Protein profile of purified plasma- and aqueous humor-derived extracellular vesicles from patients with ocular toxoplasmosis

ABSTRACT Purpose: To characterize the extracellular vesicle protein cargo in the aqueous humor and plasma of patients with ocular toxoplasmosis. Methods: Aqueous humor and plasma were collected from six patients with active ocular toxoplasmosis and six patients with cataract. Extracellular vesicles were isolated, and western blotting and mass spectrometry were performed for protein analysis. Results: All plasma samples from patients with ocular toxoplasmosis and cataract were positive for the tetraspanins CD63 and TSG101. However, the aqueous humor from patients with ocular toxoplasmosis was positive only for CD63. Sixty-seven new unreported proteins were identified in the aqueous humor and plasma of patients with the ocular toxoplasmosis and cataract. Of the 67 proteins, 10 and 7 were found only in the cataract and ocular toxoplasmosis groups, respectively. In general, these proteins were involved in immune system activation and retina homeostasis and were related to infections and retina-associated diseases. Conclusion: The distinct protein signatures between ocular toxoplasmosis and cataract may be helpful in the differential diagnosis of ocular toxoplasmosis. However, more studies are needed to better understand the role of these proteins in the pathogenesis of ocular toxoplasmosis.

Importance of biopsy sample length for cancer diagnosis during trans-perineal prostate biopsy.

OBJECTIVE: To identify the factors that determine the minimum length of biopsy sample required for accurate diagnosis. MATERIALS AND METHODS: A retrospective analysis was conducted on 1202 cases that underwent rectal ultrasound-guided trans-perineal prostate biopsy (TPB) with standardized biopsy surgical procedures and pathological evaluation. Logistic regression correlation analysis and the imbalance between groups was eliminated by propensity score matching of patients' own factors between groups (positive group and negative group). ROC curve optimal threshold analysis were performed to identify the independent factors associated with cancer detection rate and the minimum length of biopsy sample required for accurate diagnosis. RESULTS: The study included 1202 cases that underwent standardized 8-18 needle initial puncture biopsies from June 2020 to October 2023. The cancer detection rate was 40.02% (481/1202), with Gleason scores of 6, 7, 8, 9, and 10 in 164, 134, 107, 67, and 9 patients, respectively. The percentage of patients with clinical significance (International Society of Urological Pathology (ISUP) ≥ 2) was 65.90% (317/481). Multivariate analysis showed that age,prostate-specific antigen(PSA), prostate volume,positive multi-parametric magnetic resonance imaging (mp-MRI) and length of biopsy samples were significant factors (P < 0.05)。Interestingly, biopsy sample length did not correlate with the prostate volume (Pearson correlation P = 0.069). ROC curve analysis: The area under the curve AUC for sample length were 0.674 and 0.664 at before and after propensity score matching,respectively; the optimal thresholds were 12.25 mm and 11.00mm at before and after propensity score matching,respectively. CONCLUSION: The independent predictors of cancer detection rate during TPB are age, PSA, prostate volume, positive mp-MRI, and sample length. Among these, sample length is the most critical indicator affecting puncture quality, and the minimum value of biopsy sample length to be obtained is 11.00mm.

Diminished expression of the ubiquitin-proteasome system in early treatment-naïve patients with rheumatoid arthritis and concomitant type 2 diabetes may be linked to IL-1 pathway hyper-activity; results from PEAC cohort.

