Current Medicinal Chemistry - Online First

An excessive inflammatory response plays a central role in the pathogenesis of ulcerative colitis (UC), but the specific cytokines involved remain unclear. This study aimed to identify inflammatory factors associated with UC and explore the possible mechanisms of the identified targets.

Protein quantitative trait loci (pQTLs) and expression quantitative trait loci (eQTLs) for inflammatory cytokines were obtained from a genome-wide pQTL study and the eQTL consortium, respectively. Summary data for UC from the exploration and validation cohorts were derived from a genome-wide association study and the Finngen cohort. MR and colocalization analyses were conducted to identify causal associations between inflammatory cytokines and UC. Bioinformatics analyses were employed to explore the involved biological processes of candidate targets. Immunohistochemistry was used to validate the expression of these candidate targets in colon tissues.

Among all inflammatory cytokines, a significant causal association was identified between C-X-C motif chemokine ligand 5 (CXCL5) and UC. Using eQTL data, a significant genetic association was established between the mRNA expression of CXCL5 and its receptor, C-X-C motif chemokine receptor 2 (CXCR2), with UC. Colocalization analysis further supported these identified links. Differential expression analysis confirmed the dysregulation of the CXCL5/CXCR2 axis in UC patients. Enrichment and immune infiltration analysis indicated that the CXCL5/CXCR2 axis was involved in neutrophil chemotaxis and immune activation in UC. Moreover, CXCL5 expression was found to correlate with neutrophil extracellular trap (NET) formation in UC. Immunohistochemistry further confirmed the dysregulation of the CXCL5/CXCR2 axis in colon tissues of UC patients.

The CXCL5/CXCR2 axis has been implicated to play a significant role within a broader inflammatory network that includes Interleukin (IL)-17, NF-κB, and Tumor Necrosis Factor (TNF) signaling pathways. Additionally, this axis interacts with macrophages and T cells, further contributing to the complexity of inflammatory responses in UC.

There is a significant association between CXCL5/CXCR2 and UC under the MR assumption, which is potentially linked with colonic chemotaxis and activation of neutrophils. These findings highlight the potential of CXCL5/CXCR2 as a therapeutic target for UC. However, future functional studies are needed to validate these findings and explore the exact mechanisms by which CXCL5/CXCR2 influences immune cell crosstalk in UC.

Spirooxindoles have been reported to be effective anticancer drug candidates by displaying promising pre-clinical results. Therefore, to find out a lead spirocyclic oxindole template, a series of spirooxindole derivatives bearing pyrrolizidine (14a-e) and N-methyl pyrrolidine (15a-e) were synthesized using an efficient multicomponent, one-pot, and stereoselective [3+2] cycloaddition reaction and evaluated in vitro against HT29 and HCT116 human colon cancer cell lines.

The pyrrolizidine and N-methyl pyrrolidine spirooxindole derivatives were synthesised in excellent regio- and stereoselectivity using previously optimized reaction conditions. They were evaluated in vitro against cell lines HT29 and HCT116. In silico ADME profiling, molecular docking, and dynamics simulation studies were performed to ascertain the probable mode of action of the lead derivative.

The spirooxindoles were characterized using FTIR, ESI-MS, ¹H and ¹³C NMR, purity was determined by RP-HPLC, and stereochemistry was confirmed by X-ray crystallography. Compound 14a produced the best anti-colon cancer activity with IC50 values of 62.66 and 9.55 µM against HT29 and HCT116 human colon cancer cell lines, respectively. The in silico studies revealed that MDM2 protein inhibition is a probable mode of anti-colon cancer activity, supported by the data obtained in the molecular docking and molecular dynamics study.

The described [3+2] cycloaddition reaction proved to be a highly efficient and catalyst-free reaction. The in vitro cell viability assays and in silico studies revealed that more spirooxindoles can be designed with a varied degree of substitution to target colon cancer.

This study investigates the prognostic value of Epidermal Growth Factor (EGF) family genes in Cervical Cancer (CC) and experimentally validates the role of AREG in the progression of CC.

Transcriptome and clinical data of CC were obtained from the TCGA database. We constructed a prognostic model using LASSO Cox regression analysis based on candidate EGF family genes. Multiple bioinformatics approaches were employed to analyze functional pathways and immune characteristics. The biological function of Amphiregulin (AREG) was validated through in vitro experiments, including colony formation, CCK8 proliferation assay, wound healing assay, transwell assay, macrophage polarization analysis using the co-culture system, and in vivo subcutaneous tumor formation in nude mice. Combination therapy with anti-AREG and anti-PD-L1 antibodies was evaluated in a murine C57BL/6 model.

