搜索结果：Experimental and molecular pathology

Lung metastasis is a leading cause of mortality in patients with breast cancer, especially triple-negative breast cancer (TNBC), where current therapies are limited and toxic. Hedyotis diffusa injection (HDI) is used clinically as an adjuvant, but its efficacy and mechanisms against lung metastasis remain unclear. To investigate the antitumour and antimetastatic activities of HDI and its active constituent, asperulosidic acid (ASPA), identify its molecular targets, and assess its potential synergy with gefitinib. The antiproliferative, proapoptotic, antimigratory, and anti‑invasive effects of HDI and ASPA were evaluated in 4T1, MDA‑MB‑231, and MCF‑7 cells via CCK‑8, colony formation, flow cytometry, wound‑healing, and Transwell assays. Anti‑tumor efficacy, anti‑lung metastatic effects, and systemic toxicity were assessed in a BALB/c 4T1 orthotopic breast tumor model. Mechanistic studies employed data‑independent acquisition (DIA) proteomics, Western blotting, immunohistochemistry, UPLC-MS, network pharmacology, molecular docking, and microscale thermophoresis. The synergistic activity of HDI with gefitinib was examined in TNBC cells. UPLC-MS identified 1112 constituents in HDI, with deacetylasperulosidic acid, D-(+)-malic acid, and asperulosidic acid among the most abundant compounds. HDI dose- and time-dependently inhibited the proliferation of 4T1, MDA-MB-231, and MCF-7 cells, reduced colony formation, induced apoptosis, and suppressed migration and invasion. In the orthotopic 4T1 breast cancer model, HDI significantly decreased primary tumor growth and lung metastatic burden in a dose-dependent manner, while showing lower systemic toxicity than cisplatin. DIA proteomics identified PTK2B and CCN2 as key downregulated proteins and revealed suppression of focal adhesion-related signaling. Mechanistic validation showed that HDI inhibited PTK2B phosphorylation and CCN2 expression, blocked KRAS-p38 MAPK signaling, reversed EMT, and regulated extracellular matrix remodeling by increasing TIMP2 and decreasing MMP2. Functional activation or overexpression of PTK2B and CCN2 partially weakened the inhibitory effects of HDI, supporting their involvement in HDI action. Asperulosidic acid, a major iridoid glycoside in HDI, reproduced the antitumor and antimetastatic effects of HDI, directly bound PTK2B with a Kd of 56.7 μM, and suppressed the PTK2B/CCN2-KRAS-p38 axis. HDI also synergized with gefitinib in MDA-MB-231 cells and further inhibited migration, invasion, EMT, and PTK2B/CCN2-associated signaling. HDI inhibits breast cancer growth and lung metastasis by dual targeting of PTK2B and CCN2, thereby suppressing KRAS-p38 MAPK signaling, reversing EMT, and modulating TIMP2/MMP2-mediated extracellular matrix remodeling. Asperulosidic acid is identified as an important active constituent contributing to these effects. The favorable safety profile of HDI and its synergistic interaction with gefitinib support its potential application as a multitarget adjuvant therapy for metastatic breast cancer, particularly TNBC.

Ovarian cancer (OC) comprises multiple histotypes with distinct mechanisms, molecular features, and clinical behavior. We used Mendelian randomization (MR) to map histotype-stratified metabolic pathways and connect them to drug targets, establishing a translatable target-metabolic node-histotype risk chain. We built a multi-stage MR framework using Integrative Epidemiology Unit (IEU) OpenGWAS summary statistics. After screening 1400 plasma metabolites against overall ovarian cancer in UK Biobank and Ovarian Cancer Association Consortium (OCAC) with KEGG enrichment, we traced a prespecified amino acid/energy-nitrogen axis using histotype-stratified univariable MR and pathway-restricted multivariable MR. We then performed cis drug-target MR for PPARG, DPP4, ABCC8/KCNJ11, and SLC5A2, integrated triangulation, colocalization, and mediation analyses, and experimentally interrogated the prioritized PPARG/ABCC8-KCNJ11-lactate-invasive mucinous ovarian cancer (IMOC) triangle. Screening nominated 55 and 72 metabolites in UK Biobank and OCAC, respectively (IVW p < 0.05), highlighting amino-acid nitrogen and central-carbon metabolism. Univariable Mendelian randomization (UVMR) showed marked heterogeneity: alanine increased low-grade serous ovarian cancer (LGSOC) risk, glutamate was protective for endometrioid OC, and lactate-related traits most consistently implicated the low-grade/borderline serous lineage. In multivariable Mendelian randomization (MVMR), tryptophan and lactate levels emerged as independent risk nodes for serous low-grade plus low malignant potential (LG + LMP). Drug-target MR prioritized PPARG as protective (OR = 0.18) and ABCC8/KCNJ11 as risk-increasing (OR = 7.50) for IMOC, with opposite target &#x2192; lactate effects supporting a directionally symmetric target-lactate-IMOC triangle. Experimental perturbation in mucinous ovarian cancer models produced concordant reciprocal changes in lactate and malignant phenotypes, extending this triangle biologically. This integrative MR framework delineates histotype-specific metabolic drivers and links them to actionable targets, providing a roadmap from genetic prioritization to mechanistic and translational validation.

