Search Results (810)

Kaposi’s Sarcoma Herpesvirus (KSHV) is the etiological agent of Kaposi’s Sarcoma (KS), which induces metabolic stress in infected host cells. KSHV reprograms host metabolic pathways for efficient viral replication and infectious virion production. Here, we report a time-course global metabolomics study conducted in iSLK.BAC16 cells to compare latent and lytic KSHV infection. Our data show that amino acid, central carbon, and nucleotide metabolic pathways are highly dysregulated upon reactivation. During lytic KSHV infection, pathway enrichment analysis identifies purine and pyrimidine metabolism as the top two most significantly impacted and dysregulated pathways. Further experiments have shown that nucleotide metabolism is required during lytic KSHV infection to produce maximal infectious virus. Treatment with the FDA-approved drug, methotrexate (MTX), a folate antagonist that decreases nucleotide metabolism by reducing tetrahydrofolate cofactors, significantly reduced KSHV copy number and late lytic viral gene expression upon reactivation compared to controls. Additionally, titers of cell-free supernatants from MTX-treated lytic samples showed a significant reduction in infectious virion production. Furthermore, MTX significantly decreased the viral titer of murine herpesvirus 68 (MHV-68), a model virus to study gammaherpesvirus. Overall, our study demonstrates that metabolic inhibition during lytic gammaherpesvirus infection decreases productive infection and hence serves as a potential therapeutic antiviral target. Full article

Liriomyza trifolii is a global pest of vegetables and ornamental plants and a major invasive species in China. Long-term irrational use of chemical insecticides has reduced control efficacy, driving interest in plant-derived alternatives. Bitter gourd, Momordica charantia, extracts show potential against various pests, but their effects on L. trifolii are unknown. To address this, host suitability tests were first conducted and revealed that L. trifolii exhibited significantly higher adaptability to kidney bean than to bitter gourd in terms of both oviposition and feeding, and failed to complete its life cycle on bitter gourd. Further treatment with ethanol extracts of bitter gourd leaves demonstrated dose-dependent adulticidal activity; at the LC₅₀ concentration, the extract not only reduced feeding punctures and oviposition, but also significantly decreased egg hatching and larval survival, while showing no significant impact on pupation or emergence. To elucidate the underlying molecular mechanisms, integrative transcriptomic and metabolomic analyses were subsequently performed. Transcriptomics identified 254 differentially expressed genes (DEGs) enriched in ribosome, oxidative phosphorylation, and peroxisome pathways, and upregulated DEGs included a vitellin-degrading protease and cytochrome P450. Meanwhile, metabolomics detected 272 differential metabolites (DEMs): upregulated metabolites were linked to purine metabolism, while downregulated ones were associated with cysteine/methionine and arginine/proline metabolism. Integrated analysis revealed “Biosynthesis of amino acids” as the sole common pathway, with S-adenosyl-L-homocysteine, N-succinyl-LL-2,6-diaminoheptanedioate, and glutamine synthetase (Ltr05G008090) showing significant correlations. These findings suggest that reprogramming of amino acid and nitrogen metabolism may participate in the response and could serve as a candidate response pathway. Accordingly, this study may offer a theoretical basis for the future development of botanical insecticides based on bitter gourd leaf extract against L. trifolii. Full article