We identified 116 EGF family-related genes associated with CC progression and established a prognostic model. High-risk and low-risk groups showed distinct functional enrichment patterns and immune characteristics. AREG emerged as a key prognostic factor, with significantly elevated expression in CC cells. Knockdown of AREG suppressed CC cell proliferation, migration, and invasion, potentially through modulating Epithelial-Mesenchymal Transition (EMT). AREG promoted M2 macrophage polarization, fostering an immunosuppressive tumor microenvironment. Anti-AREG antibody treatment demonstrated antitumor effects in vitro and in vivo, synergizing with anti-PD-L1 therapy to significantly inhibit tumor growth and reverse EMT.

Our findings establish the first EGF family-based prognostic model for CC and reveal AREG's dual role in promoting EMT and reshaping the immune microenvironment. The observed synergy between AREG inhibition and PD-L1 blockade provides mechanistic insights for overcoming immunotherapy resistance. Limitations include retrospective data analysis and a lack of multi-omics validation.

Our study establishes a robust EGF family gene-based prognostic model for CC patients and identifies AREG as a promising therapeutic target. These findings provide new insights for CC prognosis assessment and treatment strategies.

The global incidence of colon cancer is rising, highlighting the need for complementary therapeutic approaches using natural products such as citral. A self-nano-emulsifying drug delivery system incorporated with citral (CIT-SNEDDS) was formulated, and prior studies have demonstrated its potent antiproliferative effects on colon cancer cell lines.

The apoptosis-inducing ability of CIT-SNEDDS treatment on SW620 cells was evaluated using Acridine Orange/Propidium Iodide (AO/PI) assay, Annexin V-FITC assay, and cell cycle analysis by flow cytometry. Scratch assay and migration, and invasion assays were performed to assess its anti-metastatic effects.

The cytotoxicity assay results showed that SNEDDS with citral (CIT-SNEDDS) significantly reduced cell viability in a dose-dependent manner compared to free citral and SNEDDS without citral. Acridine orange/propidium iodide staining and Annexin V assay results confirmed apoptosis in CIT-SNEDDS-treated cells. Cell cycle analysis indicated that CIT-SNEDDS induced arrest at the S and G2/M phases, which may contribute to apoptosis initiation. The scratch and trans-well assays demonstrated a reduction in SW620 cell migration and invasion capabilities following CIT-SNEDDS treatment, suggesting a potent anti-metastatic effect.

The ability of CIT-SNEDDS to induce apoptosis, disrupt the cell cycle, and inhibit cellular migration in cancer cells aligns with the goals of targeted cancer therapies, which aim to selectively eradicate cancer cells while minimizing effects on healthy tissue.

These findings highlight the therapeutic potential of CIT-SNEDDS for enhancing the efficacy of citral as an anti-tumor and antimetastatic agent for colorectal cancer, warranting further in vivo and preclinical studies to optimize its application.

Teixobactin (TX) is a new class of antibiotics with a unique structure and strong efficacy against gram-positive bacteria. It is a “head-to-side-chain” cyclodepsipeptide with considerable potential as a lead molecule for creating novel antibiotics to combat multidrug-resistant pathogens.

In this study, we systematically design, synthesize, and evaluate modified Teixobactin analogs (TX1-TX5) for antimicrobial activity. This study presents a novel peptide derived from linearized Teixobactin, with amino acid substitutions at aa1 (N-Me-D-Phe-OH), aa5 (H-L-allo-Ile-OH), and the exclusion of L-allo-Enduracididine at aa10, synthesized using solid-phase peptide synthesis. We employed various software tools, including Molinspiration and SwissADME, to estimate the pharmacokinetic features of the synthesized TX analogs. Molecular docking studies were performed using AutoDock Vina, and PyMOL and Biovia Discovery Studio visualizer were utilized to visualize the protein-ligand interactions. The molecular structures of TX and TX analogs were modeled using the Sinapsis software.

