Acute and chronic cannabidiol treatment: In vitro toxicological aspects on human oral cells.

Cannabidiol is gaining increasing interest for its potential anti-inflammatory, immunomodulatory, and antineoplastic effects. The purpose of this study is to investigate the biological effects of acute and chronic CBD administration on gingival fibroblasts and oral keratinocytes. Viability, morphology, migration, apoptosis and cell cycle, and expression of related genes (p53, BCL2, p21, and BAX) and of endocannabinoid system receptors (CB1, CB2 and GPR55) with real-time PCR and DNA damage with phospho-γ-H2AX immunofluorescence detection were analyzed. Concentrations between 100 µM and 0.001 µM were used: 50 µM (toxic dose), 25 µM (viability promoter), and 1 µM (nontoxic), were selected for subsequent chronic analysis. Acute treatment reveals significant effects than chronic, in particular in fibroblasts: concentrations ≥50 µM are highly cytotoxic, with increased apoptosis and reduced migration. Cell death correlates with increased p53 and BAX, followed by arrest in G0/G1 phase, with elevated p21 levels, suggesting a time- and dose-dependent damage. An increase in H2AX phosphorylation was observed with 25 µM and 50 µM, while 1 µM was biocompatible. Keratinocytes showed less cytotoxic effect than fibroblasts. Induced cell damage was dose- and time-related, with less damage after chronic treatment. Further investigations are needed with longer time frames to evaluate CBD dose- and time-dependent effects to identify an effective therapeutic dose.

Predictors of an informative or actionable cytological diagnosis on repeat breast aspiration after an insufficient aspirate: a multicentre retrospective review.

An insufficient/inadequate diagnosis on fine-needle aspiration cytology (FNAC) of the breast is not an uncommon diagnostic dilemma. This study aims to review the rate and clinical features predicting an informative or actionable diagnosis on repeating breast aspiration after an insufficient aspirate. METHODS: Unsatisfactory/insufficient/inadequate or equivalent breast aspirates were retrieved from the involved institutions, and those with a repeat aspiration performed within 365 days were included. Clinical and radiological information were retrieved. Available cytological slides were reviewed. RESULTS: Totally 539 paired aspirates were retrieved, with 61.2% (n=330/539) and 10.9% (n=59/539) cytological diagnosis being informative (not insufficient) and actionable (not insufficient nor benign) on repeat aspiration. Younger age (p=0.005) was associated with an informative diagnosis and prior radiotherapy (p=0.097) and insufficient aspirates performed under free-hand (p=0.097) trended with an actionable diagnosis. Radiological findings of calcification (p=0.026) and hyperechogenicity (p=0.045), a small lesion size on initial (p=0.037) and repeat (p=0.059) radiological assessment and interval size increment (p=0.019) correlated with informative/actionable diagnoses. Cytomorphological parameters, except for a trend with crushing artefact (p=0.063), do not correlate with the cytologic diagnosis of the repeat aspirate. CONCLUSIONS: Repeating breast FNAC on patients after an insufficient diagnosis yields an informative ('sufficient') result in over 60% of cases. Small lesions with calcification, hyperechogenicity and/or interval size increment are more likely to yield diagnostic results on repeat aspiration and indicate select patients suitable for repeat FNAC over more invasive procedures. The lack of associations with cytomorphological parameters cautions against overinterpretation of insufficient breast aspirates.

Adequacy of cytology and small biopsy samples obtained with rapid onsite evaluation (ROSE) for predictive biomarker testing in non-small cell lung cancer.

Complete biomarker workup of non-small cell lung cancer (NSCLC) specimens is essential for appropriate and timely clinical management decisions. This can be challenging to achieve from small cytology and histology specimens, with increasing numbers of molecular and immunohistochemical biomarkers required. We conducted a 5 year retrospective audit of cases at our institution to assess the diagnostic and biomarker testing adequacy rates, particularly those specimens obtained with rapid onsite evaluation (ROSE), performed by a cytopathologist and a cytology scientist or pathology trainee, including all endobronchial ultrasound guided transbronchial needle aspirations (EBUS-TBNA), CT guided lung fine needle aspirations (FNA) and CT guided lung core biopsies. A total of 5,354 cases were identified, of which 92.2% had sufficient material for diagnosis. Of the 1506 cases identified with a recorded diagnosis of lung adenocarcinoma or NSCLC, not otherwise specified, 1001 (66.5%) had biomarker testing requested. Sufficient material was available in 89.5% of cases for a complete biomarker workup which included EGFR and KRAS mutational testing (all cases), ALK, ROS1 and PD-L1 immunohistochemistry (all cases), and ALK and ROS1 FISH (as required). For EGFR and KRAS mutational testing across both cytology and histology specimens, 99% of cases were sufficient. Of the samples in which a complete biomarker workup was unable to be performed, approximately half were only insufficient due to inadequate numbers of tumour cells for PD-L1 immunohistochemistry. Excluding PD-L1 IHC, 952 (95.1%) of samples obtained with ROSE were sufficient for the remainder of the testing requirements. Next generation sequencing using a 33 gene custom AmpliSeq panel was achieved in up to 72% of cases. In conclusion, small cytology and histology specimens obtained with ROSE are suitable for predictive biomarker testing in NSCLC, although attention needs to be paid to obtaining sufficient cells (>100) for PD-L1 immunohistochemistry.