High-altitude environments expose aquatic organisms to complex environmental challenges, including fluctuations in dissolved oxygen levels, water temperature, and other habitat conditions. The freshwater snail Cipangopaludina cathayensis has limited dispersal ability and is strongly associated with local habitat conditions, making it a suitable species for investigating biological responses to long-term environmental variation. In this study, high-altitude and low-altitude populations of Cipangopaludina cathayensis were compared using biochemical assays, shell elemental composition analysis, transcriptomics, and metabolomics to explore altitude-associated physiological and molecular responses. Environmental monitoring showed that the high-altitude habitat exhibited lower water temperatures, greater thermal variability, and significantly higher dissolved oxygen availability than the low-altitude habitat, indicating distinct environmental conditions between the two populations. The high-altitude population exhibited significantly higher superoxide dismutase (SOD) and pyruvate kinase (PK) activities than the low-altitude population, whereas malondialdehyde (MDA) content and total ATPase (T-ATPase) activity showed no significant differences between populations. Shell microstructure and elemental analyses revealed altitude-associated differences in biomineralization. Transcriptomic analysis identified differentially expressed genes involved in intracellular transport, including KIF10, KIF11, KIF13, KIF17, MYO3, and MYO7A, as well as apoptosis-associated genes, including CASP2, CASP3, and BCL-2. Integrated transcriptomic and metabolomic analyses further highlighted purine metabolism, D-amino acid metabolism, insulin resistance, and alanine, aspartate and glutamate metabolism as shared enriched pathways between the two populations. These findings indicate that Cipangopaludina cathayensis may cope with high-altitude environments through coordinated regulation of antioxidant defense, intracellular transport, apoptosis-associated pathways, and metabolic homeostasis. Full article

Background/Objectives: Watercress (Nasturtium officinale R.Br.) is a glucosinolate-rich cruciferous vegetable with reported cardioprotective properties. However, previous human studies have relied on targeted clinical and biochemical biomarkers, limiting insight into its broader metabolic effects. This exploratory study investigated plasma metabolomic changes associated with watercress supplementation in adults with low-to-moderate cardiovascular risk using untargeted proton nuclear magnetic resonance (¹H-NMR) spectroscopy. Methods: In this randomized, single-blind, placebo-controlled pilot trial (Thai Clinical Trials Registry: TCTR20251119004), 26 participants aged 40–59 years received dried watercress capsules (8 g/day; approximately 195 mg glucosinolates/day) or placebo for 28 days. Fasting plasma samples collected at baseline and Day 28 underwent untargeted ¹H-NMR profiling. Partial least squares-discriminant analysis (PLS-DA) assessed group discrimination, while Kyoto Encyclopedia of Genes and Genomes (KEGG)-based pathway enrichment and topology analyses identified perturbed metabolic pathways. Results: A total of 209 plasma metabolites were identified. PLS-DA demonstrated modest discrimination between groups (Q² = 0.268), with 27 discriminant metabolites (variable importance in projection > 1.5) involving amino acid, nucleotide, carbohydrate, and gut microbial metabolism. Eighteen metabolic pathways were significantly perturbed, particularly galactose and fructose/mannose metabolism (p < 0.001), together with glycerolipid, purine, glycolysis/gluconeogenesis, and tryptophan metabolism. Watercress supplementation reduced metabolites associated with oxidative DNA damage, inflammatory kynurenine metabolism, and microbial co-metabolism, while increasing glycine and dimethylglycine and altered gut microbial activity. Conventional biomarkers showed a significant decrease in low-density lipoprotein cholesterol but no consistent effects on other lipids. Conclusions: Watercress supplementation was associated with coordinated metabolic alterations across multiple pathways, generating mechanistic hypotheses for its antioxidant and cardiometabolic effects. These findings are exploratory and warrant confirmation in larger, adequately powered clinical trials. Full article