Antimicrobial susceptibility tests against Staphylococcus aureus, Bacillus subtilis, E. coli, Pseudomonas sp., Aspergillus niger, and Fusarium sp. identified novel TX analogs exhibiting strong bactericidal and fungicidal activity at 80 μg/mL. Bacterial cell wall lysis assays confirmed significant cell wall breakdown upon TX analog treatment. Glucose assay results indicate reduced glucose uptake in bacterial cells treated with TX analogs. Docking studies revealed that the synthesized TX analogs exhibited good binding affinity, ranging from -5.0 to -12.5 kcal/mol, compared with bacterial and fungal proteins, as well as the Delafloxacin and Ketoconazole standards. Density Functional Theory (DFT) computations were employed to investigate chemical reactivity descriptors.

In-vitro studies indicated that TX1 and TX3 showed excellent bactericidal activity by forming inhibition zone diameters (mm) from 6.49 ±0.31 to 11.50 ±0.59 at 70 and 80 μg/mL concentrations against Staphylococcus aureus, Bacillus subtilis, E. coli, and Pseudomonas sp., compared to the standards Streptomycin (+ve) and DMSO (-ve). The TX2, TX3, and TX5 exhibit excellent fungicidal activity with inhibition zone diameters (mm) from 7.23 ±0.25 to 10.23 ±0.30 at 70 and 80 μg/mL concentrations against Aspergillus niger and Fusarium sp., compared to the standards ketoconazole (+ve) and DMSO (-ve). The bacterial cells treated with TX1 displayed more dead cells than the control in all bacterial strains, indicating excellent cell lysis.

Mass, ¹H NMR, and HPLC analysis characterized the synthesized fine TX analogs (TX1-TX5). The DFT and docking studies' electronic characteristic calculations predicted that halogenated (TX1, TX2, and TX4) and methoxy (TX3) substituted analogs have higher stability and electrophilicity, making them suitable agents for antimicrobial activity.

Acute respiratory distress syndrome (ARDS) is a life-threatening condition associated with high mortality and morbidity. However, targeted therapies that effectively improve patient outcomes remain limited. Exosomes play pivotal roles in intercellular communication and epigenetic regulation.

This study aimed to identify exosome-related differentially expressed genes (EXORDEGs) in whole blood associated with ARDS and to explore their potential mechanistic roles in the disease.

Two gene expression datasets (GSE32707 and GSE66890) were retrieved from the Gene Expression Omnibus for comprehensive bioinformatics analysis. Analytical approaches included Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses, protein-protein interaction network construction using the STRING database, and immune infiltration profiling via single-sample gene set enrichment analysis in relation to hub genes.

We identified 21 EXORDEGs, primarily enriched in biological processes such as endothelial cell development and apoptosis. Four hub genes—PI3, EEF1A1, ANAPC1, and PSMD2—were robustly associated with ARDS, with PSMD2 showing the most pronounced differential expression. Immune infiltration analysis revealed significant disparities in nine immune cell populations between ARDS and control samples.

The results of this comprehensive bioinformatics analysis identified four EXORDEGs—PI3, EEF1A1, ANAPC1, and PSMD2—with important roles in acute respiratory distress syndrome.

This study first systematically identified EXORDEGs in ARDS, discovering four hub genes and their associations with immune cells. The hub genes may be implicated in endothelial injury, inflammation, and immune dysregulation. These findings provide novel insights into ARDS pathogenesis and highlight potential therapeutic targets for further investigation. Given the disease heterogeneity, our findings primarily reflect common molecular characteristics, while the specific features of different etiological subtypes require further investigation.

Lung cancer remains a major global health burden with high mortality rates and limited therapeutic options. Natural flavonoids, particularly those derived from Cassia species, have shown immunomodulatory and anticancer potential. This study investigates the therapeutic promise of selected Cassia-derived flavonoids targeting key lung cancer-associated proteins: Prostaglandin-endoperoxide synthase 2 (PTGS2), Mast/stem cell growth factor receptor (KIT), and Xanthine dehydrogenase (XDH).

Eight flavonoids were selected based on literature and database-reported bioactivity. Target prediction was performed using SwissTargetPrediction and STITCH, followed by pathway enrichment via STRING and KEGG databases. Molecular docking was conducted using AutoDock Vina against PTGS2 (PDB: 5IKQ), KIT (1N5X), and XDH (4U0I). Top-ranked complexes underwent 100 ns molecular dynamics (MD) simulations with GROMACS to assess binding stability, RMSD, and conformational behavior. Drug-likeness, ADME, and toxicity profiles were evaluated using SwissADME and ProTox-II. Standard drugs (Trametinib, Nivolumab, Erlotinib) were used for comparison.

