Detecting microsatellite instability (MSI) plays a key role in the management of endometrial cancer (EC), as it is a critical predictive biomarker for Lynch syndrome or immunotherapy response. A pressing need exists for cost-efficient, broadly accessible tools to aid patient for universal testing. Herein, we investigate the value of ZOOMit diffusion kurtosis imaging (DKI) and diffusion weighted imaging (DWI) based on preoperative pelvic magnetic resonance imaging (MRI) images in assessing MSI in EC.

Preoperative MRI examination including ZOOMit DKI and DWI of 81 EC patients were retrospectively analyzed. The apparent diffusion coefficient (ADC), mean kurtosis (MK), mean diffusivity (MD) and the largest tumor size based on MRI images, as well as patients' clinicopathological features were compared and analyzed according to different microsatellite statuses.

Of the 81 patients, 59 (72.8%) who were microsatellite stability (MSS) and 22 (27.2%) who were MSI. Interobserver agreement for the quantitative parameter measurements was excellent (ICC 0.78-0.98). The ADC and MD values were significantly lower, while Ki-67 proliferation level and MK values were significantly higher in the MSI group compared to those of the MSS group. The parameters of MD and MK were independent predictors for determining MSI, and their combination showed better diagnostic efficacy with an area under the receiver operating characteristic curve (AUROC) of 0.860 (95% confidence interval, 0.765, 0.927), although there was no significant difference compared to each individual parameter.

The microstructural heterogeneity assessment of ZOOMit DKI allowed for characterizing MSI status in EC. Within the current universal MSI testing paradigm, DKI may provide added value as a potential noninvasive imaging biomarker for preoperative assessment of MSI tumors, thereby facilitating clinical decision-making.

Oral antibiotics damage gut epithelium while treating infections. Goji berry derives from Lycium barbarum L., which is employed as a dietary and tonic herb with the effect of promoting intestinal health. The spermidine compound lycibarbarspermidine L (LBS L) in it can promote intestinal epithelial growth. The purpose of the research is to examine the reparative effects of LBS L against antibiotic-stimulated gut epithelial damage, as well as the processes associated with T-UCR. Intestinal epithelial cells were stimulated with antibiotics to induce injury. The impact of LBS L and T-UCR-related mechanisms was evaluated through cell assays. Ultimately, a rat model of intestinal injury was developed by administering antibiotics. We found that LBS L reversed the intestinal damage caused by antibiotics through modulating uc.141A/miR195-3p. A reduction in intestinal damage was observed in rat models. In conclusion, LBS L ameliorates antibiotic-induced intestinal epithelial cell damage in rats by modulating uc.141A.

Drug delivery technology has emerged as an effective strategy to enhance the anti-tumor potential of camptothecins. In this study, we constructed an enzyme-activated targeted drug release system employing a stable valine-alanine linker in combination with a highly cytotoxic camptothecin derivative, AZ'0132. The prodrug, named HC07, was successfully delivered to the tumor site via eHSP90, where it underwent specific cleavage, thereby enhancing its therapeutic window and reducing ocular toxicity. In order to further improve its metabolic properties, we innovatively combined HC07 with plasma endogenous albumin to construct albumin-binding prodrugs HC08 and HC09 for the first time. These prodrugs demonstrated prolonged circulation times in vivo and significantly improved tumor selectivity and anti-tumor efficacy. This work presents a promising eHSP90-based long-circulating drug delivery system, providing new avenues for the design of future anti-cancer drugs.

As a novel clinical therapeutic technique, micro-focused ultrasound (MFU) has garnered significant attention in the field of skin rejuvenation in recent years. Although clinical studies have demonstrated the marked efficacy of MFU in improving skin laxity, the specific histological and molecular mechanisms of its rejuvenation effects on the skin remain unclear.

The abdominal skin of Bama minipigs was treated with MFU, specific treatment parameters employed both the 3 mm and 4.5 mm therapeutic head with the following settings: 25 Hz/1 min/6.63 W, 25 Hz/2.5 min/6.63 W, 25 Hz/1 min/1.32 W, and 10 Hz/1 min/6.63 W, with adjacent areas serving as controls. The thickness of the dermis, fat layer, and superficial musculoaponeurotic system (SMAS) layer was measured by ultrasound imaging at baseline, immediately after treatment, and at 14, 30, 90 days posttreatment. The histopathological, immunohistochemical, and transcriptomic changes of each time point were studied, to compare protein content and gene expression of matrix metalloproteinase (MMP) enzymes, transforming growth factor β (TGF-β), epidermal growth factor (EGF), Ki67, type I collagen, and elastin.

MFU induced dermal thickening, which was sustained up to 90 days. Immediately posttreatment (0 days), the parameters 4.5 mm, 10 Hz, 1 min, 6.63 W showed an average thickening of 0.16 mm, with a growth rate of 14.48 ± 3.64%, which was significantly higher than the 0% growth rate observed in the control group (p < 0.05). With the parameters of 3.0 mm, 25 Hz, 1 min, 1.32 W, the dermis thickened by an average of 0.69 mm at 90 days posttreatment, with a growth rate of 39.33 ± 14.34%. In contrast, the control group showed an increase of about 0.18 mm, with a growth rate of 0.92 ± 13.25% (p < 0.05). A prolonged treatment duration (2.5 min) and higher power levels (6.63 W) did not exhibit statistically different effects on the promotion of dermal thickness in this study. MFU also promoted collagen production in the SMAS layer in the long term. After treatment with parameters 3.0 mm, 25 Hz, 1 min, 1.32 W, the SMAS thickened by 0.12 mm at 30 days, while the control group showed an increase of 0.03 mm (p < 0.05). At 90 days, the SMAS thickened by approximately 0.17 mm, whereas the control group decreased by 0.04 mm (p < 0.05). Following treatment with parameters 3.0 mm, 10 Hz, 1 min, 6.63 W, the SMAS thickened by 0.11 mm at 30 days, with the control group showing an increase of 0.06 mm (p < 0.05). Immunofluorescence staining indicated a significant increase in type I and III collagen, MMP1, MMP3, TGF-β and Ki67 at 14 and 30 days after MFU treatment. The gene expression of MMP family members, TGF-β, EGF, type I collagen, and elastin fibers was markedly upregulated at 14, 30, and 90 days.

MFU treatment can stimulate the proliferation of collagen fibers in the dermis. It can also promote the proliferation of collagen in the SMAS layer. However, the effect of MFU treatment on fat is minimal due to limited energy and depth penetration.