Diabetic retinopathy (DR) and retinal aging, though arising from distinct causes, share converging mechanisms-including oxidative stress, chronic inflammation, mitochondrial dysfunction, and AGE accumulation-that compromise retinal integrity. These overlaps suggest common gene expression patterns, and highlight the contribution of disease-associated or accelerated aging processes to diabetes-induced retinal injury. We systematically retrieved DR- and aging-related human genes from public genetic databases, identified their overlap, and focused on those involved in key metabolic pathways (AGEs, oxidative stress, lipid metabolism, and autophagy). Genes were then cross-validated across multiple databases and filtered by ocular expression to ensure relevance to retinal pathology. Our findings show that although DR and retinal aging arise from distinct etiologies, they converge on four principal metabolic pathways-oxidative stress, AGE accumulation, lipid dysregulation, and impaired autophagy-that collectively drive similar vascular, neuronal, and inflammatory injury within the retina. Shared genes with high ocular expression reinforce the biological relevance of these pathways, while multi-pathway hub genes appear to function as central regulators that integrate redox imbalance, metabolic disruption, and proteostatic failure. These results provide a unified molecular perspective of retinal degeneration and support the potential development of therapeutic approaches designed to simultaneously target age- and diabetes-associated retinal pathology. This review suggests that DR and retinal aging, although initiated by distinct triggers, converge on shared metabolic pathways-oxidative stress, AGE accumulation, lipid dysregulation, and autophagy impairment-mediated by genes expressed in ocular tissues. Within these intersecting pathways, shared hub genes emerge as central control nodes that may amplify molecular dysfunction and represent potential therapeutic entry points. By mapping these molecular intersections, this study may provide a unified mechanistic perspective of retinal degeneration and support the development of dual-purpose strategies aimed at preventing or mitigating both DR and aging-related retinal decline. These findings highlight potential translational opportunities for targeting shared metabolic networks; however, they should be interpreted as hypothesis-generating and require further experimental and clinical validation to establish causal relationships and therapeutic relevance.

Gastric cancer (GC) is one of the main causes of cancer-related global mortality. The emergence of drug resistance and toxicity in current therapies highlights the need for novel treatment strategies. Quercetin, a natural flavonoid, has demonstrated anticancer activity; however, its molecular mechanism, particularly its effect on key targets in GC, remains underexplored. A comprehensive in silico and in vitro methods were used to elucidate the anticancer potential of Quercetin. Network pharmacology analysis was used to identify potential GC-related targets, followed by molecular docking and 200 ns molecular dynamics (MD) simulations to evaluate the binding affinity and stability of the Quercetin-target complex. In vitro experiments, including gene expression analysis and fluorescence binding assays, were conducted using AGS gastric cancer cells to validate the computational findings.nsulin-like growth factor 1 (IGF1) emerged as a key hub gene associated with GC progression. Molecular docking predicted a favorable interaction between Quercetin and IGF1, with a docking score of -&#x2009;6.3 kcal/mol and multiple hydrogen-bond interactions. MD simulations confirmed the stability of the Quercetin-IGF1 complex, with reduced RMSD values (0.48 nm vs. 0.63 nm for unbound IGF1), favorable free energy profiles, and stable hydrogen bonding. In vitro studies demonstrated a significant downregulation of IGF1 mRNA expression (p&#x2009;<&#x2009;0.001) and a dose-dependent inhibition of IGF1 activity by Quercetin. The integration of network pharmacology, computational modeling, and experimental validation suggests that Quercetin may modulate the IGF1 signaling axis and influence IGF1-associated pathways in gastric cancer. The stable binding and significant inhibitory effect observed suggests that Quercetin may interrupt IGF1-mediated signaling pathways involved in tumor growth and survival. This study identifies Quercetin as a potential modulator of IGF1-associated signaling pathways with significant therapeutic promise for gastric cancer. The findings provide mechanistic insights supporting the further development of Quercetin as a targeted therapy for IGF1-driven malignancies.