OBJECTIVE: Based on the recent evidence of IL-1 inhibition in patients with rheumatoid arthritis (RA) and concomitant type 2 diabetes (T2D), we evaluated the synovial tissue expression of IL-1 related genes in relationship to the ubiquitin-proteasome system and the effects of insulin on ubiquitinated proteins in fibroblast-like synoviocytes (FLSs). METHODS: The synovial expression of IL-1 pathway genes was compared in early (< 1 year) treatment-naïve RA patients with T2D (RA/T2D n = 16) and age- and sex-matched RA patients without T2D (n = 16), enrolled in the Pathobiology of Early Arthritis Cohort (PEAC). The synovial expression of ubiquitin in macrophages and synovial lining fibroblasts was also assessed by Immunohistochemistry/immunofluorescence and correlated with synovial pathotypes. Finally, FLSs from RA patients (n = 5) were isolated and treated with human insulin (200 and 500 nM) and ubiquitinated proteins were assessed by western blot. RESULTS: Synovial tissues of RA/T2D patients were characterised by a consistent reduced expression of ubiquitin-proteasome genes. More specifically, ubiquitin genes (UBB, UBC, and UBA52) and genes codifying proteasome subunits (PSMA2, PSMA6, PSMA7, PSMB1, PSMB3, PSMB4, PSMB6, PSMB8, PSMB9, PSMB10, PSMC1, PSMD9, PSME1, and PSME2) were significantly lower in RA/T2D patients. On the contrary, genes regulating fibroblast functions (FGF7, FGF10, FRS2, FGFR3, and SOS1), and genes linked to IL-1 pathway hyper-activity (APP, IRAK2, and OSMR) were upregulated in RA/T2D. Immunohistochemistry showed a significant reduction of the percentage of ubiquitin-positive cells in synovial tissues of RA/T2D patients. Ubiquitin-positive cells were also increased in patients with a lympho-myeloid pathotype compared to diffuse myeloid or pauci-immune-fibroid. Finally, in vitro experiments showed a reduction of ubiquitinated proteins in RA-FLSs treated with a high concentration of insulin (500 nM). CONCLUSIONS: A different IL-1 pathway gene expression was observed in the synovial tissues of early treatment-naïve RA/T2D patients, linked to decreased expression of the ubiquitin-proteasome system. These findings may provide a mechanistic explanation of the observed clinical benefits of IL-1 inhibition in patients with RA and concomitant T2D.

Lymphangiosis Carcinomatosa of the Liver and Extrahepatic Bile Duct Due to Gastric Cancer: A Case Report and Literature Review.

A 78-year-old man was diagnosed with advanced poorly differentiated gastric adenocarcinoma, presenting with jaundice and diffuse thickening of the extrahepatic bile duct. No obstructive biliary sites or liver masses were observed. The serum concentrations of proteins induced by the absence of vitamin K or antagonist-II were markedly high. Samples of the extrahepatic bile duct and liver were obtained by endoscopic examination. The patient was diagnosed with lymphangiosis carcinomatosa of the liver and extrahepatic bile duct but died 28 days after hospitalization. As the disease progresses rapidly with uncharacteristic imaging findings, biopsy samples should be obtained early using several diagnostic tools.

The Role of Liquid Biopsy in Neuroblastoma: A Scoping Review.

BACKGROUND: Neuroblastoma (NBL), is the most common, non-CNS solid tumor of childhood. This disease presents with unique biological and clinical challenges necessitating accurate diagnosis, prognosis assessment, treatment, and vigilant monitoring. Liquid biopsy is an upcoming, innovative, and non-invasive diagnostic modality. It has the potential to detect tumors and perform therapeutic monitoring through the analysis of circulating biomarkers in blood, urine, saliva, and other bodily fluids. METHODOLOGY: This scoping review offers an in-depth exploration, of the current landscape of liquid biopsy-based biomarkers in NBL. The review looks at the clinical implications, prevalent challenges, and future outlook of their clinical applications in NBL. The scoping review adhered to the guidelines of the PRISMA extension for scoping reviews, known as PRISMA-ScR, as the skeletal framework. The review involved comprehensive searches for liquid biopsy-based biomarkers in NBL across multiple databases, including PUBMED, EMBASE, SCOPUS, and WEB of Science, without restrictions. RESULTS: The scoping review process uncovered a significant body of literature (n = 201) that underwent meticulous scrutiny, ultimately leading to the final selection of studies (n = 15). The liquid biopsy biomarkers included circulating tumor cells (CTCs), circulating tumor DNA (ctDNA), exosomes, and other entities in bodily fluids. Their evaluation focused on associations with clinical outcomes such as overall survival, event-free survival, and risk stratification in NBL. CONCLUSION: Our findings highlight the potential of liquid biopsy biomarkers to revolutionize NBL diagnosis and therapeutic monitoring. This rapidly evolving frontier in pediatric oncology suggests significant advancements in precision medicine for the management of NBL.