Crohn's Disease (CD), an inflammatory bowel disorder, is influenced by genetic, immune, and environmental factors. The present study highlights the pioneering role of circular RNAs (circRNAs) in the etiology of CD, with a specific focus on hsa_circ_0005255 and its regulatory role. Utilizing both bioinformatic and experimental approaches, we exposed the mechanistic and therapeutic significance of hsa_circ_0005255 within the pathophysiological framework of CD. Our findings revealed a significant underexpression of hsa_circ_0005255 in tissue samples from CD patients and in DSS-induced CD mouse models. The overexpression of hsa_circ_0005255 markedly mitigated inflammatory responses, as indicated by decreased serum levels of tumor necrosis factor-alpha, interleukin-1 beta, and interleukin-6, and reduced histopathological indications of inflammation in colonic tissues. It substantially improved the integrity of the epithelial barrier, evidenced by the upregulation of Zonula Occludens-1 expression and the reduction of apoptosis in colonic epithelial cells. Furthermore, this regulatory effect extended to the enhancement of epithelial cell proliferation and autophagy, characterized by the elevated expression of Ki-67, microtubule-associated protein 1A/1B-light chain 3 II, and Beclin-1, along with the suppression of cleaved caspase-3 and sequestosome 1. Mechanistically, hsa_circ_0005255 functioned as a competitive endogenous RNA, absorbing miR-23a-3p and thereby regulating Nuclear Receptor Coactivator 3. This investigation not only broadens our understanding of the involvement of circRNAs in CD pathogenesis but also identifies hsa_circ_0005255 as a potent biomarker and therapeutic target.

Granular cell tumours (GCTs) are rare and typically benign neoplasms that can arise in various parts of the body, including the oesophagus. These tumours are considered to originate from Schwann cells and are characterized by their granular cytoplasm. Oesophageal GCTs (EGCTs) are particularly uncommon and often present with non-specific symptoms, making their diagnosis challenging. The present case report documents the case of a 68-year-old male patient who was admitted to the Affiliated Hospital of Jiujiang University (Jiujiang, China) after the incidental discovery of a submucosal oesophageal mass during a routine health examination. Upon diagnostic evaluation, including endoscopy and histopathological analysis, an EGCT was identified. Endoscopic ultrasonography revealed submucosal protrusion of the lower oesophagus, where endoscopic submucosal dissection (ESD) was subsequently performed. Histopathological staining and immunohistochemistry (IHC) indicated the presence of a GCT with positivity for S-100. The patient successfully underwent ESD without any complications. Although EGCTs are typically benign, they have the potential for malignant transformation and therefore require careful evaluation. The present case highlights the importance of differentiating EGCTs from other submucosal oesophageal lesions through histological and IHC methods. In particular, early detection and appropriate management are crucial for optimal patient outcomes. In conclusion, EGCT is a rare entity with a generally favourable prognosis when detected early. The present case underscores the importance of considering EGCTs in the differential diagnosis of oesophageal submucosal lesions and outlines the role of endoscopic and histopathological evaluation in their management.

Oral potentially malignant disorders may precede the development of oral cancer, and biomarkers are being investigated for their risk assessment. We aim to provide updated information on tissue biomarkers related to the risk of malignant transformation (MT) in patients with oral leukoplakia (OL) published during the last four years.

A search strategy was developed using the electronic databases PubMed, EBSCO, and Cochrane Library to search for articles related to this topic (published from 2020 to 2024) (CRD42024527395).

Of the 1385 articles identified, 14 were included, with a combined sample of 1479 patients of whom 216 suffered MT. There were 30 different biomarkers evaluated of which Bmi-1, CD3/CD8, Ki-67, p16, and DNA ploidy were evaluated in more than one study. Of the 9 studies with multivariate analysis, Bmi-1, Tipe-2, copy number alteration, DcR2, and Ki-67, PTHrP, podoplanin (PDPN), and BubR1/Mad2 biomarkers presented independent significant value on MT. In one study, adding DNA ploidy status to the grade of dysplasia increased the model's predictive power.

Although heterogeneity continues to exist, this systematic review reconfirms the role of biomarkers previously described such as PDPN and DNA ploidy and some new additional biomarkers.

Vitamin A (VitA) is an essential nutrient, affecting many cell functions, such as proliferation, apoptosis, and differentiation, all of which are important for the regeneration of various tissues. In this study, we investigated the effects of a VitA-enriched diet on the regeneration of the urothelium of the urinary bladder in mice after cyclophosphamide (CP)-induced injury. Female mice were fed VitA-enriched and normal diet for 1 week before receiving an intraperitoneal injection of CP (150 mg/kg). Urinary bladders were removed 1 and 3 days after CP. On Day 1, RNA sequencing showed that VitA upregulated two Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathways: the cell cycle and the PI3K-Akt pathway. This was confirmed by qPCR, which showed significantly increased expression of the Itga3 and Areg genes. In addition, the effect of VitA on the proliferation of urothelial cells was analyzed by immunohistochemistry of Ki-67, which confirmed an increased proliferation rate. No significant effects of the VitA-enriched diet were observed on the expression of apoptosis-related genes and on differentiation-related markers of superficial urothelial cells. Our results suggest that a VitA-enriched diet improves early urothelial regeneration after CP-induced injury by promoting cell proliferation.

Rheumatoid arthritis (RA) is a systemic autoimmune disease. Circular RNA_0001715 (circ_0001715) has been demonstrated to be involved in the progression of cancers. This study aimed to discuss the function of circ_0001715 on the development of RA. Tumor necrosis factor-α (TNF-α) was used to active human RA fibroblast-like synoviocytes (FLS) (HFLS-RA) cells. The role of circ_0001715 in the progression of RA was determined by cell counting kit-8, flow cytometry, enzyme-linked immunosorbent assay. The target of circ_0001715 was predicted using the circinteractome database and validated by the luciferase reporter assay. The relative protein expression of toll-like receptor (TLR)-4/nuclear factor-kappa B (NF-κB) axis was detected by western blot. Moreover, a collagen-induced arthritis (CIA) mouse model was constructed through the secondary immunization. The role of circ_0001715 in vivo was determined by hematoxylin and eosin (H&E), Safranin O, TRAP, ELISA and western blot. Increased levels of circ_0001715 were discovered in tissues from RA patients, TNF-α-induced HFLS-RA cells, and synovial tissues of CIA-induced mice. Knockdown of circ_0001715 decreased proliferation and inflammation, but promoted apoptosis of RA both in vitro and in vivo. Additionally, miR-326 was predicted as the target of circ_0001715, which was confirmed by the luciferase reporter assay. Knockdown of miR-326 reversed the results of proliferation, apoptosis and inflammation resulted from the circ_0001715 knockdown. Mechanically, knockdown of circ_0001715 reduced the expression of TLR-4/NF-κB axis, which were rescued by the downregulation of miR-326. Circ_0001715 sequestered miR-326 to regulate the growth, apoptosis and inflammation of HFLS-RA cells via TLR-4/NF-κB axis.