Epicatechin and Hispidulin showed the strongest binding affinities with PTGS2 (−9.04 kcal/mol) and XDH (−8.22 kcal/mol), respectively, with stable RMSD profiles. Chrysoeriol demonstrated the highest binding to KIT (−8.68 kcal/mol), outperforming Nivolumab (−6.03 kcal/mol). All selected flavonoids displayed acceptable pharmacokinetic profiles and low predicted toxicity. MD simulations confirmed the dynamic stability of key complexes.

Cassia-derived flavonoids represent promising multi-target candidates for lung cancer therapy, particularly through modulation of PTGS2, KIT, and XDH. Their favorable interaction profiles and safety predictions warrant further experimental and in vivo validation.

Non-Small Cell Lung Cancer (NSCLC) treatment is often challenged by drug resistance. The antihypertensive drug telmisartan has shown anti-tumor potential, but its underlying mechanism remains unclear. Ferroptosis, a newly identified form of cell death, may serve as a promising therapeutic target. The objective is to investigate whether telmisartan inhibits NSCLC by inducing ferroptosis and to elucidate its underlying mechanism.

in vitro cell assays and in vivo mouse models were used, along with molecular biology techniques, to evaluate the effects of telmisartan on NSCLC and its mechanism of action.

Telmisartan significantly suppressed NSCLC cell proliferation and tumor growth. Mechanistic studies revealed that telmisartan induced ferroptosis by inhibiting the nuclear translocation of Nuclear Factor Erythroid 2-Related Factor 2 (NRF2) and downregulating Glutathione Peroxidase 4 (GPX4) expression. The anti-tumor effect of telmisartan was reversed by ferroptosis inhibitors.

Telmisartan can inhibit the proliferation of NSCLC cells in vitro and in vivo and induce cell ferroptosis. Telmisartan can also inhibit the nuclear translocation of NRF2, thereby affecting the expression of GPX4.

Telmisartan inhibited NSCLC by inducing ferroptosis via the NRF2/GPX4 axis, offering a new therapeutic strategy and potential clinical application for NSCLC treatment.

Neutrophils play a key role in host immune defense. At present, neutrophils in chronic bone infections exhibit significant heterogeneity but functional alterations that remain poorly understood.

A rat model of chronic bone infection induced by Methicillin-Resistant Staphylococcus Aureus (MRSA) was established. Bone marrow cells were analyzed using scRNA-seq with Gene Ontology (GO) and pathway enrichment analysis. Differentially Expressed Genes (DEGs) were identified to assess neutrophil dysfunction, validated by immunofluorescence staining and ROS quantification.

MRSA-induced chronic bone infection was confirmed by Gram and H&E staining, which showed bacterial colonization and inflammation. Neutrophils from infected rats showed downregulated immune-related genes (e.g., Clec7a, Ccr5) and upregulated immunosuppressive factors (e.g., Nfkbia, IL10ra). Enrichment analysis showed that immune responses and neutrophil functions were inhibited. Immunofluorescence showed neutrophil polarization towards N2 phenotype and reduced Reactive Oxygen Species (ROS) production in the infection group.

This study established a rat model of MRSA-induced chronic bone infection and identified 7 neutrophil subsets via scRNA-seq analysis, with the NeuP2ry10 subset showing the most significant changes. Neutrophils displayed decreased chemotaxis, phagocytosis, and ROS production, along with elevated anti-inflammatory gene expression, suggesting functional suppression and a shift toward an immunosuppressive state.

Chronic bone infection drives neutrophil polarization toward an N2 anti-inflammatory phenotype, reducing antimicrobial capacity and promoting infection persistence. Targeting neutrophil function may offer new therapeutic strategies for chronic bone infection.

The objective of the present study was to explore the bi-directional causal relationship between IPF and diabetes (type 1 diabetes and type 2 diabetes)/diabetic nephropathy/glycemic traits [fasting glucose and glycated hemoglobin (HbA1c)]/fasting blood insulin through MR analysis.

A bi-directional two-sample Mendelian randomization (MR) study design was adopted to evaluate the causal relationship between IPF and diabetes (type 1 diabetes and type 2 diabetes), diabetic complications (diabetic nephropathy) and glycemic traits [fasting blood glucose, glycated hemoglobin (HbA1c), fasting insulin] in a European population. Genome-wide association study summary data was obtained. The inverse variance weighted (IVW) method with a fixed-effects model was used to estimate the primary causal effects. The causal effects are represented by reporting odds ratios (OR) and their corresponding 95% confidence intervals (CI). The robustness of results was assessed using the MR-Egger and Weighted Median methods.

