Combinatorial Chemistry & High Throughput Screening - Online First

DNA microarray synthesis enables the large-scale and precise generation of DNA sequences for genomic research, data storage, and synthetic biology. However, the order of nucleotide addition significantly affects synthesis efficiency and accuracy. This study aims to model DNA microarray in situ synthesis as a traveling salesman problem (TSP) and to develop an optimized synthesis strategy.

A mathematical model for in situ microarray synthesis was established, and both greedy algorithms and a simulated annealing algorithm were applied to optimize the nucleotide addition order. The performance of these approaches was evaluated by comparing the number of synthesis cycles required at different sequence scales, ranging from 10 × 10 nt to 10000 × 120 nt arrays.

The optimized synthesis schemes effectively reduced the total number of synthesis cycles. At the 10 × 10 nt scale, simulated annealing reduced cycles by 40.65% compared to the traditional scheme and by 8.52% compared to the greedy algorithm. At larger scales (100 × 100 nt to 10000 × 120 nt), cycle reductions ranged from 33.80% to 37.26%, with simulated annealing outperforming the greedy algorithm by 2.68% to 3.42%. These reductions translated into significant savings in synthesis time, reagent consumption, and overall cost.

The simulated annealing–based optimization strategy demonstrates clear advantages in improving DNA microarray synthesis efficiency while reducing material usage and waste, thereby enhancing cost-effectiveness. Such improvements offer practical benefits for applications, including gene editing, drug development, and DNA data storage.

Endometriosis (EMs), a prevalent disorder characterized by pelvic pain and infertility, affects numerous bodily systems and markedly diminishes life quality. Yiqi Huoxue formula (YQHXF) has demonstrated promising therapeutic efficacy. However, its active substances and underlying mechanisms remain ambiguous.

An innovative methodological framework incorporating NP, transcriptomics, proteomics, and molecular biology was utilized to investigate the active components and mechanisms of YQHXF. A network pharmacological analysis was conducted to identify the targets, biological processes, and pathways associated with YQHXF’s effects on EMs. Additionally, transcriptomics, proteomics, and molecular biology techniques were applied for further mechanistic exploration at both the gene and protein levels.

The findings suggest that YQHXF prevents EMs by modulating critical cellular processes—proliferation, invasiveness, adhesion, and apoptosis—within ectopic endometrial cells. The integration of network pharmacology, multi-omics, and molecular biology confirmed that this regulation occurs via key targets (EGFR, AKT1, FOS, MAPK3, NFKB1) and associated pathways.

This study employed an integrated approach combining transcriptomics, proteomics, and molecular biology to analyze the effects of YQHXF on EMs. A total of 180 direct targets, 128 indirect targets, and 19 pathways related to YQHXF’s anti-EMs effects were preliminarily identified. Based on these findings, it is proposed that YQHXF potentially achieves its remedial outcomes by influencing the proliferation, invasiveness, adhesion, and apoptosis of ectopic endometrial cells through the regulation of EGFR, AKT1, FOS, MAPK3, and NFKB1.

The findings suggest that YQHXF prevents EMs by regulating critical cellular processes, including the proliferation, invasiveness, adhesion, and apoptosis of ectopic endometrial cells.

Damp retention in the middle-jiao syndrome (DRMS), a common manifestation in Traditional Chinese Medicine (TCM), results from stagnation of damp pathogens in the middle jiao and impaired transport of food and fluids. Given the complex pathogenesis of DRMS, this study aimed to investigate its biological mechanisms using an advanced analytical approach.

A DRMS rat model was established based on three etiological factors: dietary disorders, depletion of vital qi, and excessive external dampness. Model validity was assessed via small intestinal carbon propulsion rate and histopathological examination. Urine metabolomics, using ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS), systematically characterized metabolic profile changes and potential biomarkers.

DRMS rats exhibited significantly reduced small intestinal propulsion, along with varying degrees of edema, disorganized tissue structures, and inflammatory cell infiltration in gastric, renal, and small intestinal tissues. Metabolomic analysis identified 52 differential metabolites as potential DRMS biomarkers, primarily involved in phenylalanine, tyrosine, and tryptophan biosynthesis, phenylalanine and tyrosine metabolism, the citrate cycle, and cysteine and methionine metabolism pathways. Metabolic correlation networks further validated the model’s accuracy.

The identified metabolites and pathways provide insight into the mechanisms underlying DRMS, complement existing TCM research, and offer a foundation for further studies. However, the findings are currently limited to the rat model and require human validation.

This study successfully established a DRMS animal model under clinically relevant TCM conditions and demonstrated the utility of metabolomics in elucidating DRMS mechanisms, providing experimental evidence for TCM syndrome characterization and advancing understanding of its etiology.

Huoxue San (HXS) is a traditional Chinese medicinal formulation widely used to treat bone fractures and blood stasis. Comprising seven herbs—Siphonostegia chinensis Benth, Kochia scoparia (L.) Schrad, Scutellaria barbata D.Don, Polygonum cuspidatum Sieb. et Zucc, Arisaema erubescens (Wall.) Schott, Phellodendron chinense Schneid, and Eupolyphaga sinensis Walker—HXS has been administered at Nanjing Chinese Medicine Hospital for over 50 years. It is effective in promoting fracture healing, supporting soft tissue repair, and rarely causing adverse reactions such as skin allergies. The present study aimed to elucidate the molecular mechanisms underlying HXS’s therapeutic effects.

Ultrahigh-performance liquid chromatography–quadrupole time-of-flight mass spectrometry (UHPLC–Q-TOF MS) was used to identify HXS components absorbed into the bloodstream. Network pharmacology, molecular docking, and molecular dynamics simulations were then conducted to explore the active ingredients and their regulatory mechanisms in fracture healing and blood stasis.

Transdermal absorption tests identified 20 active compounds from HXS. Network pharmacology analyses using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform highlighted vanillic acid, demethyleneberberine, palmatine hydrochloride, luteolin, apigenin, and wogonin as key active ingredients. Molecular dynamics simulations further validated the stability, conformational changes, and interactions of these compounds with their target proteins.

Analysis of the transdermal absorption samples revealed 291 potential active targets for HXS in treating fractures and blood stasis, of which 159 were common to both conditions. Protein–protein interaction (PPI) network analysis identified core targets including AKT1, ALB, EGFR, STAT3, and CTNNB1. Molecular docking confirmed strong binding interactions between HXS compounds and these core targets, while molecular dynamics simulations validated the stability and mechanistic plausibility of these interactions.

This study provides a systematic elucidation of HXS’s molecular mechanisms in fracture healing and blood stasis. Identification of active compounds, core targets, and their interactions offers a scientific basis for the therapeutic effects of HXS and supports the rational development of herbal-medicine-based interventions for fracture management and blood stasis treatment.

This study aimed to develop and validate a Cuproptosis-Related Gene (CRG) signature for predicting Biochemical Recurrence-Free Survival (BCRFS) and characterizing the Tumor Immune Microenvironment (TIME) in Prostate Cancer (PCa).

Transcriptomic and clinical data were collected from TCGA (n=405) and GEO (GSE70770, n=203). Consensus clustering based on 10 CRGs defined molecular subtypes. Differentially expressed genes between clusters were subjected to LASSO Cox regression to construct a prognostic signature in the TCGA cohort, followed by validation in GEO and combined cohorts. Quantitative real-time polymerase chain reaction (qRT-PCR) and Immunohistochemistry (IHC) were conducted for experimental validation.

Two CRG-based subtypes were identified, characterized by distinct clinicopathological features, immune checkpoint expression, and BCRFS. A six-gene signature (CALML5, MMP11, UBE2C, ANPEP, TMEM59L, COMP) stratified patients into high- and low-risk groups with significantly different BCRFS (log-rank P<0.001). The model showed good predictive accuracy (AUCs 0.717–0.837 at 1 year, 0.728–0.771 at 3 years, 0.683–0.695 at 5 years) and remained independent of clinicopathological factors. High-risk patients exhibited elevated immune/stromal scores, altered immune infiltration, and higher immune checkpoint expression. qRT-PCR confirmed upregulation of CALML5, MMP11, UBE2C, and COMP in PCa cell lines, while IHC validated differential protein expression of all six genes between PCa and BPH tissues (all P<0.05).

This six-gene CRG signature predicts BCRFS and reflects immune heterogeneity in PCa. Its integration into prognostic models may guide personalized management and inform immunotherapy strategies, warranting further validation in prospective clinical studies.

This study initially identified two cuproptosis-related molecules based on the expression patterns of cuproptosis-related genes. In addition, we developed a new cuproptosis-related molecular signature with great predictive performance for BCRFS and tumor immune environment using six DERRGs (including CALML5, MMP11, UBE2C, ANPEP, TMEM59L, COMP). These findings would be conducive to a deeper cognition of the potential mechanism of cuproptosis of PCa.

Icteric hepatitis remains a significant clinical challenge. The integration of Chinese Herbal Medicine (CHM) with conventional Western Medicine (WM) is becoming increasingly common; however, its overall efficacy and safety profile requires systematic evaluation. This study aims to conduct a meta-analysis to assess the safety and effectiveness of using CHM in conjunction with WM to treat icteric hepatitis.

We conducted a systematic search of PubMed, EMBASE, Cochrane Library, Scopus, Web of Science, and major Chinese databases (CNKI, Wanfang, VIP, CBM) from inception to December 2023 for Randomized Controlled Trials (RCTs) comparing CHM plus WM with WM alone for icteric hepatitis. Two reviewers independently performed study selection, data extraction, and quality assessment using the Cochrane Risk of Bias 2.0 tool. Meta-analysis was performed using RevMan 5.4 software.