Leukemia is the most common type of serious, life-threatening cancer that requires the immediate initiation of therapy. Ascorbic acid (AsA), commonly known as Vitamin C, has been gaining attention due to its antioxidant activity as a potential treatment for human malignancies. In this study, the THP-1 monocytic cell line was treated with two doses of AsA: a low dose (L-AsA, 2.5 µg/mL) and a high dose (H-AsA, 5 µg/mL), while the K562 lymphocytic cell line was treated with two doses of AsA: a low dose (L-AsA, 4 µg/mL) and a high dose (H-AsA, 8 µg/mL). After a 24-h incubation period, all cells were exposed to different doses of X-radiation (2, 4, 8 Gy). The viability of THP-1 and K562 treated by AsA was assessed using the MTT assay. Additionally, we evaluated apoptosis, autophagy, proliferation, cell cycle progression, hypoxia-inducible factor (HIF-1), malondialdehyde (MDA), and total antioxidant capacity (TAC). Our study demonstrated that AsA, in combination with X-radiation, induced significant apoptosis and notably reduced Ki67 levels in human leukemia THP-1 cells. Furthermore, X-radiation caused DNA damage, leading to cell cycle arrest at the G0/G1 phase in THP-1 cells. Moreover, AsA significantly reduced HIF-1 levels, which are essential for the survival of tumor cells in hypoxic conditions. We also found that the administration of AsA in combination with X-radiation had a synergistic and dose-dependent effect on THP-1 and K562 cells. Notably, the combination of L-AsA with 2 Gy X-radiation showed a more pronounced effect than 8 Gy X-radiation alone. These results suggest that AsA has promising anti-proliferative, pro-apoptotic, and autophagic effects on leukemic cells. Furthermore, the dose of X-radiation may be reduced when combined with AsA in an effort to minimize its potential side effects.

Muscle satellite (stem) cells (MSCs) reside in skeletal muscle and are essential for myogenesis. Thus, MSCs are widely used in cultured meat research. This study aimed to identify substances that promote MSC proliferation and differentiation while maintaining their intrinsic properties, with the long-term goal of replacing fetal bovine serum (FBS) for bovine MSC cultivation. Therefore, this study evaluated the effects of six growth factors (TGF-β, HGF, PDGF, insulin, IGF-1, and EGF) and the cytokine IL-2 on bovine MSCs. Each factor was applied during the proliferation and differentiation of MSCs, and the proliferation rate, differentiation rate, and expression of relevant mRNA and proteins were analyzed. Insulin most effectively promoted MSC proliferation and differentiation. Specifically, insulin increased cell proliferation rates, proliferation markers Ki67 and PCNA expressions, and markers of differentiation, such as myotube formation and creatine kinase activity, alongside the expressions of MYOD, MYOG, and MYH. Furthermore, insulin suppressed low FBS-induced reductions in proliferation and differentiation markers. This study suggests insulin can promote MSC proliferation and differentiation and reduce FBS usage. Thus, this study provides a potential means of cultivating MSCs on a large scale for cultured meat production.

Tendon ruptures are common musculoskeletal injuries associated with prolonged healing and complications such as adhesion formation and rerupture. Despite advancements in treatment strategies, full functional recovery remains a challenge. Growth factors (GFs) like insulin-like growth factor-1 (IGF-1) and platelet-derived growth factor-BB (PDGF-BB) play key roles in tendon repair and may have synergistic effects when applied together. To support tendon healing, a bioactive electrospun polymer scaffold made of Degrapol® (DP) was developed, incorporating IGF-1, PDGF-BB, or both. A range of in vitro and in vivo analyses were performed to assess scaffold structure, cell behavior, gene expression, metabolism, and biomechanical and adhesion outcomes three weeks post-surgery. Interestingly, the combined application of IGF-1 and PDGF-BB did not simply amplify individual effects but showed a complex interaction. Depending on the parameter and time point, the combination led to either enhanced or reduced responses compared to single-factor treatments, indicating a synergistic modulation rather than a purely additive effect. These findings suggest that the combination of IGF-1 and PDGF-BB can modulate key cellular and molecular processes in tendon regeneration, making this approach a promising strategy to improve tendon healing.

BackgroundThe involvement of microglia is likely to be pivotal in the pathogenesis of Alzheimer's disease (AD) by modulating the deposition of amyloid-β (Aβ) plaques. The deletion of Dedicator of cytokinesis 8 (DOCK8) has a protective effect in mouse with neurodegenerative diseases.ObjectiveTo explore the underlying mechanism of DOCK8 in AD.MethodsIn present study, we first the detected the expression of DOCK8 in the hippocampal tissue of APP/PS1 mice. Then, the expression of DOCK8 was knocked down in the hippocampal tissue of APP/PS1 mice, and the effects of DOCK8 down-regulation on cognitive function, the microglia migration around Aβ plaques, and the cell division cycle 42 (Cdc42)/p38 mitogen-activated protein kinase (MAPK) signaling pathway were detected. Next, the effects of DOCK8 knockdown on Aβ-induced migration and activation of BV-2 cells as well as the MAPK signaling pathway were detected. Finally, the transcriptional regulation of DOCK by transcription factor 3 (ATF3) was detected by a dual luciferase reporter assay.ResultsDOCK8 expression exerts a significant upregulation in the hippocampus of APP/PS1 mice. However, following the DOCK8 knockdown, there was a significant recovery in the results of the behavioral tests and a notable reduction in microglial expression. Moreover, the high expression of DOCK8 mediated by ATF3 successfully triggered the Cdc42/p38 MAPK signaling pathway, thereby enhancing the migration and recruitment of microglia towards senile plaques, accelerating the production of Aβ plaques.ConclusionsATF3-mediated high expression of DOCK8 accelerates the production of Aβ plaques, and participates in the pathogenesis of AD.

This study aimed to develop an interpretable machine learning model that accurately predicts Ki-67 expression in breast cancer (BC) patients using a combination of dynamic-contrast enhanced magnetic resonance imaging (DCE-MRI) radiomics and clinical-imaging features. A total of 195 BC patients, including 201 lesions, were enrolled retrospectively. These lesions were randomized into training and testing set (7:3). The correlation between clinical-imaging features and Ki-67 expression was analyzed via univariate analysis and binary logistic regression, leading to the development of a Clinical-imaging model. Radiomics features were extracted based on the early and delayed phases of DCE-MRI. These features were screened by Pearson correlation coefficient and recursive feature elimination (RFE). The logistic regression classifier was used to develop the Radiomics model. The clinical imaging and radiomics features were combined to form a Combined model. The Shapley Additive Explanation (SHAP) algorithm was employed to explain the optimal model, and the AUC was used to assess the model's performance. Ki-67 expression was markedly different from the internal enhancement pattern and necrosis among the imaging features. Compared to the Clinical-imaging model (AUC = 0.682), the AUCs of the Radiomics and the Combined models in the training set were 0.797 and 0.821, respectively. Clinical-imaging, Radiomics, and Combined models had AUCs of 0.666, 0.796, and 0.802 in the test set. Based on the IDI results, the combined model outperformed the Clinical-imaging and Radiomics models in the training set by 11.8% and 2.1%, respectively. They increased by 11% and 1.74% in the test set. SHAP analysis showed that ph2-original-shape-surface volume ratio was the most important feature of the model. Based on clinical-imaging features and DCE-MRI radiomics, the interpretable machine learning model can accurately predict the expression of Ki-67 in BC. Combining the SHAP algorithm with the model improves its interpretability, which may assist clinicians in formulating more accurate treatment strategies.

