Positive Expression Research Articles

Immunohistochemical expression of ephrin receptors in neuroendocrine neoplasms: a case-series of gastroenteropancreatic neuroendocrine neoplasms and a systematic review of the literature.

Erythropoietin-producing hepatocellular (EPH) receptors are the largest known family of tyrosine kinases receptors (TKR) in humans, implicated in cell proliferation, adhesion, migration, tumor angiogenesis, invasion and metastasis. The aim of the present study is to assess the expression of EPHs in neuroendocrine neoplasms (NENs). Immunohistochemical staining of specimens of 30 patients with gastroenteropancreatic and lung NENs was performed for EPH-A1, EPH-A2, EPH-A4, EPH-A5 protein expression, in addition to ki-67 multiplication index and programmed death-ligand 1. Additionally, we performed a systematic review of the available literature in three different databases reporting on the expression of EPH in all neuroendocrine neoplasms. Positive expression was seen in 16/19 (84%) specimens for EPH-A1, 15/23 (65%) for EPH-A2, 21/24 (88%) for EPH-A4, 24/26 (92%) for EPH-A5. EPH-A1 was expressed in 9/9 pancreatic, 3/4 small intestine, but not in one lung NEN, EPH-A2 in 5/10 pancreatic, 3/4 small intestine and lung, and in one of each of gastric, appendix, colorectal, and cervical NENs, respectively. EPH-A4 showed positive expression in 9/11 pancreatic, 4/4 small intestine, 3/3 lung specimens and EPH-A5 in 10/11, 4/4 and 4/4, respectively. Data retrieved from the systematic review of the literature in combination with the data from the present study are suggestive of a frequent EPH expression in pituitary, thyroid, lung and gastroenteropancreatic NENs, yet, with varying expressions of the single receptor subtypes. EPHs may have a role in NEN tumorigenesis, prognosis as well as a role in the evolving molecular-targeted therapies.

Cervical angiofibroma of soft tissue: A rare case report with literature review.

Angiofibroma of soft tissue (AFST) is a rare benign fibrous tumor recently included in the 2020 WHO classification of soft tissue and bone tumors. Currently, there are limited reports on AFST, and pathologists lack sufficient understanding of its clinical and pathological characteristics. There is scarce literature available on AFST in the cervical region. We presented a case of a 49-year-old woman who was admitted to our hospital for emergency treatment due to vaginal bleeding and fatigue. Ultrasound revealed a 63 × 27 mm hypoechoic mass extending from the cervical opening to the external opening, with a hemoglobin level of 58.0 g/L on blood routine. The tissue exhibited a pale-yellow mucinous appearance with distinct tissue boundaries and a fibrous capsule under a microscope. HE staining revealed spindle-shaped fibroblast-like cells with a consistent morphology, thin-walled branching small blood vessels, and dilated blood vessels. Regions with abundant cells and areas with sparse cells were alternately distributed and migrated gradually. Immunohistochemical analysis indicated positive expression of P53, desmin, progesterone receptor, estrogen receptor, epithelial membrane antigen, vimentin, CD68, CD163 in the tumor, but negative expression of P16, S-100, smooth muscle actin, CD117, CD10, STAT-6. CD34 was negative in the tumor cells but positive in the vascular endothelium. The Ki-67 index value was 5%. Pathological examination confirmed a soft tissue angiofibroma of the cervix. Emergency hysteroscopic surgery was performed following infusion of 3 units of packed red blood cells. A local excision of the cervical mass was performed. A 1-month follow-up ultrasound showed no abnormal mass in the cervical canal, and there have been no signs of recurrence to date. Cervical angiofibroma of soft tissue is a rare tumor with a benign clinical manifestation, minimal local recurrence, and no significant metastatic potential. Treatment primarily involves local resection with a focus on achieving negative surgical margins. By presenting this case, we aim to enhance the diagnostic and differential diagnostic capabilities of pathologists in identifying uterine tumors and preventing misdiagnosis.

Clinicopathological correlations in 38 cases of gastroenteropancreatic high-grade neuroendocrine neoplasms.