Diagnostic performance of shear wave elastography in thyroid nodules with indeterminate cytology: A systematic review and meta-analysis.

Purpose: Thyroid nodules classified as indeterminate in previous fine-needle aspiration cytology often necessitate additional evaluation to determine their histology, while shear wave elastography (SWE) offers an alternative option in this regard. The objective of this study was to assess the diagnostic effectiveness of SWE in evaluating indeterminate nodules. Methods: The PubMed, EMBASE, and Web of Science databases were searched from 1st January 1970 to 1st March 2023. The studies were reviewed and the data was extracted by two separate reviewers. A Bayesian bivariate model was utilized to quantitatively synthesize the diagnostic accuracy and yield of the studies in R. Results: A total of seven studies, involving indeterminate thyroid nodules undergoing SWE were included, and the overall malignancy rate was 34.1% (307/900). The summarized estimates of sensitivity and specificity were 0.792 (95% credible interval [CI], 0.727-0.850) and 0.845 (95% CI, 0.797-0.887), respectively. The summarized estimate for the diagnostic odds ratio (DOR) was 17.8 (95% CI, 14.0-22.6). Summarized receiver operating characteristic (SROC) plots indicated a trade-off between sensitivity and specificity, and the estimate of AUC was 0.866 (95% CI, 0.834-0.895). The summary estimates for positive and negative likelihood ratios were 4.67 (95% CI, 3.98-5.85) and 0.26 (95% CI, 0.23-0.28), respectively. Conclusions: The overall accuracy of SWE remains satisfactory in indeterminate thyroid nodules. However, it should be noted that the available data are still extremely limited, and more studies or guidelines are required to provide further insights.

The Similarities and Differences between Liquid-based and Conventional Methods in Evaluation of Thyroid Fine Needle Aspiration Cytologies.

Aims: Thyroid nodules are one of the most frequent medical issues in endocrinology in our country and around the world. The appropriate evaluation of the nodule is critical in the management of patients. Fine needle aspiration cytology (FNAC) is one of the most accurate tools for evaluating these nodules. Conventional and liquid-based (LB) methods are available for thyroid FNAC. In this paper, we aim to determine the best cytological method for the evaluation of thyroid fine needle aspiration materials. Settings and Design: Fine needle aspiration (FNA) materials obtained and prepared by conventional and LB methods from 310 patients were evaluated. These slides were compared in terms of 12 cytological parameters in a three-tiered system, and the kappa coefficient was calculated. Methods and Material: Two slides were prepared from FNA samples, and the rest of the material was left in protective solution for LB cytology. Surepath was used as a LB technique. Conventional slides (CS) were left to dry in the air and stained with May Grunwald Giemsa. Slides prepared by two different techniques were compared in 12 cytomorphological parameters, and Kappa coefficients and correlations were calculated. Statistical Analysis Used: Kappa statistics. Results: In comparison of LB and CS, the highest kappa coefficient was in chromatin texture (ĸ:0.738) and inflammatory cells (ĸ: 0.482). On other parameters, fair or poor agreement was observed. Conclusions: Although there are some superiorities to LB cytology, co-application of these two methods is more favorable in thyroid fine needle aspiration.

Application of algorithmic cytomorphological assessment and immunocytochemistry with the international system for reporting serous fluid cytopathology on pericardial fluid cytology.