Early brain injury (EBI) following subarachnoid hemorrhage (SAH) involves a complex interplay of endoplasmic reticulum (ER) stress, mitochondrial dysfunction, oxidative imbalance, neuroinflammation and endothelial dysregulation that collectively shape neuronal vulnerability. Experimental strategies capable of modulating these interconnected stress responses remain incompletely characterized. Dexpanthenol (DEX), a pantothenic acid derivative involved in cellular energy metabolism, has been shown to influence redox balance and inflammatory signaling in experimental brain injury models, but its association with coordinated EBI-related molecular responses after SAH has not been fully explored. In this study, adult male Wistar rats were assigned to Sham, DEX, SAH or SAH&#x2009;+&#x2009;DEX groups. SAH was induced by autologous arterial blood injection into the cisterna magna and DEX (500&#xa0;mg/kg/day, intraperitoneal) was administered for seven consecutive days. Histopathology, immunohistochemistry and RT-qPCR were used to assess neuronal injury, ER stress markers, mitochondrial apoptotic signaling, redox regulation, inflammatory mediators, endothelial regulators and Sirtuin-1 (Sirt1) expression. SAH was associated with marked neuronal degeneration accompanied by activation of ER stress, apoptotic and inflammatory pathways, endothelial dysregulation and reduced expression of Sirt1 and Glutathione Peroxidase 4 (GPX4). DEX administration was associated with lower expression of ER stress, inflammatory and apoptotic markers, together with partial recovery of GPX4 and a non-significant upward trend in Sirt1 expression, compared with untreated SAH animals. These findings support the view that multiple injury-related pathways remain concurrently altered during the subacute post-hemorrhagic period and are associated with DEX treatment.

Antiviral immunity profoundly impacts host metabolism, which can, in turn, modulate immune responses and influence disease pathology. The liver orchestrates systemic bile acid (BA) metabolism, a pathway disrupted in chronic liver diseases such as viral hepatitis. BAs are increasingly recognized for their immunomodulatory properties. Thus, improved understanding of the interplay between immunity and BA metabolism may reveal novel therapeutic avenues. Using lymphocytic choriomeningitis virus (LCMV) as a model, we investigated the interplay between chronic virus infection, BA metabolism, and immunity. Chronic LCMV infection increased BA levels and shifted circulating and liver BA composition toward host-derived, conjugated BAs. Hepatic BA transport and synthesis genes were broadly downregulated, in part depending on CD8+ T cells. Pharmacological inhibition of the main hepatic transporter of conjugated BAs, NTCP (Slc10a1), increased hepatic damage, while combined genetic disruption of the BA transporters Slco1a1, Slco1a4, and Slco1b2, responsible for the hepatic reuptake of unconjugated BA, reduced liver pathology and impaired antiviral CD8+ T cell responses. These findings reveal a reciprocal interplay between BA metabolism and CD8+ T cells, expanding our understanding of adaptive immunity in viral hepatitis. They also highlight how immunometabolic changes in liver disease may affect adaptive immune responses against infections.

Those affected by Type 2 diabetes (T2DM) often encounter microvascular harm, and within this context, the kidneys are especially prone to damage. Among the approaches to manage such microvascular complications, Shenqi Tangluo Pill (SQTLW), a traditional Chinese herbal formulation, is commonly utilized in clinical settings for preventing and addressing microvascular issues linked to T2DM. Nonetheless, the detailed impacts and working principles of this herbal formulation in alleviating kidney damage caused by diabetes are still not clearly understood. The aim was to confirm the kidney-protective impacts of SQTLW on T2DM mice, as well as to clarify the fundamental mechanisms related to intestinal microbiota and the microbe-derived metabolite trimethylamine N-oxide (TMAO), especially under the condition of intestinal flora depletion induced by a combination of broad-spectrum antibiotics (ABx). Diabetic db/db mice received sustained SQTLW intervention over an 8-week period. Following the 8-week intervention, evaluations were performed on fasting blood glucose levels (FBG), body mass, kidney function, and histopathological changes in renal tissue. Additionally, the makeup of gut microbial communities and the circulating concentrations of metabolites linked to TMAO were assessed. To assess the integrity of the intestinal barrier, histopathological examination of colonic tissue and expression profiles of zonula occludens protein-1 (ZO-1) and Occludin were performed. Expression levels of NOD-like receptor pyrin domain-containing 3 (NLRP3), Smad family member 2/3 (Smad2/3), fibronectin (FN), and &#x3b1;-smooth muscle actin (&#x3b1;-SMA) in renal tissue were examined using immunodetection methods. Next, db/db mice were given a combination of ABx and SQTLW treatments to further examine if the therapeutic outcome is dependent on the regulation of gut microorganisms. SQTLW notably lowered FBG and body weight in db/db mice, ameliorated renal dysfunction, and alleviated histopathological injury of renal tissue, with a notable dose-response relationship. 16S rRNA gene sequencing-based profiling revealed that SQTLW substantially altered the compositional structure of distinct bacterial genera from the phyla Bacteroidota and Firmicutes, particularly Muribaculaceae and Lachnospiraceae. Colonic histopathology and molecular detection indicated that SQTLW effectively preserved intestinal barrier architecture. Targeted metabolomics via LC-MS/MS demonstrated that SQTLW significantly modulated TMAO-related plasma metabolites, including TMAO, choline, and creatinine. Moreover, SQTLW significantly downregulated renal expression of NLRP3 and Smad2/3, thereby attenuating fibrosis-related injury. Importantly, further experiments confirmed that depletion of gut microbiota by ABx partially attenuated the renoprotective effects of SQTLW in db/db mice. SQTLW confers protective effects against renal injury secondary to T2DM, potentially via modulation of gut microbial composition, preservation of intestinal barrier function, and regulation of TMAO-related metabolic pathways.