AB062. Investigating miR-21 as a non-invasive prognostic biomarker in glioma patients.

BACKGROUND: Glioma prognosis remains a challenge due to its high recurrence and resistance to treatment. Diagnosis and follow-up in resource-constrained regions often lead to significant patient attrition. Serum microRNAs (miRNAs) are seen to be aberrantly expressed in malignancies can be found in tumor tissues and peripheral samples. This offers a pathway for non-invasive liquid biopsies. miR-21 is an established biomarker for glioma prognosis, which needs to be validated in our population. METHODS: We collected 89 intraoperative tumor tissue samples, and 42 pre- and post-operative serum samples from glioma patients, ten control tissues, eight healthy serum samples and analysed for miR-21 expression through quantitative polymerase chain reaction (qPCR) analysis. Correlational analysis with molecular markers isocitrate dehydrogenase (IDH), Ki-67, ATRX, p53, and survival curves through the Kaplan-Meier method were calculated in high and low miR-21 expression groups. The hazard ratio was quantitatively determined using Cox regression analysis, considering both univariate associations and multivariate correlations with clinical parameters. RESULTS: miR-21 expression in tissue was significantly upregulated with increase of glioma grades (P<0.001) and in patients above 50 years (P=0.003) age group. Whereas no gender bias was seen in its expression pattern. Its expression did not show any correlation with tumor volume (r=0.22, P=0.08). A similar expression pattern of miR-21 was observed in serum samples of glioma. IDH-wildtype (P=2.06e-03) and high Ki-67 (P=2.50e-03) patient group showed significant upregulation of miR-21 expression compared to IDH-mutant and low Ki-67 group. Patients with low miR-21 expression had significantly longer overall survival (OS) than patients with high miR-21 expression (P =0.006). Similarly, quantitative hazard analysis indicates that patients in the high expression group have 2.77 times higher risk of mortality [95% confidence interval (CI): 0.19-0.92], in comparison to patients in the low expression group (P=0.008). CONCLUSIONS: Our findings validate the utility of miR-21 as a prognostic serum biomarker to help diagnose and assess treatment response in advancing glioma grades, within our population.

AB085. Trans-cortical trans-ventricular approach to pineal tumors: a case series.

BACKGROUND: Attributing to its deep-seated location, surrounded by significant vessels, surgical management of pineal gland lesions is still considered a challenge. Various surgical approaches are employed to access these lesions, based on patient-specific risks and benefits. Each approach has its merits and demerits. In this study, we aim to narrate our experience with trans-cortical trans-ventricular (TT) approach for pineal tumor resection. We aim to outline the procedure details, safety, efficacy, and treatment outcome of TT. CASE DESCRIPTION: This is a single-center, consecutive case series. All patients with pineal gland tumors who underwent surgical intervention, namely biopsy or resection using TT approach from 2000 to 2023 were included. Data for the patient characteristics, intraoperative details and complications were collected from the hospital's database. Mean [standard deviation (SD)] and frequency (proportions) were calculated for continuous and discrete variables, respectively. We identified 13 patients, mean age 24 (SD: 13) years in our case series. Of them, 8 (61.5%) were male. Most common presenting complains were headaches (69%), nausea/vomiting (38.5%), seizure (23%), and visual deficit (23%). Most patients, 60%, had high grade tumor and average size of tumors were 43.5 mm (SD: 18.45 mm). Pilocytic astrocyotma (23%) and pineal parenchymal tumor of intermediate differentiation (23%) were the most common diagnoses. In total, nine patients had pineal gland lesion biopsy done using TT approach along with the cerebrospinal fluid (CSF) diversion. Of them, four had tumor resection done using the same approach. Whereas four patients had primary excision done using TT approach. There were no intra-operative complications. Two patients had post-operative seizures which were treated with anti-epileptics. We did not identify any long-term sequalae attributed to this approach. CONCLUSIONS: We presented our data regarding the safety, efficacy, and outcomes of trans-cortical trans-ventricular approach for pineal tumor surgical management. Utilizing this novel approach for pineal lesion resection can be a great addition to surgeons' armamentarium. This unique approach allows to access the tumor for biopsy/resection and perform CSF diversion procedure, simultaneously. Moreover, the same incision can be used for the second/redo surgery.

