping cytological characteristics.
Endoscopic ultrasound guided fine needle aspiration/biopsy (EUS-FNA/FNB) is the main diagnostic tool for solid pancreatic lesions. However, it is associated with a limited diagnostic yield. We aimed to assess the optimal needle passes number of FNA\FNB needles to achieve maximum diagnostic yield. We performed a retrospective study including all patients who were diagnosed with pancreatic adenocarcinoma by EUS-FNA/FNB. The diagnostic yield was reported for all needles according to the number of needle passes. Overall, 227 patients underwent EUS-FNA/FNB. FNBs in 85 patients (37.4%), and FNA in 142 patients (62.5%). The needle number passes in the FNB was 1.46 ± 0.70 versus 2.11 ± 1.12 in the FNA group (p < 0.0001). One, two, and three needle passes yielded a diagnosis rate of malignancy in 76.8%, 68.4% and 70% of cases respectively for histology, while for cytology, the yield was 78.6%, 89.4% and 100%, respectively. For FNB needle types, the acquire needle outperformed the other needle, as the histological yield for one pass was 78.6%, 80% for two passes, and 57.1% for three passes, while for cytology, the yield was 79.3%, 90%, and 100% for one, two and three passes, respectively. For the FNA needle, the cytological yield was 88.7% for one pass, and higher for two and three passes. The number of needle passes for FNB (maximum three passes) was lower than FNA needles (3-4 passes) to obtain an optimal diagnostic yield.
Cytology laboratories handle a substantial number of effusion fluid samples, and preparation of cell block with immunohistochemistry (IHC) has become an integral part. The routinely used plasma thrombin (PT) method is expensive, and a cost-effective alternative is the need of the hour. This study aimed to find the level of agreement between the sodium alginate (SA) and PT methods for cellularity, cytomorphology, and artefacts. A total of 76 samples received in the cytology laboratory were evaluated. The cell blocks were prepared using the PT (routine) and SA methods from the cell sediment. Haematoxylin and Eosin sections from cell blocks were evaluated. Two cytopathologists scored the quality in a blind fashion by assessing cellularity, cytomorphology, and artefacts. The study showed comparable performance between the two methods in terms of cellularity and cytomorphology. Satisfactory cellularity was observed in 93.4% of cases with PT and 96.0% with SA, while well-preserved cytomorphology was noted in 84.2% and 72.3% of cases, respectively. However, a notable difference was seen in the background of the SA cell block section in the form of a gel-like artefact. A few IHC markers were attempted only on the SA cell block, and the results were excellent. Overall, the two techniques performed similarly. However, artefacts were present in the SA method, which did not affect the diagnosis. IHC was feasible and diagnostically interpretable on SA cell blocks. For routine cytology diagnostics, sodium alginate cell blocks can be a viable alternative to plasma thrombin cell blocks in resource-poor settings.
Screening high-risk populations for lung cancer with low-dose computed tomography (LDCT) reduced lung cancer mortality. Endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) is a well-established technique for the diagnosis of peripheral pulmonary lesions (PPLs) during bronchoscopy. Moreover, rapid on-site evaluation (ROSE) improves the diagnostic yield and accuracy of EBUS-TBNA. Nevertheless, it remains unclear whether ROSE is an efficacious diagnostic tool for PPLs. Therefore, this meta-analysis aimed to assess the diagnostic accuracy of ROSE for PPLs during EBUS bronchoscopy. A comprehensive search of the PubMed, Embase, and Cochrane Library databases was conducted to identify studies that evaluated the diagnostic accuracy of ROSE for PPLs during EBUS bronchoscopy. Our study included articles that evaluated the performance of ROSE against a reference standard. Additionally, we examined journal articles that provided sufficient data to construct a 2 × 2 table on a per-lesion basis. To estimate the overall sensitivity and specificity, a meta-analysis was conducted using a bivariate random-effects model. Thirteen studies with 1841 PPLs were included. The meta-analysis yielded a pooled sensitivity of 88.1% and a pooled specificity of 91.7%. A subgroup analysis for studies that enrolled patients who underwent CT produced a pooled sensitivity of 94.2% and a pooled specificity of 96.9%. Furthermore, accuracy of ROSE sampled by EBUS using a guide sheath showed a pooled sensitivity of 88.5%. The ROSE technique demonstrated high sensitivity and specificity for lung cancer detection during EBUS bronchoscopy for PPLs. Furthermore, ROSE might have higher sensitivity among patients who underwent CT.