Background/Objectives: Ambient fine particulate matter (PM_2.5) poses significant health risks to older adult populations, yet the specific contributions of its chemical constituents, particularly non-phthalate and non-organophosphate ester compounds, remain poorly understood. This study aimed to elucidate the mechanistic links between PM_2.5-bound ester exposures and metabolic pathway alterations in elderly individuals. Methods: A total of 258 elderly residents aged 60 years or older from Fuzhou, China, were recruited. Personal PM_2.5 exposure was monitored over 72 h using UPAS V2 samplers, with chemical components analyzed via gas chromatography–mass spectrometry (GC–MS). Plasma metabolomic profiling was conducted using liquid chromatography–mass spectrometry (LC–MS), and metabolic pathway enrichment was performed using MetaboAnalyst 5.0. Linear regression models adjusted for covariates (age, sex, BMI, lifestyle factors) assessed associations between ester exposures and metabolite abundance. Results: The mean PM_2.5 concentration was 38.06 μg/m³, with ester compounds dominating the chemical composition. Twenty high-concentration non-target esters were prioritized for analysis. PM_2.5 ester exposure was associated with alterations in key metabolic pathways, including steroid biosynthesis, glycolysis/gluconeogenesis, glycerophospholipid metabolism, and purine/pyrimidine metabolism. When interpreted alongside prior epidemiological evidence, these alterations represent putative links to increased risks of insulin resistance, cardiovascular dysfunction, and metabolic syndrome—relationships that require confirmation in prospective cohort studies and controlled toxicological experiments. Conclusions: Putatively annotated PM_2.5-bound ester compounds, particularly non-regulated subclasses, are associated with systemic metabolic alterations in older adults, coincident with perturbations in steroid and lipid metabolism. While these findings are exploratory and hypothesis-generating, they highlight the need to incorporate specific ester profiles into PM_2.5 risk assessments and develop targeted interventions for vulnerable aging populations. Full article

Alcohol consumption has been increasingly associated with alterations in the gut microbiota and its metabolic activity; however, evidence regarding microbiota-derived metabolites remains fragmented. This systematic review aimed to synthesize current evidence on the effects of alcohol consumption on gut microbiota-derived metabolites in humans and non-human primates. The review was conducted according to Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines and included studies published between 2012 and 2026. Searches were performed in PubMed, Web of Science, Scopus, and ScienceDirect. Study quality was assessed using the Newcastle–Ottawa Scale for human studies and the Systematic Review Centre for Laboratory Animal Experimentation (SYRCLE) Risk of Bias tool for non-human primate studies. Twelve studies met the inclusion criteria, comprising four non-human primate studies and eight human studies. Alcohol exposure was consistently associated with metabolomic alterations across multiple biological matrices. Recurrent findings included reductions in short-chain fatty acids, alterations in tryptophan-derived metabolites, changes in phenolic and aromatic amino acid-related compounds such as hippuric acid, and disturbances in bile acid and purine metabolism. Findings regarding microbial diversity and taxonomic composition were more heterogeneous, with several studies reporting reduced abundances of Faecalibacterium and related butyrate-producing taxa. Studies evaluating abstinence suggested partial recovery of both microbial and metabolomic alterations. Overall, the available evidence suggests that alcohol consumption is associated with alterations across several microbiota-related metabolic pathways, highlighting candidate metabolites that may contribute to alcohol-related pathophysiology and serve as potential translational biomarkers. Full article

A bioactive polysaccharide (CDP-D1) was isolated from the bulbs of Cistanche deserticola and purified by DEAE anion-exchange chromatography, yielding a purity of 95.5% and a recovery rate of 33.1%. It exhibits an irregular amorphous structure, with a porous and loose surface, and displays distinct shear-thinning behavior. Its molecular weight (MW) is 142.52 kDa, and its polydispersity index (Mw/Mn) is 1.77, confirming its heterogeneity. Gas chromatography–mass spectrometry (GC-MS) and nuclear magnetic resonance (NMR) analysis identified it as a heteropolysaccharide. Functional studies indicate that CDP-D1 mediates a protective effect against lipopolysaccharide (LPS)-induced inflammation in RAW264.7 macrophages. CDP-D1 restored LPS-induced decreases in cell viability to near-normal levels and suppressed pro-inflammatory signaling by downregulating TLR4 mRNA, key cytokines (IL-1, IL-6, TNF-α), and their cascade amplification. Metabolomic analysis (PLS-DA) validated the reliability of the model and identified 420 metabolites. LPS induced metabolic suppression, whereas CDP-D1 reversed this trend. Key pathways regulated by CDP-D1 include glutathione metabolism and amino acid/nucleoside/purine metabolism, partially restoring metabolic homeostasis. These findings provide preliminary evidence that CDP-D1 modulates inflammatory responses in LPS-stimulated macrophages, potentially through regulation of oxidative stress and metabolic reprogramming, and warrant further investigation into its molecular mechanism of action. Full article