AIMS: The international system for reporting serous fluid cytopathology (ISRSFC) set forth a five-tiered reporting system with comprehensive validation on pleural and peritoneal fluid cytology. An algorithmic approach for cytomorphological assessment and immunocytochemistry was also described in ISRSFC. Limited data on pericardial fluid are supportive but would benefit from further investigation. METHODS: Consecutive pericardial fluid cytology over a 4-year period was reviewed by multiple board-certified pathologists according to the ISRSFC. Cytomorphology and immunocytochemistry were assessed sequentially, with respective diagnostic performances computed and compared. Literature review was performed. RESULTS: In total 358 specimens, including 53 with immunocytochemistry available, were reviewed. There were 137 benign and 221 malignant (MAL) cases. The risks of malignancy were 23.5% non-diagnostic (ND), 29.2% negative for malignancy (NFM), 56.0% atypia of undetermined significance (AUS), 82.6% suspicious for malignancy (SFM) and 99.2% (MAL) for cytomorphological assessment, improving to 23.5% (ND), 29.1% (NFM), 56.8% (AUS), 78.9% (SFM) and 99.3% (MAL) incorporating immunocytochemistry. Ten cases (2.8%) received a change in diagnosis after review of immunocytochemistry. All revisions of diagnostic category were appropriate upgrades/downgrades referenced against clinical information. Cytomorphological typing was accurate for adenocarcinoma (n=81/83, 97.6%), while other carcinomas and lymphomas required immunocytochemistry. Certain subcategories within AUS and SFM pertaining to bland indeterminate epithelial cells or mucinous material were not seen for pericardial fluid. CONCLUSIONS: The ISRSFC shows robust diagnostic performance for pericardial fluid cytology. For pericardial effusion, disease composition and applicable cytological subcategories differ from its peritoneal and pleural counterparts. Incorporating immunocytochemistry by an algorithmic approach improves diagnostic accuracy. Cytomorphology is accurate for identifying adenocarcinomas, but further typing necessitates immunocytochemistry is necessary.

HER2/ERBB2 overexpression in advanced gallbladder carcinoma: comprehensive evaluation by immunocytochemistry and fluorescence in situ hybridisation on fine-needle aspiration cytology samples.

AIMS: Advanced gallbladder carcinoma (AGBC) carries a poor prognosis with dismal survival. There are no data regarding HER2/ERBB2 expression in AGBC. This study evaluated the overexpression of HER2/ERBB2 in cytological aspirates from AGBCs to identify potential patients for whom anti-HER2 targeted therapies can benefit. METHODS: This prospective, case-control study was performed on 50 primary AGBC cases. A detailed cytomorphological assessment, followed by immunocytochemistry (ICC) for HER2/ERBB2, was performed on AGBC cell blocks. A similar number of age-matched and gender-matched resected chronic cholecystitis specimens were included as controls. Fluorescence in situ hybridisation (FISH) was performed in equivocal cases. RESULTS: A total of 10 (20%) cases showed positive (3+), 19 (38%) equivocal (2+) expression and 21 (42%) were negative on HER2/ERBB2 ICC. None of the equivocal cases demonstrated HER2 amplification by FISH. Among the controls, none showed positive (3+) immunoexpression, 23 (46%) demonstrated equivocal expression and 27 (54%) were negative. On statistical analysis, HER2/ERBB2 overexpression was significantly associated with AGBC compared with the controls. Of all the clinical, radiological and cytomorphological parameters, the predominant papillary or acinar arrangements of the tumour cells were significantly associated with HER2/ERBB2 overexpression. CONCLUSIONS: This is the first study to evaluate the expression of HER2/ERBB2 on cytological aspirates in AGBC using ICC and FISH. HER2/ERBB2 overexpression(20%) was significantly associated with AGBC. Furthermore, predominant papillary or acinar arrangements of tumour cells in the cytological smears were significantly associated with HER2/ERBB2 overexpression. They can serve as potential predictors of HER2/ERBB2 overexpression to select AGBC patients for anti-HER2 targeted therapies.

Evaluation of 1,030,482 Cervical Smear Results in Brazilian Population.