Endoscopic ultrasonography-guided fine needle aspiration (EUS-FNA) is widely used for diagnosing pancreatic neoplasms, contributing to improved therapeutic strategies for pancreatic cancer. This aim of this study is to evaluate the diagnostic utility, sample adequacy, and limitations of EUS-FNA cytology for pancreatic lesions. These were further confirmed after surgical resection and histopathological evaluation. We retrospectively included patients referred to two Teaching Hospitals in southern Iran for EUS-FNA due to suspected pancreatic cancer, who later underwent surgical resection or biopsy and eventually had a histopathological confirmed diagnosis. The cytological smears from EUS-FNA were compared with the final histology reports to assess diagnostic performance. Thirty patients were included in the final analysis. EUS-FNA cytology showed sensitivity of 90.00% [95% CI: 68.30, 98.77], specificity of 80.00% [95% CI: 44.39, 97.48], negative predictive value (NPV) of 80.00% [95% CI: 50.89, 93.92], positive predictive value (PPV) of 90.00% [95% CI: 72.09, 96.91], and overall accuracy of 86.67% [95% CI: 69.28, 96.24]. Two false-positive diagnoses (hydatid cyst and chronic pancreatitis) and two false-negative diagnoses (malignancies attributed to sampling errors) were observed. Immunohistochemical tests confirmed 8 out of 9 diagnoses made by cytology. Notably, all EUS-FNA samples provided adequate material for conclusive diagnosis. These results support the high importance of the diagnostic performance of EUS-FNA on solid pancreatic lesions, even without Rapid On Site Evaluation (ROSE), given that the sample is of adequate size for testing. Despite a few false negative and false positive diagnoses, EUS-FNA should be considered the first-line diagnostic tool for pancreatic lesions assessment.
Risk stratification of indeterminate thyroid nodules, particularly Bethesda III, remains a major diagnostic challenge in thyroid cytology. We evaluated whether architectural subtyping of three-dimensional (3D) follicular cell groups provides additional diagnostic information in indeterminate thyroid cytology. A total of 107 thyroid fine-needle aspiration (FNA) cases (97 Bethesda III and 10 Bethesda IV) with histopathological follow-up were retrospectively analysed. Cytology smears were examined for 3D follicular groups, which were classified as spherules or cell clusters based on defined morphologic criteria. Ancillary cytologic features were also recorded. Conventional multivariable statistical analyses and receiver operating characteristic analyses were performed to assess diagnostic performance. Among Bethesda III cases, spherules were mostly found in non-neoplastic lesions and were not associated with malignancy, demonstrating a strong negative predictive value. In contrast, cell clusters were significantly associated with neoplastic outcomes (p < 0.001) and were identified as an independent predictor of neoplasia (OR: 8.03, 95% CI: 2.90-22.18). Absence of colloid and presence of isolated cells were also associated with neoplastic outcomes. A morphology-based scoring scheme showed a strong association with histopathological outcome (AUC: 0.895 for malignancy). Although the number of Bethesda IV cases was limited, concordant trends were observed. Architectural subtyping of 3D follicular groups represents a practical and reproducible method for risk stratification in Bethesda III thyroid cytology. True spherules strongly favour benignity, whereas cell clusters are associated with increased risk of neoplasia. Combined assessment of architectural and background cytologic features may facilitate the interpretation of indeterminate thyroid nodules, particularly where molecular testing is not readily available.