Objectives: The purpose of this study was to investigate the protective effects and underlying mechanisms of fucoidan on contrast-induced acute kidney injury (CI-AKI) in mice, focusing on the TLR4/NF-κB and Nrf2/GPX4 pathways. Methods: Five-week-old male ICR mice were randomly divided into normal control, contrast model, low-dose (100 mg/kg), and high-dose (300 mg/kg) fucoidan groups. Renal index, biochemical markers, histopathology, oxidative stress indicators, inflammatory cytokine levels, and the expression of TLR4/NF-κB, Nrf2/HO-1, and ferroptosis-related proteins were assessed. Untargeted metabolomics followed by KEGG pathway enrichment was also performed. Results: Our results showed that contrast successfully established the CI-AKI model, as evidenced by an increased kidney index, abnormal biochemical parameters, severe renal pathological damage, oxidative stress imbalance, inflammatory activation, ferroptosis, and metabolic disturbances. Fucoidan dose-dependently improved kidney index and biochemical markers, alleviated pathological injury, enhanced antioxidant capacity, suppressed inflammation and ferroptosis, and reversed metabolic pathway disorders (e.g., purine and glycerophospholipid metabolism), with the high dose showing more pronounced effects. Conclusions: Fucoidan could effectively ameliorate CI-AKI, and its effects are closely associated with the inhibition of the TLR4/NF-κB pathway, activation of the Nrf2/HO-1 pathway, regulation of ferroptosis-related proteins, and improvement of key metabolic disturbances, suggesting a new research direction for the prevention of CI-AKI. Full article

Gut lactic acid bacteria are emerging as potential targets for modulating host purine metabolism and alleviating hyperuricemia-related disorders. This study aimed to isolate purine-degrading lactic acid bacterial strains from porcine intestine and systematically evaluate their probiotic potential through both in vitro and in vivo approaches. A strain designated Pedio6-1 was isolated and identified as Pediococcus acidilactici based on 16S rRNA sequencing. In vitro assays demonstrated that P. acidilactici Pedio6-1 exhibited nearly complete adenine clearance (approaching 100%) and a total purine clearance rate of 30.73%, along with strong tolerance to acidic conditions, bile salts, and gastrointestinal enzymes. To further assess its in vivo efficacy, a high-fat diet-induced obese mouse model was employed. After 8 weeks of intervention, Pedio6-1 supplementation significantly reduced the final body weight and liver index, and markedly decreased hepatic guanine levels (p < 0.05), with a trend toward lower total purine content compared to the obese control group. Mechanistically, Pedio6-1 treatment significantly downregulated the hepatic mRNA expression of pro-inflammatory cytokines IL-1β and TLR4 (p < 0.05). Serum untargeted metabolomics revealed that Pedio6-1 treatment shifted the metabolic profile toward that of normal diet-fed mice, with differential pathways predominantly enriched in porphyrin metabolism and amino acid biosynthesis and metabolism. Notably, key metabolites with antioxidant and metabolic regulatory functions, including bilirubin, biliverdin, glutathione, citrate, L-cystathionine, and 4-pyridoxic acid, were significantly elevated following treatment, while N-acetylornithine and methylmalonate ester were decreased, indicating coordinated remodeling of oxidative stress defense, vitamin B6 homeostasis, and energy metabolism. Collectively, these findings indicate that Pedio6-1 not only possesses direct purine-degrading activity in vitro, but also ameliorates purine metabolic disturbances, inflammation, and oxidative stress in obese mice, likely through the modulation of porphyrin and amino acid metabolic pathways, highlighting its promise as a functional probiotic candidate for managing obesity-associated purine metabolic disorders. Full article