While Cisplatin remains a cornerstone of oncological intervention, its induction of systemic oxidative stress arranges a deleterious cascade that compromises male reproductive homeostasis. Selenium nanoparticles (SeNPs) may protect against oxidative stress and capping them with Arab gum (AG) could enhance their therapeutic efficacy. This study investigated the comparative gonadoprotective potential of Selenium Nanoparticles (SeNPs) and Arab gum (AG)-coated SeNPs (AG-SeNPs) against Cis-induced testicular dysfunction. Forty-five male Wistar rats were randomized into nine experimental groups: four baseline controls (Normal, AG, SeNPs, and AG- SeNPs) and five treatment groups receiving AG, SeNPs, AG-SeNPs, or a physical mixture of AG&#x2009;+&#x2009;SeNPs following Cis administration. Assessment parameters included the gonadosomatic index, biochemical markers of oxidative stress (CAT, GPX, MDA), and pro-inflammatory cytokines (TGF-beta, TNF-&#x3b1; and IL-6). Testicular integrity was further evaluated via histopathology, PCNA protein expression (cellular proliferation), and DNA fragmentation analysis using the Comet assay and apoptotic markers (Caspase-3 and Cytochrome C release). Administration of AG-SeNPs demonstrated superior gonadoprotective efficacy compared to uncoated SeNPs or physical mixtures. AG-SeNPs significantly restored redox homeostasis, via augmenting antioxidant enzyme activities (CAT, GPX) and reducing lipid peroxidation (MDA). Furthermore, AG-SeNPs significantly downregulated inflammatory signaling (TGF-beta, TNF-&#x3b1; and IL-6) and mitigated DNA damage. The treatment preserved genomic stability and inhibited the intrinsic apoptotic pathway by suppressing Cytochrome C release and Caspase-3 activation. Histological examination confirmed the restoration of seminiferous tubular architecture and enhanced cellular proliferation (PCNA expression). These findings suggest that AG-coating enhances the therapeutic index of SeNPs, likely due to improved bioavailability and synergistic antioxidant properties. AG-SeNPs represent a promising nanomedicine-based strategy for mitigating the gonadotoxic side effects of cisplatin-based chemotherapy.

Extrahepatic cholangiocarcinoma (eCCA) is characterized by marked molecular heterogeneity and limited therapeutic options. MicroRNAs (miRNAs) are key post-transcriptional regulators of cancer-related pathways, but their contribution to tumor adaptation in physiologically relevant models remains poorly understood. Three-dimensional (3D) tumor spheroids better mimic in vivo conditions than conventional two-dimensional (2D) cultures. We compared miRNA expression profiles in two eCCA cell lines (Sk-ChA-1 and Mz-ChA-1) grown as monolayers (2D) or multicellular tumor spheroids (3D). MiRNA profiling was performed using NanoString technology. Predicted targets were analyzed by over-representation analysis, and selected miRNAs and genes were validated by RT-qPCR and ELISA-based assays. 3D growth induced extensive miRNA remodeling, with distinct (54 deregulated in Sk-ChA-1 and 29 in Mz-ChA-1 cells) and partially overlapping signatures (miR-1283, miR-577, and miR-2113). Among the shared miRNAs, predicted targets included DUSP10 and RBFOX1, while in spheroids, cell-specific multiple miRNAs converged on shared targets (TNRC6B, SMARCAD1, ATG14, HMGA2, and CLOCK) displaying inverse expression patterns. The transcriptional program impacted MAPK signaling, enhanced EMT, and activated stress-adaptive networks but attenuated proliferation in 3D Sk-ChA-1 cells, while Mz-ChA-1 cells retained a more epithelial and proliferative profile. In this context, we point out the involvement of miR-19b-3p using anti-miR transfection experiments. Our findings reveal a miRNA-driven regulatory landscape associated with 3D growth in eCCA, linking tumor architecture to signaling rewiring and cellular plasticity, and highlight potentially druggable candidate targets and pathways to investigate as candidates using inhibitors or gene therapy-based interventions.