In the present paper, we tried to build a basic convolutional neural network model and a transfer learning model on it to distinguish follicular adenoma (FA) and follicular carcinoma (FC) of thyroid in fine needle aspiration cytology (FNAC). We selected histopathology proven cases of FA (25 cases) and FC (29 cases). In each case, the best May Grunwald Giemsa stained smear was selected and 10 to 11 representative microphotographs were taken by Olympus microscope (Olympus DP74) in 40× objectives from the most representative area of the smear. There were total 347 microphotographs in FA and 229 microphotographs of FC. The images were divided into training (406 images, 70%), validation (89 images, 15%) and test set (81 images, 15%). The basic (scratch) model of convolutional neural network (CNN) was built in Python 3.11.11. We subsequently, also used a transfer learning model by a pre-trained MobileNetV2 in the same images. We ran the model in Jupyter notebook for 15 epochs with 13 steps in each epoch. The sensitivity and specificity of the base model were 72.41% and 88.46%, respectively. The accuracy, precision and F1 score were 82.71%, 77.78% and 75.00%. The area under the curve (AUC) of receiver operating characteristic (ROC) was 0.85. The sensitivity and specificity of the transfer learning model are 82.76% and 92.31%, respectively. The accuracy, precision and F1 score were 88.89%, 85.71% and 84.21%. The area under the curve (AUC) of receiver operating characteristic (ROC) is 0.94. We built a CNN base model and transfer learning model (MobileNetV2) and successfully distinguished FA and FC in thyroid FNAC. To the best of our knowledge, this is the first study of CNN in thyroid follicular tumours.
The COVID-19 pandemic severely disrupted healthcare delivery worldwide, including cytopathology services essential for timely diagnosis and management of malignancies. This study evaluated the pandemic's impact on cytopathology specimen trends at the National Health Laboratory Service (NHLS) Universitas Academic Hospital laboratory in central South Africa. A retrospective analysis was conducted on adult non-gynaecological cytology specimens from March 2018 to April 2024, divided into pre-COVID-19, COVID-19, and post-COVID-19 eras. Specimens were categorised into fine needle aspirates (FNAs) and other non-gynaecological specimens, and stratified by anatomical site and diagnosis (benign, malignant, and suspicious for malignancy). Unsatisfactory, paediatric, and inadequate samples were excluded. Time series analysis was used for comparisons. Specimen volumes declined significantly during the COVID-19 era (p < 0.001) and did not recover in the post-pandemic era. Although the proportion of FNAs remained consistent, specific trends were observed, including a marked decline in breast FNAs and benign diagnoses, with a concurrent increase in malignant and cases that were suspicious for malignancy. Respiratory tract FNAs increased during the COVID-19 era, reflecting a shift in clinical priorities. Cytopathology services in central South Africa experienced sustained reductions in the volume of specimens received during and after COVID-19. Increased malignant diagnoses may reflect delayed presentations and diagnostic backlogs created by the lockdown periods. These findings highlight the importance of resilient diagnostic services during health crises, as well as the importance of relying on more cost-effective and faster diagnostic tools, such as cytopathology.
This study aimed to analyze the morphological characteristics of cells observed in pleural effusion cytology specimens using computer-assisted image analysis (CAIA). We examined 166 pleural effusion cytology specimens obtained for suspected lung cancer, including 80 negative, 22 suspicious and 64 positive cases. Whole-slide images (WSIs) were generated from Papanicolaou-stained specimens, and image analysis was performed using virtual slide cytology image analysis software. A scatter plot was then created, with the x-axis representing the area of each cell or cell cluster and the y-axis representing the maximum nuclear area within that cluster. The resulting plot type and the positive rate (PR) were subsequently evaluated. Four distinct plot types were defined from the scatter plots: small cluster type (S-type), horizontal type (H-type), vertical type (V-type) and diagonal type (D-type). Overall, S-type and H-type patterns were commonly observed in non-cancer or cytologically negative cases, whereas V-type and D-type patterns predominated in cytologically positive cases. In suspicious cases, S-type and V-type plots each accounted for roughly half of the samples. The PR was significantly higher in cytologically positive cases relative to non-cancer and cytologically negative cases (p < 0.0001). In pleural cytology specimens from patients with lung cancer, the degree of cell aggregation and nuclear overlap is an important factor for distinguishing benign from malignant lesions. Moreover, although AI-based image analysis has advanced rapidly in recent years, this study emphasizes the continued value of CAIA approaches that employ algorithms readily interpretable by humans.