Proton therapy spares normal tissues better than photon therapy, potentially reducing toxicity while maintaining tumor control. However, challenges remain due to variations in proton dose distribution, particularly at the distal edge of the spread-out Bragg peak (SOBP); for organs at risk, such variation is crucial. We evaluated the biological effects by comparing two positions of the proton profile, the entrance plateau (EP) and SOBP, in a murine model and investigated distinct metabolomic and proteomic signatures. Mice exposed to the EP beam segment (LET_d = 0.8 keV/µm) exhibited less weight reduction than their SOBP-irradiated counterparts (LET_d = 2.6 keV/µm). Two hours post-irradiation, the SOBP caused more severe DNA damage in the hippocampus and thalamus. Hematoxylin and eosin staining revealed eosinophil aggregation in both groups, with more surviving neurons in the EP group. Metabolomic profiles differed more between the EP and SOBP groups at 2 h than at 3 days post-irradiation. Relative to EP, SOBP irradiation at 2 h increased fructose-1,6-bisphosphate (FBP), dihydroxyacetone phosphate (DHAP), and inosine but decreased prostaglandin F2α; subsequently, proteomic analysis at day 3 showed that calcium signaling, NF-κB, and endocytosis pathways were enriched in the SOBP group. Combined multi-omics analysis further demonstrated that SOBP irradiation significantly activated the pentose phosphate pathway, purine metabolism, and phospholipase D signaling, while concurrently suppressing arachidonic acid metabolism. Our findings underscore the need for early detection of proton-induced brain toxicity and demonstrate that the higher-LET SOBP segment causes more severe damage than the EP. Targeting these dysregulated multi-omics pathways may offer a promising strategy for mitigating radiation-induced brain injury during proton therapy. Full article

Background/Objectives: The acute kidney injury (AKI) to chronic kidney disease (CKD) transition has been associated with a failed-repair proximal tubule (FR-PT) cell state. Hyperuricemia is an established CKD risk factor, but whether FR-PT cells have a coordinated metabolic signature reproducibly detectable across mouse models and human kidney disease remains unresolved. Methods: We assembled a pre-registered meta-analysis of 16 public mouse-kidney metabolomic cohorts (383 samples; five model classes; three injury phases) through a three-path harmonization pipeline. Convergent validation drew on KPMP single-nucleus RNA-seq (78,480 proximal tubule cells), two-sample Mendelian randomization of serum urate (102 instruments) against AKI/CKD/eGFR GWAS, and cross-cohort human plasma metabolomics across 2058 CKD/DKD patients. Results: Mouse meta-analysis identified uric acid (g = +2.70) and AICAR (g = +1.50) as the only cross-class conserved metabolites, with uric acid peaking during the AKI-to-CKD transition and returning to baseline in mouse CKD due to uricase clearance. Cross-class overlap between model classes was low (mean pairwise Jaccard = 0.109, below the pre-registered 0.20 threshold), invoking the pre-registered M-S1 stop rule: model-specific metabolic responses dominate, and the conserved signal is a small two-metabolite core superimposed on these largely model-specific programs. KPMP failed-repair PT cells exhibited transcriptional patterns consistent with coordinated four-arm purine-pathway dysregulation (de novo synthesis ↑, AMPK ↑, MOCOS ↑, URAT1 ↓), with this transcriptional pattern observed in both AKI and CKD donors (Spearman ρ = +0.900). Mendelian randomization was consistent with a possible causal contribution of serum urate to AKI/CKD/eGFR risk, with colocalization evidence at GCKR (PP.H4 = 1.000). Cross-cohort human plasma profiling across 2058 patients confirmed the systemic detectability of 29 purine-pathway metabolites in uricase-deficient humans (a translational-plausibility check, not validation of cellular source). Conclusions: The integrated data support a model in which FR-PT-associated purine-pathway reprogramming may contribute to the elevated urate signal observed during AKI-to-CKD transition, with uric acid as the candidate metabolic readout. Differences in uricase activity between mice and humans may help explain why chronic-phase urate signals are attenuated in mice but persist in humans. This work nominates a candidate cell-state metabolic readout of the AKI-to-CKD transition as a hypothesis for prospective testing; it does not establish causation, and therapeutic translation would require dedicated interventional studies. Full article