Real-time ultrasound elastography in the diagnosis of newly identified thyroid nodules in adults: the ElaTION RCT.

Background: Strain and shear wave elastography which is commonly used with concurrent real-time imaging known as real-time ultrasound shear/strain wave elastography is a new diagnostic technique that has been reported to be useful in the diagnosis of nodules in several organs. There is conflicting evidence regarding its benefit over ultrasound-guided fine-needle aspiration cytology alone in thyroid nodules. Objectives: To determine if ultrasound strain and shear wave elastography in conjunction with fine-needle aspiration cytology will reduce the number of patients who have a non-diagnostic first fine-needle aspiration cytology results as compared to conventional ultrasound-only guided fine-needle aspiration cytology. Design: A pragmatic, unblinded, multicentre randomised controlled trial. Setting: Eighteen centres with a radiology department across England. Participants: Adults who had not undergone previous fine-needle aspiration cytology with single or multiple nodules undergoing investigation. Interventions: Ultrasound shear/strain wave elastography-ultrasound guided fine-needle aspiration cytology (intervention arm) - strain or shear wave elastography-guided fine-needle aspiration cytology. Ultrasound-only guided fine-needle aspiration cytology (control arm) - routine ultrasound-only guided fine-needle aspiration cytology (the current standard recommended by the British Thyroid Association guidelines). Main outcome measure: The proportion of patients who have a non-diagnostic cytology (Thy 1) result following the first fine-needle aspiration cytology. Randomisation: Patients were randomised at a 1 : 1 ratio to the interventional or control arms. Results: A total of 982 participants (80% female) were randomised: 493 were randomised to ultrasound shear/strain wave elastography-ultrasound guided fine-needle aspiration cytology and 489 were randomised to ultrasound-only guided fine-needle aspiration cytology. There was no evidence of a difference between ultrasound shear/strain wave elastography and ultrasound in non-diagnostic cytology (Thy 1) rate following the first fine-needle aspiration cytology (19% vs. 16% respectively; risk difference: 0.030; 95% confidence interval -0.007 to 0.066; p = 0.11), the number of fine-needle aspiration cytologies needed (odds ratio: 1.10; 95% confidence interval 0.82 to 1.49; p = 0.53) or in the time to reach a definitive diagnosis (hazard ratio: 0.94; 95% confidence interval 0.81 to 1.10; p = 0.45). There was a small, non-significant reduction in the number of thyroid operations undertaken when ultrasound shear/strain wave elastography was used (37% vs. 40% respectively; risk difference: -0.02; 95% confidence interval -0.06 to 0.009; p = 0.15), but no difference in the number of operations yielding benign histology - 23% versus 24% respectively, p = 0.70 (i.e. no increase in identification of malignant cases) - or in the number of serious adverse events (2% vs. 1%). There was no difference in anxiety and depression, pain or quality of life between the two arms. Limitations: The study was not powered to detect differences in malignancy. Conclusions: Ultrasound shear/strain wave elastography does not appear to have additional benefit over ultrasound-guided fine-needle aspiration cytology in the diagnosis of thyroid nodules. Future work: The findings of the ElaTION trial suggest that further research into the use of shear wave elastography in the diagnostic setting of thyroid nodules is unlikely to be warranted unless there are improvements in the technology. The diagnostic difficulty in distinguishing between benign and malignant lesions still persists. Future studies might examine the role of genomic testing on fine-needle aspiration samples. There is growing use of targeted panels of molecular markers, particularly aimed at improving the diagnostic accuracy of indeterminate (i.e. Thy3) cytology results. The application of these tests is not uniform, and their cost effectiveness has not been assessed in large-scale trials. Study registration: This study is registered as ISRCTN (ISRCTN18261857). Funding: This award was funded by the National Institute for Health and Care Research (NIHR) Health Technology Assessment programme (NIHR award ref: 12/19/04) and is published in full in Health Technology Assessment; Vol. 28, No. 46. See the NIHR Funding and Awards website for further award information.