Eight RCTs involving 645 participants were included. The combined therapy group demonstrated a significantly higher clinical efficacy rate (RR = 1.22, 95% CI [1.11, 1.34], P < 0.0001) compared to the WM alone group. The combined therapy also resulted in greater improvements in liver function, with significant reductions in ALT (WMD = -58.33 U/L, 95% CI [-87.75, -28.91]), AST (WMD = -47.11 U/L, 95% CI [-69.83, -24.39]), TBIL (WMD = -48.27 μmol/L, 95% CI [-67.48, -29.06]), and DBIL (WMD = -31.30 μmol/L, 95% CI [-45.16, -17.44]). Furthermore, the integrated approach led to lower levels of inflammatory markers (IL-6, CRP, TNF-α) and faster symptom resolution. There was no significant difference in the incidence of adverse events between the two groups (RR = 0.83, 95% CI [0.44, 1.57], P = 0.56).

The pooled evidence suggests that adding CHM to standard WM treatment can enhance therapeutic outcomes by improving liver function and reducing systemic inflammation. The synergistic effects may be attributed to the multi-target pharmacological properties of the herbs used. However, the findings are limited by the high risk of bias and significant heterogeneity across the included studies.

The adjunctive use of CHM with WM appears to be an effective and safe strategy for treating icteric hepatitis. Nonetheless, due to methodological weaknesses in the primary studies, these results should be interpreted cautiously. High-quality, rigorously designed RCTs are needed to confirm these findings.

Metabolic Dysfunction-Associated Steatohepatitis (MASH) is a growing global health concern, with only one FDA-approved therapy currently available. Acetyl-CoA carboxylase (ACC) inhibition has emerged as a promising strategy, yet effective and clinically translatable inhibitors remain limited. This study aimed to identify potential ACC inhibitors for MASH via drug repurposing.

A small-molecule library was screened using structure-based virtual screening, and candidate compounds were validated in a free fatty acid-induced MASH cell model. Intracellular triglyceride (TG) and aspartate aminotransferase (AST) levels were measured, while quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to evaluate lipid metabolism-related gene expression. Molecular dynamics simulations were conducted to further evaluate binding stability.

Lanatoside C was identified as the most potent candidate. In vitro studies revealed significant reductions in TG and AST levels, downregulation of lipogenesis-related genes (SREBP1, FASN, ACC), and upregulation of fatty acid oxidation genes (CPT1A, ACOX1, FABP1). Molecular dynamics simulations confirmed the stable binding of Lanatoside C to ACC.

These findings indicate that Lanatoside C exerts dual regulatory effects on lipid metabolism by suppressing fatty acid synthesis and enhancing oxidation. As an FDA-approved cardiac glycoside, Lanatoside C’s known pharmacological profile supports its potential repositioning for MASH, although further in vivo studies and mechanistic validation are warranted.

Lanatoside C demonstrates promise as a repurposed ACC inhibitor for MASH treatment, offering a cost-effective repurposing strategy to advance therapeutic options for MASH.

The key toxicological constituents and mechanisms of Epimedii Folium and its formulations, such as Xianling Gubao Capsules (XLGB) and Zhuanggu Guanjie Pills (ZGGJ), remain insufficiently understood, particularly when used in combination. The objective of this study is to investigate the hepatotoxic effects and mechanisms of Epimedii Folium and its formulations, XLGB and ZGGJ, using network toxicology, molecular docking, and in vitro validation.

Potential hepatotoxic components and targets of Epimedii Folium, XLGB, and ZGGJ were screened from multiple databases. PPI networks were constructed, and GO/KEGG enrichment analyses were performed. Molecular docking was used to assess the binding affinities between key components and core targets. In vitro validation was conducted using HepG2 cells to assess cell viability and ROS levels through CCK-8 and HCS assays, respectively.

This study confirms that Sagittatoside A, Epimedin B, and Icariside I are the primary hepatotoxic constituents of Epimedii Folium, capable of targeting core pathways involving KDR, AR, PTGS2, F7, and DPP4. Furthermore, Sagittatoside A and Icariside I significantly elevated ROS levels. The toxic constituents of XLGB and ZGGJ overlapped with those of Epimedii Folium, and Bavachinin and Neobavaisoflavone from PCL were found to exert synergistic hepatotoxic effects. Neobavaisoflavone enhanced the hepatotoxicity of Epimedin B and Icariside I, while Bavachinin showed synergistic toxicity when combined with Sagittatoside A.

Molecular docking confirmed strong binding affinities between these compounds and their targets. In vitro experiments demonstrated that Sagittatoside A and Icariside I significantly increased ROS levels. The compound formulations XLGB and ZGGJ shared similar hepatotoxic components and mechanisms. Additionally, Bavachinin and Neobavaisoflavone from PCL synergistically enhanced the hepatotoxicity of Epimedii Folium monomers, providing a modern scientific basis for evaluating compatibility principles in traditional Chinese medicine.

This study comprehensively elucidates the hepatotoxicity and synergistic toxic effects of Epimedii Folium and its formulations XLGB and ZGGJ, offering a modern scientific rationale to guide the safe formulation and compatibility of traditional Chinese medicine.

Longlutong Decoction (LLTD) is a Chinese traditional prescription used for coronary heart disease (CHD). The present study aimed to illuminate the mechanisms of LLTD treatment on CHD.

The therapeutic effect of LLTD on CHD was investigated using a CHD rat model. The chemical components of LLTD were identified, following which network pharmacology approaches were utilized to identify active components and disease-related targets. GO and KEGG analyses were conducted to explore potential molecular mechanisms. Finally, the molecular mechanism of LLTD treatment of CHD was verified.

Histopathological assessment revealed markedly attenuated myocardial injury severity in the medicated groups when compared to the model group. Moreover, 81 potential active ingredients were identified in LLTD, with 645 overlapping targets between component targets and disease targets. Network analysis identified Pinocembrin, Magnoflorine, Jatrorrhizine as key active ingredients, and AKT1, TNF, IL-6, STAT3, and Bcl-2 as primary core targets. A total of 1792 biological processes were affected according to GO analysis, and 187 pathways were identified through KEGG analysis. Finally, molecular docking and experimental results validated that LLTD could alleviate cardiomyocyte injury in CHD by regulating the primary core targets.

This study indicates that LLTD may achieve systematic modulating of the signaling network through a “network pharmacology” model, which provides valuable insights for the development of multi-target therapies targeting the complex pathological mechanism underlying CHD.

LLTD may exert cardioprotective effects by regulating inflammatory responses, apoptosis, and oxidative stress.

One of the most dangerous illnesses in the world today is hepatitis B virus (HBV) infection, which mainly affects the liver and can cause cirrhosis, hepatocellular carcinoma (HCC), and chronic infection. The primary goal of this study was to determine whether genotype polymorphisms at particular locations of the IL-18 promoter region may affect the host susceptibility to HBV infection in the North Indian population.

Genetic polymorphism of the IL-18 gene in the promoter region at positions -607 and -137 was performed in the North Indian population (100 controls and 100 HBV patients) using the PCR-RFLP method. Genotypic, allelic, and haplotype frequencies were compared using SHEsis software.

There were no significant differences in individual genotype or allele frequencies at positions -607 and -137 between HBV patients and controls. However, there were statistically significant differences with high frequencies of the -607A/-137C haplotype in HBV patients (p=0.010), whereas the -607C/-137C haplotype was more prevalent in controls (p=0.001), indicating a protective effect.

The individual SNPs did not show significant association, but specific haplotypes of the IL-18 promoter region may influence the risk of HBV infection. These results align partially with previous studies and suggest that haplotype-based analysis provides improved insight into genetic susceptibility.

The present study indicates that a double mutation (polymorphism) -607A/-137C in the IL-18 gene promoter region may contribute to the onset of HBV infection, while a single nucleotide polymorphism (-607C/-137C) may provide less susceptibility to HBV and may have a protective impact.

Migraine, a primary headache disorder, is the third disabling disease of neurological disorders worldwide. The pathological mechanism underlying migraine remains poorly understood. Lipid metabolism may be related to migraine pathophysiology. We aimed to investigate the causal relationship between plasma lipids and migraine or its subtypes, including migraine with aura (MA) and migraine without aura (MO), and explore whether the circulating cytokines serve as mediators in the pathway from plasma lipids to migraine.

A two-step Mendelian randomization (MR) approach was used to assess the mediating effect. The summary genetic data for 179 lipid species were obtained from were derived from a genome-wide association studies (GWAS) summary dataset encompassing 7,174 individuals. The summary genetic data for 91 circulating cytokines were obtained from genome-wide pQTL mapping data. The summary genetic data of GWAS related to migraine and its subtypes were derived from the FinnGen Release 10 database. The MR Analysis methods included the inverse-variance-weighted (IVW), MR-Egger, and weighted median.

The risk of migraine was reduced mediated by CD5 with phosphatidylinositol (18:1_18:2), sphingomyelin (d34:1), and sphingomyelin (d38:1). The risk of migraine and MA was reduced mediated by CD6 with sphingomyelin (d40:1) and sphingomyelin (d42:2). The risk of migraine and MA was increased mediated by CD6 with phosphatidylethanolamine (O-16:1_18:2).

CD5 and CD6 were found to be related to migraine. CD5 and CD6 may affect migraine by immunological dysregulation-induced neuroinflammation. Six plasma lipids are associated with two cytokines, indicating that lipid metabolism participates in neuroinflammation, and T cells may be part of it. Plasma levels of lipids were associated with the risk of migraine. The circulating cytokines may serve as mediators in the pathway from plasma lipids to migraine.

CD5 and CD6 appeared to mediate the causal relationship between plasma lipids and migraine or MA, including four kinds of sphingomyelin, one phosphatidylinositol, and one phosphatidylethanolamine.

Sleep disorders (SD) affect approximately 25% of the global population. Traditional Chinese Medicine (TCM) formulas have been shown to alleviate SD by modulating endogenous melatonin. This study used data mining, network pharmacology, and meta-analysis to identify key herbal pairs from TCM formulas and the mechanism of action.

Literature was retrieved from PubMed, Web of Science, Embase, Cochrane Library, CNKI, Wanfang Data Information Site, China Science and Technology Journal Database, and SinoMed. R was used for frequency and association rule analysis, SPSS for clustering, and Cytoscape, STRING, Gene Ontology, and KEGG enrichment analyses were utilized to explore targets, protein-protein interactions, and pathways. A meta-analysis using the Metan command was performed to assess the optimal herbal pairs for SD treatment.