(1) Objective: To characterize structural and functional alterations of gut microbiota in patients with newly diagnosed active pulmonary tuberculosis (ATB), screen differential bacterial taxa associated with Mycobacterium tuberculosis (MTB) infection, and explore tuberculosis-related microbial metabolic alterations via predictive functional profiling. (2) Methods: Fresh morning fecal samples were collected from 33 treatment-naive patients with newly diagnosed ATB (ATB group) and 30 healthy controls (HC group). 16S rRNA gene high-throughput sequencing was performed to compare intergroup differences in gut microbial α/β diversity, taxonomic composition, and predicted KEGG functional profiles. Receiver operating characteristic (ROC) curve analysis was conducted to evaluate the internal discriminative ability of candidate differential genera in this single small cohort. (3) Results: The ATB group showed significantly lower gut microbial α-diversity than healthy controls (all P < 0.05). Principal coordinate analysis (PCoA) based on Bray-Curtis distances combined with permutational multivariate analysis of variance (PERMANOVA) revealed significant overall dissimilarity of gut microbial community structure between the two groups (R² = 0.31, p = 0.005). At the phylum level, the relative abundances of Firmicutes and Bacteroidetes were higher, while Proteobacteria was less abundant in ATB patients relative to HC. At the genus level, Streptococcus, R. gnavus and Parabacteroides were significantly enriched in ATB patients, whereas Bifidobacterium, Pseudomonas, Megamonas and Faecalibacterium were depleted. Linear discriminant analysis effect size (LEfSe) analysis uncovered group-specific signature taxa, with pro-inflammatory genera Streptococcus and R. gnavus markedly enriched in the ATB group. ROC analysis yielded area under the curve (AUC) values of 0.836 for Streptococcus and 0.805 for R. gnavus. Predictive KEGG functional analysis demonstrated obvious intergroup differences in microbial metabolism, with purine and pyrimidine nucleotide metabolism pathways significantly upregulated in the ATB group. (4) Conclusions: Treatment-naive patients with ATB exhibited reduced gut microbial diversity in this small cohort. The enrichment of Streptococcus and R. gnavus, as well as the upregulation of purine and pyrimidine metabolic pathways, are all associated with active MTB infection. Full article

Background/Objectives: Reproductive inefficiency remains a major contributor to heifer culling and reduced herd longevity in beef systems. This study examined the molecular basis of fertility by analyzing granulosa cells and follicular fluid from Angus–Simmental crossbred heifers classified as fertile or subfertile. Methods: Granulosa cells and follicular fluid were collected for RNA sequencing and metabolomic analysis. Differential expression, network, and gene–metabolite integration analyses identified genes, metabolites, and pathways associated with fertility differences between groups. Results: We identified 90 differentially expressed genes from the granulosa cells, including CXCL12 and HOXD family members, CALCRL, DNER, GADD45G, immune-related genes, and nine metabolites differentially abundant in follicular fluid, including parabanic acid, pimelic acid, α-tocopherol, glycine, and arachidonic acid. Network analysis revealed extensive rewiring in the network from the subfertile heifers for both genes and metabolites as compared to the fertile heifers. The gene–metabolite integration revealed a high degree of connectivity between arachidonic acid and α-tocopherol, correlating with immune and signaling genes, and between CXCL12 and carbohydrate intermediates. Pathway over-representation analysis highlighted carbohydrate and purine metabolism in fertile heifers, and aminoacyl-tRNA biosynthesis, alanine/aspartate/glutamate metabolism, and Hippo signaling in the subfertile heifers. ABC transporters, ferroptosis, mineral absorption, and glyoxylate/dicarboxylate metabolism were common pathways identified in both groups. Integrative granulosa-follicular fluid analysis revealed coordinated gene–metabolite rewiring in subfertility, including functional candidates (CXCL12, CALCRL, DNER; α-tocopherol, arachidonic acid, fucose, parabanic acid) and fertility-associated pathways. Conclusions: These signatures provide novel targets and pathways underlying beef heifer fertility. Full article

