Purpose: Huatan Huoxue decoction (HTHXD) is effective in treating polycystic ovary syndrome (PCOS); however, its underlying mechanisms of action remain unclear. Methods: Gene targets were obtained through Mendelian randomization (MR) analysis using druggable genes from public databases and genome-wide association study data for PCOS from the FinnGen R12 database. These targets were integrated with the active ingredients and protein targets of HTHXD retrieved from the TCMSP and BATMAN-TCM databases for network pharmacology analysis. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were conducted. Core targets were validated using molecular docking. Experimental validation was performed using a letrozole-induced PCOS rat model treated with HTHXD, with key targets assessed via quantitative PCR (qPCR) and immunohistochemical staining. Results: MR analysis identified 131 proteins causally related to PCOS, of which 52 with strong associations remained after quality control. Network pharmacology analysis revealed 549 gene targets and 243 active ingredients in HTHXD. The intersection of these datasets identified four core targets for HTHXD in PCOS treatment (CA2, CASP3, SNCA, and TSPO). Molecular docking identified tanshindiol B, 4-methylenemiltirone, and danshenol B in Danshen and crocetin in leech as key active components. HTHXD substantially ameliorated polycystic ovarian morphology and reduced corpora lutea in PCOS model rats. CA2 expression was significantly upregulated in PCOS ovarian tissue (P < 0.0001), and markedly reduced after HTHXD treatment, consistent with immunohistochemical findings. Conclusion: CA2 may serve as a potential candidate gene associated with the therapeutic response to HTHXD in PCOS, with tanshindiol B and crocetin as major active components.
This study provides real-world evidence by comparing the long-term clinical outcomes of percutaneous coronary intervention (PCI) versus medical therapy alone in patients with chronic coronary syndrome (CCS) diagnosed non-invasively, given the lack of evidence in this population. This retrospective cohort study (2013-2018) utilized data from a medical center’s Cardiovascular Disease Databank, which was linked to national mortality records in Taiwan. Propensity score matching (PSM) and stabilized inverse probability of treatment weighting (IPTW) were applied to adjust for baseline differences between the PCI and medication-only groups. Major adverse cardiovascular events (MACE), including all-cause mortality, non-fatal myocardial infarction (MI), and revascularization, were compared between groups. After 1:4 propensity score matching, a total of 4,369 patients were included (PCI, N=1,058; medication only, N=3,311). Adjusted Cox proportional hazards models showed that the PCI group had a higher hazard of MACE (aHR, 1.38 [95% CI, 1.22-1.56]), primarily driven by increased revascularization (aHR, 1.65 [95% CI, 1.39-1.95]), compared to the medication-only groups. No significant differences were observed in all-cause mortality (aHR, 1.13 [95% CI, 0.96-1.34]) or MI (aHR, 1.08 [95% CI, 0.77-1.52]). Furthermore, the mean total healthcare cost during the index year was US$3,232 (SD = 9,896) for the medication-only group, compared with US$10,931 (SD = 8,781) for the PCI group. Among CCS patients diagnosed non-invasively, the medication-only group demonstrated comparable all-cause mortality and MI rates, but a significantly lower risk of revascularization compared with the PCI group, while incurring only one-third of the healthcare costs. Clinicians should carefully evaluate the benefits and risks of stent placement in CCS patients.
The tubers of Lepidium meyenii (commonly known as maca) have been used in the Andean cuisine for ages. The study aims to determine the behavior of the extract from the red phenotype of maca in the in vitro gastrointestinal tract model and also to define the overall impact of the extract on the microbiota. Glucosinolates, lepidilines, and a derivative of β-carboline alkaloid, i.e. 1-methyltetrahydro-β-5,6-hydride-carboline-3-carboxylic acid, as representatives of one of the main groups of metabolites were selected for the quantitative analysis after in vitro digestion at every stage of the digestion model. The results showed that lepidilines remained highly stable throughout the digestive simulation, whereas glucosinolates and the β-carboline derivative were substantially degraded, under the influence of the enzymes and bacteria applied in the “large intestine”. In addition, the red maca extract exhibited prebiotic activity by promoting the growth of selected lactic acid bacteria, with Lactococcus lactis showing the most pronounced response, suggesting that maca-derived preparations may hold promising potential as ingredients in functional foods and microbiota-targeted nutritional formulations.