About half the population will have lumps in their thyroid if examined by an ultrasound scan but may not know they have one. About one in twenty people will feel a thyroid lump in their neck at some time in their life, with about one in twenty of those being malignant. Currently, the recommended way of getting a diagnosis of thyroid nodules is by using ultrasound to guide a needle to get cells from the lump, called ultrasound-guided fine-needle aspiration cytology. These cells are examined to determine the cause of the lump. If there are enough cells, Doctors can then make a diagnosis of whether the lump is benign or malignant. If not, patients will undergo another ultrasound-guided fine-needle aspiration cytology. One in five ultrasound-guided fine-needle aspiration cytologies are non-diagnostic with an overall false-positive rate of approximately 24%. This means one in five patients, with benign disease, may undergo unnecessary diagnostic operations. Thyroid surgery carries risks of complications, which could be avoided if we had better ways to diagnose which patients actually need an operation. We conducted a randomised trial, ElaTION, to determine if a new technology called strain and shear wave elastography, commonly known as real-time elastography, would be better at helping the radiologist take a sufficient sample of cells and reduce the number of non-diagnostic results, reducing the number of fine-needle aspiration cytologies required to make a definitive diagnosis. Nine hundred eighty-two patients were recruited between 2015 and 2018 and followed up until the end of the trial. Patients were randomised into two groups: 489 patients received the standard ultrasound-guided fine-needle aspiration cytology alone, and 493 patients received ultrasound-guided fine-needle aspiration cytology + shear wave elastography. Ultrasound shear/strain wave elastography did not reduce non-diagnostic cytology at first fine-needle aspiration cytology or improve the likelihood of determining whether the lump is benign or malignant. The results of ElaTION do not support the use of shear wave elastography-fine-needle aspiration cytology in the diagnosis of thyroid nodules.

The potential of circulating cell-free RNA in CNS tumor diagnosis and monitoring: A liquid biopsy approach.

Early detection of malignancies, through regular cancer screening, has already proven to have potential to increase survival rates. Yet current screening methods rely on invasive, expensive tissue sampling that has hampered widespread use. Liquid biopsy is noninvasive and represents a potential approach to precision oncology, based on molecular profiling of body fluids. Among these, circulating cell-free RNA (cfRNA) has gained attention due to its diverse composition and potential as a sensitive biomarker. This review provides an overview of the processes of cfRNA delivery into the bloodstream and the role of cfRNA detection in the diagnosis of central nervous system (CNS) tumors. Different types of cfRNAs such as microRNAs (miRNAs), long noncoding RNAs (lncRNAs) and circular RNAs (circRNAs) have been recognized as potential biomarkers in CNS tumors. These molecules exhibit differential expression patterns in the plasma, cerebrospinalfluid (CSF) and urine of patients with CNS tumors, providing information for diagnosing the disease, predicting outcomes, and assessing treatment effectiveness. Few clinical trials are currently exploring the use of liquid biopsy for detecting and monitoring CNS tumors. Despite obstacles like sample standardization and data analysis, cfRNA shows promise as a tool in the diagnosis and management of CNS tumors, offering opportunities for early detection, personalized therapy, and improved patient outcomes.

Resection versus biopsy in patients with glioblastoma (RESBIOP study): study protocol for an international multicentre prospective cohort study (ENCRAM 2202).