Data mining identified 77 commonly used herbs, revealing four advantageous herbal pairs: PAEONIAE RADIX ALBA (PRA)-BUPLEURI RADIX (BR), COPTIDIS RHIZOMA (CR)-CINNAMOMI CORTEX (CC), PORIA (PA)-BUPLEURI RADIX (BR), and ZIZIPHI SPINOSAE SEMEN (ZSS)-MARGARITIFERA CONCHA (MC). Network pharmacology showed that (PRA-BR)-SD, (PA-BR)-SD, (CR-CC)-SD, and (ZSS-MC)-SD targeted CACNA1D, GRIN2A, AGT, and ATP1A1 via prion diseases, nicotine addiction, neuroactive ligand-receptor interaction, and cardiac muscle contraction pathways, respectively.

Research shows that CACNA1D could regulate Ca²⁺ inward flow, avoid mitochondrial dysfunction in prion diseases, and reduce ROS generation, thus indirectly maintaining MT levels and sleep. GRIN2A as an amygdala hub gene closely related to daily smoking, combining brain transcriptome analysis and tobacco consumption GWAS data. The sleep regulation mechanism of MT relies on the neuroactive ligand-receptor pathway. As a neuroactive ligand, MT triggers sleep-promoting physiological responses by activating the G-protein-coupled receptors MT1 and MT2 and transmitting “night” signals to the relevant neural networks. Insufficient MT secretion or circadian rhythm disruption might lead to abnormal blood pressure rhythms accompanied by sympathetic overactivation, increasing the risk of insomnia and cardiovascular disease. ATP1A1 is a key molecule in the maintenance of electrochemical gradients in cardiac myocytes through the modulation of the Na⁺/K⁺ homeostasis affects myocardial excitability, calcium kinetics, and contractile function.

Meta-analysis and network pharmacology suggest that the PA-BR pair might offer superior efficacy by modulating membrane potential and nicotine addiction pathways, targeting GRIN2A, GRIN1, GRIN3A, and GRIN2B to regulate melatonin levels.

Observational studies have linked diabetes with cataracts, but they cannot fully elucidate the underlying causes and mechanisms. This investigation aims to evaluate the causal relationship between genetically predicted diabetes and cataract risk utilizing Mendelian randomisation (MR) techniques.

We identified single nucleotide polymorphisms (SNPs) with a significant threshold of P < 5×10^-8 as instrumental variables from genome-wide association study datasets pertaining to Type 1 (finn-b-E4_DM1, n=189,113), Type 2 diabetes (finn-b-E4_DM2, n=215,654), and cataract (ukb-b-8329, controls=136,388, cataract=14,254). Various Mendelian randomisation methods were employed, including inverse-variance weighted (IVW), MR-Egger, weighted median, simple mode (SM), and weighted mode analyses. Additionally, sensitivity analyses were conducted to assess the robustness of the findings, encompassing tests for heterogeneity, pleiotropy, and leave-one-out assessments. A multivariable (MVMR) approach was used to account for potential confounders, such as obesity (IEUA-92, controls = 47468, obesity = 2896), smoking (ukb-a-16, n = 337030), and alcohol consumption (IEUA-1283, n = 112,117).

The analysis included 12 SNPs, which were derived from loci specifically associated with Type 1 diabetes and known to govern immune-inflammatory and metabolic pathways. The genetically-predicted Type 1 diabetes was found to elevate cataract risk significantly (OR=1.003, 95% CI: 1.001–1.005, P=0.001). The results of the sensitivity analyses corroborated the robustness of these findings, showing no significant heterogeneity (Cochran Q, P value = 0.73) or pleiotropy (MR-Egger intercept, P value = 0.38). Furthermore, multivariable MR demonstrated that the impact of diabetes on cataract risk remained significant after adjustment for multiple lifestyle factors.

We provide novel MR evidence that Type 1 diabetes causally increases the risk of cataract through the synergistic activity of immune dysregulation, chronic inflammation, and metabolic disturbance, with immune-metabolic crosstalk as the primary driver.

T1D causally increases the risk of cataract through the disruption of immune-inflammatory and metabolic pathways. Targeting immune-metabolic interactions may offer novel therapeutic strategies for preventing diabetic cataracts.

This study investigated the anti-ovarian cancer (OC) effects of Shuangzi Powder (SZP) and its regulatory impact on the tumor microenvironment.

This study employed systems biology approaches, integrating molecular docking and experimental validation, to explore the pharmacological mechanisms of SZP in OC treatment. To identify potential bioactive compounds and target genes of SZP, network pharmacology, protein–protein interaction network analysis, Gene Ontology (GO) analysis, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment were conducted.

Among the 11 bioactive ingredients identified in SZP, 1,767 potential therapeutic targets were predicted, while 2,637 differentially expressed genes were found to be associated with OC. KEGG pathway analysis revealed significant enrichment in pathways related to cancer, apoptosis, the PI3K-Akt signaling pathway, and the PD-L1/PD-1 checkpoint pathway. Treatment of A2780 cells with β,β-Dimethylacrylshikonin (DMAS) inhibited cell viability, migration, and invasion. Moreover, DMAS downregulated the expression of cell cycle- and apoptosis-related genes (CCNB1, CHEK1, CCNE1, and PARP1) and upregulated the immune checkpoint gene PD-L1.

These findings indicate that multiple components, targets, and pathways are involved in OC treatment by SZP.

DMAS, one of the bioactive ingredients of SZP, was predicted and preliminarily validated to exert inhibitory effects on OC cells, mainly through the regulation of the cell cycle, apoptosis, and immune response, as demonstrated by molecular docking and experimental analyses.

Y-box binding protein 1 (YBX1), an RNA-binding protein capable of recognizing the 5-methylcytosine (m5C), plays a role in the development and progression of various cancers. In this study, we aim to investigate the functional mechanism of YBX1-mediated m5C modification in Bladder Cancer (BCa).

The impact of YBX1 on glycolysis and biological functions in BCa cells was evaluated through a set of in vitro experiments. The underlying mechanisms involving YBX1, Transmembrane 4 L six family 1 (TM4SF1), and β-catenin/C-myc in BCa and their relationship were investigated using RNA immunoprecipitation (RIP), m⁵C-RIP, Actinomycin D, and luciferase reporter gene assays.

BCa cells exhibited elevated expression levels of YBX1 compared to human transitional bladder epithelial cells. YBX1 knockdown inhibited BCa cell proliferation, migration, and invasion while also attenuating glycolytic activity, as evidenced by reduced glucose uptake, lactic acid production, and ATP synthesis. Mechanically, we found that YBX1-dependent m⁵C modification promoted the stability of TM4SF1 mRNA, thereby upregulating TM4SF1 expression and subsequently activating the β-catenin/C-myc signaling. Furthermore, we discovered that overexpression of β-catenin could reverse the inhibitory effects of TM4SF1 silencing on proliferation and glycolysis in BCa cells.

This study has refined the mechanism of BCa progression, but the clinical significance and in vivo functions of the YBX1/TM4SF1 axis still require further verification.

YBX1 stabilizes TM4SF1 mRNA via m⁵C modification in BCa, activating β-catenin/c-Myc signaling to drive tumor growth and glycolysis. This reveals a novel therapeutic target for BCa.

Barberry Root (Sankezhen, SKZ), a traditional Uyghur herb from Xinjiang, China, has been shown to alleviate diarrhea-predominant irritable bowel syndrome (IBS-D); however, its molecular mechanisms remain unclear. This study aimed to systematically predict SKZ’s therapeutic targets and pathways for IBS-D using computational and experimental integration.

Active SKZ compounds and targets were sourced from TCM-Suite, BATMAN-TCM, and related databases. IBS-D targets were identified via DisGeNET and GeneCards, etc. Protein-Protein Interaction (PPI) networks, Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed. Molecular docking and 100-ns Molecular Dynamics (MD) simulations validated compound-target stability. In vitro (LPS-induced RAW264.7 macrophages) and in vivo (IBS-D model rats, isolated intestinal segments) experiments verified SKZ’s effects.

Fifteen bioactive compounds and 85 overlapping targets were identified, with four key compounds [(R)-Reticuline, Ferulic acid 4-O-glucoside, Magnoflorine, SW 7] and 15 hub targets (e.g., ESR1, EGF, ALB) prioritized. Enrichment analyses linked targets to inflammation and intestinal motility pathways. Docking showed strong binding affinities (<-8.0 kcal/mol), and MD simulations confirmed stability. SKZ suppressed inflammatory mediators, downregulated CHAT/C-FOS/5-HT3R/5-HT4R mRNA, and antagonized acetylcholine/barium chloride-induced intestinal contractions.

The findings highlight SKZ’s synergistic role in ameliorating IBS-D via multi-pathway regulation, consistent with existing research on inflammation and neurotransmission, though limitations include the need for further validation of individual compounds.

SKZ exerts synergistic therapeutic effects on IBS-D by ameliorating inflammation and regulating neurotransmission and intestinal motility, potentially via NF-κB/MAPK, COX-2/PGE2, cholinergic/5-HT, and calcium/potassium channel pathways, forming a multidimensional network.

Sarcopenia (Sar) is an age-related loss of muscle mass and function. Propolis, a natural product with anti-inflammatory properties, may help prevent Sar, but its active components and mechanisms remain unclear.

Network pharmacology identified intersecting targets of propolis ethanol extract (PEE) and Sar. PPI and CTP networks highlighted key compounds and targets, verified by molecular docking. In vitro, apigenin (Ap), the predicted main compound, was tested on D-galactose-induced senescent C2C12 myoblasts via cell viability and Western blotting.