Diagnosis and treatment of gastroenteropancreatic high-grade neuroendocrine neoplasms (GEP-HG-NENs), particularly G3 well-differentiated neuroendocrine tumours (NETs) and poorly differentiated neuroendocrine carcinomas (NECs) relies on histopathological morphology, immunohistochemistry, and molecular biological markers, which are lacking especially in cases with ambiguous histomorphology. In this study to contribute to the development of more targeted treatment strategies, we examined various immunohistochemical and molecular biological markers and their association with clinicopathological features in GEP-HG-NENs. We included 38 patients with GEP-HG-NENs in this study, with their retrospective follow-up data. The expression of tumour protein p53 (TP53), RB transcriptional corepressor 1 (RB1), somatostatin receptor 2 (SSTR2), clusterin (CLU), and marker of proliferation Ki-67 (MKI67) was immunohistochemically analysed. KRAS proto-oncogene, GTPase (KRAS) and B-Raf proto-oncogene, serine/threonine kinase (BRAF) V600E expression was evaluated using quantitative real-time polymerase chain reaction (qRT-PCR). The relationships between immunohistochemical and molecular biological markers and clinicopathological characteristics were examined using a Cox risk regression model, receiver operating characteristic (ROC) curve, and Kaplan-Meier survival analyses. SSTR2, RB, TP53, and CLU expression differed between NET G3 and NECs, with variations among the NET G3 and small- and large-cell NEC (SCNEC and LCNEC, respectively) groups (p < 0.05). The median MKI67 proliferative index was approximately 40% and 70% in G3 NETs and NECs, respectively. The NET G3 group exhibited a median survival of 25 months, indicating a relatively better prognosis than that of the NECs group (median survival, 11 months). Both Kaplan-Meier survival analysis and the Cox risk regression model indicated a statistical correlation among treatment methods, CLU expression, and prognosis (p < 0.05). The BRAF V600E mutation rate was 32.4% in G3 NETs and SCNEC, demonstrating a significant difference between both types (p = 0.0086). Furthermore, ROC curve analysis highlighted the diagnostic significance of the positive expression of the immunohistochemical markers CLU, SSTR2, and RB in identifying NET G3. To guide more suitable treatment strategies, it is essential to develop and apply valuable and more targeted immunohistochemical and molecular pathological markers for a comprehensive analysis.

Comparative Immunohistochemical Analysis of Craniopharyngioma and Ameloblastoma: Insights into Odontogenic Differentiation.

Histopathological similarities between craniopharyngioma (CP) and ameloblastoma (AB) have long been recognized, particularly the shared features of palisading columnar epithelium and stellate reticulum-like areas. This study aimed to investigate potential odontogenic differentiation in CP akin to AB using immunohistochemical odontogenic markers. We analyzed AMELX, ODAM, and CK19 expression in 44 cases (20 CP and 24 AB). While AMELX and ODAM showed diffuse strong positive expression in both tumors with no significant statistical differences, CK19 expression was notably higher in CP. The markers AMELX and ODAM associated with odontogenic differentiation exhibited similar profiles in both tumors due to shared similar embryological origins. Notably, CK19, a biomarker of odontogenic epithelium, showed even higher expression, suggesting distinct pathways. These findings offer insights into tumor biology and may aid in diagnostic and therapeutic approaches.

Investigation of PLGF, sFlt-1 expression in Plasenta Previa, Plasenta Accreta and Normotensive Plasentas in the 3rd Trimester of Pregnancy

Background: We aimed to immunohistochemically examine the expression level of Placental growth factor (PLGF) and Soluble FMS-like tyrosine kinase-1 (sFlt-1) proteins in normotensive, Placenta accreta (PA) and Placenta previa (PP). Materials and Methods: Three groups were created for the study: 20 Nomotensive placentas, 20 PP and 20 PA from women diagnosed with placenta samples. 4-6 μm thick sections were taken from the placentas, PLGF and sFlt-1 immunostaining were applied to the obtained sections, and their expression intensities and localizations in the tissues were determined. Results: As a result of our observations, normotensive placenta group; sFlt-1 expression was observed in hofbauer cells and syncytial nodes and PLGF positive expression was observed in nuclei of muscle cells in some tunica media region. PP and PA group; sFlt-1 expression was positive in decidua cells, hofbauer cells and dilated vascular endothelium. PP and PA group; negative PLGF expression was observed in syncytial nodes and positive PLGF expression was observed in hofbauer cells and endothelial cells. There was a significant difference between normotensive, PP and PA groups (p&amp;lt;0.05). Conclusions: It can be thought that sFlt-1 and PLGF may be important signal stimulators and markers in the trophoblastic degeneration, apoptotic cells, increase in angiogenesis and imbalance in implanta-tion seen in cases of PP and PA.