In the forward MR analysis, the IVW method revealed a significant causal effect of IPF on type 2 diabetes (OR=1.031, 95% CI: 1.011-1.052). Similar estimates were obtained through the Weighted Median method. However, no significant causal effects were observed on type 1 diabetes, diabetic nephropathy, fasting blood glucose, HbA1c, and fasting insulin, respectively (p>0.05). We performed the reverse MR analysis using similar methods to the forward MR approach. MR analysis only showed a significant causal association of fasting insulin with IPF risk, with an OR of 3.576 (95% CI: 1.958-6.531).

Genetically determined IPF was linked to an elevated risk of type 2 diabetes. The inverse MR analysis indicated that there was no causal impact of genetically predicted type 2 diabetes on the IPF risk.

Genetically predicted fasting blood insulin was found to be positively associated with IPF risk.

Osteoporosis (OP) and aortic stenosis (AS) are highly prevalent age-related disorders that frequently coexist. Epidemiological studies suggest a pathological link between OP and AS beyond age, yet the molecular mechanisms underlying this bone-vascular axis remain poorly defined. This study aimed to identify shared genes and pathways contributing to the comorbidity of OP and AS.

Publicly available AS and OP transcriptomic datasets were retrieved from the GEO database. Weighted gene co-expression network analysis (WGCNA) and differential gene expression (DEG) analysis were conducted to identify disease-associated genes. Candidate hub genes were screened through protein-protein interaction (PPI) network analysis using twelve network topology algorithms. High-confidence genes were obtained by intersecting candidates with AS-related genes from the Comparative Toxicogenomics Database (CTD). Independent cohorts were used to validate candidate genes, and least absolute shrinkage and selection operator (LASSO) regression was performed to assess their diagnostic potential.

WGCNA revealed 665 shared genes enriched in immune and inflammatory processes, cell adhesion, and glycosaminoglycan biosynthesis. PPI network analysis identified 32 candidate hub genes, and integration with CTD yielded 15 high-confidence genes. Validation across independent datasets confirmed dysregulated expression of CD4, GZMB, and SDC1 in both AS and OP samples. ROC analysis demonstrated high diagnostic accuracy of these genes, with a combined AUC of 0.94.

These findings highlight immune and inflammatory pathways as convergent mechanisms driving both AS and OP. The hub genes CD4, GZMB, and SDC1 participate in immune regulation and extracellular matrix remodeling, suggesting their involvement in the shared pathogenesis of skeletal and cardiovascular degeneration.

Integrative bioinformatics identified CD4, GZMB, and SDC1 as key genes linking OP and AS, providing potential biomarkers and therapeutic targets for managing these age-related comorbidities.

This study aimed to explore potential causal relationships between mitochondria-related genes and irritable bowel syndrome (IBS) using integrative multi-omics analysis.

Genome-wide association study data for IBS (1,480 cases and 454,868 controls) were integrated with mitochondrial gene data from DNA methylation quantitative trait loci, blood expression quantitative trait loci, and protein quantitative trait loci. Molecular trait associations with IBS were assessed through summary-based Mendelian randomization and co-localization analyses. Steiger filtering analysis was applied to identify causal directions, and candidate genes were independently replicated by two-sample MR in the FinnGen R11 cohort.

Three primary genes supported by multi-omics evidence—CASP3, GATM, and PDK1—were identified. Increased CASP3 methylation, expression, and protein were positively associated with IBS risk, indicating pro-apoptotic and pro-inflammatory mechanisms, whereas elevated GATM expression and protein were negatively associated, consistent with a protective role via creatine-mediated energy homeostasis.

Additionally, 19 genes were classified as secondary evidence genes and 5 as tertiary evidence genes. Among these, genes such as ACAD10 and MSRA were validated using FinnGen data.

This study represents the first application of multi-omics techniques to elucidate the relationship between mitochondrial genes and IBS. The findings indicate multiple candidate pathogenic genes and highlight the role of mitochondrial dysfunction in IBS pathogenesis. These findings offer new opportunities for the discovery of IBS biomarkers and the development of therapeutic strategies.