Twelve overlapping targets were identified between PEE and Sar, with TNFα and IL6 highlighted as hub targets. Network analysis determined Ap as the main active compound. Molecular docking revealed strong binding affinities of Ap with TNFα and IL6. In vitro experiments demonstrated that Ap significantly enhanced the viability and differentiation of senescent C2C12 cells, downregulated TNFα and IL6 expression, and inhibited JAK2 and STAT3 phosphorylation, indicating suppression of the JAK-STAT signaling pathway.

The findings suggest that PEE, primarily through Ap, alleviates Sar by targeting inflammatory pathways and suppressing JAK-STAT signaling, thereby promoting muscle regeneration. The integration of network pharmacology, molecular docking, and in vitro validation provides mechanistic insights supporting the therapeutic potential of PEE in Sar. Limitations include the absence of in vivo confirmation, which warrants further animal and clinical studies to validate these effects and explore translational applications.

This study identifies Ap as the key active compound in PEE that alleviates Sar by downregulating TNFα and IL6 and inhibiting the JAK-STAT pathway. The results provide a molecular basis for the use of propolis as a natural intervention for Sar and support its development as a functional food or therapeutic agent targeting age-related muscle degeneration.

Hua-Zhuo-Ning-Fu decoction (HZD) is a traditional Chinese medicine prescription that has been clinically used by Chinese medical master Wang Xinlu for treating psoriasis. However, the specific molecular mechanisms remain unclear.

To identify the effective compounds of HZD and psoriasis-related genes, we conducted comprehensive searches in public databases, including TCMSP, SwissTargetPrediction, Gene Cards, and OMIM. Based on the degree values, core genes of HZD against psoriasis were determined. Furthermore, the affinity energy between the active compounds of HZD and their core targets was validated via molecular docking. Finally, the anti-psoriasis effects and potential mechanisms of HZD were examined in M5-stimulated HaCaT cells in vitro and IMQ-induced psoriasis mice in vivo.

Network pharmacological analysis of HZD for psoriasis treatment identified 43 active components and 243 targets. Topological and molecular docking analyses identified interleukin (IL)-6 and tumor necrosis factor-α (TNF-α) as core targets for its anti-psoriasis effects. Specifically, the docking energy of isovitexin with IL-6 was lower (-7.30 kcal/mol), and that of baicalin with TNF-α was lower (-6.70 kcal/mol). KEGG analysis revealed that the main pathway was the PI3K/AKT pathway. HZD inhibited cell viability, inflammation, and oxidative stress in M5-induced HaCaT cells. Animal experiments demonstrated that HZD alleviated psoriatic dermatitis, histopathological features, and inflammation in IMQ-induced mice with psoriatic plaques. Notably, HZD inhibited the expression of TNF-α and IL-6 and the activation of the PI3K/AKT pathway both in vivo and in vitro.

Specific upstream/downstream regulators of the PI3K/AKT axis regulated by HZD still need to be explored. Further investigation is essential to clarify the functional relationship between the predicted targets and active components.

In summary, HZD potentially mitigated inflammatory responses by targeting the TNF-α and IL-6 proteins, interfered with the PI3K/AKT pathway, and consequently drove the anti-psoriatic effect in IMQ-induced mice. Our findings provide a theoretical basis for HZD’s clinical use in psoriasis treatment.

The comorbidity of myocardial ischemia reperfusion injury (MIRI) and depression (DEP) may worsen the prognosis of coronary heart disease surgery. Currently, research on medications and therapeutic mechanisms for MIRI combined with DEP is still insufficient. This study aims to explore the relationship between DEP and MIRI, and the therapeutic effects and mechanisms of Baihe Dihuang Danshen decoction (BDDSD) on DEP combined with MIRI.

SD rats were assigned to a final experimental framework of six groups (Sham, MIRI, DEP+MIRI, BDDSD, DEP drug control, MIRI drug control). DEP was induced via 6-week chronic unpredictable mild stress (CUMS), with BDDSD administered during the final 2 weeks. MIRI was then induced by 30-minute coronary artery ligation and 2-hour reperfusion. DEP severity was assessed using behavioral tests (open field, elevated plus maze, sucrose preference, forced swimming). MIRI outcomes were evaluated via infarct size, histopathology, serum markers (LDH, IL-6, IL-1β), myocardial oxidative stress (MDA, GSH, SOD, Fe²⁺), and NADPH/FSP1/CoQ10 pathway proteins (FSP1, CoQ10, FTL, NOX2, NOX4, COX2).

Compared with the MIRI group, DEP significantly exacerbated MIRI, manifested by increased serum IL-6 and IL-1β levels, enlarged infarction area, and aggravated oxidative damage (elevated MDA/Fe²⁺, decreased SOD/GSH). Compared with the DEP+MIRI group, BDDSD intervention relieved DEP of rats, and subsequently reduced infarction area; decreased serum LDH, IL-6, and IL-1β; lowered myocardial MDA and Fe²⁺ while increasing SOD and GSH; upregulated FSP1/CoQ10/FTL; and downregulated NOX2/NOX4/COX2 expression.

DEP can aggravate inflammation and oxidative stress, promoting cardiac ferroptosis, thereby exacerbating MIRI. Our results demonstrate that BDDSD alleviates MIRI-DEP comorbidity through a dual mechanism, mitigating depressive symptoms and inhibiting myocardial ferroptosis via modulation of the NADPH/FSP1/CoQ10 pathway. Although the efficacy of BDDSD is encouraging, its dose-effect relationship and long-term safety require further study.

BDDSD effectively treats DEP-MIRI comorbidity through its dual mechanism, mitigating DEP and protecting against myocardial ferroptosis. Our study not only offers a novel therapeutic strategy for patients with DEP requiring coronary heart disease surgery but also provides new targets for developing drugs to treat MIRI combined with DEP.

Chronic atrophic gastritis (CAG) is an important stage in the occurrence and development of gastric cancer, and the morbidity of CAG is increasing year by year. Qilianshupi Decoction (QLSP) is a Chinese herbal compound which has been proved to reverse CAG, but its mechanism remains unknown. We wanted to identify the main components of QLSP by mass spectrometry and liquid phase analysis, and investigate their potential pathways for CAG treatment in combination with network pharmacology.

The main active components of QLSP were identified by liquid chromatography and mass spectrometry. Combined with network pharmacology, the targets where the drugs may act were identified and verified by animal experiments. Rats were randomly divided into control group, model group, QLSP low-dose group, QLSP medium-dose group, QLSP high-dose group and Weifuchun group. Rat CAG model was prepared by “N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) + ethanol intragastric + ranitidine feed”. After the test, gastric tissues were taken for pathological staining and immunohistochemistry.

We identified 51 prototype components of QLSP and found that QLSP treatment of CAG was closely related to p53. In animal experiments, CAG results in the decrease of E-cadherin and the increase of N-cadherin, Vimentin, p53, SMAD2 and TGF-β (p<0.05). Both QLSP and Weifuchun can increase E-cadherin and decrease N-cadherin, Vimentin, p53, SMAD2 and TGF-β (p<0.05).

QLSP, a traditional Chinese medicine formula with multi-component and multi-target characteristics, has been shown in our study to effectively regulate key EMT (epithelial-mesenchymal transition) markers and their upstream/downstream regulators. In animal experiments, QLSP successfully reversed the EMT process in CAG model rats. This finding provides new therapeutic targets for CAG treatment, though several challenges remain in clinical translation: First, rat CAG models differ from human CAG in pathological features and disease progression, and species-specific physiological and metabolic variations may limit the extrapolation of these findings. Second, network pharmacology analysis identified IL-6, alongside TP53, as another critical target of QLSP in CAG intervention. Therefore, future studies should further clarify the molecular mechanisms by which QLSP modulates EMT via IL-6-related pathways and validate its efficacy through well-designed clinical trials, ultimately providing a comprehensive understanding of QLSP's therapeutic potential in CAG.

QLSP inhibits epithelial-mesenchymal transition (EMT) in gastric mucosal epithelial cells and prevents CAG, possibly by regulating p53/TGF-β signaling pathway.

Metabolic syndrome (MetS) is a metabolic disorder characterized by the accumulation of various risk factors, including obesity, dyslipidemia, hypertension and so on. Songyang Duanwu Tea (SYT) has a high value in nutrition and health care, and it is widely used in traditional Chinese medicine for weight loss. Nevertheless, the mechanisms of SYT improving MetS remain to be elucidated. The objective of this study was to investigate the molecular targets and potential mechanisms by which SYT may improve MetS based on animal experiments and bioinformatics.

MetS model mice were established by a high-fat, high-sugar, high-salt diet (HFSSD). Obesity, dyslipidemia, hypertension, hyperuricemia and non-alcoholic fatty liver disease (NAFLD) of MetS model mice were evaluated to assess the effect of SYT on the treatment effects of MetS. The bioactive components in SYT were identified by bioinformatics and verified by HPLC-QTOF-MS. The possible molecular targets and mechanisms of action were predicted and verified using bioinformatics.

SYT (1.2 g/kg) ameliorated obesity, dyslipidemia, hypertension, hyperuricemia and NAFLD in HFSSD-induced mice. Bioinformatics results suggested that the major bioactive components in SYT include the flavonoid components apigenin, kaempferol, luteolin and quercetin, and the polyphenolic component eugenol. HPLC-QTOF-MS further validated the presence of apigenin, kaempferol, luteolin and quercetin. These 4 bioactive components are involved in the regulation of SYT to improve MetS by regulating metabolism and attenuating inflammation, and the key targets include peroxisome proliferator-activated receptor gamma (PPARG), tumor necrosis factor alpha (TNFα), interleukin 1beta (IL1B) and interleukin 6 (IL6).

SYT effectively improved the MetS model mice induced by HFSSD. The potential mechanism may regulate PPARG and attenuate inflammatory targets: TNFα, IL1B and IL6 through 4 flavonoid components: apigenin, kaempferol, luteolin and quercetin.

Metabolic dysfunction-associated steatotic liver disease (MASLD) is a global health concern, even among lean individuals. The Gan-Jiang-Ling-Zhu decoction (GZD), a traditional Chinese medicine formula, shows therapeutic potential against MASLD. This study investigated the efficacy of GZD in lean MASLD and explored its mechanisms of action.