The purpose of this research is to evaluate the predictive capabilities of a radiomics model for the Ki67 index and its correlation with prognosis in advanced gastric tubular adenocarcinoma patients.

Clinical data from 213 patients were analyzed, categorizing patients into high and low Ki67 index groups. The radiomic features of 192 patients were selected by lasso method and the model was constructed, which was validated using the TCIA dataset. Univariate and multivariate Cox regression analyses were used to further analyze clinical features associated with the prognosis of gastric cancer, radiomic models are also used to assess patient outcomes.

The radiomics model demonstrated moderate accuracy, with AUC values of 0.634, 0.666, and 0.602 for the training, validation 1, and validation 2 sets, respectively. Additionally, a significant correlation was found between the Ki67 index and radiomics scores, a higher Ki67 index was associated with improved outcomes. Kaplan-Meier analysis showed distinct survival differences between patients with high and low radiomics scores, indicating that higher scores predict better prognosis.

The radiomics model accurately predicts the Ki67 index and correlates with prognosis in advanced gastric tubular adenocarcinoma, offering valuable insights for clinical decision-making and personalized treatment strategies.

Gastric schwannoma (GS) is often misdiagnosed as gastrointestinal stromal tumors due to the high incidence of the latter. However, these two types differ significantly in pathology and biological behavior.

To evaluate the computed tomography characteristics of GS and provide insights into its accurate diagnosis.

Twenty-three cases of GS confirmed between January 2011 and December 2023 were assessed clinically and radiologically. Imaging characteristics, including tumor location, size, contour, ulceration, growth pattern, enhancement degree and pattern, cystic change, calcification, and perigastric lymph nodes (PLNs), were reviewed by two experienced radiologists.

Our sample included 18 females and 5 males, with a median age of 54.7 years. A total of 39.1% of cases were asymptomatic. GSs appeared as oval and well-defined submucosal tumors, with exophytic (43.5%) or mixed (endoluminal + exophytic; 43.5%) growth patterns. The tumors were primarily located in the gastric body (78.3%). Ulcerations were observed in 8 cases (34.5%), and PLNs were observed in 15 cases (65%). The average degree of enhancement was 48.3 Hounsfield units. Twenty cases (87%) showed peak enhancement in the delayed phase. Most GSs were homogeneous, while cystic change (13.0%) and calcification (17.4%) were rare.

GS predominantly showed gradual homogenous enhancement with peak enhancement in the delayed phase. PLNs around GS are helpful in differentiating GS from other gastric submucosal tumors.

Intravascular large B-cell lymphoma (IVLBCL) is a rare and aggressive lymphoma that can have heterogeneous central nervous system involvement and cerebrovascular complications. The development of IVLBCL can be fatal. Although relatively rare, the development of specific clinical syndromes, such as IVLBCL, has been implicated following infection and vaccination. To our knowledge, this is the first comprehensive neuropsychological evaluation assessing neurocognitive and psychological status after IVLBCL diagnosis.

The current study presents a right-handed mid-60-year-old male with a university-level education, who was diagnosed with IVLBCL following viral vector SARS-CoV-2 vaccination. He presented with a complex medical history including antiphospholipid syndrome, deafness (prior to cochlear implant), and cardiovascular complications secondary to lymphoma. Brain magnetic resonance imaging showed parietal, frontal, and cerebellar infarcts; encephalomalacia; and periventricular deep chronic ischemic changes. A comprehensive neuropsychological evaluation was completed.

In consideration of an individual with an estimated above-average baseline, his neurocognitive profile demonstrated impairments across multiple domains that were more lateralized to the non-dominant hemisphere including visual attention, visual processing speed, visuo-construction, memory, motor dexterity, and right-sided ataxia (e.g., dysmetria). Clinical elevations for depression, hopelessness, and anxiety were also found.

The current study highlights a novel cognitive profile of IVLBCL and comorbidities with the patient having more predominant nondominant hemispheric-related deficits. There was evidence of disruption to visual processing networks, largely consistent with neuroimaging lesions. The current case also describes the unique experience of an individual coping with a rare condition, especially when resulting in functional decline (e.g., loss of audition). Implications are discussed.

The methylation status of the MGMT gene promoter is recognized as a key predictive biomarker for glioblastoma patients, influencing treatment decisions and outcomes. Emerging evidence suggests that enhancer methylation may also play a role in gene regulation and is associated with various clinical parameters, genetic variants, and demographic factors. This study aimed to assess DNA methylation levels in intergenic and intragenic MGMT enhancers to investigate their relationship with MGMT promoter methylation, MGMT protein expression, and clinical and demographic characteristics in glioblastoma. We developed 18 pyrosequencing assays to analyze 54 CpGs, including 34 in intergenic and 20 in intragenic enhancers. The assays were applied to tumor cells derived from 38 glioma patients. Intragenic enhancer CpGs showed significantly higher methylation than intergenic enhancer CpGs. Intragenic enhancer methylation showed a strong negative correlation with MGMT promoter methylation. For several CpGs in intragenic enhancers, an inverse L-shaped relationship between methylation levels and MGMT expression was observed. We identified distinct associations between enhancer methylation and clinical and demographic parameters. Intergenic enhancer methylation was primarily linked to the TERT SNP rs2853669 genotype, Ki-67 expression, age, and sex, whereas intragenic enhancer methylation was associated with MGMT promoter methylation, MGMT expression, overall survival, and progression-free survival. Further studies with larger patient cohorts are needed to validate the clinical relevance of intergenic and intragenic MGMT enhancer methylation in glioblastoma.

Ki-67 scoring is of essential importance in the evaluation of breast cancer. We evaluated a Ki-67 algorithm as a decision support tool to improve accuracy for pathology residents. We retrospectively evaluated Ki-67 scores on whole slide images (WSI) obtained from 156 consecutive breast cancer patients. Two senior pathologists determined the 2.1 mm2 hotspot to be evaluated. Ki-67 scores from senior pathologists were compared with results generated by the algorithm, results from 10 pathology residents, and results from pathology residents with the assistance of the algorithm. In addition to numerical results from the algorithm, residents were also presented with a visual representation of nuclei that were counted and excluded. Statistical analysis was performed using Wilcoxon and intra-class correlation (ICC) tests. The mean Ki-67 scores from senior pathologists and the algorithm were 23 ± 18 and 24 ± 18, respectively (ICC, 0.98). Ki-67 scores from the residents were 19 ± 16 and 22 ± 16, without and with input from the algorithm, respectively. With input from the algorithm, residents' scores were significantly closer to those obtained by senior pathologists (p = 0.008). Residents modified their scores in 53.8% of the cases where 74% of the better scores were characterized by an increase in the original scores. The results obtained by the Ki-67 algorithm were highly correlated with those assessed by senior pathologists. We demonstrated that the algorithm may serve as a decision support tool for residents to align their results with those of senior pathologists.