Immunohistochemical expression of PD-L1 in esophageal carcinoma and its correlation with histopathological features

Background: Carcinoma of the esophagus poses a considerable medical and public health challenge in many parts of the world. Despite advances in screening and multimodal management of this disease, overall survival remains poor. There is an utmost need to find a marker for the establishment of targeted immunotherapy. One immunohistochemical marker that has surfaced with the advent of anti-PDL1 immunotherapy is PD-L1 which has been proven to be advantageous. This study aimed to determine PD-L1 expression and its correlation with the grade of tumor and other histopathological parameters in 50 patients. Materials and Methods: A cross-sectional study was conducted on 50 cases of histopathologically proven esophageal carcinoma. Antibody used for PD-L1 was rabbit monoclonal (CAL10) Biocare. PD-L1 expression was considered positive when a complete or partial membranous or cytoplasmic staining pattern was noted in more than 5% of tumor cells. Correlation between PD-L1 expression and clinicopathological parameters was calculated. Proper consent was taken. The findings of the study were statistically analyzed using Statistical Package for the Social Sciences (SPSS) software version 21.0, a statistical program for Microsoft Windows. Results: This study revealed that PD-L1 positivity increases as the histopathological grade of esophageal squamous cells increases. Conclusions: The expression of PD-L1 increases with grade and shows more positive expression in poorly differentiated tumors. This indicates that PD-L1 overexpression is associated with aggressive disease. Targeted therapy against PD-L1 may prove to be beneficial in such patients.

Predictors of breast cancer HER2-receptor positivity by MRI intuitive imaging features

BackgroundToday, breast cancer is the most diagnosed cancer worldwide. There are many different clinical presentations, radiological characteristics, and histological types of breast cancer. HER2 is overexpressed in a significant number of breast cancer cases reaching 20% of all breast cancers, and its overexpression is seen directly proportionate with a poor outcome and prognosis.MethodsWe started this cross-sectional research from January 2022–December 2023 on 202 breast cancer patients who had 220 lesions. The molecular subtypes of the different lesions were determined in all the included cases. Magnetic resonance imaging (MRI) studies were conducted in all included cases. The MRI parameters included conventional MRI, diffusion-weighted analysis, and dynamic post-contrast T1-weighted imaging.ResultsThe prevalence of irregular margins (P < 0.001), linear and segmental distribution (P = 0.044), heterogeneous pattern (P < 0.001), and type 2 curve was statistically significantly higher in the HER2-positive lesions. Nipple infiltration incidence showed statistically significant elevation in the HER2-positive lesions (P = 0.017). The lesions’ ADC and perilesional ADC in the HER2-positive lesions were also statistically significantly elevated. The best cutoff point of ADC to detect lesions with positive HER2 expression was > 0.885 × 10–3 mm2/s, with 65.7% sensitivity and 60% specificity, with a statistically significant value (p = 0.005).ConclusionsMagnetic resonance imaging of breast imaging is a promising noninvasive method for identifying breast tumors with the HER2 molecular subtype. Combining various radiological features by MRI may provide a conclusion for recognizing positive HER2 lesions.

Expression and its clinical significance of cell-cycle dependent kinase 1 in malignant peripheral nerve sheath tumors

To explore the role and clinical significance of cell-cycle dependent kinase 1 (CDK1) and its upstream and downstream molecules in the development of malignant peripheral nerve sheath tumor (MPNST) through the analysis of clinical tissue samples. A total of 56 tumor samples from MPNST patients ("Tianjin" dataset) who underwent surgical resection, confirmed by histology and pathology between September 2011 and March 2020, along with 17 normal tissue samples, were selected as the research subjects. MPNST-related hub genes were identified through transcriptome sequencing, bioinformatics analysis, immunohistochemistry staining, and survival analysis, and their expression levels and prognostic associations were analyzed. Transcriptome sequencing and bioinformatics analysis revealed that upregulated genes in MPNST were predominantly enriched in cell cycle-related pathways, with CDK1 occupying a central position among all differentially expressed genes. Further differential analysis demonstrated that CDK1 mRNA expression in sarcoma tissues was significantly higher than in normal tissues [based on searching the cancer genome atlas (TCGA) dataset, P<0.05]. In MPNST tissues, CDK1 mRNA expression was not only significantly higher than in normal tissues (based on Tianjin, GSE141438 datasets, P<0.05), but also significantly higher than in neurofibromatosis (NF) and plexiform neurofibromas (PNF) (based on GSE66743 and GSE145064 datasets, P<0.05). Immunohistochemical staining results indicated that the expression rate of CDK1 protein in MPNST tissues was 40.31%. Survival analysis results demonstrated that CDK1 expression was associated with poor prognosis. The survival time of MPNST patients with high CDK1 mRNA expression was significantly lower than that of the low expression group ( P<0.05), and the overall survival trend of patients with positive CDK1 protein expression was worse than that of patients with negative CDK1 expression. Additionally, differential analysis of CDK family genes (CDK1-8) revealed that only CDK1 was significantly upregulated in MPNST, NF, and PNF. Increased expression of CDK1 is associated with poor prognosis in MPNST patients. Compared to other CDK family members, CDK1 exhibits a unique expression pattern, suggesting its potential as a therapeutic target for MPNST.