The present study represents the first detailed report of Yersinia ruckeri infection in farmed rainbow trout (Oncorhynchus mykiss) from the Indian Himalayan Region, where intensification of aquaculture has elevated the pathogen load and transmission dynamics, causing occurrence of enteric redmouth disease (ERM). The study examined infections by Y. ruckeri, RTACP-14&#xa0;A (PX411334), in farmed rainbow trout from the Indian Himalayas. Affected fish displayed clinical signs such as hemorrhagic mouth, deep ulceration at caudal peduncle region, fin degeneration, lesions, black pigmentation, and erratic swimming behaviour. The isolated Y. ruckeri RTACP-14&#xa0;A strain was identified through biochemical tests and confirmed by 16&#xa0;S rRNA sequencing showing the closest evolutionary relationship (100) to Y. ruckeri strain BY-1Y. Virulence analysis revealed &#x3b2;-hemolysis, low hydrophobicity, and substantial biofilm formation under glucose-rich conditions. The cumulative mortality increased from 12.5% at day 1 to 83% by day 32 during a controlled experimental infection trial conducted in rainbow trout. Antibiotic susceptibility test showed that Y. ruckeri RTACP-14&#xa0;A was sensitive to most antibiotics, with imipenem, fosfomycin, and ciprofloxacin exhibiting the largest inhibition zones (>&#x2009;40&#xa0;mm), while bacitracin was the only antibiotic to which resistance was observed. The lowest minimum inhibitory concentrations (MICs) were recorded for oxytetracycline (0.125&#xa0;&#xb5;g mL-&#x2009;1), florfenicol (1&#xa0;&#xb5;g mL-&#x2009;1), and tobramycin (1&#xa0;&#xb5;g mL-&#x2009;1). Histopathological analysis revealed gill hyperplasia, intestinal villus necrosis, kidney damage, liver congestion, and muscle degradation, with distinct patterns distinguishing acute from chronic infections. To mitigate outbreaks of enteric red mouth disease caused by Y. ruckeri, trout farmers should adopt best management practices, including maintaining optimal water quality, minimizing handling stress, implementing strict biosecurity measures, and using antimicrobials judiciously within a regulated framework. These measures are essential for safeguarding fish health and welfare, minimizing economic losses, and ensuring the sustainability of trout farming in the region.

To evaluate whether cumin (Cuminum cyminum) oil attenuates nandrolone decanoate (ND)-induced hepatic alterations in rats. Eighty male Sprague-Dawley rats were randomized into six groups: control, cumin oil alone, ND low dose (10&#xa0;mg/kg/week), ND high dose (20&#xa0;mg/kg/week), ND low dose&#x2009;+&#x2009;cumin oil, and ND high dose&#x2009;+&#x2009;cumin oil. Cumin oil was administered orally at 400&#xa0;mg/kg/day for 4&#xa0;weeks. Outcomes included relative liver weight index, serum ALT/AST, total bilirubin, lipid profile, and blinded histopathology. Both ND doses increased ALT/AST and bilirubin levels and worsened the lipid profile compared with controls, with more pronounced and significant alterations in the high-dose ND group. Co-administration of cumin oil attenuated ND-associated elevations in liver enzymes and bilirubin, improved lipid parameters, and was associated with reduced histological damage compared with ND alone. Interestingly, cumin oil alone increased ALT/AST and lipid parameters compared with controls, although liver architecture remained unremarkable on H&E. In this rat model, cumin oil co-administration partially attenuated ND-induced hepatic biochemical, lipid, and histological alterations. However, cumin oil alone increased ALT/AST and lipid parameters despite unremarkable H&E morphology. Therefore, the present findings should be interpreted as evidence of context-dependent attenuation during ND exposure, not as proof of an independent hepatoprotective or lipid-lowering effect of cumin oil in healthy rats. Dose-ranging, safety evaluation, batch-specific chemical profiling, preparation standardization, and mechanistic studies incorporating antioxidant/oxidative stress markers such as MDA, GSH, SOD, CAT, GPx, and molecular endpoints are needed to clarify pathways and translational relevance.

