Fine needle aspiration cytology (FNAC) is a standard diagnostic tool and has several advantages which are specific to soft tissue tumors (STT). STT arises from non-epithelial and extra-skeletal tissue of the body, excluding the reticuloendothelial system, glia, and supporting tissues of organs. Primary STTs are very rare and encompass a wide range of different tumor types; therefore, the classification and diagnosis remain complex. If the tumor is diagnosed as benign, surgery may not be necessary. However, in the case of malignancy, a cytological diagnosis allows for the initiation of palliative treatment. This study was conducted to evaluate the role of FNAC in the diagnosis of STT by sub classifying them based on their cytology features and finally correlating their FNAC findings with histopathological results with special reference to immunohistochemistry (IHC) in diagnostic challenging cases. It was a hospital-based retrospective study conducted in a tertiary care center, over a period of 2 years, i.e., from July 2016 to May 2018. After obtaining approval from the ethics committee of the institution, the study was carried out. All STT cases subjected to FNAC followed by incisional, excisional, or core needle biopsy or complete resection for histopathological examination were studied. The slides of these patients were retrieved and reviewed to establish a proper diagnosis. IHC was carried out in most cases whenever there was a diagnostic dilemma. In our study, among 50 patients who presented with STT, 40 were benign, and 10 were malignant. The median age of incidence was 41, and male-to-female ratio was 0.85:1. The most common site was the trunk. There were four discordant cases, and the final evaluation of all results showed sensitivity, specificity, positive predictive value of tests, and negative predictive value of tests of FNAC as 100%, 76.92%, 92.5%, and 100%, respectively. In STT, most were benign tumors, with lipomas being the most common among them. While benign and malignant were easy to diagnose, intermediate spindle cell tumors were often missed on cytology. These intermediate tumors represent the diagnostic "grey zone." With increasing experience, STT can be diagnosed more accurately. FNAC is therefore a standard pre-operative diagnostic tool and a reliable alternative to open biopsy for diagnosing STT.

Breast cancer continues to be a leading and aggressive cancer in women. Despite improvements in early treatment, challenges such as rapid tumor proliferation, metastasis, and drug resistance persist. This research examines how the deubiquitinase ubiquitin-specific protease 5 (USP5) maintains Forkhead box M1 (FOXM1) protein stability and its impact on the advancement of breast cancer. Data from The Cancer Genome Atlas were utilized to investigate the expression patterns and interactions of USP5 and FOXM1 in breast cancer. The breast cancer cell lines were subjected to functional testing, including invasion, migration, and proliferation. The expression and interaction of USP5 and FOXM1 were examined using quantitative real-time polymerase chain reaction, Western blot, and co-immunoprecipitation analyses. Protein stability and ubiquitination assays were performed to evaluate the effect of USP5 on FOXM1 stability. USP5 stabilized the FOXM1 protein by deubiquitination. Overexpression of USP5 increased FOXM1 levels, while USP5 knockdown accelerated FOXM1 degradation. The deubiquitinating enzyme USP5 inhibited the proteasomal degradation of FOXM1, enhancing its stability. Functional assays showed that USP5 overexpression promoted breast cancer cell progression, while USP5 knockdown inhibited these malignant phenotypes. In vivo analysis showed that FOXM1 knockdown reduced tumor volume, and USP5 overexpression with FOXM1 knockdown increased tumor size. The findings suggest that USP5 promotes breast cancer progression by regulating FOXM1 stability. USP5 enhances breast cancer progression by stabilizing FOXM1 through deubiquitination.

This study aims to investigate the role of topoisomerase II binding protein 1-interacting checkpoint and replication regulator (TICRR) on the proliferation of lung adenocarcinoma (LUAD) and its potential molecular mechanism as well as to provide a new strategy to improve LUAD treatment. The expression level of TICRR in patients with LUAD was analyzed on the basis of the Cancer Genome Atlas program database, and loss- and gain-of-function experiments were conducted to validate whether TICRR promoted the proliferation of LUAD cells. Then, pathway enrichment analysis and luciferase reporter assays were performed to dissect the potential mechanism. Overexpression of TICRR in patients with LUAD was associated with poor survival (P < 0.001). In addition, overexpression of TICRR aggravated LUAD cell proliferation, which was ameliorated by TICRR depletion. In addition, TICRR could activate the myelocytomatosis oncogene (MYC) signaling pathway by regulating Bcl-2-associated X protein and Cyclin B1, which are the pivotal effectors of the MYC signaling pathway. TICRR plays a critical role in fostering the progression of LUAD by regulating the MYC signaling pathway.