Ovarian Microcystic Stromal Tumor with Significant Nestin Expression: A Unique Case.

Ovary microcystic stromal tumor (MCST) is an extremely rare subtype of sex cord-stromal neoplasm, and only 57 cases have been reported. We herein report a unique case of ovarian MCST with positive nestin expression in a 39-year-old Chinese woman. The tumor showed microcystic stromal histological structures and characteristically expressed the CD10, WT-1, and Ki67 proteins. A molecular analysis identified a point mutation (c.110C > T) in exon 3 of the CTNNB1 gene. To our knowledge, no report has described a case of ovarian MCST with positive staining for nestin protein. Our study provides new insights into the tumor biology of ovarian MCST.

Can multiplexing Immunohistochemistry for PD-L1, CD68, and CD3 improve scoring for “Keytruda®” Indication?

Abstract Introduction/Objective FDA-approved companion diagnostics immunohistochemistry scoring for PD-L1 is essential for Keytruda ® treatment in patients with NSCLC. The challenge with PD-L1 testing alone is false positive PD-L1 expression score due to alveolar macrophage infiltration. Multiplexing with PD-L1, CD68, and CD3 could help alleviate this issue. By viewing PD-L1 expression on tumor cells, CD68 expression on macrophages, and the presence of CD3+ T cells on a single slide, pathologists will be able to see the presence of an immune microenvironment (high PD-L1 expression with abundant macrophages) and the potential for response to other types of Immunotherapies (presence of T cells). Methods/Case Report Positive NSCLC FFPE blocks were sectioned and stained using Agilent Dako Omnis. Two combinations of multiplex staining were prepared for each block by staining PD-L1 (brown) alone first followed by CD3 and CD68. First slide: CD3(Vina Green), CD68(Magenta); Second slide: CD3(Magenta), CD68(Vina Green). To prevent wash out, dehydration and clearing occurred with two changes of 100% alcohol and three changes of xylene for manual cover slipping. Results (if a Case Study enter NA) Manual scoring of PD-L1 alone vs two different multiplex combinations was compared to Agilent Proscia digital AI scoring using a preliminary study set (9 positives, 2 negatives). CD68-green and CD3-magenta showed better discrimination than CD68-magenta and CD3-green at a glance. This could be due to the staining intensity and nonspecific background staining of CD68-magenta. Manual scoring was higher compared to digital scoring, which could be due to the human error of positive bias. Digital scoring was significantly lower for PD-L1 alone vs both multiplex stains. The digital scoring for the multiplex stains showed a higher scoring percentage by removing false positive macrophages. Conclusion Reflex scoring workflow is being developed to focus on difficult cases using the multiplex triple stain, i.e. clear negative and clear &amp;gt;50% with 3+ intensity. More cases will be collected and studied to further investigate the improved PD-L1 scoring with digital imaging assisted by artificial intelligence and to compare the multiplex color combinations.

Irisin suppresses PDGF-BB-induced proliferation of vascular smooth muscle cells in vitro by activating AMPK/mTOR-mediated autophagy.

Restenosis is a pivotal factor that restricts the efficacy of coronary artery bypass grafting. Inhibition of vascular smooth muscle cells (VSMCs) proliferation can improve intimal hyperplasia and lumen stenosis. Irisin, a polypeptide secreted by muscle cells, has been demonstrated to have a protective role in various cardiovascular diseases. However, the effect and mechanism of irisin on VSMCs proliferation and phenotype switching remain unclear. Cell proliferation ability was assessed using the methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay and 5-ethynyl-2'-deoxyuridine (EdU) incorporation. Cell cycle analysis was performed using flow cytometry, while expression levels of contractile and synthesis-related proteins were determined through RT-qPCR and Western blot. The VSMCs were infected with an adenovirus carrying GFP-LC3, and the proportion of cells showing positive expression was assessed. Additionally, the formation of autophagic lysosomes in cells was observed through transmission electron microscopy. In this study, we have demonstrated the inhibitory effects of irisin on the proliferation and phenotypic transition of platelet-derived growth factor-BB (PDGF-BB)-induced VSMCs. More importantly, we have discovered that irisin can activate the AMP-activated protein kinase/mammalian target of rapamycin (AMPK/mTOR) signaling pathway to mediate autophagy in PDGF-BB-induced VSMCs. The inhibitory effect of irisin on PDGF-BB-induced VSMCs proliferation was significantly attenuated by the AMPK inhibitor, Compound C. Conversely the mTOR inhibitor, rapamycin further enhanced the inhibitory effect of irisin on PDGF-BB induced VSMCs proliferation. In conclusion, our findings suggest that irisin effectively suppresses the aberrant proliferation of VSMCs following PDGF-BB stimulation by modulating autophagy levels through the AMPK/mTOR signaling pathway.