Atopic dermatitis (AD) is a chronic inflammatory disease characterized by recurrent bacterial infections that cause pruritus, erythema, xerosis and thickening. Cajanus cajan root extract prepared with 95% ethanol (EECR95) has been shown to possess antioxidant, anti-inflammatory, and antibacterial properties in previous studies. This study aimed to evaluate EECR95’s ability to improve AD by inhibiting methicillin-resistant Staphylococcus aureus (MRSA) growth and reducing inflammation. This study found EECR95 effectively inhibits MRSA growth (IC50 = 103.74 μg/mL) and reduces pro-inflammatory cytokine production in an MRSA-keratinocyte-macrophage model. LC/MS/MS analysis revealed the presence of active compounds- flavonoids, isoflavones, and flavonoid glycosides, highlighting EECR95's potential in improving atopic dermatitis in vitro.
Heroin is classified as a Schedule I controlled substance with severe legal consequences for its use, whereas liquid Brown Mixture (BM) is a legally available and commonly prescribed cough remedy in Taiwan. However, numerous cases of BM ingestion have been misinterpreted as heroin use because BM contains opium tincture, leading to similar urine test results. Owing to the rapid metabolism of heroin, detection of the parent drug and its primary metabolite, 6-acetylmorphine, in urine is difficult; thus, interpretation often relies on morphine concentration and the morphine-to-codeine (M/C) ratio, which can be inconclusive. In our BM analysis, the M/C ratio was 3.3 ± 0.4, and among 5,094 morphine-positive urine samples, 279 (5.5%) had M/C ratios between 2 and 4, suggesting that a substantial number of individuals could be misidentified. Given the lack of reliable urinary biomarkers to distinguish BM from heroin use, this study aimed to identify BM-specific indicators. BM contains antimony (Sb) potassium tartrate, an emetic agent absent in heroin formulations. Although urinary Sb levels are low, they can be quantified using highly sensitive inductively coupled plasma mass spectrometry (ICP–MS). A pharmacokinetic study involving single and multiple BM doses revealed that urinary Sb increased from ≤ 0.11 ng/mL pre-dose to 0.4 – 98.9 ng/mL post-dose. Morphine and codeine were determined using gas chromatography–mass spectrometry (GC–MS), and M/C ratios ranged from 2.0 to 14.9, showing a secondary rise at later stages. These findings suggest that urinary Sb may serve as a useful supplementary biomarker for BM ingestion, thereby enhancing the accuracy of forensic toxicological interpretation and reducing false identification of heroin use.
Cannabis sativa L. or hemp is a cosmopolitan, multi-purpose plant and area of its cultivation has been growing in recent years. Its public perception is influenced by potential to produce narcotic substances. However, hemp seed can be a valuable source of nutrients for humans and may be of particular interest in more sustainable food consumption patterns, including vegetarian and flexitarian diets. The paper presents benefits and risks of hemp seeds in case of their consumption in food. Low cannabinoids varieties are considered only. The biological activity of cannabinoids and related effects are considered out of scope of the study. Nutritional value of hemp seeds regarding content of amino acids is comparable to the value of egg whites and soya. Oil is a source of two essential fatty acids: linoleic acid and alpha-linolenic acid in a desirable ratio of 3:1. Hemp seeds are also a source of minerals, vitamins and carbohydrates, including fiber. Their most characteristic hazardous properties are ability to accumulate toxic heavy metals and produce anti-nutritional substances. Breeding hemp to reduce the anti-nutritional compounds content may be promising in view of its growing popularity as a food source.
Garambullo (Myrtillocactus geometrizans) is a Mexican fruit rich in bioactive phytochemicals. Its fermented extract (FEG), obtained after in vitro gastrointestinal digestion and fermentation, was evaluated in HT-29 and SW480 colorectal cancer cells. FEG reduced cell viability (IC50: 2985 and 3200 μg/mL, respectively) and induced apoptosis and necrosis. In 3D assays, FEG inhibited colony formation, with complete suppression in HT-29 cells. Molecular docking further suggested that butyric acid identified in FEG interacts with Caspase-3 and RIPK1. These findings support the antiproliferative and pro-death effects of FEG in colorectal cancer models.