Breast cancer is the most common cancer and a leading cause of cancer-related death among women globally. Determining which patients will benefit from chemotherapy remains challenging. Proliferative markers such as Ki-67, mini chromosome maintenance (MCM) proteins, and proliferating cell nuclear antigen (PCNA) offer valuable insights into tumor growth and treatment response. This review evaluates their clinical roles, with a focus on chemotherapy implications and emerging digital pathology techniques for marker quantification.

A narrative review was conducted by searching PubMed, Scopus, and Google Scholar for studies related to Ki-67, MCM, PCNA, breast cancer, and chemotherapy. Studies were thematically categorized into five areas. A bibliometric analysis of publications from 2000 to April 2023 was performed using the Bibliometrix R package and VOSviewer to assess research trends and thematic evolution.

Eighty studies were included in the narrative synthesis. Ki-67 is the most commonly used marker, particularly useful in predicting response to neoadjuvant chemotherapy (NAC). MCM proteins show promise for identifying proliferative potential across tumor grades, while PCNA is associated with aggressive tumor features and poor prognosis. Post-chemotherapy changes in Ki-67 levels are linked to survival outcomes. Bibliometric analysis revealed a shift in research focus from basic mechanisms to clinical applications and digital quantification.

Proliferative markers play an essential role in breast cancer management. Ki-67 remains a key predictor of chemotherapy response, while MCM and PCNA offer complementary prognostic insights. Integration of these markers with digital pathology and AI-driven tools may enhance diagnostic accuracy and personalized treatment strategies. Standardization of assessment methods is crucial for broader clinical application.

Sophisticated refinements in histopathology are evolving to improve meningioma outcome prediction. The aim of this study is to evaluate the stand-alone performance of Ki-67 and progesterone receptor (PR) algorithm scores in meningiomas and their power in predicting recurrence and disease-free survival of the patients. Whole slide images of Ki-67 and PR-stained slides from 404 meningioma cases were analyzed by a digital image viewer and analysis software Virapath-2.1, which analyzes the tumor cells by size, color, and shape. Ki-67 scores were calculated in the hotspot region that contains at least 1000 tumor cells, while PR was calculated on the whole slide. The results were compared with WHO grade, tumor recurrence and disease-free survival (DFS). Mean Ki-67 scores were 4.2 ± 3.5, 12.1 ± 10.6 and 22. ± 8.5 for grade 1, 2 and 3 tumors (p < 0.05), while PR scores were 49 ± 35, 43 ± 34 and 16 ± 30, respectively (p > 0.05). Median survival of patients based on Ki-67 values ≤13.2 % and > 13.2 % was 122 versus 60 months (p = 0.004). Prediction of recurrence based on Ki-67 score was found to have acceptable discrimination (AUC = 0.74). PR expression was not found to correlate with DFS, but recurrent tumors had lower PR scores than non-recurrent tumors (31.3 ± 33.8 vs. 49.0 ± 33.0; p = 0.03). Elevated Ki-67 levels identified by the algorithm may classify meningioma patients at high recurrence risk and inform clinical management. Although PR scores did not correlate with DFS, lower expression in recurrent tumors suggests a role in recurrence risk assessment. Larger prospective studies are needed for routine clinical practice.

Cancer of unknown primary (CUP) is a diagnosis that the primary lesion cannot be confirmed by a series of imaging, endoscopic and pathological examinations. The present study aimed to assess the clinical characteristics and survival outcomes of patients with CUP. The present retrospective observational study included patients diagnosed with malignancies confirmed as CUP using histopathology at the Oncology Department of the Fourth Hospital of Hebei Medical University (Shijiazhuang, China) from January 2009 to January 2021. Clinical and pathological data, genetic testing results, treatment modalities and median overall survival (OS) were analyzed. A total of 107 patients were included, with a mean age of 56.59 years. The median follow-up period was 48.8 months. Adenocarcinoma was the most common pathological type (38.3%), followed by squamous cell carcinoma (31.8%) and neuroendocrine carcinoma (16.8%). The median OS was 28.4 months, with 1-, 2-, 3- and 4-year OS rates of 68.2, 54.1, 48.4 and 42.3%, respectively. Imaging revealed that 31 patients (29%) had visceral metastases, and these patients had a significantly shorter median OS compared with those without visceral metastases (8.9 vs. 69 months; P=0.001). Patients who received local treatment (n=31; 29%) had significantly longer survival times than those who did not (69 vs. 17.9 months; P=0.009). Of the 107 patients, 101 (94.4%) received systemic treatment. The median OS times for different treatment groups were as follows: Chemotherapy alone, 28.4 months; chemotherapy combined with immune checkpoint inhibitors, anti-angiogenic agents or targeted therapy, not reached; no chemotherapy, 8.0 months; and untreated, 9.4 months, with significant differences observed among the groups (P=0.008). The survival outcomes of patients with CUP varied based on the presence of visceral metastasis and the treatment modalities employed. Systemic treatments, particularly those incorporating targeted therapy, appear to have the potential to improve prognosis.

The reciprocal connection between cell migration and proliferation relies on the intertwined contributions from substrate-associated and intercellular cues in the microenvironment. However, how cells perceive the substrates, make contact with their neighbors, and switch phenotypes under different trade-off conditions are still not fully understood. Here, we designed a distinct heterogeneous electric surface potential gradient of piezoelectric biomaterials to decouple the density-dependent migration/proliferation dichotomy. We found that the surface potential gradient accelerated both individual and collective cell migration but reduced proliferation through G0/G1 cell cycle arrest via the integrin/cytoskeleton signaling axis in low density. Interestingly, the initial cell density encodes the proliferative potential independent of the substrate feature. While in high density, the surface potential gradient ceased cell proliferation mainly via the E-cadherin/β-catenin signaling axis. Taken together, these results shed light on the underlying mechanism of the intertwined contributions of cell-material and cell-cell cross-links on migration and proliferation and also provide a new paradigm of materiobiology.

Pancreatic cancer is an aggressive malignancy with poor prognosis and limited treatment options. Chemotherapy remains a primary therapeutic approach, but patient responses vary significantly, emphasizing the need for reliable biomarkers. This review explores the potential role of proliferative markers, including Ki-67, PCNA, Cyclin D1, and PHH3, as predictive and prognostic indicators in pancreatic cancer management, aiming to enhance personalized treatment strategies.

We conducted a narrative review by searching Scopus, PubMed, and Google Scholar for studies focusing on Ki-67, PCNA, Cyclin D1, and PHH3 in relation to pancreatic cancer and chemotherapy. The literature was reviewed to evaluate the role of these markers in predicting chemotherapy response, tumor progression, and overall patient survival.

The review highlights the clinical significance of these markers. Ki-67 and PCNA are associated with cell proliferation, while Cyclin D1 regulates cell cycle progression and PHH3 is linked to mitotic activity. High expression levels of these markers often correlate with increased tumor aggressiveness and poorer patient outcomes. Moreover, they show promise in predicting chemotherapy response, which can inform tailored therapeutic strategies. However, challenges remain, including standardization of detection methods and determination of optimal cutoff values.