Keratin-15 high expression links with lymph node metastasis and poor survival prognosis in epithelial ovarian cancer patients

BackgroundKeratin-15 (KRT15) involves in the progression and owns prognostic values in several solid cancers, whose clinical role in epithelial ovarian cancer (EOC) is rarely reported. This study aimed to identify the association of KRT15 expression with tumor features and survival of surgical EOC patients.MethodsFormalin-fixed paraffin-embedded tumor tissues of 140 EOC patients who underwent tumor resection were retrieved for KRT15 determination using immunohistochemistry (IHC) assay.ResultsThe median (interquartile range) KRT15 IHC score was 0.0 (0.0–1.0), ranging from 0.0 to 12.0. Among all, 36.4% of patients had positive KRT15 expression (IHC score > 0) and 15.0% of patients had high KRT15 expression (IHC score > 3). KRT15 was positively related to lymph node metastasis incidence (P = 0.027), and showed a tendency to correlate to FIGO stage but without statistical significance (P = 0.052), while it was not correlated with age, other tumor features, and tumor markers. Positive KRT15 expression was linked with poor disease-free survival (DFS) (P = 0.009) and overall survival (OS) (P = 0.032). Notably, high KRT15 expression showed an even stronger relationship with worse DFS (P = 0.001) and OS (P < 0.001). After adjustment of multivariable Cox’s regression, high KRT15 expression was independently correlated with unfavorable DFS (hazard ratio (HR): 2.241, P = 0.007).ConclusionEven though KRT15 is insufficiently expressed in EOC tissues generally, its positive expression or high expression can predict the lymph node metastasis and poor survival prognosis in EOC patients who undergo tumor resection.

Correlation analysis of Ki67 changes with survival outcomes in breast cancer before and after neoadjuvant therapy based on residual cancer Burden grade

PurposeThis study aims to investigate the change of Ki67 value pre- and post-neoadjuvant therapy (NAT) and evaluate its potential value in predicting survival outcomes in different molecular subtypes of breast cancer. MethodsA total of 257 breast cancer patients who underwent NAT at Renmin Hospital of Wuhan University from July 2019 to Sep 2023 were included in this study. The Ki67 index of the patients was re-interpreted by two attending physicians, and the changes of Ki67 value pre- and post-NAT were compared. Chi-square test (χ2) and logistic regression were conducted to examine the correlation between various characteristics and the efficacy of NAT. Disease-free survival (DFS) was calculated using the Kaplan-Meier curve and compared using the log-rank test. ResultsPatients with higher histological grade, negative expression of estrogen receptor (ER) or progesterone receptor (PR), positive expression of human epidermal growth receptor 2 (HER2), higher pretreatment Ki67 index, absence of lymph node metastasis, and those with HER2 positive and triple-negative breast cancer were associated with improved efficacy of NAT. Our study identified that the optimal cut-off value for the changes in Ki67 index pre- and post-NAT related to the effectiveness of NAT was “-88.19 %” in whole chort, which was related to the aforementioned clinical characteristics. Besides, the optimal cut-off values for the luminal, HER2-enriched and triple-negative subtypes were “-91.83 %”, “-46.12 %” and “-81.67 %”, respectively. Survival analysis demonstrated that the changes in Ki67 value were significantly associated with DFS in the HER2-enriched and triple-negative subtype, but not in the luminal subtype. ConclusionsPreoperative clinicopathological features and changes in Ki67 value pre-and post-NAT can contribute to providing patients with a more accurate prognosis.

Expression Profiles of Integrin-Linked Kinase, Vascular Endothelial Growth Factor A, and Ephrin Type-A Receptor 2 in Colorectal Cancer Lymph Nodes.