Objective:Oncocytic thyroid follicular neoplasms (ONs), including oncocytic adenoma (OA) and oncocytic carcinoma (OC), are distinct follicular cell-derived tumors with unique molecular profiles. Oncocytic changes can also occur in other situations, such as papillary thyroid carcinoma (PTC) and thyroiditis. Due to overlapping cytological features, distinguishing benign from malignant ONs preoperatively remains challenging. This study examines cytological patterns and genetic alterations in ONs and aims to enhance diagnostic accuracy.Material and Methods:We retrospectively analyzed ON and oncocytic PTC (OPTC) cases from January 2018 to March 2025 at Cooper University Hospital, identifying 119 surgical resections (55 malignant, 62 benign, and two oncocytic non-invasive follicular thyroid neoplasms with papillary-like nuclear features. Sixty-five cases had fine-needle aspiration biopsy results categorized using The Bethesda System for Reporting Thyroid Cytopathology. Molecular examinations were performed in Afirma, ThyroSeq, or ThyGeNEXT/ThyraMIR. Statistical analyses included Wilcoxon Rank Sum tests, logistic regression, and Fisher’s exact tests.Results:No significant differences were observed between benign and malignant groups in terms of patient sex or tumor size; however, younger patient age was associated with malignant neoplasm within the cytology tested samples (P = 0.0300). Molecular analysis showed that 3.6% (1/28) of malignant cases were negative for tested alterations, while 33.3% (12/36) of benign cases were negative (P = 0.0035). Common mutations included eukaryotic translation initiation factor 1A X-linked (EIF1AX) (27.3%) in OCs and Harvey rat sarcoma viral oncogene homolog (HRAS) Q61R (41.2%) in OPTCs. In contrast, OA exhibited a higher frequency of chromosomal copy number alterations (33.3%) and variable other genetic changes.Conclusion:ONs pose diagnostic challenges due to significant cytological and molecular overlap between benign and malignant cases. Current thyroid molecular testing reveals a broad range of genetic alterations in ONs but lack sufficient specificity to reliably distinguish benign from malignancy in most instances. In this study, genetic alterations were detected in 66.6% (24/36) of OAs, and telomerase reverse transcriptase (TERT) mutations – typically associated with malignancy – were identified in two OAs. However, notably, certain mutations were exclusive to specific groups, such as B-Raf proto-oncogene, serine/threonine kinase (BRAF), anaplastic lymphoma kinase/echinoderm microtubule-associated protein-like 4 (ALK/EML4), and paired box 8 — peroxisome proliferator activator receptor gamma (PAX8-PPARG) in OPTCs or EIF1AX in OCs. Importantly, only 3.6% (1/28) of malignant OPTC/ONs were tested negative on molecular analysis, suggesting that ONs with negative molecular test results are more likely to be benign than malignant.

Objective:The objectives of the study are to investigate the differential metabolic paradigms and distribution of regulatory T (Tregs) cells between primary prostate cancer (PCa) and lymph node (LN) metastases.Material and Methods:Single-cell RNA sequencing analysis of primary PCa and LN metastases was employed to reveal the immune infiltration, identify Treg cell clusters, and analyze their metabolic regulation. Immunohistochemical (IHC) for FOXP3 and cluster of differentiation antigen 45 was used to verify different distribution and infiltration of Treg cells.Results:Immune cell infiltration was prominent around PCa cells, with Tregs significantly enriched in node-positive samples, suggesting an immunosuppressive microenvironment. Three Treg subsets were identified: Inhibitory Tregs, effector Tregs, and double-positive Tregs, each exhibiting distinct metabolic profiles. IHC confirmed higher Treg infiltration in LN metastases compared to primary tumors, particularly within tumor stroma.Conclusion:Tregs promote lymphatic metastasis in PCa through metabolic reprogramming, with their infiltration levels serving as a potential biomarker for metastatic risk.