A lean MASLD mouse model was established using C57BL/6 mice fed with a cholesterol-rich Paigen’s diet (PD). Following successful modeling, mice were administered GZD (1.8, 3.6, or 7.2 g/kg) or vehicle control. Body weight, food intake, and liver weight were monitored. Hepatic steatosis and lipid accumulation were assessed via H&E and Oil Red O staining, while serum enzymes were quantified biochemically. Gut microbiota composition was analyzed by 16S rRNA gene sequencing, and bile acid (BA) profiles in feces and serum were measured using UPLC-TQMS.

Twelve weeks of PD feeding induced a lean MASLD phenotype characterized by reduced body weight alongside hepatic steatosis and dyslipidemia. The GZD treatment dose-dependently ameliorated liver steatosis and lipid accumulation, with the highest dose (7.2 g/kg) showing superior efficacy. GZD restored gut microbiota balance by reducing pathogenic bacteria and enriching taxa involved in BA metabolism, leading to increased fecal excretion of secondary BAs. Conversely, serum levels of secondary BAs were significantly reduced after GZD treatment.

Our study highlights the promising role of GZD in lean MASLD, the involvement of gut microbiota and related BA metabolism that aligns with emerging evidence that gut dysbiosis and disrupted BA homeostasis are central to MASLD pathogenesis, even in lean individuals. However, the mechanistic links between specific microbial changes, BA pool composition, and hepatic outcomes remain to be elucidated.

GZD ameliorates hepatic steatosis in lean MASLD mice, an effect associated with modulation of gut microbiota composition and increased fecal excretion of secondary BAs. These findings suggest the potential of GZD as a therapeutic option for lean MASLD through gut-liver axis regulation.

Bladder cancer (BLCA) is a highly aggressive malignancy with poor prognosis. DNA replication stress-related genes (DRSGs) hold prognostic significance in multiple cancers, and their expression patterns in BLCA may reveal novel biomarkers and therapeutic targets.

This study was designed using a public database and the Cancer Genome Atlas (TCGA). Genes associated with DNA replication stress in BLCA were discovered by analyzing data from the TCGA and GEO databases using bioinformatics tools. The prognostic gene expression profiles in BLCA cell lines were analyzed using Western blotting (WB). The motility capacity of BLCA cells was evaluated using the wound healing and Transwell migration assays, while cell growth was ascertained with the CCK-8 assay.

Five DRSGs with prognostic significance were identified, and a risk score model was constructed using univariate Cox regression and the Least Absolute Shrinkage and Selection Operator (LASSO) regression algorithm. Kaplan-Meier (KM) analysis showed worse Overall Survival (OS) in the high-risk group (P < 0.05). Gene Set Enrichment Analysis (GSEA) indicated involvement in tumor-related pathways. The nomogram effectively predicted OS in both training and validation cohorts. WB and functional assays confirmed gene expression and effects on BLCA cell proliferation and migration.

This study first validates DRSGs’ prognostic value in bladder cancer, highlighting potential biomarkers and targets. Limitations include reliance on public data and in vitro tests. Future research should use multicenter cohorts and animal models to confirm clinical relevance.

This study aims to validate the hypothesis that Modified Qianzheng Powder (MQZP) exerts a protective effect on atherosclerosis (AS) by targeting macrophage-associated inflammatory pathways through a multi-target approach.

The active compounds and targets of MQZP were identified through TCMSP, HERB, and published literature. AS-related targets were extracted from disease databases. Using Venny 2.1.0, intersection targets were obtained, followed by PPI network construction and topological analysis to identify core therapeutic targets of MQZP for AS. Metascape facilitated GO and KEGG enrichment analyses. Molecular docking validated core target-compound interactions, with experimental verification of network pharmacology results.

We identified 124 active compounds and 417 potential therapeutic targets for atherosclerosis. Key bioactive constituents included cyclo(D)-Pro-(D)-Phe, aurantiamide, and beauverilide A, with TP53, SRC, STAT3, and AKT1 as core targets. GO and KEGG enrichment analyses revealed 3,417 biological processes and 238 signaling pathways. Molecular docking confirmed stable binding between core targets and compounds. In vitro, MQZP exhibited no significant cytotoxicity, effectively reducing ox-LDL-induced macrophage lipid accumulation and downregulating the levels of IL-1β, TNF-α, and MCP-1.

In summary, we found a variety of active ingredients of MOZP, which interfere with multiple targets of AS through multiple pathways. In vitro experiments verified that MOZP can reduce lipid accumulation in macrophages, reduce inflammation levels, and inhibit the IL-6/STAT3 signaling pathway, thereby exerting its anti-atherosclerosis effect.

In this paper, the molecular mechanism of MOZP against AS has been preliminarily explored; nonetheless, the therapeutic mechanism needs to be further investigated.

Chronic atrophic gastritis (CAG) is the initial phase in the carcinogenesis of gastric cancer (GC). Therefore, effective treatment for CAG is important in reducing the risk of GC progression. As an isoflavone compound, formononetin (FMN) has been identified as a potential therapeutic agent for acute gastric ulcers and GC. However, no study has reported the protective effect of FMN against CAG and its underlying mechanism. This study aimed to explore the therapeutic effects of FMN on CAG and its underlying mechanisms in vitro.

Network pharmacology was applied to predict the core targets of FMN therapy in CAG. The CAG cell model was developed using N-methyl-N-nitro-N-nitrosoguanidine (MNNG)-triggered human gastric epithelial cells (GES-1). The CCK-8 assay was applied to estimate cellular viability. The expression of inflammatory cytokines in cell supernatant was detected by ELISA. The protein levels and localization of nuclear receptor coactivator 1 (NCOA1), c-Jun, and c-Fos were evaluated using western blotting and immunofluorescence staining. Cell apoptosis was measured using flow cytometry.

Network pharmacology analysis identified c-Jun as the core target of FMN in the treatment of CAG, with biological processes primarily involving the regulation of apoptosis and inflammation. In vitro, MNNG exposure reduced GES-1 cell viability as well as increased inflammation and cellular apoptosis, and these effects were reversed by FMN treatment. In detail, FMN decreased the protein levels of NCOA1, c-Jun, and c-Fos in MNNG-triggered GES-1 cells. The activator protein-1 (AP-1) inhibitor T-5224 enhanced the effects of FMN treatment on cell viability, inflammatory response, and apoptosis in MNNG-triggered GES-1 cells.

This study employed network pharmacology analysis to identify FMN's therapeutic targets for CAG and validated the underlying mechanisms in vitro. While these results are promising, in vivo validation is required to confirm the efficacy of FMN. A comparative pharmacological evaluation against existing therapeutic agents and bioactive compounds would further elucidate FMN's therapeutic potential for CAG treatment.

FMN ameliorated the cell damage that MNNG triggered in GES-1 cells. The mechanism involved the anti-inflammatory and anti-apoptotic effects of FMN via modulation of the NCOA1/AP-1 signaling axis. The present preliminary study found FMN to exhibit a potential therapeutic effect against CAG.

With a large number of species' genomes assembled, sequence comparison has become an effective method for further studying biological classification and evolution. Traditional sequence alignment relies on predefined scoring functions, but it is computationally intensive and lacks molecular justification for scoring the differences between sequences. Therefore, we have developed a graphical representation method for DNA sequences to facilitate better sequence comparison and evolutionary analysis.

In this article, we introduce a novel method for representing DNA sequences using three-dimensional (3D) graphics. This method possesses two significant properties: (1) the graphical representation is acyclic; (2) each DNA sequence maintains a bijective relationship with its graphical representation.

Leveraging this proposed visualization method, we computed the corresponding ALE index for any DNA sequence by converting it into an L/L matrix and constructed a 12-dimensional feature vector. The feasibility of our proposed method has been validated through the construction of phylogenetic trees in four test sets: terrestrial vertebrates, hantavirus, fish and Japanese encephalitis virus.

This method enables both quantitative analysis and visual comparison of DNA sequences, providing a versatile tool for evolutionary studies.

Individual constitutions are classified into nine types in traditional Chinese medicine (TCM), and qi stagnation constitution (QSC) manifests as disrupted Qi circulation and increased susceptibility to emotional disorders and cancers. However, as a pre-disease state mainly affecting women, the biological basis of QSC and its susceptible mechanism to related diseases are still unclear. Exosomal microRNAs (miRNAs) are the stable regulators of gene expression and intercellular communication, and analysis of miRNAs enables us to understand the QSC better. This study profiles plasma exosomal miRNAs in QSC and balanced constitution (BC) females via high-throughput sequencing, aiming to identify the potential biomarkers of QSC and reveal its biological basis and the mechanism of its susceptible disease.

In this cross-sectional observation, female college students were recruited according to the criterion of QSC and BC in Classification and Determination of Constitution in TCM. Exosomal miRNAs were isolated from peripheral blood plasma and then profiled using high-throughput sequencing. Differentially expressed miRNAs (DEMs) were identified with fold change > 2 and P < 0.05, and screened as biomarkers to construct the receiver operating characteristic (ROC) curve. The diagnostic values of these biomarkers in different types of cancers were also validated based on the published data. Functional analysis were explored based on the predicted target genes.

Subjects with QSC showed significantly higher concentrations of albumin (ALB) and alkaline phosphatase (ALP) compared to those with BC, while there was no significant difference in baseline information and other clinical indicators between groups. A total of 54 DEMs were identified, including 30 up-regulated and 24 down-regulated miRNAs in the QSC group. The area under the ROC curve (AUC) for 7 specific up-regulated DEMs was 1.0, as well as the AUCs for therein 6 DEMs in various cancers were all above 0.9. The enriched KEGG (Kyoto Encyclopedia of Genes and Genomes) pathways included “signal transduction,” “infectious disease,” and “cancers”, and the most associating systems included immune, endocrine, and nervous systems, while the GO (Gene Ontology) function was mainly enriched in “protein binding,” “nucleus” and “transcription, DNA-templated”.