Curcuma longa, commonly known as turmeric, contains curcumin, which is its main compound and has been reported to possess a wide variety of pharmacological activities, such as anti-carcinogenic, anti-malarial, antioxidant, antibacterial, anti-mutagenic, anti-inflammatory, and immunomodulatory effects. Even though it has many strong biological properties, curcumin lacks solubility, which affects its clinical efficacy. DK1 is a curcumin analogue that has been found to possess selective cytotoxicity on breast cancer cells compared to normal breast cells; however, its effectiveness in colon cancer has yet to be validated. This study was performed to investigate the effects of DK1 on colon cancer using an in vivo model in terms of its anti-apoptotic, immunoregulatory, and antioxidant potential. The pathways affected by the DK1 treatment were also evaluated.

In this study, male BALB/c mice induced with colon cancer were utilized, and the resulting tumours and spleen were subjected to TUNEL, immunophenotyping, and several antioxidant assays, such as nitric oxide, malondialdehyde, and superoxide dismutase, as well as gene and protein expression analyses.

K1 treatment led to tumor shrinkage, an increase in apoptotic tumor cells, and elevated populations of helper and cytotoxic T cells by 5% and 3%, respectively. Besides that, the NO and MDA levels were also significantly reduced. This study also observed dysregulations in several oncogenes in the VEGF pathway, such as CMYC, iNOS, and IL-1β genes, which are involved in angiogenesis and inflammation.

The effects of DK1 treatment included antitumor and anti-inflammatory properties against the inoculated CT26 tumour. DK1 showed potential in regulating the inflammation via the VEGF pathway by the significant downregulation of TNF-α and IL-1β pro-inflammatory genes, as well as PTX3, OPN, and serpin-E1 pro-angiogenic proteins.

The results suggested that DK1 may potentially function as an immunoregulator and anti-cancer agent for colon cancer therapy.

Ischemic stroke (IS) is a major cause of death and disability worldwide. The transcriptional mechanism of neutrophil extracellular trap-related genes (NRGs) and their diagnostic potential remain unknown. This study aims to explore the mechanism of NRGs in IS through machine learning and single-cell RNA sequencing (scRNA-seq).

We conducted differential analysis and functional enrichment analysis on the GEO dataset. Machine learning algorithms were used to identify NRGs related to IS. ScRNA-seq analysis was employed to verify the expression of NRGs in different cell types, and cellchat was used to explore the interactions between cell types in the IS. The expression of Eno1 was also verified in the mouse model of middle cerebral artery occlusion (MCAO).

We identified 26 differentially expressed NRGs (DE-NRGs). The diagnostic models constructed from five DE-NRGs (ENO1, HMGB1, ILK, ORAI1, SUCNR1) demonstrated high predictive ability. Single-cell analysis revealed that NRGs were highly expressed in the IS group. The experiment verified the significant upregulation of Eno1.

This study employed machine learning and scRNA-seq to identify the DE-NRGs-related diagnostic model, providing a certain theoretical basis for IS risk stratification. More experiments are needed to verify the role of DE-NRGs in IS in the future.

This study identified DE-NRGs with diagnostic capabilities in IS and verified their high expression through scRNA and experimental methods. DE-NRGs may be potential therapeutic targets for IS.

Ischemic Stroke (IS) represents the most prevalent subtype of cerebrovascular disease, characterized by complex pathophysiological mechanisms that remain inadequately characterized, particularly concerning mitochondrial dysfunctions. These mitochondrial impairments are increasingly recognized as contributory factors in IS pathogenesis, emphasizing the need for further investigation into the underlying molecular mechanisms involved.

In this study, we integrated transcriptomic datasets from the Gene Expression Omnibus (GEO) with the comprehensive MitoCarta3.0 mitochondrial proteome inventory to elucidate the role of dysregulated Mitochondrial-Related Genes (MRGs) in IS. We employed an advanced bioinformatics and machine learning pipeline, incorporating differential expression profiling alongside network-based prioritization using CytoHubba. Rigorous feature selection was conducted through LASSO regression, Support Vector Machine (SVM), and Random Forest (RF) algorithms to derive a robust core MRG signature. Our methodology included training and validation cohorts to construct diagnostic models, which were critically evaluated via Receiver Operating Characteristic (ROC) curves, nomograms, and calibration analyses.