Objective:Ovarian cancer (OC) is the most common and deadliest cancer in women worldwide. The high incidence and mortality rates highlight the serious threat that OC poses to women’s health. Regulator of chromosome condensation (RCC1) and broad-complex, tramtrack and bric a brac ( BTB) domain-containing protein 1 (RCBTB1), which includes the RCC1 and BTB domains, is a cell proliferation-related protein. This study aims to reveal the role of RCBTB1 in OC and its possible pathway.Material and Methods:The expression of RCBTB1 in OC cells was measured by quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemistry (IHC). Transwell assay, wound-healing assay, clone formation assay, IHC, immunofluorescence, and methylthiazolyldiphenyl-tetrazolium bromide assay were used to evaluate the effect of silencing RCBTB1 in vivo and in vitro. The neurofibromin 1 (NF1)/rat sarcoma (Ras) signal axis was determined by Western Blot, qRT-PCR, and immunofluorescence.Results:The A2780 cells had increased RCBTB1 expression (P < 0.01). By suppressing Ras signaling, RCBTB1 silencing hindered the proliferation of OC cells with Kirsten rat sarcoma viral oncogene (KRAS) mutations. RCBTB1 activated protein kinase kinase Cε, which degrades NF1. RCBTB1 also plays a key role in the Ras/ extracellular regulated protein kinase (ERK) signal axis by inhibiting Ras GTPase. RCBTB1 knockdown may alleviate mitogen-activated protein kinase kinase ( MEK) inhibitor resistance in KRAS-mutated OC by inhibiting Ras/ERK signaling.Conclusion:RCBTB1 may regulate the NF1/Ras signaling axis, which is critical for OC and MEK inhibitor resistance. This research offers a unique therapeutic approach for ovaries with KRAS mutations and uncovers a previously unknown connection between RCBTB1 and NF1/Ras signaling.

Objective:This study aims to explore the modulatory effect of 15-hydroxyprostaglandin dehydrogenase (15-PGDH) protein in the Notch1 signaling pathway in cervical cancer (CC) cells and assess how this modulation affects the proliferation and migration of CC cells. Moreover, this study offers fresh perspectives on the molecular mechanisms underlying CC by thoroughly analyzing the relationship between 15-PGDH and the Notch1 signaling pathway, and investigates the therapeutic potential of 15-PGDH.Material and Methods:Human normal cervical epithelial cells and CC cell lines (human CC cell line [HeLa], human cervical squamous carcinoma cell line [Caski], and human cervical epidermoid carcinoma cells [ME180]) were selected as experimental models. Western blotting (WB) and quantitative reverse transcription polymerase chain reaction were performed to evaluate the protein and messenger RNA levels of 15-PGDH and Notch receptor 1 (Notch1) signaling pathway-related proteins (Jagged canonical Notch ligand 1 [Jagged1] and Hes family bHLH transcription factor 1 [Hes1]). Results suggested that the HeLa and Caski cells exhibited significant expression of 15-PGDH and Notch1 signaling-related proteins. A series of experiments, including WB, cell counting kit-8 assay, Transwell migration assay, and 5-ethynyl-2'-deoxyuridine assay, was conducted in the HeLa and Caski cells to obtain an extensive understanding of how 15-PGDH influences Notch1 signaling regulation. This study also utilized the 15-PGDH inhibitor SW033291 and a Notch1 overexpression vector to evaluate the effect of 15-PGDH on CC cell growth, motility, and Notch1 signaling pathway modulation.Results:Results demonstrated that in the normal human cervical epithelial cells, 15-PGDH was highly expressed, while the Notch1 signaling pathway-related proteins exhibited low expression quantities. However, in HeLa and Caski CC cells, 15-PGDH expression was significantly downregulated (P < 0.001 or P < 0.01), whereas the Notch1 signaling pathway was activated. Further studies revealed that 15-PGDH or its inhibitor influenced the stimulation of the Notch1 signaling pathway in the HeLa and Caski cells. Specifically, the 15-PGDH inhibitor SW033291 reduced 15-PGDH expression (P < 0.001 or P < 0.01) and promoted Notch signaling activation. Meanwhile, 15-PGDH upregulation suppressed Notch signaling activation. Furthermore, 15-PGDH successfully prevented the proliferation and migration of CC cells induced by Notch1 overexpression and reduced the activation of the Notch signaling pathway, as shown by the downregulation of Notch1 and its downstream effectors, Jagged1 and Hes1.Conclusion:This study highlights the role of 15-PGDH in regulating the Notch1 signaling pathway in CC cells, focusing on its effect on cell proliferation and migration. The results demonstrate that 15-PGDH suppresses CC cell proliferation and migration by downregulating the Notch1 signaling pathway. These findings provide new insights into the molecular mechanisms underlying CC and suggest 15-PGDH as a promising therapeutic target.