These 7 potential biomarkers of QSC have been confirmed to regulate oncogenic processes through epithelial-mesenchymal transition modulation and metabolic reprogramming, as well as therein 1 can also improve depression by lowering the expression of 5-hydroxytryptamine 1A receptor. The results of this study deepen the understanding of the constitutions in TCM. However, the small single-sex sample limits the application of the conclusion, and a large-scale clinical cohort including both sexes is still needed in future.

The expression of exosomal miRNAs in QSC showed unique features that have the potential to serve as biomarkers, and the related functional changes might be the biological basis for the susceptible diseases of QSC.

Depression is a common comorbidity in epilepsy, significantly impacting patients' quality of life. The hippocampus, linked to depression and neurodegeneration, is vulnerable in epilepsy. Epileptogenesis involves inflammation, oxidative stress, and neuronal damage, with the PI3K/AKT pathway playing a key role. Apigenin (API), a flavonoid in fruits and vegetables, shows neuroprotective, anti-inflammatory, and anti-apoptotic effects. This study investigates API's mechanisms in a LiCl-pilocarpine epileptic rat model, focusing on hippocampal neurogenesis and PI3K/AKT signaling as potential therapeutic targets.

We studied the effects of API and valproate (VPA) on depressive behavior and astrocytes in Lithium chloride (LiCl)-pilocarpine-induced epileptic rats. Additionally, we predicted the potential molecular targets of API for treating epilepsy using network pharmacology. Finally, we conducted in vivo experiments to validate the predicted mechanism.

In the API and VPA groups, there was a reduction in seizure frequency and seizure severity compared with the control group. The model group showed more depressive behavior than the control (CON) group, and these behaviors improved significantly after VPA and API treatment. HE staining showed that both API and VPA treatment improved LiCl-pilocarpine-induced nuclear contraction and cell swelling. Nissl staining demonstrated that Nissl vesicles in the CA3 region of the hippocampus were decreased in the model group, but the neurons were larger, more abundant, and more neatly arranged after API and VPA treatment. In the model group, the p-PI3K/PI3K and p-AKT/AKT protein ratios and PI3K, AKT mRNA expression were reduced, while brain-derived neurotrophic factor (BDNF) and glial fibrillary acidic protein (GFAP) were markedly increased. API and VPA treatment effectively reversed these changes.

API reduces seizures and depressive behaviors in LiCl-pilocarpine-induced epileptic rats, comparable to VPA API mitigates hippocampal neuronal damage, preserves Nissl bodies, and suppresses astrocyte activation via the PI3K/AKT pathway, suggesting neuroprotective and anti-inflammatory effects. While API shows promise as an antiepileptic and antidepressant agent, further studies are needed to confirm its direct modulation of PI3K/AKT and efficacy in other epilepsy models.

Our study suggests that API improves depression in rats and has anti-epilepsy activity, which may be involved in activating the PI3K/AKT pathway to protect astrocytes.

Obesity is a global health issue linked to metabolic disorders and cardiovascular diseases. Guang hawthorn (Malus doumeri) leaves have been traditionally used for medicinal purposes, but their bioactive compounds and anti-obesity potential remain underexplored.

This study extracted compounds from M. doumeri leaves using 70% ethanol and ethyl acetate. The extracts were administered to high-fat diet-induced obese rats. Serum levels of total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) were measured. The chemical composition of the extracts (HML) was analyzed using chromatography, NMR, and mass spectrometry. Network pharmacology and enrichment analyses were conducted using R and Cytoscape to identify compound-target interactions.

Rats treated with high-dose extracts showed significantly reduced TC, TG, and LDL-C levels and increased HDL-C (all p < 0.05). Three major compounds-phlorizin, sieboldin, and kumatakenin β-7-O-glucoside-were identified. A total of 272 overlapping targets and 32 core targets were found between compound-related and obesity-related gene sets. Functional analysis linked these targets to phosphorylation, apoptosis, cell proliferation, and kinase regulation.

The anti-obesity effects of M. doumeri may be mediated by modulation of the PI3K-Akt and FoxO signaling pathways, as well as proteoglycan biosynthesis. These pathways are associated with metabolic regulation and obesity-related changes.

M. doumeri leaf extracts demonstrate anti-obesity potential through multi-target and multi-pathway mechanisms, particularly via sieboldin and kumatakenin β-7-O-glucoside. These findings support their potential as natural therapeutic agents for obesity management.

Soaps are vital for preserving our health and personal hygiene since they not only eliminate germs but also rid the body of pollutants.

The current study aims to determine the physicochemical and antibacterial properties of Eucalyptus camaldulensis leaves using the agar disc diffusion technique and assess the effectiveness of different branded liquid soaps (25 mg/ml, 50 mg/ml, 75 mg/ml, and 100 mg/ml) with the Eucalyptus leaf extract against skin-infecting human pathogenic bacteria.

The combined antimicrobial susceptibility of E. camaldulensis and five liquid soaps showed an inhibition zone of 17.67±0.58, 13.33±0.58, 12.67±0.58, and 15.67±0.58 against Staphylococcus aureus, Streptococcus pyogenes, Pseudomonas aeruginosa, and Escherichia coli. The antibacterial properties of Av soap by itself did not work against S. pyogenes. Nevertheless, the extract and DI together showed a detrimental effect against S. aureus and P. aeruginosa, with no halo forming.

The absence of inhibition zones for the extract combined with DI against S. aureus and P. aeruginosa may indicate antagonistic interactions or reduced efficacy in that formulation. Overall, the data highlight the potential of E. camaldulensis to improve the antimicrobial properties of commercial soaps, though the effectiveness varies with microbial strain and formulation.

Antimicrobial activity was observed to increase with higher concentrations of the soap-extract combinations. Although liquid soap (seve) was effective against bacterial isolates, a combination of eucalyptus and aqua vera was shown to be more effective.

Obstructive sleep apnea syndrome [OSAS] is a common sleep breathing disorder accompanied by multiple organ intermittent hypoxemia. Our previous study has suggested that the expression of a lncRNA termed ENST00000592016 [lnc2016 for short] derived from plasma exosomes is remarkably elevated in OSA patients compared to the normal population, and lnc2016 can improve the diagnostic efficiency of OSA.

To unmask the role of the lnc2016 in vascular endothelial cells, targeted hypoxia is the goal of the current research.

Primary human ADSCs and HUVEC cells were cultured. CCK-8, cytometric assay, transwell, and tubular formation assay were used to determine cell viability, cell apoptosis, cell cycle, cell migration, as well as tubular formation ability.

Adipose-derived stem cells [ADSCs]-derived exosomes contained robust lnc2016. After co-culture with human umbilical vein endothelial cells [HUVECs], exosomal lnc2016 could enhance cell proliferation, DNA synthesis, migration, and tubular formation, whereas suppress cell apoptosis of HUVECs against hypoxic conditions.

Under hypoxic conditions, ADSCs secrete various reparative factors and transmit them via exosomes; among them, lnc2016 may participate in the regulation of hypoxia-induced injury through the ceRNA network, which requires further investigation.

Ln2016 can promote the cell growth, migration, DNA synthesis, and tubular formation as well as suppress the cell apoptosis of vascular endothelial cells against hypoxia in vitro.

Cranberry (Vaccinium macrocarpon) is rich in vitamins, minerals, anthocyanins, flavonoids, and phenolic acids, offering potent antioxidant activity. Polyphenols in cranberries are linked to neuroprotective effects via modulation of oxidative stress, inflammation, and signaling pathways.

This study evaluated the neuroprotective effects of cranberries on behavioral and neurochemical abnormalities induced by intracerebroventricular (ICV) quinolinic acid (QA) in Wistar rats, focusing on ERK and PI3K/AKT pathway modulation.

Thirty Wistar rats were divided into groups: control, QA (240 nM, ICV), QA + cranberry (0.5 g/kg, p.o.), and QA + high-dose cranberry (2 g/kg, p.o.). Treatments continued for 21 days. Behavioral performance was assessed via Novel Object Recognition, Morris Water Maze, rotarod, and footprint analysis. Biochemical assays measured oxidative/nitrosative stress markers, mitochondrial complex activities, and cholinergic function. Histological analysis evaluated neuronal integrity.

QA treatment impaired cognition, motor function, and mitochondrial activity, increased oxidative stress (↑MDA, ↑nitrite, ↓GSH), and induced cholinergic dysfunction. Cranberry supplementation, particularly at 2 g/kg, significantly improved memory, learning, and motor coordination, restored GSH, reduced MDA and nitrite levels, enhanced mitochondrial complexes I, II, and IV activities, and normalized cholinergic markers. Histology confirmed reduced neuronal degeneration and inflammation.

Cranberries exhibit neuroprotective effects likely via antioxidant, anti-inflammatory, and anti-excitotoxic mechanisms, promoting synaptic plasticity and neuronal survival.

Cranberries may serve as a potential natural therapeutic strategy for cognitive deficits and neurodegenerative conditions, warranting further translational studies.

The mechanisms of chemotherapy sensitivity and toxicity are complex. Metabolomics can better reflect the status of anticancer drugs, tumors, and hosts simultaneously.

Mice were implanted with human gastric cancer cells through subcutaneous xenografting, and then treated with the PF (platinum-fluorouracil) regimen, with saline serving as the control. Tumor growth was monitored by measuring tumor volume, and body weight was recorded on Days 0, 2, 4, 6, and 8. Kidney damage was assessed using H&E staining. To analyze differential responses, PF-treated mice were grouped separately according to chemotherapy sensitivity (high/medium/low via tumor response) and toxicity (high/medium/low via body weight changes). Serum metabolomics was evaluated using Mass Spectrometry.