Pancreatic malignancies present major diagnostic challenges. The gold standard for diagnosis is endoscopic ultrasound-guided fine-needle aspiration (FNA) with cytopathological assessment. Workforce shortages have driven interest in computer-assisted diagnosis (CAD) to improve efficiency. However, its overall diagnostic impact in pancreatic cytology remains unclear. We evaluated the accuracy of CAD in this setting. We conducted a systematic review and meta-analysis. Five databases were searched (January 2010-May 2025) for studies applying CAD to pancreatic FNA cytology. Two reviewers independently screened studies, with risk of bias assessed using QUADAS-2. Random-effects bivariate models generated pooled sensitivity, specificity, and summary receiver-operating-characteristic (SROC) curves. Studies were analysed according to the unit of analysis: image-level studies generated predictions from individual cytopathological images, whereas patient/case-level studies integrated multiple images per case to produce a single diagnostic output. Ten studies met eligibility criteria and provided quantitative data. At the image-level, pooled sensitivity and specificity were 91% [95% confidence interval: 86-94] and 92% [87-96], respectively, with a SROC area under the curve (AUC) of 0.962. At the patient/case-level, pooled sensitivity and specificity were 85% [69-93] and 91% [73-97], respectively, with an SROC AUC of 0.919. Heterogeneity was high (I2 = 65%-93%), driven by retrospective designs and variable reference standards. CAD tools achieve near-expert accuracy for definitive diagnoses, reducing routine screening burdens and accelerating surgical planning. However, selection bias, single-centre training and inconsistent thresholds limit generalisability. Prospective multi-centre validation using whole-slide workflows is warranted, and telepathology integration could expand low-resource access. This systematic review and meta-analysis shows that computer-assisted diagnosis for pancreatic EUS-FNA cytology achieves high accuracy in distinguishing benign from malignant lesions, with pooled sensitivity up to 91% and specificity up to 92%. Current evidence is dominated by retrospective and curated image-level studies, highlighting the need for prospective multicentre validation using whole-slide workflows before routine clinical adoption. Computer-assisted diagnosis for pancreatic EUS-FNA cytology shows promising accuracy in this systematic review and meta-analysis, but current evidence is still limited by retrospective, curated datasets. Considerable prospective multicentre validation in real-world whole-slide workflows is needed before clinical adoption.
Performance feedback aims to improve quality, yet challenges in data selection, presentation and management of human interactions persist. Interactive dashboards summarising key performance indicators (KPIs) and fostering active engagement may enhance feedback. This study presents our experience with the CytoLog application-an updated version of the pilot dashboard at Eurofins-Medserv Laboratory (EML)-and pilot results at the Cytology Laboratory, Kenézy Gyula Campus, University of Debrecen (CLUD). Data extraction, processing (including KPI calculation), and dashboard development were performed in Python and deployed as a web-based application. API endpoints were utilised to ensure secure, anonymized patient data access and user-specific dashboards. CytoLog introduces data tables, trend charts, and a quality gauge as additions to the pilot dashboard and supports quarterly updates. At EML, 1,045,034 Pap tests and 14,227 thyroid cytology cases (2018-2024) were examined, involving 11 cytopathologists (CPs) and 23 cytotechnologists (CTs). A gratifying improvement in the ASC/LSIL ratio (0.8 to 1.7) was observed, and the ASC-US/ASC-H ratio corrected between 2023 and 2024 (75.7%/24.3% to 80.3%/19.7%). The abnormal rate decreased (6.6% to 3.5%). At CLUD, 198,663 Pap tests (2020-2024) were examined, involving 3 CPs and 10 CTs. A decrease in both the abnormal rate (8.3% to 5.3%) and the ASC/LSIL ratio (6.6 to 3.2) was observed. The CytoLog application enables continuous performance monitoring while ensuring secure data management and adaptability across different laboratory environments. The improvement of the ASC/LSIL and ASC-US/ASC-H ratios at EML testify to the impact of self-directed quality improvement even without structured interventions. Awareness of metric limitations and potential bias remains essential when interpreting data-driven trends.