Cisplatin is a key treatment for head and neck squamous cell carcinoma (HNSCC), but the development of resistance severely limits its effectiveness. The molecular determinants underlying cisplatin resistance in HNSCC remain unclear. Multiple databases were used to screen the core genes related to cisplatin resistance in HNSCC patients. Tumor tissue samples from HNSCC patients were collected and the expression of FOXA2 was verified through various pathological tests to establish the correlation between FOXA2 expression and the clinical characteristics of the patients. The in&#xa0;vitro and patient-derived organoids (PDOs) models were used to verify the regulatory effect of FOXA2 on the cisplatin resistance of HNSCC. Transcriptome sequencing combined with multi-omics analysis demonstrated that LAMC2 is a downstream target of FOXA2 in regulating cisplatin resistance. Bioinformatic screening of cisplatin-resistant cohorts revealed that FOXA2 was the only gene significantly associated with poor survival outcomes in TCGA-HNSCC patients. Transcriptomic profiling and pathway enrichment analyses revealed the activation of the PI3K/AKT signaling cascade. We identified LAMC2 as a direct transcriptional target of FOXA2. Chromatin immunoprecipitation and luciferase reporter assays confirmed FOXA2 binding to the LAMC2 promoter, resulting in transcriptional activation. FOXA2-mediated upregulation of LAMC2 increased PI3K and AKT phosphorylation, and LAMC2 overexpression reversed the impaired malignant phenotypes caused by FOXA2 silencing. In xenograft models and PDO systems, FOXA2 overexpression reduced responsiveness to cisplatin, whereas FOXA2 inhibition significantly increased therapeutic sensitivity. Our research has identified a previously unrecognized regulatory axis involving FOXA2, LAMC2, and PI3K/AKT, which plays a crucial role in the progression and resistance to cisplatin in HNSCC. Therefore, targeting the FOXA2-LAMC2 axis may represent a novel therapeutic strategy to overcome cisplatin resistance in HNSCC.

Tumor-infiltrating immune cells play a fundamental role in shaping patient prognosis across many cancers. Retroperitoneal dedifferentiated liposarcomas (DDLPS) and leiomyosarcomas (LMS) are associated with poor prognosis, therefore, a deeper understanding of the immune landscape in these tumors is warranted to identify novel prognostic biomarkers and immunotherapeutic targets. In this exploratory study, we analyzed spatial organization, frequency, and proportions of CD8+ T cell (including activated, proliferating, and exhausted phenotypes), T helper, and macrophage subsets, and B cells in primary treatment-naive retroperitoneal DDLPS (n&#xa0;=&#xa0;22) and LMS tissues (n&#xa0;=&#xa0;10) using classical and multiplex immunohistochemistry. CD8+LAG-3+ T cells comprised the highest proportion of all analyzed CD8+ subsets. Their elevated density was associated with improved overall survival (OS). Moreover, higher density of CD8+Ki-67+ T cells was linked to better OS and tended to independently predict OS. Interestingly, enhanced density of regulatory T cells was associated with favorable OS. Densities and proportions of T cell subsets did not significantly differ between DDLPS and LMS. A lower density of M2-like macrophages (CD68+CD163+) was linked to better progression-free survival (PFS) and tended to serve as an independent prognostic factor. Invasive margin revealed distinct prognostic patterns compared to tumor core, where high levels of CD8+Ki-67+TCF1+ were linked to better PFS and elevated proportions of CD8+GrzB+ and CD3+T-Bet+ T cells were associated with inferior PFS. These findings highlight the clinical relevance of immune infiltration in retroperitoneal DDLPS and LMS. Moreover, they support the rationale for further exploration of the immune architecture for prognostic biomarkers and development of targeted immunotherapeutic strategies to improve the clinical outcomes of the patients.

Breast cancer is the most prevalent malignancy worldwide. Beyond genetic factors, dietary components like palmitic acid (PA) and sucrose modulate tumor progression and microenvironment dynamics. However, their combined mechanistic impacts on tumor-stroma crosstalk remain undefined. We investigated how dietary PA and sucrose affect breast cancer growth and the tumor microenvironment using a murine model. Mice received Control, high-PA (HPOD), high-sucrose (HCD), or combined (HPCD) diets. We assessed tumor growth, metabolic parameters, membrane fatty acids, histopathology, proliferative indices, and microenvironment markers (CD8, F4/80, &#x3b1;-SMA, HIF-1&#x3b1;, FASN). In vitro assays evaluated PA and fructose effects on LM3 cells and fibroblasts. HCD and HPCD markedly promoted tumor growth versus Control and HPOD. HPOD increased membrane saturated fatty acids, whereas HCD elevated &#x3c9;-6 PUFAs. Both HCD and HPCD increased macrophage infiltration (F4/80&#x2009;+), while HPCD specifically activated cancer-associated fibroblasts (&#x3b1;-SMA). HIF-1&#x3b1; protein increased across all experimental diets without mRNA alterations, suggesting post-transcriptional stabilization. FASN protein upregulation occurred in HCD and HPCD, while Fasn mRNA increased specifically in HCD. In vitro, combining PA and fructose enhanced LM3 cell proliferation and viability while reducing apoptosis. Additionally, this co-treatment specifically increased fibroblast viability. Diets rich in PA and sucrose drive breast cancer progression through distinct mechanisms. Carbohydrates transcriptionally activate FASN-mediated lipogenesis, whereas their combination with PA remodels the tumor microenvironment via CAF activation and HIF-1&#x3b1; post-transcriptional stabilization. These findings highlight specific dietary components as critical variables in cancer progression, offering potential implications for nutritional interventions and targeted therapies.