Background Colorectal cancer (CRC) is the third most frequently diagnosed cancer globally, with a high incidence of morbidity and mortality. Early diagnosis of CRC is crucial for determining appropriate treatment regimens, thereby prolonging overall survival rates and improving prognostic outcomes. Objectives This study aimed to investigate the expression profiles of specific proteins in the lymph nodes (LNs) of CRC patients, including integrin-linked kinase (ILK), vascular endothelial growth factor A (VEGFA), and ephrin type-A receptor 2 (EphA2), through immunohistochemical studies. Methods The study involved a sample of 76 patients clinically diagnosed with CRC who were referred by their specialist oncologist. Tumor staging was determined based on the TNM classification. Immunohistochemical studies were conducted to measure the expression patterns of the candidate markers (ILK, EphA2, and VEGFA). Expression levels were scored as negative, low, or high. Results The highest percentage of CRC patients were diagnosed with conventional adenocarcinoma, predominantly in stages II and III. Of the 76 CRC tissue specimens, the majority (46, 60.52%) tested negative for lymphatic invasion, while the remaining 30 (39.47%) tested positive. According to the TNM classification, 14 samples had N1 (one invaded LN), and 16 had N2 (two or more invaded LNs). Furthermore, the percentage of patients with low and high EphA2 expression was significantly higher (p<0.0001) compared to the negative expression controls. Regarding ILK, 15 cases (50.0%) showed negative expression, while an equal number displayed positive expression. Additionally, the group with low VEGFA expression was statistically significantly higher (p=0.01) compared to the negative control. Conclusion The expression levels of EphA2, ILK, and VEGFA were found to be higher in LNs with lymphatic invasion compared to those with negative expression, highlighting the role of these proteins in CRC progression.

Macrophage hyperfunction as a cellular trigger for fibroplastic changes in internal organs in pneumoconiosis among miners

Introduction. The significant prevalence of anthracosilicosis and silicosis, characterized by the rapid development of pneumofibrosis in coal mine workers, is a serious socially significant problem worldwide. A key component of the pathogenesis of pneumoconiosis is the macrophage reaction to fibrogenic dust, which is a complex immunological response, with prolonged and intense exposure leading to chronic inflammation in various organs and systems, fibrosis and a progressive decrease in their functions. A deep understanding and evaluation of the macrophage reaction as a potential biomarker for the diagnosis of lung damage in miners will allow us to develop more effective strategies to combat dust damage to organs and systems. The study aims to consider macrophage hyperfunction as a trigger mechanism for fibroplastic changes in internal organs in pneumoconiosis in miners. Materials and methods. Using light microscopy, specialists conducted extended cytological, histological and immunohistochemical studies of bronchial smears and autopsy material (fragments of bronchi, lungs, heart, liver and kidneys) of 50 miners of Kuzbass coal enterprises who died simultaneously as a result of a man-made disaster. Morphometric analysis of histological structures with measurement of linear dimensions was used for a comprehensive assessment of pathomorphological changes. Results. There was a sharp increase in the number and size of actively phagocytic alveolar macrophages loaded with dust particles in the lungs of miners. Pathological changes in internal organs were characterized by pronounced fibrosis in the peribronchial and perivascular zones, stroma with deposition of dust particles and a pronounced macrophage reaction. In a group of miners with more than 15 years of experience in the central and distal parts of the lungs, in the intermuscular zones of the myocardium, the portal tracts of the liver, and the central segments of the kidneys, there was a constant increase in the expression of immunohistochemical markers CD14, CD34, actin and vimentin, which can be used as diagnostic markers for the identification of macrophage inflammation and fibrosis, will allow to assess the degree of severity of the pathological process. A characteristic feature of the miners' long experience of dust exposure was the presence of progressive pronounced perivascular sclerosis extending beyond the vascular histione. Experts found fibroblastic cells with positive expression of CD14, CD31 and CD34 in the sclerotic zones. Limitation. The absence of cytogenetic and electron microscopic examination, which is expected in the future in conjunction with the expansion of the range of immunohistochemical research methods. Conclusion. The results of extended pathomorphological and cytological studies emphasize the importance of an integrated approach to the diagnosis of occupational diseases in the coal industry. The use of a combination of these methods will allow timely detection of macrophage inflammation and fibrous process in dust pathology, specific changes at the cellular and tissue levels, as well as to assess the severity of the pathological process. Ethics. The study of autopsy material was based on a secondary examination of blocks and ready-made histological micro-preparations (glasses) of the material of the Bureau of Forensic Medical examination of Novokuznetsk, Osinniki, Prokopyevsk. The studies of pathomorphological material were carried out strictly on the basis of Federal Law No. 323-FZ dated 11/21/2011 "On the Basics of Protecting the Health of Citizens in the Russian Federation", in particular Article 67 "Conducting pathological anatomical autopsies", Federal Law No. 8-FZ dated 12.01.1996 "On Burial and Funeral Business" (Article 5, pp. 1, 2), as well as in accordance with the Order of the Ministry of Health of the Russian Federation dated 06.06.2013 No. 354n "On the procedure for conducting pathological anatomical autopsies", Order of the Ministry of Health of the Russian Federation dated 03/24/2016 No. 179n "On the Rules for conducting pathological anatomical studies".