BACKGROUND: The objective was to determine the prevalence of abnormal cytologic smears in a large population of a Brazilian city. METHODS: Retrospective study of cervical cytology results performed at a private laboratory in São Paulo - Brazil. A total of 1,030,482 cytology tests were performed between January 2010 and December 2015. RESULTS: Among the satisfactory cytologies, we observed abnormal results in 8.9% (91,371). Analyzing the proportion of exams with altered results over the years, there was no change in these rates. We observed that the proportion of abnormal exams decreased with increasing age. Atypical squamous cells of undetermined significance, possibly non-neoplastic (ASC-US) cytology was the most common abnormality found in the general population with 73.19%, followed by low-grade intraepithelial lesion (LSIL) with 20.5%; these 2 cytological abnormalities add up to 93.69 % of all abnormal results. There was an increase in ASC-US with advancing age, a decrease in prevalence of LSIL, especially after the age of 30 and high-grade intraepithelial lesion (HSIL) shows two peaks of elevation, respectively in the age groups of 30-39 years and over 70 years. CONCLUSION: Women under the age of 30 showed a higher prevalence of cytological abnormalities, however most of these changes are low-grade lesions. In older women, cytological abnormalities are largely high-grade lesions, requiring greater care to prevent progression to cancer.

Comparison of small biopsy and cytology specimens: Subtyping of pulmonary adenocarcinoma.

Objectives: The aims of this study was to investigate the use of cytologic samples for subclassification of lung adenocarcinoma and the cytologic-histologic correlation in lung adenocarcinoma subtypes using small samples. Methods and Methods: Cytological characteristics of lung adenocarcinoma subtypes were summarized by a literature review. Cytology samples from 115 patients with lung adenocarcinoma confirmed by small biopsies were classified by subtype. The diagnostic concordance of subtypes between biopsy and cytology samples was assessed. Results: Among the 115 cases, 62 (53.9%) had acinar predominant pattern, 16 (13.9%) were papillary predominant pattern, 29 (25.2%) had solid predominant pattern, 3 (2.6%) had lepidic predominant pattern, and 5 (4.3%) had micropapillary predominant pattern. All corresponding cytologic samples were classified into five subtypes based on cytomorphology features, with concordance rates of 74.2% (46 patients) in c-acinar subtype, 56.3% (nine patients) in c-papillary subtype, 24.1% (seven patients) in c-solid subtype, 66.7% (two patients) in c-lepidic subtype, and 40% (two patients) in c-micropapillary subtype. Collectively, the cytology and small biopsy concordance rate was approximately 57.4%. Conclusion: Subtyping of lung adenocarcinoma using cytologic specimens is challenging and the consistency rate varies with the subtype. Acinar predominant tumors have an excellent cytologic-histologic correlation compared to tumors with predominant solid or micropapillary pattern. Evaluating cytomorphologic features of different lung adenocarcinoma subtypes can reduce the false-negative rate of lung adenocarcinoma, particularly for the mild, atypical micropapillary subtype, and improve diagnostic accuracy.

Características clínicas, epidemiológicas y quirúrgicas de pacientes con afecciones nodulares tiroideas / Clinical, Epidemiological and Surgical Characteristics of Patients with Thyroid Nodular Conditions

Introducción: En las últimas décadas la incidencia del cáncer tiroideo en el curso de la enfermedad nodular se ha incrementado debido a las novedosas técnicas de diagnóstico; sin embargo, la tasa de mortalidad se ha mantenido muy baja. Objetivo: Evaluar las características clínicas, epidemiológicas y quirúrgicas de pacientes con afecciones nodulares tiroideas. Métodos: Se realizó un estudio descriptivo observacional de cohorte prospectivo, longitudinal con los pacientes operados de afecciones tiroideas durante el período comprendido entre enero del 2008 y diciembre del 2018. El universo y la muestra quedaron constituidos por 467 pacientes que cumplieron con los criterios de inclusión. Resultados: Predominaron el sexo femenino (89,5 por ciento) y el grupo etario de 45-60 años (29,5 por ciento). Asociaron comorbilidades 338 pacientes y algún factor de riesgo de malignidad (6,2 por ciento). Un total de 174 pacientes manifestaron síntomas y 264 mostraron algún signo. Predominaron los reportes ecográficos (TI-RADS) y citológicos (Bethesda) tipo II (54,3 por ciento) y (55,5 por ciento), respectivamente. La hemitiroidectomía fue el procedimiento más realizado (59,9 por ciento) y la disfonía la complicación más encontrada (1,9 por ciento). Conclusiones: El diagnóstico oportuno del cáncer tiroideo en el curso de una enfermedad nodular contribuye a individualizar todas las decisiones terapéuticas atendiendo a las características de cada paciente y sus circunstancias(AU)