Proliferative markers such as Ki-67, PCNA, Cyclin D1, and PHH3 hold potential as predictive and prognostic tools in pancreatic cancer management. Their integration into clinical practice could improve the accuracy of treatment decisions and enhance patient outcomes. Further research and validation are necessary to overcome existing challenges and optimize their application in personalized oncology.

Intelligent hydrogels are promising in constructing scaffolds for the controlled delivery of drugs. Here, a dual thermo- and pH-responsive hydrogel called PCG [poly ( N -isopropyl acrylamide-co-itaconic acid)/chitosan/glycerophosphate (PNI/CS/GP)] was established as the carrier of 5-fluorouracil (5-FU) for triple-negative breast cancer (TNBC) treatment. The PCG hydrogel was fabricated by blending synthesized [poly ( N -isopropyl acrylamide-co-itaconic acid), pNIAAm-co-IA, PNI] with CS in the presence of GP as a crosslinking agent. The interaction between PCG hydrogel compositions was characterized by Fourier transforms infrared, NMR spectroscopy, and scanning electron microscopy. The PCG hydrogel presented an interconnected and porous structure with similar pore size, rapid swelling/deswelling rate in response to both temperature and pH change, and biocompatibility, upon which it was proposed as a great drug carrier. 5-FU had a dual thermo- and pH-responsive controlled release behavior from the PCG hydrogel and displayed an accelerated release rate in an acidic pH environment than in a neutral pH condition. The application of 5-FU-loaded PCG hydrogel exhibited a more promoted anticancer activity than 5-FU against the growth of TNBC cells both in vitro and in vivo . The outcomes suggested that the PCG hydrogel could be an excellent platform for local drug-delivery systems in the clinical therapy of TNBC.

Aberrations in primary cilia expression and intraflagellar transport (IFT) protein function have been implicated in tumourigenesis. This study explores the relationship between the alteration of primary cilia and tumourigenesis by investigating primary cilia expression and the role of IFT20 in regulating matrix metalloproteinase 9 (MMP-9) expression in oral squamous cell carcinoma (OSCC) cell lines.

The frequency and length of primary cilia were determined in OKF6-TERT2 cells, HSC-2 cells, and HSC-3 cells using immunofluorescence. Additionally, primary cilia presence in non-proliferating OSCC cells was examined. OSCC cells were treated with either small interfering RNA (siRNA) negative control or siRNA targeting IFT20 for functional analysis. mRNA expression levels of IFT20 and MMP-9 were quantified using quantitative reverse transcription polymerase chain reaction (qRT-PCR).

Results showed that HSC-2 cells exhibit abundant primary cilia when cultured in low serum media (2% serum) for 48 h, followed by serum starvation for over 72 h. No significant changes in cilia expression were observed in HSC-3 cells compared to OKF6-TERT2 cells. Ciliated cells were found in non-proliferating HSC-2 and HSC-3 cells. OSCC cells showed longer cilia than OKF6-TERT2 cells, indicating ciliary abnormalities. Changes in ciliation and cilium length of OSCC cells were accompanied by increased expression of IFT20, an intraflagellar transport protein crucial for the primary cilia assembly. However, IFT20 knockdown did not affect MMP-9 at the mRNA level in these cells.

This study reveals the differences in primary cilia expression among OSCC cells. Furthermore, the increased abundance and elongation of primary cilia in OSCC cells are accompanied by elevated expression of IFT20. Nonetheless, IFT20 did not affect MMP-9 mRNA expression in OSCC cells.

Here, we describe SPoRTS, an open-source workflow for high-throughput spatiotemporal image analysis of fluorescence-based ratiometric biosensors in living spheroids. To achieve this, we have implemented a fully automated algorithm for the acquisition of line intensity profile data, ultimately enabling semi-quantitative measurement of biosensor activity as a function of distance from the center of the spheroid. We demonstrate the functionality of SPoRTS via spatial analysis of live spheroids expressing a ratiometric biosensor based on the fluorescent, ubiquitin-based cell-cycle indicator (FUCCI) system, which identifies mitotic cells. We compare this FUCCI-based SPoRTS analysis with spatially quantified immunostaining for proliferation markers, finding that the results are strongly correlated.

Brain-machine interface performance can be affected by neuroinflammatory responses due to blood-brain barrier (BBB) damage following intracortical microelectrode implantation. Recent findings suggest that certain gut bacterial constituents might enter the brain through damaged BBB. Therefore, we hypothesized that damage to the BBB caused by microelectrode implantation could facilitate microbiome entry into the brain. In our study, we found bacterial sequences, including gut-related ones, in the brains of mice with implanted microelectrodes. These sequences changed over time. Mice treated with antibiotics showed a reduced presence of these bacteria and had a different inflammatory response, which temporarily improved microelectrode recording performance. However, long-term antibiotic use worsened performance and disrupted neurodegenerative pathways. Many bacterial sequences found were not present in the gut or in unimplanted brains. Together, the current study established a paradigm-shifting mechanism that may contribute to chronic intracortical microelectrode recording performance and affect overall brain health following intracortical microelectrode implantation.

The increasing number of type 2 diabetes mellitus (T2DM) patients leads to higher rates of morbidity and mortality related to lung cancer.

To investigate the utility of the proliferating cell nuclear antigen Ki-67 in patients with lung adenocarcinoma in situ (AIS) complicated by T2DM.

One hundred patients with AIS and T2DM (group A), 100 patients with AIS alone (group B), and 60 patients with benign lung lesions (group C) admitted to the Department of Thoracic Surgery and Endocrinology of the First Affiliated Hospital of Soochow University from November 2021 to December 2022 were enrolled. Ki-67 expression was compared among the groups.

Group A had significantly higher levels of fasting plasma glucose (FPG), total cholesterol (TC), total triglyceride, low-density lipoprotein cholesterol, glycosylated hemoglobin (HbA1c), and insulin than groups B and C (P < 0.01). Meanwhile, group B had higher insulin levels than group C (P < 0.01). Group A exhibited a significantly higher average Ki-67 positivity rate than group B (P < 0.01). The Ki-67 positivity rate in group A was 86.87%, while the positivity rate in group B was 77%. Ki-67 was positively correlated with FPG (P < 0.01) and HbA1c levels (P < 0.01). Ki-67, FBG, insulin, HbA1c, high-density lipoprotein cholesterol and TC were independent factors for patients with AIS complicated by T2DM.

Ki-67 expression was higher in patients with AIS complicated by T2DM than in patients with AIS alone. Therefore, detecting the Ki-67 level might assist in the diagnosis of AIS in patients with T2DM.