Platinum-Fluorouracil (PF) chemotherapy significantly reduced tumor weight in mice, although it also induced notable body weight loss and renal toxicity compared to controls. Serum metabolomic analysis revealed significant differences between PF and control groups, involving metabolites like deoxymethylmycin and dehydrocorticosterone, associated with AMPK and cortisol synthesis/secretion pathways. Further comparisons highlighted: (1) High- vs. low-sensitivity subgroups differed significantly in metabolites, such as palmitoyl-CoA and indoleacetic acid (linked to AGE-RAGE, insulin resistance, and AMPK pathways). (2) High- vs. low-toxicity subgroups displayed significant metabolic differences, including methylguanosine and methylcytidine (implicated in ferroptosis, ether lipid, and fatty acid metabolism pathways).

The PF regimen effectively inhibits the growth of subcutaneous tumors in nude mice, while causing varying levels of sensitivity and toxicity in tumor chemotherapy. These observed effects of sensitivity and toxicity are linked to underlying metabolic mechanisms.

Cancer is a major public health concern, and conventional treatments like surgery, chemotherapy, and radiotherapy are associated with several disadvantages, including chemoresistance, toxicity, and economic burden to the family of cancer patients. Thus, discovery of novel agents of natural agents to reduce these side effects is crucial. A series of studies have shown anthraquinones as a promising adjuvant in enhancing the effectiveness of standard cancer therapies.

This review explores the anticancer potential of anthraquinones and their role in enhancing standard chemotherapy.

Various freely available databases, including PubMed, Scopus, Google Scholar and Web of Science were searched for updated and relevant information on anthraquinones and their use as an adjuvant with standard chemotherapeutic agents.

In this article, we looked at the recent developments in the utilization of anthraquinones as adjuvants in chemotherapy. Further, we have elaborated the mechanism of action that anthraquinones target to chemosensitize the drug-resistant cancer cells.

This review provides updated information on emerging role and their potential to be utilized as adjuvants in augmenting the efficacy of conventional cancer therapies.

Pressure injury (PI) severely affect the quality of life of patients. Infection and inflammation are key factors contributing to the progression of PI; therefore, their inhibition plays a crucial role in preventing the worsening of the condition. Huanglian Jiedu Decoction (HLJDD), as a typical traditional Chinese medicine with heat-clearing and detoxicating effects, has good broad-spectrum antibacterial and anti-inflammatory effects. It is commonly used in the treatment of clinically infected external wounds. However, the therapeutic effects of HLJDD on PI remains unclear.

The extract of HLJDD was prepared using the water extraction and alcohol precipitation method. Sixty male SD rats were randomly divided into five groups (n = 12/group): control group, model group, normal saline group (negative control group), iodophor group (positive control group), and HLJDD group (test group). Except for the control group, magnet clamping and the Staphylococcus aureus inoculation method were used to construct the model of stage 3 PI infection wound in the other groups. After irrigating the wound, the healing rate, bacterial concentration, concentrations of IL-1, IL-6, and TNFα, and the protein expression levels of TLR2, MyD88, and NF-Bp65 were examined. Skin and ocular mucosal irritation tests were conducted to evaluate the safety of the topical application of HLJDD.

Rats treated with HLJDD exhibited an improved wound healing rate, along with reduced bacterial concentration on the wound surface and a significant decrease in the content of inflammatory cytokines (IL-1β, IL-6, and TNF-α). The protein expression levels of TLR2, MyD88, and NF-κBp65 were down-regulated after the administration of HLJDD. The prepared HLJDD did not cause any irritation.

HLJDD can promote the healing of PI wounds and has a protective effect on PI through its anti-inflammatory and anti-bacterial properties.

ABCA family proteins regulate cholesterol transport, which affects cancer-related processes such as membrane dynamics and tumor progression. However, their roles in gastric cancer (GC) remain unclear.

This study systematically investigated the expression profiles, prognostic significance, and immune-related roles of ABCA family genes in GC using data from The Cancer Genome Atlas (TCGA). Kaplan-Meier and Cox regression analyses assessed survival relevance, while logistic regression and ROC curves evaluated clinical associations and diagnostic value. Immune infiltration and gene correlation were analyzed via ssGSEA and Pearson correlation. TIDE and “oncoPredict” were used to estimate immunotherapy response and chemotherapy resistance. Gene Set Enrichment Analysis (GSEA) identified related signaling pathways. Quantitative PCR validated ABCA expression in cell lines.

Several ABCA genes (e.g., ABCA1, ABCA2, ABCA7, ABCA13) were upregulated, while others (e.g., ABCA8, ABCA9) were downregulated in GC tissues. Expression levels correlated with pathological stage, grade, and lymph node metastasis. ABCA1, ABCA3, ABCA4, ABCA6, ABCA8, and ABCA9 were identified as independent prognostic factors. Nomogram models showed good predictive performance. High ABCA expression was associated with increased infiltration of multiple immune cells and co-expression with immune checkpoint genes. TIDE analysis indicated lower predicted ICI response, and ABCA levels correlated with resistance to cisplatin, 5-FU, and paclitaxel. GSEA revealed enrichment in ECM-receptor interaction, cell adhesion, autophagy, and PI3K-Akt pathways.

ABCA genes exhibit distinct expression and prognostic patterns in GC and are closely linked to tumor immunity and drug resistance, supporting their potential as biomarkers and therapeutic targets.

This study aimed to explore the mechanism of moxibustion in the knee by combining osteoarthritis metabolomics and network pharmacology.

A rat knee osteoarthritis (KOA) model was established by intra-articular injection of papain. The efficacy of moxibustion in KOA rats was evaluated by swelling degree, pathological progress, and mobility loss of knee joint. On this basis, the metabolic mechanism of moxibustion in relieving knee osteoarthritis was analyzed by metabolomics analysis.

Moxibustion significantly reduced joint swelling and inflammation in the knee joint of KOA rats. Sixteen metabolites and nine metabolic pathways were found to be associated with the mechanism of action of moxibustion in metabolomics analysis results. According to network pharmacology, 3186 KOA disease targets, 158 drug targets, and 89 intersecting targets were obtained. The key targets included MAPK-3, AKT-1, RELA, MAPK-8, MAPK-14, etc. Signal pathways were found to be involved in mechanisms of moxibustion in knee osteoarthritis, such as alanine, aspartate, and glutamate metabolism, cysteine and methionine metabolism, and arginine and proline metabolism.

At present, the mechanism of moxibustion for KOA is not completely clear, but it is certain that its effect is related to the effect produced by heat and radiation. In addition, the aromatic substances produced during the combustion of moxa leaves have anti-inflammatory, antioxidant, and immune-enhancing effects on KOA.

The mechanism of moxibustion in knee osteoarthritis may involve alanine, aspartate, and glutamate metabolism, cysteine and methionine metabolism, arginine and proline metabolism, amino tRNA biosynthesis, and D-glutamine and D-glutamate metabolism signaling pathways with MAPK-3, AKT-1, RELA, MAPK-8, and MAPK-14 as core targets. More precise mechanisms need to be verified by further systematic molecular biology experiments.

Sepsis is a critical illness with high morbidity and mortality, particularly due to gastrointestinal complications. Despite improvements in therapeutic strategies, effective pharmacological treatments remain lacking. Banxia Xiexin Decoction (BXD), a traditional Chinese formula, has shown potential in regulating intestinal function. This study aimed to investigate the therapeutic effects and underlying molecular mechanisms of BXD in sepsis-induced intestinal injury, focusing on the PINK1/Parkin pathway.

Human intestinal epithelial cell (HIEC) injury induced by lipopolysaccharide (LPS) and a cecal ligation and perforation (CLP) rat model of sepsis were used. Experimental groups received BXD at varying doses, while PINK1 knockdown HIECs were used to assess mechanistic pathways. ELISA was employed to measure IL-6, IL-1β, IFABP, and DAO levels. Pathological changes were assessed by H&E staining, while tight junction proteins (ZO-1, Occludin), TOM20, mitochondrial membrane potential, and autophagy markers (PINK1, Parkin, LC3, p62) were analyzed via immunohistochemistry, immunofluorescence, flow cytometry, Western blot, and RT-PCR.

BXD treatment significantly reduced IL-6, IL-1β, DAO, and IFABP levels compared with controls. It restored ZO-1 and Occludin expression, improving intestinal mucosal barrier function. In septic rats, BXD enhanced TOM20 expression, preserved mitochondrial membrane potential, and upregulated the PINK1/Parkin-mediated mitophagy pathway. These effects collectively reduced inflammation, mitochondrial dysfunction, and intestinal damage.

Findings suggest that BXD exerts protective effects against sepsis-induced intestinal injury by reducing systemic inflammation and promoting mitochondrial homeostasis through the activation of PINK1/Parkin-mediated mitophagy.

BXD alleviates intestinal mucosal damage and systemic inflammation in sepsis, offering a promising therapeutic approach by targeting mitochondrial autophagy.

The intricate relationship between the gut microbiome and myopia is increasingly recognized, underscoring the need to explore its causal dynamics. Despite emerging evidence, the influence of Gut Microbiota (GM) on ocular development remains underexplored.

This study utilized Mendelian Randomization (MR) to investigate the causal impact of GM on the development of myopia. Instrumental variables (IVs) were identified from Genome-Wide Association Studies (GWAS), focusing on genetic variants significantly associated with microbiome composition. A comprehensive array of MR techniques was applied to ensure a robust estimation of causal effects and to adjust for potential confounders and pleiotropy.

The Inverse-Variance Weighted (IVW) method was used to identify significant associations between GM and myopia. Increased risk of myopia was linked to the class Betaproteobacteria (OR=1.01, 95% CI 1.004-1.017, P=0.003), the order Burkholderiales (OR=1.009, 95% CI 1.001-1.016, P=0.02), the family Oxalobacteraceae (OR=1.005, 95% CI 1.001-1.01, P=0.023), and several genera including Eubacterium xylanophilum group (OR=1.007, 95% CI 1.001-1.013, P=0.033), and Bifidobacterium (OR=1.005, 95% CI 1-1.01, P=0.038). Protective effects were noted for the order Mollicutes RF9 (OR=0.994, 95% CI 0.99-0.999, P=0.014), the genus Allisonella (OR=0.996, 95% CI 0.993-0.999, P=0.019), the genus Lachnospiraceae UCG001 (OR=0.994, 95% CI 0.989-1, P=0.045), and the family Enterobacteraceae (OR=0.991, 95% CI 0.982-1, P=0.047) and order Enterobacteriales (OR=0.991, 95% CI 0.982-1, P=0.047). Sensitivity analyses further confirmed the robustness of these findings.