Introduction: In recent decades, the incidence rates of thyroid cancer in the course of nodular disease has increased due to novel diagnostic techniques; however, the mortality rate has remained very low. Objective: To evaluate the clinical, epidemiological and surgical characteristics of patients with nodular thyroid disease. Methods: A descriptive, observational, of prospective cohort, longitudinal and observational study was conducted with patients operated on for thyroid disorders during the period from January 2008 to December 2018. The study universe and sample consisted of 467 patients who met the inclusion criteria. Results: The female sex (89.5 percent) and the age group 45-60 years (29.5 percent) predominated. Comorbidities were present in 338 patients, as well as some risk factor for malignancy in 6.2 percent. A total of 174 patients manifested symptoms and 264 showed some sign. There was a predominance of echography (TI-RADS) and cytology (Bethesda) type II reports, accounting for 54.3 percent and 55.5 percent, respectively. Hemithyroidectomy was the most performed procedure (59.9 percent), while dysphonia was the most encountered complication (1.9 percent). Conclusions: Timely diagnosis of thyroid cancer in the course of nodular disease contributes to individualizing all therapeutic decisions considering the characteristics of each patient and their circumstances(AU)

Correlation of immediate prevalence of cervical precancers and cancers with HPV genotype and age in women with ASC-US/hrHPV+: a retrospective analysis of 2292 cases.

AIMS: To stratify the risk of cervical precancers (high-grade squamous intraepithelial lesion (HSIL) and adenocarcinoma in situ (AIS)) and cancers (squamous cell carcinoma (SCC) and adenocarcinoma) based on distinct high-risk human papillomavirus (hrHPV) genotypes as well as age groups among women with atypical squamous cells of undetermined significance (ASC-US) and hrHPV+results. METHODS: In total, 2292 cases of ASC-US/hrHPV+ with immediate follow-up biopsy results were included in the study for prevalence analysis. RESULTS: Overall, 12.2% women with ASC-US /hrHPV+ had HSIL+ while 0.22% had AIS+ lesions. The HPV-16+ group (31.6%) showed significantly higher prevalence of HSIL+ squamous lesions than other genotype groups (p<0.0001). The prevalence of SCC is significantly higher in HPV-16+ (1.8%) or HPV-18/45+ (1.1%) group than women in other genotype groups (0.1%) (p<0.0001). The HPV-18/45+ group (1.7%) showed significantly higher prevalence of AIS+ glandular lesions than other genotype groups (p=0.003). In addition, SCC prevalence was significantly higher in age over 50 group than that in age under 50 group (1.2% vs 0.2%, p=0.012). CONCLUSION: Women with ASC-US/hrHPV+ are at significant risk of cervical precancers and cancers; notably, HPV-16+ group has a higher risk of HSIL squamous lesions and SCC while HPV-18/45+ group has a higher risk of AIS+ glandular lesions. In addition, the older patient group (>50 years) has a significantly higher risk of SCC. Therefore, HPV genotyping as well as patient age need to be considered in the clinical management of patient.

Cytomorphological characterisation of angioimmunoblastic T cell lymphoma: a case-control study.

AIMS: Angioimmunoblastic T cell lymphoma (AITL) is often misdiagnosed in cytology. Hence, the present study was conducted to identify the distinctive cytomorphological features of AITL in lymph node fine-needle aspirates (LN-FNA). METHODS: This was a 4-year retrospective case-control study. Cases included LN-FNAs from patients with histopathologically confirmed AITL. The controls included LN-FNAs from patients with histopathologically confirmed reactive lymphoid hyperplasia (RLH; n=25). Eleven cytomorphological features were assessed in all the aspirates; the strength of association was determined by OR, Cramer's V and multiple correspondence analysis (MCA). RESULTS: Of a total of 22 cases of AITL reported on histopathology, 19 adequate aspirates from 14 patients (63.6%) were available for review. On univariate analysis, 5 of 11 cytomorphological variables were found to be significant for AITL; however, on MCA, 3 of these parameters, viz absence of tingible body macrophages (OR=0.014; V=0.74), presence of atypical lymphoid cells (OR=10.8; V=0.41) and singly scattered epithelioid cells (OR=19.3; V=0.31), were found to be the strongest predictors of AITL. CONCLUSIONS: The absence of tingible body macrophages, presence of atypical lymphoid cells and singly scattered epithelioid cells in polymorphic LN-FNAs are significant cytomorphological predictors of AITL in comparison with RLH. Knowledge of these diagnostic predictors, supplemented by clinicoradiological correlation and appropriate ancillary studies, can help diagnose AITL on aspiration cytology.

Reference standards for gene fusion molecular assays on cytological samples: an international validation study.