Respiratory epithelial cells can survive direct infection by influenza viruses, and the long-term consequences of that infection have been characterized in a subset of proximal airway cell types. The impact on the cells that survive viral infection in the distal lung epithelia, however, is much less well-characterized. Utilizing a Cre-expressing influenza B virus (IBV) and a lox-stop-lox tdTomato reporter mouse model, we identified that alveolar type 2 (AT2) pneumocytes, a progenitor cell type in the distal lung, can survive viral infection. We show that survival of infection is associated with transcriptional dysregulation compared to bystander AT2 pneumocytes from the same lung. Furthermore, ex vivo experiments revealed a significant reduction in proliferation rates in survivor AT2 pneumocytes compared to matched, non-directly infected bystander cells. Our findings not only enhance our understanding of the AT2 pneumocyte response to IBV infection but could also have broader implications for the mechanisms of respiratory epithelial repair post-viral infection.

Alveolar type 2 (AT2) pneumocytes are a cell type critical for repair of the distal lung after an injury, such as a viral infection. After epithelial damage, AT2 pneumocytes proliferate for both self-renewal and differentiation into type I pneumocytes to repopulate the epithelium. Theoretically, some of the long-term lung sequelae associated with viral infections could be the result of inappropriate AT2 behavior. Here, the authors report that during an influenza B virus infection, some of the actively infected AT2 pneumocytes can ultimately eliminate all traces of the viral RNA and persist in the host long term. As a consequence of having been infected, however, the cells display an altered transcriptional profile and decreased proliferative capacity. These data together suggest a mechanism for how an acute viral infection can have long-term impacts on the pulmonary system.

Lung cancer remains a leading cause of cancer-related mortality worldwide. Our study investigates the involvement of the PD-L1/MALAT1/miR-200a-3p axis in lung tumor progression using a murine model of lung carcinogenesis. Lung tumors were induced in rats, which were divided into groups and sacrificed at different stages of tumor development. A histopathological examination was performed to assess tumor progression. Immunohistochemistry was applied to evaluate the expression of Ki-67 and programmed death-ligand 1 (PD-L1). The level of carcinoembryonic antigen (CEA) and expression analysis of metastasis-associated lung adenocarcinoma transcript 1 (MALAT1), miR-200a-3p, and zinc finger E-box binding homeobox 1 (ZEB1) were evaluated for each stage of induction. Immunohistochemical analysis demonstrated a progressive upregulation of the proliferative marker Ki-67 and the immune checkpoint protein PD-L1 during the induction process, indicative of enhanced tumor proliferation and immune evasion. Additionally, CEA levels revealed a progressive increase across induction stages, with a significant increase in advanced tumor stages, highlighting its clinical relevance as a biomarker for lung cancer progression. Expression analysis revealed dynamic upregulation of MALAT1 and downregulation of miR-200a during lung tumor induction, which correlated with advanced tumor stages and elevated PD-L1 expression, suggesting that the negative correlation between MALAT1 and miR-200a is involved in the development of lung tumors. ZEB1 expression exhibited a notable increase in the advanced stages of induction, consistent with its association with aggressive lung cancer. Our findings underscore the interplay between molecular pathways involved in lung tumor development and the potential diagnostic and therapeutic implications of the PD-L1/MALAT1/miR-200a-3p axis.

Lineage plasticity and histologic transformation from prostate adenocarcinoma to neuroendocrine (NE) prostate cancer (NEPC) occur in up to 15% to 20% of patients with castration-resistant prostate cancer (CRPC) as a mechanism of treatment resistance and are associated with aggressive disease and poor prognosis. NEPC tumors typically display small cell carcinoma morphology with loss of androgen receptor (AR) expression and gain of NE lineage markers. However, there is a spectrum of phenotypes that are observed during the lineage plasticity process, and the clinical significance of mixed histologies or those that co-express AR and NE markers or lack all markers is not well defined. Translational research studies investigating NEPC have used variable definitions, making clinical trial design challenging. In this manuscript, we discuss the diagnostic workup of metastatic biopsies to help guide the reproducible classification of phenotypic CRPC subtypes. We recommend classifying CRPC tumors based on histomorphology (adenocarcinoma, small cell carcinoma, poorly differentiated carcinoma, other morphologic variant, or mixed morphology) and IHC markers with a priority for AR, NK3 homeobox 1, insulinoma-associated protein 1, synaptophysin, and cell proliferation based on Ki-67 positivity, with additional markers to be considered based on the clinical context. Ultimately, a unified workup of metastatic CRPC biopsies can improve clinical trial design and eventually practice.

 This study aimed to examine the suppressive effect of the natural antioxidant vitamin C (VC) against submandibular gland toxicity induced by copper oxide nanoparticles (CuO-NPs).

 Three groups of 30 mature male albino rats (4 weeks old) weighing between 150 and 200 g were selected. The rats were randomly assigned for 6 weeks to receive: intraperitoneal injection (IP) of vehicle (control group); IP of 2.5 mg/kg body weight (bw) of CuO-NPs (CuO-NPs group); and IP of 2.5 mg/kg bw of CuO-NPs, combined with a daily oral dose of 100 mg/kg bw of VC in drinking water via gavage (CuO-NPs/VC group). The rats were euthanized, and their submandibular glands were dissected for histological evaluation, including hematoxylin and eosin staining and immunohistochemistry for Ki-67 and caspase-3.

 The area expression for Ki-67 and caspase-3 was statistically analyzed using GraphPad Prism. Following analysis of variance analysis, Tukey's post hoc was used for multiple comparisons. The significance level was set at p < 0.05.

 CuO-NPs caused significant cytotoxic effects on submandibular salivary gland cells in albino rats. This led to an increase in Ki-67 and caspase-3 levels compared with the control group. VC administration improved tissue histology and reduced Ki-67 and caspase-3 levels in the VC/CuO-NPs group compared with rats treated with CuO-NPs alone.

 The study revealed significant cytotoxic effects of CuO-NPs on the submandibular salivary gland of albino rats. VC effectively mitigated these toxic effects, suggesting its potential as a readily available antioxidant.

Effective treatment of infected wounds remains a challenge due to the rise of antibiotic-resistant microorganisms. The development of advanced materials with strong antimicrobial properties is necessary to address this issue. In this study, a unique composite of electrically modified bacterial cellulose (EBC) with allantoin (ABC) and zein was developed by dipping diffusion method. Morphological structural analysis revealed a uniform distribution of zein and aligned fibers, confirming the synthesis of the ABC-Zein composite. The formation of ABC-Zein was further confirmed by attenuated total reflection-Fourier transform infrared (ATR-FTIR), which displayed additional peaks corresponding to EBC, indicating the incorporation of zein into ABC. X-ray diffraction (XRD) analysis of ABC-Zein demonstrated a similar crystalline structure with EBC. The ABC-Zein showed mechanical integrity (tensile strength: 1.15 ± 0.21 MPa), thermal stability (degradation temperature: 290 °C), porous structure (porosity: 40.23 ± 0.21 %), and hydrophilic (water contact angle: 53.3 ± 5.3°) properties. Furthermore, the antimicrobial agent terpinen-4-ol (T4O), derived from tea tree oil, was incorporated into the ABC-Zein composite. Biological studies confirmed the antimicrobial efficacy (Staphylococcus aureus inhibition: 88.5 ± 7.19 %) and biocompatible (cell viability: 84.95 ± 5.6 %, hemolysis: 4.479 ± 0.39 %) nature of the T4O-ABC-Zein composite. The combined effects of the aligned fiber structure, zein protein, and antimicrobial T4O significantly enhanced infected wound healing by day 7, promoting inflammatory response, granular tissue formation, cell proliferation, and angiogenesis. By day 14, T4O-ABC-Zein facilitated complete wound healing, with reepithelization, collagen I deposition, and downregulation of CD 31, Ki67, and α-SMA. Overall, the innovative T4O-ABC-Zein composite, with an aligned fiber structure, improved biocompatibility, and antimicrobial properties, holds significant potential for the treatment of infected wounds.