Association Between Recurrence of High-grade Squamous Intraepithelial Lesions of the Uterine Cervix and p16, C-myc and PIK3CA Proteins-A Single-center Retrospective Study.

Cervical high-grade squamous intraepithelial lesions (HSIL) are one of the common types of cervical cancer precancerous changes, and HPV16/18 positivity is a risk factor for HSIL recurrence. By detecting the expression of relevant markers in the lesion tissue of recurrent patients, it is helpful for the diagnosis of HPV16/18 positivity and can provide a basis for disease recurrence risk assessment. Therefore, this study analyzed the relationship between p16, C-myc, PIK3CA proteins and HPV16/18 positivity in recurrent cervical HSIL patients. By examining the p16, C-myc, and PIK3CA proteins in the cervical lesion tissue of 180 HSIL recurrent patients who underwent examination in the hospital from January 2020 to December 2022, this study analyzed the relationship between p16, C-myc, and PIK3CA proteins and HPV16/18 positivity. PIK3CA expression detection found that the proportion of positive expression of p16, C-myc, and PIK3CA in HPV16/18 (+) patients was significantly higher than that in HPV16/18 (-), and the expression of HPV16/18 in HSIL patients was significantly positively correlated with p16, C-myc, and PIK3CA. Meanwhile, a prediction model F was constructed based on binary logistic regression analysis data with good fit, and through ROC curve analysis. It was found that p16, C-myc, PIK3CA, and logistic model F can effectively predict HPV16/18 (+), with model F having the best diagnostic performance.

Early detection of cervical tumor cell of origin through Oncogene-induced senescent HPV-positive cells.

Human Papilloma Virus (HPV) is an oncogenic virus and is the most common cause of cervical cancer. HPV has been shown to induce senescence. Cellular senescence is involved in cancer progression and tumorigenesis. Identification and isolation of cells of tumor origin before tumorigeneses is an important step in cancer prevention and treatment. This study aimed to investigate the early cervical atypical senescent cytological preneoplastic change in non-menopausal women. Cervical smears of 121 patients were randomly selected and included in the study which cytopathologically diagnosed as atypical squamous cells of undetermined significance (AS-CUS) in correlation to HPV status, parakeratosis (PK), p16 immunostaining, enlarged Squamous cells nuclei (ES) and inflammatory cells infiltration (ICI). Results revealed that out of the total 121 patients, 32 cases (26%) were positive for high-risk HPV (HR-HPV), 26 cases (22%) were positive for low-risk HPV (LR-HPV) and 63 (52%) were negative for HPV. HPV infections were significantly associated with age groups (p<0.026), PK (p = 0.043), p16 (p = 0.001), ES (p = 0.002) and ICI (p = 0.049). The positive immunostaining expression of p16 was only noticed in two HR-HPV patients. ES cells were found in 9.5% of HPV-negative cases, 27% of LR-HPV cases and 40.5% of HR-HPV cases. High PK cell positivity was seen only in HR-HPV. High ICI scores were seen in 40.6% of HR-HPV patients, 26.9 % of LR-HPV and 17.4 % of negative HPV patients. It was concluded that high PK positivity, high ICI score, positive p16 immunostaining and ES were correlated with HR-HPV in non-menopausal women. These findings could provide potential diagnostic clues for HPV-harboring senescent cells as a strategy for reducing HPV risk of cervical cancer development and identifying the cell of tumor origin, which could be beneficial for improving the utility of senolytic agents and immunotherapy in clinical practice.