AIMS: Gene fusions assays are key for personalised treatments of advanced human cancers. Their implementation on cytological material requires a preliminary validation that may make use of cell line slides mimicking cytological samples. In this international multi-institutional study, gene fusion reference standards were developed and validated. METHODS: Cell lines harbouring EML4(13)-ALK(20) and SLC34A2(4)-ROS1(32) gene fusions were adopted to prepare reference standards. Eight laboratories (five adopting amplicon-based and three hybridisation-based platforms) received, at different dilution points two sets of slides (slide A 50.0%, slide B 25.0%, slide C 12.5% and slide D wild type) stained by Papanicolaou (Pap) and May Grunwald Giemsa (MGG). Analysis was carried out on a total of 64 slides. RESULTS: Four (50.0%) out of eight laboratories reported results on all slides and dilution points. While 12 (37.5%) out of 32 MGG slides were inadequate, 27 (84.4%) out of 32 Pap slides produced libraries adequate for variant calling. The laboratories using hybridisation-based platforms showed the highest rate of inadequate results (13/24 slides, 54.2%). Conversely, only 10.0% (4/40 slides) of inadequate results were reported by laboratories adopting amplicon-based platforms. CONCLUSIONS: Reference standards in cytological format yield better results when Pap staining and processed by amplicon-based assays. Further investigation is required to optimise these standards for MGG stained cells and for hybridisation-based approaches.

Conventional cytologic diagnosis of human papillomavirus-induced anal intraepithelial neoplasia: The experience of a referral center in Rio de Janeiro, Brazil.

BACKGROUND: Screening of anal cancer is rarely available or performed in Brazil. This study analyzes the diagnostic performance of conventional cytology (CC) in the prevention of anal cancer in a coloproctology and gynecology outpatient clinics in a public hospital in Rio de Janeiro, Brazil. METHODS: From 2005 to 2017, 1066 conventional cytological samples were collected. We analyze the causes of unsatisfactory samples (11.3%) and compare the cytological diagnoses of 83 samples from persons living with HIV and persons not living with HIV and in specific situations, using as the gold standard high-resolution anoscopy or histopathology in cases biopsied within 6 months after cytology. RESULTS: The sensitivity of cytology with diagnosis of ASC-US for detection of anal intraepithelial neoplasia of any grade was 85%, specificity was 41%, positive and negative predictive values were 64% and 75%, respectively, and positive and negative likelihood ratios were 1.46 and 0.35, respectively. CONCLUSION: Conventional cytology available in resource-limited settings is a simple, noninvasive, low-cost method that proved feasible for outpatient screening of precursor lesions of the anal canal.

Nationwide differences in cytology fixation and processing methods and their impact on interlaboratory variation in PD-L1 positivity.

Programmed death ligand-1 (PD-L1) immunostaining, which aids clinicians in decision-making on immunotherapy for non-small cell lung cancer (NSCLC) patients, is sometimes performed on cytological specimens. In this study, differences in cytology fixation and cell block (CB) processing between pathology laboratories were assessed, and the influence of these differences on interlaboratory variation in PD-L1 positivity was investigated. Questionnaires on cytology processing were sent to all Dutch laboratories. Information gathered from the responses was added to data on all Dutch NSCLC patients with a mention of PD-L1 testing in their cytopathology report from July 2017 to December 2018, retrieved from PALGA (the nationwide network and registry of histo- and cytopathology in the Netherlands). Case mix-adjusted PD-L1 positivity rates were determined for laboratories with known fixation and CB method. The influence of differences in cytology processing on interlaboratory variation in PD-L1 positivity was assessed by comparing positivity rates adjusted for differences in the variables fixative and CB method with positivity rates not adjusted for differences in these variables. Twenty-eight laboratories responded to the survey and reported 19 different combinations of fixation and CB method. Interlaboratory variation in PD-L1 positivity was assessed in 19 laboratories. Correcting for differences in the fixative and CB method resulted in a reduction (from eight (42.1%) to five (26.3%)) in the number of laboratories that differed significantly from the mean in PD-L1 positivity. Substantial variation in cytology fixation and CB processing methods was observed between Dutch pathology laboratories, which partially explains the existing considerable interlaboratory variation in PD-L1 positivity.