Current studies pictured the enteric nervous system and macrophages as modulators of neuroimmune processes in the inflamed gut. Expanding this view, we investigated the impact of enteric neuron-macrophage interactions on postoperative trauma and subsequent motility disturbances, i.e., postoperative ileus. In the early postsurgical phase, we detected strong neuronal activation, followed by transcriptional and translational signatures indicating neuronal death and synaptic damage. Simultaneously, our study revealed neurodegenerative profiles in macrophage-specific transcriptomes after postoperative trauma. Validating the role of resident and monocyte-derived macrophages, we depleted macrophages by CSF-1R-antibodies and used CCR2-/- mice, known for reduced monocyte infiltration, in POI studies. Only CSF-1R-antibody-treated animals showed decreased neuronal death and lessened synaptic decay, emphasizing the significance of resident macrophages. In human gut samples taken early and late during abdominal surgery, we substantiated the mouse model data and found reactive and apoptotic neurons and dysregulation in synaptic genes, indicating a species' overarching mechanism. Our study demonstrates that surgical trauma activates enteric neurons and induces neurodegeneration, mediated by resident macrophages, introducing neuroprotection as an option for faster recovery after surgery.

Stomach adenocarcinoma (STAD) is a common malignant tumor with high morbidity and mortality. Major histocompatibility complex (MHC) is an important component of the immune system responsible for antigen presentation. However, no studies have yet reported on the relationship between major histocompatibility complex-related differentially expressed genes (MHCRDEGs) and the survival prognosis of STAD. The aim of this study is to explore the relationship between MHCRDEGs and survival prognosis in STAD patients.

Using The Cancer Genome Atlas (TCGA) database, we screened for differentially expressed MHCRDEGs, and a survival prognosis model was constructed based on these genes. We generated training and validation samples from the TCGA and Gene Expression Omnibus (GEO) datasets to enhance the robustness of our findings. The predictive effects of the model were assessed using Kaplan-Meier (KM) survival curve analysis, receiver operating characteristic (ROC) curve analysis, calibration analysis and decision curve analysis (DCA), with statistical significance reported as P values. The differences in the expression of key MHCRDEGs between different subgroups of TCGA and GEO databases were analyzed. Finally, a multifactorial survival prognostic model was constructed by combining MHC score (MHCs), and quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was used to verify the expression of key genes.

We identified five key MHCRDEGs: MKI67, MYB, SERPINE1, TRIM31, and HAVCR1. In the first prognostic model, the KM curves demonstrated a highly statistically significant difference in predicting overall survival (OS) in patients (P<0.001). The ROC curves indicated that the model showed relatively low accuracy in predicting 1-year [area under curve (AUC) =0.616], 3-year (AUC =0.644), and 5-year (AUC =0.619) occurrence. Furthermore, calibration analysis and DCA suggested that the model's predictions of OS were consistent with the actual patient survival, with the 5-year prognostic model exhibiting the best clinical utility. In the TCGA and GEO datasets, most of the key genes showed significant expression differences between the STAD/GEO and normal groups (P<0.001). Finally, the predictive model constructed by combining MHCs with clinicopathological staging demonstrated good predictive accuracy with optimal clinical utility at 5 years, with specific accuracy metrics provided as part of our results, and validated their expression via qRT-PCR in cell lines (MKI67: P=0.01, MYB: P=0.02, SERPINE1: P=0.02, TRIM31: P=0.02, HAVCR1: P<0.0001).

In this study, the expression and distribution of MHCRDEGs in STAD were analyzed by various methods, and a clinical prediction model of STAD was constructed using MHCRDEGs. The validity of this model confirms the feasibility of MHCRDEGs as prognostic markers for STAD, elucidating their potential clinical implications in guiding treatment strategies for this disease.

Splicing factors control exon inclusion in messenger RNA, shaping transcriptome and proteome diversity. Their catalytic activity is regulated by multiple layers, making single-omic measurements on their own fall short in identifying which splicing factors underlie a phenotype. Here, we propose splicing factor activity can be estimated by interpreting changes in exon inclusion. We benchmark methods to construct splicing factor→exon networks and calculate activity. Combining RNA-seq perturbation-based networks with VIPER (virtual inference of protein activity by enriched regulon analysis) accurately captures splicing factor activation modulated by different regulatory layers. This approach consolidates splicing factor regulation into a single score derived solely from exon inclusion signatures, allowing functional interpretation of heterogeneous conditions. As a proof of concept, we identify recurrent cancer splicing programs, revealing oncogenic- and tumor suppressor-like splicing factors missed by conventional methods. These programs correlate with patient survival and key cancer hallmarks: initiation, proliferation, and immune evasion. Altogether, we show splicing factor activity can be accurately estimated from exon inclusion changes, enabling comprehensive analyses of splicing regulation with minimal data requirements.

Proteasomes degrade intracellular proteins. Different proteasome forms were identified. Proteasome inhibitors are used in cancer therapy, and novel drugs directed to specific proteasome forms are developed. Breast cancer (BC) therapy depends on the subtype of the tumor, determined by the expression level of Ki67, HER-2, estrogen and progesterone receptors. Relationships between the presence of specific proteasome forms and proteins that determine the BC subtype remain unclear. Here, using gene expression data in 19,145 tumor samples from 144 datasets and tissues from 159 patients with different subtypes of BC, we investigated the association between the activity and expression of proteasomes and levels of BC subtype markers.

Bioinformatic analysis of proteasome subunit (PSMB1-10) gene expression in BC was performed. Proteasome heterogeneity in BC cell lines was investigated by qPCR. By Western blotting, proteasome composition was assessed in cells and patient tissue lysates. Proteasome activities were studied using fluorogenic substrates. BC molecular subtypes were determined by immunohistochemistry.

BC subtypes demonstrate differing proteasome subunit expression pattern and strong PSMB8-10 co-correlation in tumors. A significant increase in chymotrypsin- and caspase-like proteasome activities in BC compared to adjacent tissues was revealed. The subunit composition of proteasomes in tumor tissues of BC subtypes varied. Regression analysis demonstrated a positive correlation between proteasome activities and the expression of Ki67, estrogen receptors and progesterone receptors.

BC subtypes demonstrate differences within the proteasome pool. Correlations between the proteasome activity, hormone receptors and Ki67 indicate possible mutual influence. Obtained results facilitate development of novel drug combinations for BC therapy.