This study provides causal evidence for the “Microbiome-Gut-Eye Axis” in myopia development, identifying specific gut microbiota that influence myopia risk. These findings suggest potential for microbiota-targeted interventions, warranting further research in diverse populations.

The findings support the “Microbiome-Gut-Eye Axis” as a potential factor in myopia pathogenesis and highlight microbiota-targeted interventions as novel therapeutic strategies for managing myopia. This study lays the groundwork for further research on how modifying GM can influence eye health and offers new perspectives on preventive health strategies.

Qishen Huoxue Granule (QHG), a classical Traditional Chinese Medicine prescription, can reduce septic cardiomyopathy in clinic. However, the mechanism of QHG remains unclear. This study aims to investigate the mechanism and effect of QHG-contained serum (QHG-CS) on sepsis-induced cardiomyopathy (SICM).

QHG was administered to Wistar rats via gavage to obtain QHG-CS. The chemical constituents of QHG-CS were identified via UPLC-Q-TOF-MS. In vitro, rat cardiomyocytes H9c2 cells isolated from embryonic BD1X rat heart tissue, and septic myocardial injury model was established by inducing H9c2 cells with lipopolysaccharide (LPS). Cell viability was assessed through CCK-8. Protein expression was determined using western blot, and gene expression was measured using real-time quantitative PCR. Cell autophagy was investigated by detecting LC3 expression using flow cytometry and immunofluorescence. In addition, three inhibitors, A779 (MasR), wortmannin (PI3K) and rapamycin (mTOR) were used to localize the potential therapeutic targets.

QHG-CS significantly improved the survival of septic cardiomyocytes (p<0.0001). The expression of autophagy-related markers Beclin1, ATG5, and LC3II/I was increased in LPS-induced cardiomyocytes, which could be inhibited by QHG-CS. QHG-CS upregulated the mRNA expression of MasR, PI3K, and AKT, as well as the phosphorylation of PI3K, AKT, and mTOR. Moreover, A779 markedly lowered mRNA levels of MasR, PI3K, and mTOR, while wortmannin decreased mRNA levels of PI3K and mTOR, whereas rapamycin only suppressed mTOR phosphorylation.

By inhibiting excessive autophagy through upregulation of the MasR/PI3K-AKT-mTOR pathway, QHG can alleviate sepsis-induced cardiomyocyte damage. This study provides novel perspectives for the management of sepsis-induced cardiac damage.

Chenopodium album Linn. is a nutritionally and pharmacologically significant herb that generally grows in the winter season along with other crops. It is rich in fibers, protein, minerals (Mg, Ca, Fe, K, P, and others), vitamins (ascorbic acids, thiamine, riboflavin, and others), and several other biologically active chemical components like flavonoids, saponins, steroids and many more. In this article, the authors briefly describe and assess the chemistry and pharmacology of this nutritionally significant plant.

This study is based on several literature searches conducted via Google Scholar, Research Gate, PubMed, and many other online sources.

Due to its richness with bioactive phytochemicals, it has become a valuable functional food. C. album has several medicinal properties like antioxidant, antimicrobial, anti-arthritic, anti-diabetic, anti-infection, anti-ulcer, and many others. Even after its rich nutritional values, chemical compositions, and a broad spectrum of pharmacological properties, this is a highly ignored herb worldwide. Therefore, extensive research and awareness regarding the functional role of this herb is needed.

This study aimed to investigate the modulation of intestinal flora by electroacupuncture in a murine ulcerative colitis (UC) model, with a focus on analyzing microbial taxa and identifying key regulatory targets and pathways.

A UC model was established in mice using 5% dextran sodium sulfate (DSS). Electroacupuncture was applied at bilateral “Shangjuxu” (electrostimulation) and “Tianshu” (manual acupuncture) points from days 5–9, while the mesalazine group received 0.5 g/kg/day via gavage. Disease activity index (DAI), colon length, and histopathology (hematoxylin-eosin staining) were evaluated. Intestinal flora composition was analyzed via 16S rDNA sequencing.

Electroacupuncture significantly reduced DAI scores on days 7 and 9 (P < 0.05; P < 0.01) compared to the model group, improved colon morphology, and reduced inflammation. Linear discriminant analysis and Wilcoxon tests revealed an increased abundance of Roseburia and elevated alpha diversity in the electroacupuncture group. Functional prediction demonstrated suppressed RNA transport and glycerophospholipid metabolism in the model group (P < 0.05), which were significantly enhanced post-electroacupuncture (P < 0.01).

Electroacupuncture restored beneficial taxa (e.g., Roseburia) and microbial diversity, suggesting gut homeostasis modulation. Enhanced lipid metabolism and RNA transport pathways may underlie its anti-inflammatory and mucosal repair effects.

Electroacupuncture alleviates UC by modulating the structure and function of intestinal flora, with Roseburia and associated metabolic pathways identified as key targets. These findings highlight the potential of electroacupuncture as a UC therapy.

Atherosclerosis (AS) is a leading cause of cardiovascular diseases, characterized by lipid accumulation in arterial walls. The gene Ephx2, which encodes soluble epoxide hydrolase (sEH), is implicated in AS development, but its precise mechanisms and therapeutic potential are not fully understood.

This study aimed to analyze gene expression data from low-density lipoprotein receptor knockout (LDLR^−/−) and LDLR^−/−sEH^−/− mice to identify significant genes associated with AS.

A directed compound-protein interaction network was constructed based on these genes and related pathways from the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. In the end, through resistance distance (RD) between any two nodes in this network, the Independent Cascade (IC) model was applied to explore Ephx2 mechanisms in AS, such as important Adverse Outcome Pathways (AOPs).

Several AOPs were identified as critical in AS treatment via Ephx2. The key AOPs included inflammatory response and cytokine release, cholesterol deposition and oxidation, disruption of plaque stability, smooth muscle cell proliferation and migration, and platelet activation and coagulation. Within the top AOPs of inflammatory response and cytokine release, potential target genes were identified, such as Mapk3, Pik3cd, Gnai2, Mapk10, Arnt, and RhoA. Critical paths from Ephx2 to these target genes were established, suggesting mechanisms by which Ephx2 may influence AS pathogenesis.

By defining the AS network and corresponding RD, this study elucidates potential mechanisms by which Ephx2 affects AS through specific KEGG pathways, AOPs, and target genes. These findings enhanced the understanding of AS pathogenesis and highlighte potential targets like Mapk3 for developing therapeutic strategies in AS prevention and treatment.

Astragaloside (AST), a natural saponin extracted from Astragalus membranaceus (Fisch.) Bunge., has been consistently utilized in the treatment of rheumatoid arthritis (RA). N6-methyladenosine (m6A), the most prevalent modification of mRNA, is associated with the progression of various diseases, including RA. Nonetheless, the effects of AST on m6A modification in RA remain to be elucidated.

The MH7A cell model was established through induction with TNF-α. The effects of AST on the expression levels of WTAP, BAX, BCL2, and TRAIL-DR4 were evaluated utilizing immunofluorescence, RT-qPCR, and Western blot analysis. Furthermore, CCK-8 and flow cytometry were used to assess MH7A cell viability, cell cycle, apoptosis, and proliferation. Then, the m6A modification of TRAIL-DR4 was elucidated via MeRIP-qPCR.

The optimal dose administration time was 50 μg/mL at 48 h. AST not only reduced the expression levels of WTAP, BCL2, BAX, TRAIL-DR4, and the m6A modification level of TRAIL-DR4 but also significantly enhanced apoptosis in MH7A cell, while inhibiting cell viability and proliferation. Furthermore, AST was capable of reversing the effect on MH7A cell proliferation and apoptosis induced by WTAP overexpression.

This study elucidates the protective role of AST on MH7A cells by attenuating m6A/WTAP-mediated apoptosis, offering novel insights into the mechanisms of AST.

Transcription factors (TF) are the central regulatory hubs of signaling pathways in eukaryotic cells. Here, we explored the abnormal expression of TF in septic-induced lung injury by sequencing.

The levels of target proteins were detected using Western Blot and Elisa. Cell function was evaluated using CCK8 and transwell assays. A double luciferase reporter assay was performed to detect interactions between target molecules.

We found that TF glioma-associated oncogene (GLI) family zinc finger 3 (GLI3) was abnormally low expressed in a lipopolysaccharide (LPS) induced acute lung injury (ALI) cell model. In an in vitro model, GLI3 overexpression promoted the proliferation and migration and inhibited apoptosis of lung epithelial cells in LPS-induced inflammatory environment. Importantly, GLI3 overexpression inhibited the secretion of inflammatory factors IL-1β, IL-6, and TNF-α. Additionally, miR-143-3p inhibited the expression of GLI3. MiR-143-3p inhibitor alleviated the cell damage caused by LPS, while knocking down GLI3 counteracted this effect, indicating that miR-143-3p downregulated GLI3 and inhibited its anti-inflammatory effect. Secreted frizzled related protein-1 (SFRP1) was upregulated in LPS-treated cells and SFRP1 promoter interacted with GLI3, suggesting that SFRP1 was a target of TF GLI3. Co-transfection with GLI3 knockdown and SFRP1 overexpression plasmids attenuated the secretion of inflammatory factors IL-1β, IL-6, and TNF-α caused by GLI3 knockdown in LPS-treated cells, indicating that SFRP1 plays an anti-inflammatory role as a GLI3 target in the ALI cell model.

miR-143-3p caused degradation of GLI3 mRNA and thus inhibited the transcription of SFRP1, leading to decreased proliferation and increased levels of inflammatory factors, providing new potential targets for the clinical diagnosis and treatment of ALI.