Objective:Pregnancy-induced hypertension (PIH) is a common complication during pregnancy and is closely associated with vascular endothelial cell damage and nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3)-mediated pyroptosis. This study aimed to investigate whether mitophagy alleviates vascular endothelial cell damage in PIH by inhibiting NLRP3-mediated pyroptosis. The regulatory mechanisms of pyroptosis-related pathways were systematically investigated by establishing a cellular model of PIH and incorporating mitophagy intervention.Material and Methods:An Nω-nitro-L-arginine methyl ester (L-NAME)-induced gestational hypertension model was established, and the cell samples were grouped as follows: Control group (Control), L-NAME-induced gestational hypertension group (L-NAME), mitochondrial autophagy inhibition group (L-NAME+ 3-methyladenine [3-MA]), and mitochondrial autophagy activation group (L-NAME+ rapamycin [Rapa]). Cell viability was assessed through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, lactate dehydrogenase (LDH) levels were measured to evaluate cell damage, and reactive oxygen species (ROS) kits were used to quantify ROS accumulation. Cell death was evaluated using terminal deoxynucleotidyl transferase dUTP nick end labeling staining to detect apoptotic cells. Immunofluorescence, Western blot analysis, and quantitative real-time polymerase chain reaction were performed to assess the expression levels of proteins and genes associated with mitophagy (e.g., microtubule-associated protein 1 light chain 3 and sequestosome 1) and those linked to pyroptosis (e.g., NLRP3, gasdermin D (GSDMD), cysteinyl aspartate-specific proteinase 1 (caspase-1), interleukin (IL)-1β, and IL-18). The role of NLRP3 in pyroptosis regulation through mitochondrial autophagy was further examined using NLRP3 small interfering RNA (siNLRP3) transfection experiments.Results:L-NAME treatment substantially decreased vascular endothelial cell viability, elevated LDH release and ROS levels, and upregulated pyroptosis-related proteins (NLRP3, GSDMD, and caspase-1) and inflammatory factors (IL-1β and IL-18). The inhibition of mitochondrial autophagy with 3-MA further enhanced pyroptosis and aggravated cell damage, and its activation with Rapa reduced pyroptosis, improved cell survival, and decreased LDH release and ROS levels. NLRP3 silencing (siNLRP3) significantly inhibited pyroptosis and alleviated the cell damage caused by 3-MA. Meanwhile, Rapa enhanced the protective effect of NLRP3 silencing.Conclusion:This study demonstrates that mitophagy can effectively alleviate the vascular endothelial cell damage associated with PIH by inhibiting NLRP3-mediated pyroptosis. The findings provide new theoretical support for the treatment of PIH and suggest potential intervention targets.

ObjectivePancreatic cancer is the cancer type with the highest mortality rate worldwide, and despite advances in treatment, molecular biomarkers are needed for both early diagnosis for developing targeted therapies and improving survival rates in this challenging malignancy. In our study, the contributions of programmed death-ligand 1 (PD-L1) expression to the determination of pancreatic cancer subtypes and patient prognosis and its impact on survival were investigated.Material and MethodsParaffin-embedded tissues from 92 patients diagnosed with pancreatic cancer were included in this study. Tumor-infiltrating lymphocytes (TILs) and lymphocytes in the stromal area within the tumor borders were scored as stromal TILs; lymphocytes in tumor islands were scored as intratumoral TILs. Staining in each area was scored as a percentage, and staining with a score of 5 or more in tumor and immune cells for PD-L1 was scored as positive.ResultsAfter staining, a score of 5 or more with tPD-L1 staining was used to identify one patient as having micropapillary adenocarcinoma, three patients as having ductal adenocarcinoma, and four patients as having signet ring cell carcinoma. When the clinical parameters and outcomes were compared, a statistically significant difference was found between the histopathologic type of signet ring cell carcinoma and poor differentiation and positivity of PD-L1 expression (P < 0.05). Survival was significantly influenced by tumor location, histopathological subtype, degree of differentiation, PD-L1 expression, and tumor size, with tumor size being the most critical factor (P < 0.05).ConclusionOur findings suggest that PD-L1 positivity is notably prevalent in signet ring cell carcinoma of the pancreas and is strongly associated with poor survival outcomes. Given these results, further studies with larger patient cohorts are warranted to validate these observations and explore potential therapeutic implications.