This study evaluated the diagnostic performance of high-resolution whole slide imaging (WSI) for primary cytological diagnosis across a broad range of specimen types and preparations. In a single-centre, retrospective validation study, 88 archived cytology cases representative of routine clinical practice were scanned at 40× equivalent magnification (0.23 μm/pixel) using the Hamamatsu NanoZoomer S360MD Slide scanner system. Specimens included gynaecological and non-gynaecological exfoliative and fine needle aspirate samples, prepared as smears, ThinPreps, cytospins or cell blocks. WSI were each reviewed independently by two expert cytopathologists with minimal prior digital cytology reporting experience, blinded to original light microscopy (LM) diagnoses. Discordant cases underwent LM review. Diagnostic concordance and agreement were assessed using percentage concordance and Cohen's κ (unweighted and weighted, for non-gynaecological cases only). A post-study questionnaire captured qualitative feedback. Of the 88 cases, 65 showed concordance between digital and LM diagnoses. Amongst the remaining 23 cases, 15 demonstrated diagnostic discrepancy by LM. Excluding these, the digital-LM concordance rate was 95.1%. When all cases were included, concordance ranged from 89.5% for within-observer analysis to 89.8% when compared with a majority LM diagnosis. Agreement analysis demonstrated substantial to near-perfect digital-LM agreement (unweighted κ = 0.86; weighted κ = 0.83-0.95), improving further following exclusion of diagnostically discrepant cases (κ = 0.89-0.97). Inter-observer agreement was lower than intra-observer agreement for both digital and LM comparisons. Feedback indicated that image quality was generally good. Challenges included visualising thick smear preparations, three-dimensional cell clusters, sparse atypical cells and screening gynaecological (cervical) cytology cases. All participants expressed openness to adopting digital cytology. High-resolution WSI demonstrated strong diagnostic concordance with traditional LM across a wide range of cytological specimen types and preparations. Despite limited prior experience, digital cytology was considered acceptable and feasible, supporting its integration into routine clinical practice, with appropriate training and quality assurance.
Epstein-Barr virus (EBV)-positive nodal T- and natural killer (NK)-cell lymphoma (EBV+ NTNKL) is a recently characterised rare malignancy arising from cytotoxic T or NK cells, typically presenting as nodal disease in adults. This lymphoma was recently recognised in the 5th edition of the World Health Organization Classification of Haematolymphoid Tumours. A comprehensive review of the existing literature revealed no cytological descriptions of this disease. Here, we present the first description of cytological observations from a touch smear of EBV+ NTNKL. In this case, the cytological features of EBV+ NTNKL included centroblastoid nuclei, resembling diffuse large B-cell lymphoma (DLBCL), along with abundant cytoplasm containing azurophilic granules observable with May-Giemsa staining. These cytoplasmic features are consistent with the cytotoxic T/NK-cell lineage of the tumour cells. In the cytological diagnosis of EBV+ NTNKL, it is critically important to understand the distinguishing features that differentiate it from morphologically similar DLBCL. It is also necessary to differentiate EBV+ NTNKL from extranodal NK/T-cell lymphoma, which is an important differential diagnosis in the evaluation of EBV-associated lymphomas. Recognition of the specific cytomorphological features of this lymphoma is of practical significance when fine-needle aspiration cytology, frequently used as an initial diagnostic step for lymph node lesions, is employed and when touch smear cytology is performed to complement histological findings.
The aim of this study was to evaluate the concordance of in-house epidermal growth factor receptor (EGFR) mutation testing using the Biocartis Idylla (Idylla) platform compared with next-generation sequencing (NGS) for formalin-fixed, paraffin-embedded (FFPE) and liquid based cytology (LBC) samples. A secondary objective was to investigate the role of rapid on-site evaluation (ROSE) in optimising sample adequacy, ensuring sufficient material is collected to facilitate comprehensive molecular testing in cases of lung adenocarcinoma. This retrospective study used data collected on patients between November 2022 and September 2024, a total of 180 samples, in which EGFR mutations were tested for using MiSEQ Next generation sequencing (NGS) platform or the Idylla Console. The source of the data was from the Royal Cornwall Histopathology Department. Among 180 samples tested, 14 (7.78%) expressed various EGFR mutations by either NGS and/or the Idylla platform. Concordance was observed in 12 of the 14 cases (86%). However, NGS identified 2 mutations in which Idylla was unable to identify. Whilst Idylla was able to identify one EGFR Exon 21 L858R mutation in a sample in which NGS was unable to test due to insufficient material. LBC samples consistently demonstrated a higher sample quality for testing. With significantly lower CQ values on average 20.94 compared to 22.28 than that of FFPE samples (p = 0.00151 3.s.f). The Idylla Console demonstrates high concordance with NGS and effectively complements molecular diagnostics in resource and time-constrained settings. 5.5% of cases tested using the NGS resulted in insufficient or failed testing suggesting a need for multi-platform testing. LBC samples, particularly with the utilisation of ROSE, enhance testing quality and efficiency, offering a robust alternative to FFPE samples.