Circadian rhythms regulate fundamental aspects of physiology and behavior. The disruption of circadian rhythms has been associated with metabolic disorders, cardiovascular disease, obesity, and impaired immune function. Feeding behavior and gut microbiota represent two essential nodes within this regulatory network, as both are highly responsive to environmental cues such as diet composition and sleep-wake cycles. Research has shown that high-fat diets, excessive fructose, and chronic alcohol consumption perturb peripheral circadian rhythms and impair metabolic pathways, contributing to obesity and insulin resistance. Conversely, dietary interventions such as time-restricted feeding and macronutrient-specific entrainment can restore rhythmic gene expression and prevent metabolic pathologies. The gut microbiota plays a central role in mediating these effects through its diurnal oscillations in community composition and metabolite production, particularly short-chain fatty acids, which act as systemic cues for host circadian clocks. Disruption of microbial rhythmicity, whether through irregular eating schedules, jet lag, or environmental pollutants, can exacerbate circadian misalignment and metabolic dysfunction. Recent findings highlight a bidirectional relationship between the host circadian system and the microbiota, underscoring their interdependence in maintaining metabolic homeostasis and gastrointestinal integrity. Although preclinical models have produced mechanistic insights, the translation to humans remains limited by methodological variability and incomplete characterization of signaling pathways. This review synthesizes the current knowledge on diet-microbiota-circadian interactions and discusses potential therapeutic strategies, including dietary modulation and natural products, to mitigate circadian misalignment and its associated health risks.
Dietary strategies and plant-based fermented foods have attracted increasing interest for supporting gastrointestinal health, particularly under conditions of drug-induced mucosal stress. Soybean fermentation broth (SFB), a fermented soybean-derived product, may possess enhanced nutritional and functional properties through fermentation-associated generation of bioactive peptides and antioxidant metabolites. This study evaluated the gastroprotective effects of SFB in an indomethacin-induced gastric injury model in rats and investigated the underlying mechanisms. Indomethacin markedly induced gastric ulcer formation (16.61 ± 1.38 mm2), whereas SFB pretreatment significantly attenuated ulcer development in a dose-dependent manner, reducing ulcer area to 6.66 ± 0.98, 3.89 ± 0.63, and 1.46 ± 0.32 mm2 in low-, medium-, and high-dose groups, respectively (p < 0.05 vs. Vehicle). Histopathological examination revealed that SFB preserved epithelial integrity and reduced submucosal edema. SFB pretreatment significantly decreased lipid peroxidation, restored gastric glutathione content, and enhanced antioxidant enzyme activities, including glutathione peroxidase (GPx), glutathione reductase (GRd), superoxide dismutase (SOD), and catalase (CAT). Furthermore, SFB suppressed pro-inflammatory mediator expression and favorably modulated cyclooxygenase-1 (COX-1) and COX-2 expression, accompanied by restoration of prostaglandin E2 (PGE2) production. These findings indicate that SFB protects gastric mucosa against indomethacin-induced injury through coordinated regulation of oxidative stress, inflammatory signaling, and prostaglandin-mediated mucosal defense, supporting its potential as a functional fermented soybean product.
Mass spectrometry imaging (MSI) has become an indispensable tool in metabolomics for visualizing the spatial distribution of biomolecules within tissues. Due to its direct, ambient analysis capability, desorption electrospray ionization (DESI-MSI) has evolved from a rapid screening tool into a precise and reliable quantitative platform. We delve into its technical principles and particularly review the criticality of sample preprocessing, highlighting the paradigm shift from a "no-preprocessing" approach to one of "optimized preprocessing." The paper details how factors like sample preservation, choice of embedding agents, washing, drying, and on-tissue chemical derivatization all impact data quality. We also discuss how the addition of standards and internal standards can mitigate matrix effects and signal variability, enabling accurate quantitative analysis. Finally, this article looks ahead to the future of DESI-MSI, including its combination with emerging technologies such as tissue expansion mass spectrometry imaging to achieve single-cell-level spatial resolution and open up new possibilities for metabolomics research.