Bifocal endometriomas involving a Pfannenstiel incision. Clinical case

Endometrioma of the anterior abdominal wall (EAAW) is a rather rare variant of extragenital endometriosis, which in most cases occurs after obstetrical and gynecological procedures. EAAW presented predominantly as a single tumour‑like mass, and multiple ectopias were observed in only 1.9—5.6% of cases, exclusively after Pfannenstiel laparotomy. Here we present a clinical case of a 37‑year‑old patient who complained of the large tumour‑like nodules along the postoperative anterior abdominal wall scar, accompanied by severe cyclic, catamenial pain. Additionally, the patient noted an increase in tumour size during menstruation. Thirty‑three months ago, she underwent an elected cesarean section for obstetric indications. Based on ultrasonography and computed tomography scans, the presence of two EAAW in the corners of the postoperative scar was established: 46 × 32 × 31 mm and 14 × 18 × 13 mm, respectively. Both lesions were excised out without damaging their integrity. The fascial defect was replaced by synthetic polypropylene mesh. The diagnosis of EAAW was finally confirmed based on pathological (presence of endometrial glands and cytogenic stroma) and immunohistochemical (positive membrane expression of CD10 in cytogenic stroma, intense cytoplasmic expression of CK7 in endometrial glands, marked nuclear expression of progesterone (PR) and estrogen (ER‑α) receptors in endometrial glands and cytogenic stroma, proliferative activity index Ki‑67 — 2%) studies. At a follow‑up after 19 months, the patient was asymptomatic; according to physical examination and ultrasound scan, there was no evidence of recurrence. Abdominal wall endometriosis is a rare condition. Clinicians should be aware of this pathology, especially in women presenting with a painful mass near the scar of a previous obstetrical and gynecological surgery. Surgery is the best treatment modality for endometrioma, whereas its optimal volume is considered to be R0 resection with preservation of endometriomas’ integrity. The final diagnosis of EAAW requires pathological and immunohistochemical confirmation.

Prognostic Significance of Programmed Cell Death-Ligand 1 Expression in Tobacco Associated Oral Squamous Cell Carcinoma: An Immunohistochemical Based Cross-Sectional Study.

Advancements in immuno-oncology have dramatically transformed cancer treatment. Immunotherapy, targeting immune check point proteins, notably Programmed cell death ligand 1 (PD-L1) and its receptor Programmed Cell Death-1 (PD-1) which modulate the activity of immune response in Head and Neck squamous cell carcinomas (HNSCC), is an area of much research. The immunohistochemical expression of PD-L1 in cancer cells has been proposed as a predictive biomarker for selecting candidates for immunotherapy. Thus, the present study was undertaken to study the expression of PD-L1 in the primary tumour cells and evaluate its correlation with various clinicopathological parameters and prognosis in tobacco associated oral squamous cell carcinomas (OSCC). Expression of PD-L1 was investigated in 75 surgically resected cases of OSCC by immunohistochemistry and its association with different clinicopathological features and prognosis was analysed. PD-L1 protein was detected in 68% (51 cases) of cases. Tumour stage (p = 0.04), lymph node (LN) metastasis (p < 0.01) and moderate to marked tumour infiltrating lymphocytes (TILs) (p < 0.05), significantly correlated with the PD-L1 expression in the primary tumour. PD-L1 expression did not show a significant association with overall survival (OS) rate, however, patients with positive PD-L1 expression showed a poorer survival rate. Patients exhibiting nodal positivity had the worst prognosis (p < 0.005). These data demonstrated a significant association of ≥ 5% PD-L1 expression in the primary tumour and the presence of LN metastasis, moderate to marked TILs and advancing tumour stage, thus, making it a plausible immunotherapeutic target molecule in OSCC patients.

CyCadas: accelerating interactive annotation and analysis of clustered cytometry data.

Single cell profiling by cytometry has emerged as a key technology in biology, immunology and clinical-translational medicine. The correct annotation, which refers to the identification of clusters as specific cell populations based on their marker expression, of clustered high-dimensional cytometry data, is a critical step of the analysis. Its accuracy determines the correct interpretation of the biological data. Despite the progress in various clustering algorithms, the annotation of clustered data still remains a manual, time consuming and error-prone task. We developed a user-friendly cluster annotation and differential abundance detection tool that can be applied on data generated with Self Organizing Map clustering algorithms, thus simplifying the annotation process of datasets that consist of hundreds or thousands of clusters. We present Cytometry Cluster Annotation and Differential Abundance Suite (CyCadas), a semi-automated software tool that facilitates cluster annotation in cytometry data by offering both visual and computational guidance. CyCadas addresses the critical need for efficient and accurate annotation of high-resolution clustered cytometry data, significantly reducing the time needed to perform the analysis compared to both manual gating approaches and manual annotation of clustered data. The tool features a user-friendly interface, visual tools enabling data exploration and automated threshold estimation to separate negative and positive marker expression. It facilitates the definition and annotation of cell phenotypes among multiple clusters in a tree-based data structure. Finally, it calculates the abundance of various cell populations across the conditions with statistical interpretation. It is an ideal resource for researchers aiming to streamline their cytometry workflow. CyCadas is available as open source at: https://github.com/DII-LIH-Luxembourg/cycadas.