Cervical cancer remains a major global health concern. Liquid-based cytology (LBC) is a cornerstone of screening, but its diagnostic accuracy for high-grade lesions, particularly in specific populations such as Southern China, requires ongoing evaluation. To evaluate the diagnostic accuracy of LBC for high-grade squamous intraepithelial lesion (HSIL) and squamous cell carcinoma (SCC) in a large Chinese cohort, and to analyse associated human papillomavirus (HPV) genotype patterns and diagnostic pitfalls. Retrospective analysis of 518,519 LBC records (2014-2024), focusing on 1758 cases diagnosed as HSIL, HSIL suspected SCC, or SCC. The detection rate for HSIL/SCC was 0.34%. Histopathological follow-up was available for 1315 cases. The concordance rates between cytology and histology were 44.5% for HSIL and 78.5% for SCC. All 52 cases labelled "HSIL suspected SCC" were confirmed as invasive SCC (positive predictive value [PPV] = 100%). Among discordant cases, 21.9% of cytological HSIL were upgraded to SCC on histology, while 67 cases originally diagnosed as HSIL/SCC were reclassified as other malignancies (e.g., adenosquamous carcinoma). HPV genotypes 16, 58, 52, and 33 were predominant (70.0% in HSIL, 81.0% in SCC). A small proportion (2.3%) of high-grade lesions were HPV-negative. LBC demonstrates high specificity for SCC and useful predictive value for the "HSIL suspected SCC" category. However, diagnostic challenges remain, including under-recognition of invasive carcinoma in HSIL cases and misclassification of non-squamous malignancies. These findings underscore the importance of cytological expertise and the complementary role of HPV testing within screening programs.
The objective of this study was to investigate the protein expression of PD-L1 in the pleural fluid of lung adenocarcinoma patients with malignant pleural effusion. Additionally, we aimed to analyse the association between PD-L1 expression and the mutational status of ten driver genes: EGFR, ALK, ROS1, BRAF, KRAS, NRAS, HER2, RET, PIK3CA, and MET. A total of 161 cytological specimens were collected from patients that had been diagnosed with lung adenocarcinoma at the Fourth Hospital of Hebei Medical University between January 2021 and September 2024. The cytologic samples were tested for tumour PD-L1 expression using a VENTANA PD-L1 (SP263) assay. EGFR, ALK, ROS1, BRAF, KRAS, NRAS, HER2, RET, PIK3CA, and MET mutations in the fresh cytological samples were detected using an amplification refractory mutation system and an ABI 7500 RT-qPCR system. Among 161 pleural fluid cytological specimens, 24.2% (39/161) presented a PD-L1 tumour proportion score (TPS) of ≥ 50%, whereas 39.1% (63/161) presented a TPS ranging from 1% to 49%. Additionally, 36.7% (59/161) demonstrated a TPS of < 1%. The mutation status analysis of 160 pleural fluid cytological specimens revealed EGFR mutations in 75 cases (46.9%), no mutations in 35 cases (21.9%), KRAS mutations in 20 cases (12.5%), ALK mutations in 9 cases, BRAF mutations in 7 cases, MET mutations in 3 cases, ROS1 mutations in another set of 3 cases, and other types of mutations identified in an additional 8 cases. The expression level of PD-L1 in pleural fluid cytological samples from patients with EGFR mutations was not significantly different from that in those from patients with no mutations (p = 0.473). In contrast, the expression levels of PD-L1 in patients with KRAS, ALK, and BRAF mutations were significantly different from those in patients with no mutations (p = 0.045; p = 0.007; p = 0.01). Our findings suggest that PD-L1 immunohistochemistry is effective for evaluating pleural fluid cytological specimens and that PD-L1 expression is significantly higher in lung adenocarcinoma patients with malignant pleural effusions associated with the KRAS, ALK and BRAF mutations.