Trifolium repens (white clover) is rich in bioactive volatiles with therapeutic potential. This study developed an advanced hybrid ultrasound-infrared-microwave hydro-distillation (UIMHD) method for efficient essential oil (EO) isolation and investigated its active sesquiterpene, pseudoionone, for multi-mechanistic neuroprotection in diabetic neuropathy (DN). The optimized UIMHD system synergistically combined ultrasound cavitation, infrared heating, and microwave hydro-distillation, enhancing yield and reducing time versus conventional extraction. Gas chromatography-mass spectrometry (GC-MS) profiling identified pseudoionone as the dominant constituent. Pharmacological evaluations were performed in alloxan-induced diabetic mice to assess antidiabetic, anti-inflammatory, neuroprotective, and anticonvulsant activities. Cytokine modulation (TNF-α, IL-6, IL-10), oxidative stress markers (CAT, GSH, TBARS), and strychnine-induced seizure assays were conducted to elucidate mechanisms. The UIMHD technique increased T. repens essential oil (TR-EO) yield by 62.5% and shortened isolation time by 50%. Pseudoionone (49.16%) exhibited potent hypoglycemic and insulin-restorative effects, alleviated thermal hyperalgesia and tactile allodynia, and improved antioxidant defenses. Both the TR-EO and pseudoionone significantly down-regulated TNF-α and IL-6 while up-regulating IL-10, indicating anti-inflammatory cytokine balance restoration. Additionally, pseudoionone delayed strychnine-induced tonic seizures, suggesting functional involvement of glycinergic pathways. Strong antibacterial and antibiofilm activities were also observed against multidrug-resistant Shigella dysenteriae. The advanced UIMHD method provides a sustainable platform for isolating bioactive volatiles from T. repens. Pseudoionone emerges as a multi-target natural agent mitigating diabetic neuropathy via TNF-α/IL-6/IL-10 modulation, oxidative- stress restoration, and strychnine-sensitive GlyR potentiation.
Metabolomics studies small-molecule metabolites to provide insights into health and disease, supporting early diagnosis and personalized medicine. Advances in mass spectrometry and nuclear magnetic resonance (NMR) have expanded its use in metabolic, cancer, and cardiovascular diseases. In pediatrics, high-resolution NMR metabolomics has been instrumental in identifying age-related metabolic changes during early childhood and their associations with growth, nutrition, and disease risk. However, a comprehensive review of its clinical applications and future potential remains limited. This review highlights how utilizing specific NMR pulse sequences, such as CPMG and NOESY, allows for precise and non-destructive metabolic profiling of diverse biofluids, supported by minimal sample preparation and high-throughput automated analysis. Data processing tools like NMRProcFlow and MetaboAnalyst facilitate spectral preprocessing, statistical analysis, and biological interpretation, streamlining metabolomics workflows. Clinically, NMR-based metabolomics has elucidated metabolic alterations in pediatric growth, prematurity, nutrition-related sensitizations, allergic diseases, lipid metabolism, infectious conditions, and neurobehavioral disorders. In particular, metabolomics has been applied to identify specific metabolic signatures underlying the molecular mechanisms of childhood allergic asthma. Despite limitations in detecting low-abundance metabolites, NMR's ability to preserve sample integrity and integrate multi-omics data, especially gut microbiota-derived metabolites, shows great promise in advancing precision pediatric medicine, early disease screening, and personalized therapeutic strategies.
Metabolomics is the most recent and final discipline among the post-genomic, multiomics fields. It encompasses key technologies in systems biology, providing an overview of low-molecular-weight metabolites while examining disturbances to offer pathophysiological explanations. The two key analytical tools in metabolomics are nuclear magnetic resonance spectroscopy and mass spectrometry, which includes gas chromatography-mass spectrometry and liquid chromatography-mass spectrometry. Metabolomic research relies on a three-stage data generation process: the first stage involves precise analytical chemistry to produce data; the second stage employs multivariate statistical analysis; and the last stage uses big data as a reference for comparison to establish biochemical, metabolic, and pathophysiological correlations, thereby providing possible interpretations. The initiative of precision medicine starts with cancer as the first disease target. This review discusses and illustrates the application of metabolomics in precision medicine, choosing two types of cancer as example. Warburg effect and metabolic reprogramming are particularly discussed. By providing global, top-down, and less biased information, metabolomics enables precision medicine.
To our knowledge, this is the first case reported in literature, regarding an increase in tumoral biomarkers during a treatment with Semaglutide. A 61-year-old patient was referred to our gynecological consultation due to suspected adnexal cysts following an MRI performed for a functional defecation disorder. The patient was on Semaglutide treatment. Gynecological examination and transvaginal ultrasound were normal, with no evidence of adnexal cysts. Tumor biomarker testing revealed a significant increase in CA 19-9 and a mild elevation in CA-125. In an asymptomatic patient, these findings raised suspicion of a possible neoplastic origin involving the pancreas or biliary tract. However, all further investigations yielded normal results. At a follow-up laboratory check one month later, CA 19-9 levels had decreased from 717 to 62 kU/L. A subsequent evaluation two months later confirmed the normalization of tumor biomarkers. Despite the elevation of CA19-9 during pancreatitis, which could be induced by Semaglutide, it is important to recognize that not all patients will experience this side effect. Moreover, an increase in tumor biomarkers can occur even in the absence of a clinically detectable disease, such as pancreatitis or cancer. Further research is needed to explore the potential relationship between Semaglutide and elevated CA 19-9 or CA-125 levels.