Study on influencing factors of positive peritoneal cytology and its relationship with prognosis in patients with endometrial cancer / 肿瘤研究与临床

Objective:To investigate the risk factors of positive peritoneal cytology (PPC) in patients with endometrial cancer and the impact of PPC on patients' prognosis.Methods:The clinicopathological data of 202 patients who underwent initial surgical treatment and were diagnosed with endometrial cancer by postoperative pathology at Qilu Hospital of Shandong University from January 2015 to December 2019 were retrospectively analyzed, and the peritoneal fluid of patients were sent intraoperatively for cytological liquid-based smear examination. Logistic regression was used to perform univariate and multivariate analyses of PPC in the whole group of patients and the early-stage patients; Univariate analysis of the progression-free survival in the whole group of patients and the early-stage patients was performed by Kaplan-Meier method and compared by log-rank method, and multivariate analysis of the progression-free survival in the whole group of patients and the early-stage patients was performed by Cox proportional hazards model.Results:Of 202 patients, 183 (90.6%) had negative peritoneal cytology (NPC) and 19 (9.4%) had PPC; 180 patients (89.1%) were stage Ⅰ-Ⅱ and 22 (10.9%) were stage Ⅲ-Ⅳ; 180 patients (89.1%) had early-stage endometrial cancer. Deep myometrial infiltration ( OR = 3.57, 95% CI 1.02-12.45, P = 0.046) and lymph node metastasis ( OR = 7.16, 95% CI 1.70-30.23, P = 0.007) were independent risk factors for PPC in patients with endometrial cancer; deep myometrial infiltration was an independent risk factor for PPC in patients with early-stage endometrial cancer ( OR = 6.22, 95% CI 1.22-31.73, P = 0.028). The 3-year PFS rates for the whole group of patients with PPC and NPC were 72.9% and 92.7%, and the difference was statistically significant ( P = 0.001); the 3-year PFS rates for early-stage patients with PPC and NPC were 82.5% and 96.2%, and the difference was statistically significant ( P = 0.002). PPC was an independent risk factor for PFS in the whole group of patients with endometrial cancer ( HR = 4.80, 95% CI 1.14-20.17, P=0.032); PPC was also an independent risk factor for PFS in patients with early-stage endometrial cancer ( HR = 8.85, 95% CI 1.96-39.93, P = 0.005). Conclusions:Deep myometrial infiltration is an independent risk factor for PPC, and PPC is an independent risk factor for PFS in patients with endometrial cancer. Routine cytological examination of peritoneal fluid is recommended in patients with endometrial cancer.

Detection of meningeal carcinomatosis by metagenomic next-generation sequencing and copy number variation analysis of cerebrospinal fluid / 中华神经科杂志

Objective:To evaluate the significance of copy number variation (CNV) and metagenomic next-generation sequencing (mNGS) of cerebrospinal fluid (CSF) in the diagnosis of meningeal carcinomatosis (MC).Methods:Ten patients with MC diagnosed in the Department of Neurology of Peking Union Medical College Hospital from March 2022 to June 2022 were consecutively enrolled in this study. The patients were diagnosed according to the criteria of the Chinese expert consensus on the diagnosis of MC by the Chinese Society of Infectious Diseases and Cerebrospinal Fluid Cytology, and the diagnosis of MC was confirmed by CSF cytology. The control group included 10 patients who were diagnosed as autoimmune encephalitis or viral encephalitis. CSF mNGS and CNV analysis were performed simultaneously in all the patients.Results:Of the 10 patients with MC, 6 had lung adenocarcinoma, 4 had breast cancer. CSF mNGS and CNV analysis detected large CNV in 8 of 10 patients with MC, including 4 patients with breast cancer and 4 patients with lung cancer. The results of pathogenic microorganism analysis of CSF mNGS in all the patients were negative. Meanwhile, large CNV was not detected in the control group.Conclusions:CSF CNV can serve as a diagnostic marker for MC. The combination of mNGS and CNV analysis has demonstrated a high positive rate in the diagnosis of MC. The dual-omics analysis of pathogenic microorganisms and CNV has been proposed as a potential strategy to further expand the clinical utility of CSF mNGS in the realm of auxiliary diagnosis.