INTRODUCTION: There are no guidelines or prospective studies defining the optimal surgical treatment for glioblastomas in older patients (≥70 years), for those with a limited functioning performance at presentation (Karnofsky Performance Scale ≤70) or for those with tumours in certain locations (midline, multifocal). Therefore, the decision between resection and biopsy is varied, among neurosurgeons internationally and at times even within an institution. This study aims to compare the effects of maximal tumour resection versus tissue biopsy on survival, functional, neurological and quality of life outcomes in these patient subgroups. Furthermore, it evaluates which modality would maximise the potential to undergo adjuvant treatment. METHODS AND ANALYSIS: This study is an international, multicentre, prospective, two-arm cohort study of an observational nature. Consecutive patients with glioblastoma will be treated with resection or biopsy and matched with a 1:1 ratio. Primary endpoints are (1) overall survival and (2) proportion of patients that have received adjuvant treatment with chemotherapy and radiotherapy. Secondary endpoints are (1) proportion of patients with National Institute of Health Stroke Scale deterioration at 6 weeks, 3 months and 6 months after surgery; (2) progression-free survival (PFS); (3) quality of life at 6 weeks, 3 months and 6 months after surgery and (4) frequency and severity of serious adverse events. The total duration of the study is 5 years. Patient inclusion is 4 years; follow-up is 1 year. ETHICS AND DISSEMINATION: The study has been approved by the Medical Ethics Committee (METC Zuid-West Holland/Erasmus Medical Center; MEC-2020-0812). The results will be published in peer-reviewed academic journals and disseminated to patient organisations and media. TRIAL REGISTRATION NUMBER: NCT06146725.

Comparison Between Endobronchial-Guided Transbronchial Biopsy and Computed Tomography-Guided Transthoracic Lung Biopsy for the Diagnosis of Central Pulmonary Lesions.

BACKGROUND: Lung cancer is one of the most common malignant tumors at present. This study aimed to compare the diagnostic accuracy, complication rates, and predictive values of computed tomography (CT)-guided percutaneous transthoracic needle biopsy (PTNB) and electronic bronchoscopy-guided transbronchial lung biopsy (TBLB) for patients with central pulmonary lesions (CPLs) with a diameter ≥ 3 cm. METHODS: We retrospectively included 110 patients with CPLs with a diameter ≥ 3 cm who underwent preoperative PTNB and TBLB examinations and ultimately underwent surgery to remove CPLs and obtained pathological results. Detailed information was collected in order to compare whether there was a difference between two groups. Data were processed using SPSS software (Version 26.0; IBM Corp). Data were compared by t-test or chi-square test. p < 0.05 was considered statistically significant. RESULTS: All patients underwent surgical treatment at the department of thoracic surgery and obtained a final pathological diagnosis. The rate of positive predictive value (PPV) was comparable between the two methods, and the negative predictive value (NPV) was significantly higher in the PTNB group compared with the TBLB group (p < 0.05). In addition, PTNB was more sensitive and accurate than TBLB (p < 0.05). However, the PTNB group had a higher probability of complications, and TBLB was a relatively safer examination method. CONCLUSION: PTNB demonstrated a higher accuracy and sensitivity than TBLB in the treatment of CPLs with a diameter ≥ 3 cm, but the complication rates of PTNB are relatively high. These methods exhibited different diagnostic accuracies and therefore should be selected based on different medical conditions.

Evaluation of Hepatic Shear Wave Elastography to Assess Liver Fibrosis in Biliary Atresia Patients and Its Correlation with Liver Histology and Surgical Outcomes: A Prospective Observational Study.

Introduction The native liver survival in biliary atresia (BA) depends on various factors, and one of the crucial factors is the rate of progression of liver fibrosis after portoenterostomy, but there is no reliable investigation to assess it. This study evaluated shear wave elastography (SWE) to detect liver fibrosis in BA patients and assess its utility during follow-up. Materials and Methods This was an observational study; SWE was done preoperatively and postoperatively at 3 and 6 months. The SWE values were analyzed to determine their correlations with preoperative liver histology as well as with postoperative SWE variation between different postoperative outcomes. Results Twenty-one patients were included in the study; the preoperative SWE values were strongly correlated with liver biopsy grading ( p < 0.001). At the 3 months postoperatively, SWE was done for 18 children: 12 in group A (patent bilioenteric drainage on hepatobiliary iminodiacetic acid scan) and 6 (nonpatent) in group B; mean SWE value was 12.8 and 17.3 kPa, respectively ( p < 0.001). Ten children from group A underwent SWE 6 months postoperatively, and the mean value was 13.23 kPa. Conclusion The SWE values correlate with liver histology grading, suggesting a reliable alternative to biopsy. Additionally, the baseline SWE values and their trend during follow-up can provide information on the disease's progression.