This study investigated how defatted silkworm pupae meal (DSPM) modulates flesh quality in pearl gentian grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂). A total of 360 size-uniform fish were randomly assigned to four dietary treatments for 8 weeks: D0, the fishmeal (FM) based control diet; and D1, D2, and D3, in which DSPM substituted 25%, 50%, and 100% of dietary FM, respectively, with three replicates per treatment and 30 fish per replicate. Compared with D0, D1 and D2 improved final body weight, weight gain rate, specific growth rate, and feed conversion ratio, whereas D3 reduced survival and feed utilization. For flesh quality, D2 showed the most favorable phenotype, characterized by higher crude protein, collagen, flavor-associated amino acids, pH, texture attributes, and muscle fiber density, together with lower ether extract, cooking loss, freezing loss, and shear force. DSPM substitution also increased unsaturated and polyunsaturated fatty acid proportions and improved lipid health indices. Moderate substitution, especially D2, enhanced oxidative stability by increasing SOD and CAT activities and reducing MDA accumulation. Non-targeted metabolomics combined with qPCR analysis of selected flesh quality-related genes suggested that these improvements were associated with osmotic regulation, collagen remodeling, purine metabolism, membrane lipid homeostasis, nutrient sensing, myogenesis, and antioxidant defense. Complete FM substitution impaired muscle structure, water retention, and redox balance. Thus, 50% FM substitution with DSPM effectively optimized grouper flesh quality in this study. Full article

Diffuse intrinsic pontine glioma (DIPG) is a rare and fatal pediatric brainstem malignancy for which effective treatment options are lacking. Cerebrospinal fluid (CSF) analysis can reveal intrinsic alterations and characteristic metabolic profiles of the tumor microenvironment. In this study, the proteome and metabolome of CSF from DIPG patients were comprehensively analyzed to identify potential biomarkers and the pathways involved. Functional annotation and pathway enrichment analyses were performed using the GO (Gene Ontology) and KEGG (Kyoto Encyclopedia of Genes and Genomes) databases. Bioinformatics methods were used to comprehensively analyze the proteomic and metabolomic results to identify key differentially expressed proteins, metabolites, and potential signaling pathways involved in DIPG. In total, 885 DEPs (differentially expressed proteins) were identified in cerebrospinal fluid from DIPG patients, of which 54 were upregulated and 831 were downregulated, primarily originating from the cytoplasm and cell membrane. Among the top 20 upregulated proteins, URB1 (nucleolar pre-ribosomal-associated protein 1) had the greatest statistical significance, while the remaining proteins were mostly immunoglobulin fragments. GO enrichment analysis revealed that the downregulated proteins were enriched primarily in cellular processes, metabolic processes, and binding functions. KEGG analysis revealed that upregulated proteins were significantly enriched in complement and coagulation cascades, whereas downregulated proteins were primarily associated with endocytosis and certain microbial infections. A total of 1372 metabolites were identified, of which 40 were differentially expressed: 24 were upregulated, and 16 were downregulated. Pathway analyses of the differentially expressed metabolites revealed that they were primarily related to purine metabolism and tyrosine metabolism. The multiomics analysis revealed that purine metabolism is particularly important in DIPG. Full article