Atherosclerosis, a major complication of diabetes mellitus, involves complex metabolic and inflammatory disruptions that remain inadequately addressed by current therapies. Ferulic acid (FA), a plant-derived polyphenol, possesses antioxidant and anti-inflammatory characteristics, yet its role in diabetic atherosclerosis remains underexplored. In the current study, we explore the anti-atherosclerosis effects of FA against diabetes-induced atherosclerosis in rats. Forty male Wistar rats received a high-fat diet, streptozotocin, and vitamin D3 to induce atherosclerosis. Rats were divided into control, atherosclerotic, and treated with FA (50&#x2009;mg/kg, daily for 7&#x2009;weeks as preventive and 3&#x2009;weeks as therapeutic) or rosuvastatin (5&#x2009;mg/kg, daily for 7&#x2009;weeks). Body weight, biochemical, molecular, and histopathological analyses were conducted. Atherosclerotic rats exhibited significant hyperglycemia, dyslipidemia, inflammation, and vascular injury. FA administration obviously improved body weight, fasting and postprandial glucose levels, lipid profile, and cardiovascular indices. Moreover, FA significantly reduced serum oxLDL, MDA, and NO levels, while it significantly increased serum activities of both SOD and GPX, besides GSH levels. FA treatment significantly downregulated NF-&#x3ba;B p65, iNOS, and MMP-9 aortic protein levels. Additionally, serum pro-inflammatory cytokines (TNF-&#x3b1; and MCP-1) were reduced, while serum levels of anti-inflammatory IL-10 were improved in FA-treated atherosclerotic rats. Serum miR-27 and miR-29 expression levels were significantly modulated after treatment with FA. The overall results suggested that FA confers multi-targeted protection against diabetic atherosclerosis through modulation of metabolic, oxidative stress, inflammatory, and epigenetic pathways. These findings highlight its potential as a complementary or preventative approach in the management of vascular diseases associated with diabetes.

Acral lentiginous melanoma (ALM) is characterized by a low mutational burden, frequent chromosomal rearrangements, and profound epigenetic dysregulation, distinguishing it from ultraviolet (UV)-induced melanoma. Among the epigenetic regulators, Enhancer of Zeste Homolog 2 (EZH2), the catalytic component of the Polycomb Repressive Complex 2 (PRC2), plays a central role in chromatin compaction and transcriptional repression through trimethylation of histone H3 on lysine 27 (H3K27me3). EZH2 overexpression or hyperactivation contributes to tumor progression, immune evasion, and therapeutic resistance. Recent multi-omic studies have highlighted the importance of EZH2 in regulating melanoma plasticity, immune modulation, and metabolic reprogramming. In ALM, where canonical oncogenic mutations such as BRAF V600E and NRAS Q61 are less frequent, EZH2-driven epigenetic mechanisms may play an even more dominant role in tumor initiation and progression. Pharmacological inhibitors of EZH2, including tazemetostat, have shown promise in preclinical melanoma models by restoring antigen presentation, enhancing CD8+ T-cell infiltration, and reversing transcriptional programs associated with immune resistance. This review aims to summarize the role of EZH2 in the molecular pathogenesis of ALM, emphasizing its contributions to epigenetic regulation, tumor plasticity, and immune escape, and discusses emerging therapeutic strategies targeting EZH2-mediated pathways to improve outcomes for this aggressive melanoma subtype.

Per- and polyfluoroalkyl substances (PFASs) are widely used in numerous industrial processes and consumer products and are now ubiquitously distributed in the environment, thereby creating multiple routes of human exposure. Their physicochemical properties confer high persistence, bioaccumulation potential, and toxicity, which have raised increasing concern about their long-term impacts on human health. Because of its central role in xenobiotic uptake, metabolism, transport, and excretion, the liver is considered a major target organ of PFAS toxicity. Over the past few years, a growing body of epidemiological, in vivo, and in vitro evidence has linked PFAS exposure to a broad spectrum of hepatic abnormalities, including liver injury, cholestatic liver injury and bile acid dysregulation, metabolic dysfunction-associated steatotic liver disease (MASLD), and hepatocellular carcinoma (HCC). In addition to legacy long-chain PFASs, short-chain congeners and emerging alternatives such as GenX and 6:2 Cl-PFESA have also shown considerable hepatotoxic potential. This review summarizes current evidence on the contribution of PFAS exposure to liver disease, with particular attention to human relevance and the mechanisms underlying PFAS-induced hepatotoxicity, including oxidative stress, inflammatory activation, disruption of the gut-liver axis and enterohepatic circulation, lipid metabolic reprogramming, and impairment of bile acid homeostasis. Remaining knowledge gaps and future perspectives are also highlighted to support mechanistic understanding and improve PFAS-related liver risk assessment.