Mutational profiling of 103 unresectable pancreatic ductal adenocarcinomas using EUS-guided fine-needle biopsy.

Background and Objective: Pancreatic ductal adenocarcinoma (PDAC) is among the most lethal cancers, with a 5-year survival rate of around 9%. Only 20% are candidates for surgery. Most unresectable patients undergo EUS-guided fine-needle biopsy (EUS-FNB) for diagnosis. Identification of targetable mutations using next-generation sequencing (NGS) is increasingly requested. Data on feasibility of EUS-FNB for NGS and knowledge regarding mutational profile of unresectable PDAC are scarce. We evaluated the "technical yield" of EUS-FNB for NGS in unresectable PDAC: relative fraction of diagnostic EUS-FNBs meeting technical criteria. We also investigated the "molecular yield": relative fraction of EUS-FNBs included in NGS containing sufficient DNA for detection of at least one mutation. Furthermore, we determined the relative frequency of cancer-associated mutations in unresectable PDAC. Patients and Methods: Formalin-fixed and paraffin-embedded EUS-FNBs diagnostic of unresectable PDAC and fulfilling these criteria were included (n = 105): minimum 3-mm2 tissue, minimum of 2-mm2 tumor area, and minimum 20% relative tumor area. NGS was performed using Ion GeneStudio S5 Prime System and Oncomine™ Comprehensive Assay v.3 including 161 cancer-related genes. Results: Technical yield was 48% (105/219) and molecular yield was 98% (103/105). Most frequently mutated genes were KRAS (89.3%) and TP53 (69.9%), followed by CDKN2A (24.3%), ARID1A (9.7%), SMAD4 (7.8%), TSC2 (7.8%), and CCND3 (6.8%). Conclusion: EUS-FNB for NGS of unresectable PDAC is feasible. Our technical criteria for NGS, using leftovers in formalin-fixed and paraffin-embedded blocks after routine pathology diagnosis, were met by around half of EUS-FNBs. Almost all EUS-FNBs fulfilling the technical criteria yielded a successful NGS analysis.

The factors that influence the diagnostic accuracy and sample adequacy of EUS-guided tissue acquisition for the diagnosis of solid pancreatic lesions.

Background and Objectives: EUS-guided tissue acquisition (EUS-TA) is the preferred method to acquire pancreatic cancer (PC) tissues. The factors associated with false-negative outcomes and inadequate samples should be explored to gain an understanding of EUS-TA. Methods: The patients who underwent EUS-TA for suspected solid PC but whose results were false-negative were analyzed. The PC patients who underwent EUS-TA with true-positive results on the first day of every month during the study period were selected as the control group. The factors influencing diagnostic accuracy and sample adequacy were explored. Results: From November 2017 to January 2022, 184 patients were included in the false-negative group, and 175 patients were included in the control group. Multivariate logistic regression demonstrated that the recent acute pancreatitis [odds ratio (OR): 0.478, 95% confidence interval (CI): 0.250-0.914, P = 0.026] and high echo component within the tumor (OR: 0.103, 95% CI: 0.027-0.400, P = 0.001) were independently associated with false-negative EUS-TA results. Meanwhile, using fine-needle biopsy (FNB) needles (OR: 2.270, 95% CI: 1.277-4.035, P = 0.005), more needle passes (OR: 1.651,95% CI: 1.239-2.199, P = 0.005), large tumor size (OR: 1.053, 95% CI: 1.029-1.077, P < 0.001), and high CA-19-9 level (OR: 1.001, 95% CI: 1.000-1.001, P = 0.019) were independently associated with true-positive EUS-TA outcomes. Three needle passes are needed to achieve optimal EUS-TA outcomes. Tumor location in the body/tail (OR: 1.38, 95% CI: 1.01-1.72; P = 0.04), needle passes ≥3 (OR: 1.90; 95% CI: 1.22-2.56; P < 0.001), and using the FNB needle (OR: 2.10; 95%: 1.48-2.85; P < 0.001) were independently related to sample adequacy. Conclusion: Numerous factors were identified to be associated with the diagnostic accuracy and sample adequacy of EUS-TA.