Transmission electron microscopy in oral cytopathology: a pilot study / Microscopia eletrônica de transmissão em citopatologia bucal: um estudo piloto

Introduction: Cytopathology is a collection method that allows cell analysis through the different techniques. The oral mucosa exfoliated cells observation demonstrates morphological, biochemical and/or molecular aspects depending on the type of processing of the sample. Aim: This study tested the use of oral cytopathology associated with transmission electron microscopy (TEM) to observe the morphology of cells, mainly in relation to the cell nucleus, the cytoplasmic membrane, and cell junctions. Materials and Methods: Exfoliated epithelial cells from the oral mucosa were analyzed by TEM from individuals exposed to tobacco and alcohol, with leukoplakia or with a histopathological diagnosis of squamous cell carci-noma. Results: The cytoplasmic cell-cell junctions in the malignant samples lost the characteristic irregular pattern formed by the numerous interdigitations and the junctional process of normal cells and started to present a straight cytoplasmic surface. The nuclei of cells from leukoplakia and squamous cell carcinoma samples showed heterogeneous staining, while non-lesional cells were homogeneous. Discussion: The analysis of oral cytopathological smears by TEM contributes to the un-derstanding of the changes that occur during the process of malignancy of the oral mucosa, especially with regard to the cytoplasmic membrane and intercellular junctions. Conclusion: TEM may be a good analytical method to investigate morphological changes in exfoliated cells of the oral epithelium.

Introdução: A citopatologia é um método de coleta que permite a análise celular por meio de diferentes técnicas. A observação das células esfoliadas da mucosa bucal demonstra aspectos morfológicos, bioquímicos e/ou moleculares dependendo do tipo do processamento empregado. Objetivo: Este estudo testou o emprego da técnica de citopatologia bucal associada à microscopia eletrônica de transmissão (MET) para observar a morfologia das células, principalmente com relação à membrana citoplasmática, as junções celulares e ao núcleo da célula. Materiais eMétodos: Células epiteliais esfoliadas da mucosa bucal foram analisadas por MET de indivíduos expostos a tabaco e álcool, apresentando leucoplasia ou com diagnóstico de carcinoma espinocelular. Resultados:As junções citoplasmáticas célula-célula nas amostras malignas perderam o padrão irregular característico formado pelas inúmeras interdigitações e o processo juncional das células normais e passaram a apresentar uma superfície citoplasmática reta. O núcleo das células das amostras de leucoplasia e do carcinoma espinocelular apresentou coloração heterogênea, enquanto as células não lesionais foram homogêneas. Discussão: A análise de esfregaços citopatológicos bucais por MET contribui para o entendimento das alterações que ocorrem durante o processo de malignidade da mucosa bucal, principal-mente no que diz respeito à membrana citoplasmática e as junções intercelulares. Conclusão: A MET pode ser um bom método analítico para investigar alterações morfológicas em células esfoliadas do epitélio bucal.

Consideraciones actuales sobre el diagnóstico y tratamiento del cáncer tiroideo

Introducción: Desde los inicios de este siglo se ha producido un notable incremento mundial de la tasa de incidencia del cáncer de tiroides, el cual generalmente tiene un curso larvado y asintomático. Objetivo: Profundizar en el conocimiento de los aspectos novedosos del diagnóstico oportuno y tratamiento personalizado del cáncer tiroideo. Desarrollo: El cáncer tiroideo es la enfermedad maligna más frecuente del sistema endocrino. En las últimas décadas, su incidencia se ha incrementado aceleradamente, aunque la mortalidad se ha mantenido baja. El descubrimiento y desarrollo de nuevas técnicas de imágenes, inmunológicas y moleculares, han permitido estudiar en profundidad la neoplasia de la tiroides. Esto ha favorecido avanzar en los aspectos que más han modificado la nueva actitud respecto al diagnóstico oportuno y su tratamiento. Conclusiones: En años recientes, los avances de las investigaciones básicas, clínicas y traslacionales (aplicación real de los conocimientos básicos en la práctica clínica), han transformado antiguos conceptos relacionados con el cáncer tiroideo y han dotado de nuevas herramientas para el diagnóstico oportuno y tratamiento personalizado.

Introduction: Since the beginning of this century there has been a notable increase worldwide in the incidence rate of thyroid cancer, which generally has a latent and asymptomatic course. Objectives: To deepen the knowledge of the novel aspects of timely diagnosis and treatment of thyroid cancer. Development: Thyroid cancer is the most frequent malignant disease of the endocrine system. In recent decades, its incidence has increased rapidly, although mortality has remained low. The discovery and development of new imaging, immunological and molecular techniques have made it possible to study thyroid neoplasm in depth. This has favored advancing in the aspects that have most modified the new attitude regarding timely diagnosis and its treatment. Conclusions: In recent years, advances in basic, clinical and translational research have transformed old concepts related to thyroid cancer and have equipped with new tools for timely diagnosis and personalized treatment.