Our analysis identified a seven-gene signature comprising DNAJA3, ACSL1, HSDL2, ECHDC2, ECHDC3, ALDH2, and PDK4, which demonstrated significant correlation with activated CD8⁺ T-cell and natural killer cell infiltration. Furthermore, integrative network analyses revealed intricate regulatory interactions among MRGs, microRNAs, and transcription factors. Notably, drug-target predictions indicated Bezafibrate as a promising therapeutic agent for modulating mitochondrial homeostasis in the context of IS.

These findings offer a novel framework for ischemic stroke diagnosis and therapy, yet their computational derivation underscores the need for thorough experimental validation of MRGs and drug candidates, along with the integration of diverse clinical data to confirm their real-world applicability.

Our findings underscore mitochondrial dysfunction not only as a critical factor in IS pathogenesis but also as a viable therapeutic target. The identified MRG signature presents potential for clinical application in diagnostic and pharmacological strategies aimed at ameliorating ischemic injury. This study highlights the translational significance of systems biology approaches within cerebrovascular medicine, warranting further mechanistic exploration of mitochondrial-immune interactions in stroke pathology.

Gliomas are aggressive brain tumors with a poor prognosis and high recurrence. Oridonin, a traditional Chinese medicine, has shown potential in treating various cancers, but its role in glioma treatment, especially in modulating Epithelial-Mesenchymal Transition (EMT), remains underexplored.

We identified 371 potential target genes of Oridonin using various bioinformatics databases. Enrichment analyses, including Differential Expression Analysis, Gene Set Enrichment Analysis (GSEA), and Weighted Gene Co-expression Network Analysis (WGCNA), were performed to link these targets to glioma characteristics. in vitro experiments validated Oridonin's impact on EMT-related gene expression in glioma cells.

Enrichment analyses identified 19 common genes between Oridonin and glioma targets, with 12 EMT-related core genes. KEGG enrichment highlighted PI3K-Akt, MAPK pathways, and glioma pathways, while DO enrichment included high-grade gliomas. CCK8 assay showed Oridonin IC50 values of 6.92 μM for H4 and 10.54 μM for SW1783 glioma cell lines. WB results indicated increased E-Cadherin and decreased Vimentin, N-Cadherin, and Snail expression after Oridonin treatment. PPI network and single-cell transcriptome analyses identified key genes linked to glioma progression and immune cell infiltration.

Oridonin may inhibit glioma progression by targeting EMT-related pathways like PI3K-Akt and MAPK. The upregulation of E-Cadherin and downregulation of Vimentin, N-Cadherin, and Snail suggest a reversal of the EMT process. Future work should validate these effects in vivo and explore Oridonin's ability to cross the blood-brain barrier.

Oridonin may provide a novel therapeutic approach for glioma by targeting EMT-related pathways, offering a foundation for further clinical investigation.

Neuroinflammation, primarily mediated by activated microglia, is a significant contributor to neurodegenerative diseases, such as Alzheimer's and Parkinson's disease. Quercetin (QCT), a dietary flavonoid, has demonstrated anti-inflammatory and neuroprotective properties; however, the detailed molecular mechanisms behind these effects remain unclear. This study aimed to investigate the anti-inflammatory actions of QCT, particularly focusing on its potential to suppress the activation of microglia and subsequent neuroinflammation.

BV2 microglial cells were stimulated with lipopolysaccharide (LPS) to induce an inflammatory response and were subsequently treated with various concentrations of QCT. Cell viability was assessed using the MTT assay. Levels of pro-inflammatory cytokines (IL-6, TNF-α) and nitric oxide (NO) were quantified through ELISA and Griess reaction methods, respectively. Western blot analysis was conducted to examine inducible nitric oxide synthase (iNOS), NF-κB, IκBα, and phosphorylated IκBα protein expressions. In silico approaches, including protein-protein interaction (PPI) network analysis and molecular docking, were employed to explore potential molecular mechanisms involving NF-κB signaling pathways.

Treatment with QCT significantly reduced the secretion of IL-6 (96%) and TNF-α (87%), as well as NO production (42%), in a dose-dependent manner. Western blot results demonstrated a marked reduction in iNOS expression and inhibition of NF-κB activation through reduced phosphorylation of IκBα following QCT treatment. Molecular docking indicated a strong binding affinity between QCT and IKKβ, suggesting inhibition of the NF-κB pathway.