The role of EUS in the diagnosis of early chronic pancreatitis.

The diagnosis of early chronic pancreatitis (ECP) is challenging due to the lack of standardized diagnostic criteria. EUS has been considered a sensitive diagnostic modality for chronic pancreatitis (CP), with advancements in technique such as EUS-guided fine needle aspiration and biopsy (EUS-FNA/FNB) being developed. However, their role in the diagnosis of ECP remains unelucidated. This review thereby aimed to provide an overview of the clinical landscape of EUS in the field of ECP.

Castration-resistant prostate cancer monitoring by cell-free circulating biomarkers.

Background: Prostate cancer is the second leading cause of male cancer-related deaths in Western countries, which is predominantly attributed to the metastatic castration-resistant stage of the disease (CRPC). There is an urgent need for better prognostic and predictive biomarkers, particularly for androgen receptor targeted agents and taxanes. Methods: We have searched the PubMed database for original articles and meta-analyses providing information on blood-based markers for castration-resistant prostate cancer monitoring, risk group stratification and prediction of therapy response. Results: The molecular markers are discussed along with the standard clinical parameters, such as prostate specific antigen, lactate dehydrogenase or C-reactive protein. Androgen receptor (AR) alterations are commonly associated with progression to CRPC. These include amplification of AR and its enhancer, point mutations and splice variants. Among DNA methylations, a novel 5-hydroxymethylcytosine activation marker of TOP2A and EZH2 has been identified for the aggressive disease. miR-375 is currently the most promising candidate among non-coding RNAs and sphingolipid analysis has recently emerged as a novel approach. Conclusions: The promising biomarkers have the potential to improve the care of metastatic prostate cancer patients, however, they need further validation for routine implementation.

CT-guided transperineal biopsy for prostate cancer in the absence of rectal access.

This case report presents CT-guided transperineal biopsy as an alternative method for diagnosis of prostate cancer in a patient with anorectal stenosis. A 69-year-old male had a history of anorectal surgeries. Conventional transrectal biopsy was unfeasible due to anorectal stenosis. The CT-guided transperineal biopsy was successfully performed using a cranio-caudal puncture technique, revealing adenocarcinoma. After the biopsy, the patient received appropriate hormone therapy and radiation therapy. This case report highlights the feasibility and safety of CT-guided transperineal biopsy for the patients with anorectal complications.

Liquid biopsy: Comprehensive overview of circulating tumor DNA (Review).

Traditional tumor diagnosis methods rely on tissue biopsy, which can be invasive and unsuitable for long-term monitoring of tumor dynamics. The advent of liquid biopsy has notably improved the overall management of patients with cancer. Liquid biopsy techniques primarily involve detection of circulating tumor cells (CTCs) and circulating tumor DNA (ctDNA). The present review focuses on ctDNA because of its significance in tumor diagnosis, monitoring and treatment. The use of ctDNA-based liquid biopsy offers several advantages, including non-invasive or minimally invasive collection methods, the ability to conduct repeated assessment and comprehensive insights into tumor biology. It serves crucial roles in disease management by facilitating screening of high-risk patients, dynamically monitoring therapeutic responses and diagnosis. Furthermore, ctDNA can be used to demonstrate pseudo-progression, monitor postoperative tumor status and guide adaptive treatment plans. The present study provides a comprehensive review of ctDNA, exploring its origins, metabolism, detection methods, clinical role and the current challenges associated with its application.