RNA metabolic dysregulation is a key pathological mechanism underlying the onset and progression of Alzheimer's disease (AD), involving multiple aspects such as abnormal RNA splicing, loss of function in RNA-binding proteins, dysregulation of non-coding RNAs, and impaired nuclear-cytoplasmic transport. In recent years, the emergence of epigenome research has revealed the critical role of RNA chemical modifications in regulating RNA metabolism at the post-transcriptional level. N4-acetylcytidine (ac4C) is the only known RNA acetylation modification in eukaryotes and is specifically catalyzed by N-acetyltransferase 10 (NAT10). The ac4C modification is widely found in tRNA, rRNA, and mRNA, and by influencing RNA stability, translation efficiency, and ribosome assembly, it participates in various biological processes such as the cell cycle, differentiation, aging, and stress responses. In AD, the ac4C modification profile undergoes significant changes, involving GABAergic synapses, the PI3K-AKT signaling pathway, and various lncRNAs. Although indirect evidence from progeria and tumor models suggests that via &#x3b1;-tubulin acetylation and intersect with AD pathology via the p53 pathway and regulation of autophagy, these mechanisms currently lack direct experimental validation in NAT10 may participate in axonal transport through &#x3b1;-tubulin acetylation and intersect with AD pathology via the p53 pathway and autophagy regulation, these mechanisms currently lack direct experimental validation within the AD system. This article systematically summarizes the molecular basis and regulatory networks of ac4C modification, integrates existing evidence and unresolved questions regarding its role in AD, and explores its potential value as a diagnostic biomarker and therapeutic target, with the aim of providing guidance for future research in this field.

Hydroxysafflor Yellow A (HSYA), the major bioactive component from Carthamus tinctorius L., exerts significant protective effects against myocardial ischemia-reperfusion injury (MIRI). Mitophagy is pivotal in the pathological process of MIRI, yet the specific molecular mechanism underlying HSYA-mediated mitophagy regulation remains unclear. This study aimed to investigate the association between HSYA treatment and mitochondrial autophagy in murine MIRI and to explore the potential mechanistic role of the SIRT1-FOXO3-BNIP3 signaling pathway using functional loss-of-function and rescue experiments. These findings may provide preliminary evidence supporting the clinical translational potential in MIRI therapy. Mouse myocardial ischemia-reperfusion injury (MIRI) model and oxygen-glucose deprivation/reoxygenation (OGD/R)-induced AC16 cardiomyocyte injury models were established. Metabolomics, molecular docking, and surface plasmon resonance (SPR) techniques were combined to screen the potential targets of HSYA. The SIRT1 inhibitor EX527 and SIRT1 siRNA were used to verify the underlying mechanism. Cardiac function, myocardial infarct size, mitochondrial function, the expression of autophagy-related proteins, and protein-protein interaction were detected and analyzed. Compared with the MIRI group, HSYA significantly improved cardiac function in mice, as evidenced by increased left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) (p < 0.01), attenuated ST-segment elevation, and improved myocardial perfusion. HSYA also markedly reduced myocardial infarct size (p < 0.01) and serum levels of CK-MB, LDH, and cTnI (all p < 0.01) and ameliorated myocardial histopathological damage and mitochondrial ultrastructural integrity. Mechanistic studies revealed that HSYA significantly upregulated the expression of SIRT1, FOXO3, BNIP3, Beclin-1, and the LC3II/I ratio while downregulating p62 expression (p < 0.01), consistent with enhanced mitophagy-related activity. Furthermore, these protective effects were markedly attenuated upon SIRT1 inhibition or siRNA-mediated silencing, whereas HSYA intervention partially reversed these alterations. Additionally, co-immunoprecipitation (Co-IP) and pull-down assays demonstrated that HSYA promoted protein-protein interactions between SIRT1-FOXO3, FOXO3-BNIP3, and BNIP3-LC3B. These findings highlight that HSYA is associated with improved cardiac function, enhanced mitophagy-related activity, and upregulated SIRT1-FOXO3-BNIP3 signaling, providing robust experimental evidence for its clinical translational application in MIRI treatment.