The findings indicated QCT to exert potent anti-inflammatory effects in LPS-stimulated BV2 cells by modulating key proteins involved in the NF-κB signaling pathway. Specifically, the docking results implied QCT’s direct interaction with the catalytic subunit IKKβ, inhibiting IκBα phosphorylation and subsequent NF-κB activation. The results have been found to be consistent with previous literature, reinforcing QCT's role in reducing neuroinflammation through specific molecular targets and pathways. Further in vivo studies are necessary to validate the findings.

Quercetin effectively suppressed neuroinflammation in microglial cells through inhibition of the NF-κB signaling pathway, reducing levels of critical pro-inflammatory mediators. The outcomes have highlighted the potential of quercetin as a preventive nutraceutical for neurodegenerative diseases, necessitating future in vivo investigations to confirm its therapeutic efficacy.

The causality between thyroid disorders and Gastroesophageal Reflux Disease (GERD) remains to be deciphered. This two-sample Mendelian Randomization (MR) study was performed to elucidate the causal association between GERD and thyroid diseases and functions.

Summary statistics for GERD were retrieved from a published GWAS dataset deposited in the Integrative Epidemiology Unit OpenGWAS database. Thyroid hormone level data were obtained from the ThyroidOmics Consortium, and genetic variants associated with thyroid disorders were sourced from the FinnGen Project. MR statistical analyses used the Inverse Variance Weighted (IVW) algorithm, followed by various sensitivity and reliability analyses. Odds Ratio (OR) and beta coefficient (β) with 95% Confidence Interval (CI) were estimated for categorical and continuous outcomes, respectively. The significant causal association was determined based on a Bonferroni-corrected threshold of p-value < 0.0021 (calculated as 0.05/24 trait pairs).

The findings of MR analysis tend to favor the causality of GERD for hyperthyroidism (IVW: OR = 1.517, 95% CI: 1.164 to 1.978, p = 2.04E-03) but not the other thyroid disorders. The reverse MR estimates suggested that thyroid disorders may not affect the susceptibility of GERD. Moreover, genetic proxied GERD was significantly negatively associated with circulating Thyroid Stimulating Hormone (TSH) level (IVW: β = -0.048, 95% CI: -0.078 to -0.019, p = 1.17E-03), whereas the causality of this enteropathy on Free Triiodothyronine (FT3), Free Thyroxine (FT4), Total Triiodothyronine (TT3), FT3/FT4 ratio, and TT3/FT4 ratio (and vice versa) is unfounded.

This MR study indicates that the genetic liability to GERD is significantly detrimental to hyperthyroidism risk and the homeostasis of TSH.

The findings suggest that effective GERD management could mitigate hyperthyroidism risk.

Diffuse large B-cell lymphoma (DLBCL) pathogenesis is poorly understood, with limited causal evidence linking circulating inflammatory proteins (CIPs) and metabolites to disease risk. Observational studies face challenges from confounding and reverse causation, while existing Mendelian Randomization (MR) analyses lack bidirectional designs and multi-omics integration.

A bidirectional two-sample MR design was applied using inverse-variance weighting (IVW). Genetic instruments for 91 CIPs derived from Olink proteomic data (14,824 participants). DLBCL genetic associations (1,050 cases; 314,193 controls) were obtained from FinnGen (R10 release). Data for 1,091 blood metabolites and 309 metabolite ratios were sourced from the GWAS Catalog.

Ten CIPs exhibited causal effects on DLBCL. Risk-increasing proteins included: IL-10 (OR=1.46, 95%CI=1.05-2.03), TSLP (1.37,1.01-1.84), IL-17C (1.34,1.05-1.72), NRTN (1.30,1.02-1.66), OPG (1.29,1.01-1.66), and MCP1 (1.26,1.04-1.52). Protective proteins included: CD40 (0.82,0.67-1.00), CXCL9 (0.78,0.61-0.98), CD5 (0.77,0.61-0.97), and MCP3 (0.76,0.58-0.99). Reverse causation was absent for 7 proteins. Mediation analysis revealed 17.2% (p=0.048) of CD5’s protective effect was mediated by 1-methylhistidine.

These findings establish CIPs as causal factors in DLBCL pathogenesis and identify metabolite-mediated pathways as novel mechanistic links. The bidirectional design and multi-omics integration overcome key limitations of prior research, though statistical power for some mediation tests was limited by metabolite GWAS sample sizes.

Plasma inflammatory proteins causally influence DLBCL risk, partially mediated by metabolites. This underscores metabolite pathways as potential targets for therapeutic intervention.