Phthalates are ubiquitous environmental contaminants that raise particular concern for children, who generally experience higher exposure levels and are more susceptible to environmental pollutants. To understand potential molecular mechanisms, we conducted a cross-sectional study of 256 Taiwanese children aged 8-10 years from two population-based cohorts: the Taiwan Birth Panel Study and the Taiwan Early-Life Cohort. Serum phosphocholine-containing lipids were profiled by liquid chromatography coupled with triple quadrupole mass spectrometry and associated with urinary levels of 12 phthalate metabolites. Multiple linear regression models, adjusted for demographic and socioeconomic factors, revealed that mono-(2-ethyl-5-hydroxyhexyl) phthalate (MEHHP), an oxidative metabolite of di(2-ethylhexyl) phthalate (DEHP), exhibited the strongest associations with lipid alterations. Specifically, higher urinary MEHHP was positively associated with several diacyl-phosphatidylcholines and lyso-phosphatidylcholines, while inversely related to sphingomyelins and ether-linked phosphatidylcholines. These lipid signatures suggest disturbances in lipoprotein metabolism, peroxisome proliferator-activated receptor-related signaling, and sphingolipid balance, offering mechanistic clues to previously observed links between phthalate exposure and hepatic, cardiovascular, and neurological outcomes. Overall, this study identifies lipid perturbations associated with metabolites of DEHP exposure in children and provides molecular insights into how phthalates may disrupt lipid metabolic homeostasis during early life.
The gut microbiota plays a crucial role in host digestion and immune regulation; however, most existing research relies on fecal sampling, leaving uncertainty regarding whether fecal microbiota can accurately reflect the symbiotic microbial components at distinct gut sites. To address this gap, this study employed 16S rRNA gene sequencing to systematically analyze the microbial composition of three spatial compartments in the rat colon: luminal contents, mucus layer, and epithelial tissue. The results revealed notable differences in microbial communities across the three compartments. The luminal contents, which were closely associated with the fecal microbiota, exhibited high microbial abundance, encompassing gram-positive bacteria, gram-negative bacteria, aerobic bacteria, and obligate anaerobes. By contrast, the mucus layer and epithelial tissue were predominantly enriched in gram-negative bacteria and obligate anaerobes, with a marked scarcity of aerobic bacteria. Further analysis indicated that the ACE and Chao1 indices were significantly higher in mucus samples than in luminal samples, whereas tissue samples showed reduced richness but relatively stable diversity. Additionally, Proteobacteria were rarely detected in the lumen but were significantly enriched in the mucus and tissue. These findings suggest that the oxygen content in the intestinal lumen is significantly higher than that of the host-derived mucus layer and epithelial tissue and that this oxygen gradient drives the spatial partitioning of the gut microbiota. Collectively, this study clarifies the spatial distribution characteristics of the gut microbiota and highlights oxygen availability as a key factor in regulating microbial adhesion and colonization.
Bifidobacterium longum is a probiotic that resides in the intestine, and extracellular vesicles (EVs) released by microorganisms have garnered attention in recent years for their role in regulating host health. This study found that triacylglycerol, cholesteryl esters, and free fatty acid (18:0 and 16:0) are primarily composed of lipids in B. longum-derived EVs by lipidomics assay. Such lipid composition facilitates the entry of EVs into ARPE-19 cells and protects the cells from oxidative death induced by blue light exposure. Transcriptomic analysis revealed that treatment with B. longum-derived EVs can regulate phosphoinositide 3-kinase and mTOR, thereby enhancing the survival capacity of blue light-induced ARPE-19 cells. Intestinal microbiota is strongly associated with host disease, and this study provides a strategy suggesting that intestinal probiotic EVs might potentially be used in the future to protect eyes from damage caused by blue light sensitivity through the gut-eye axis.
Hepatocellular carcinoma (HCC) remains a leading cause of cancer-related deaths worldwide. Although advanced oral anticancer drugs, such as sorafenib and lenvatinib, are available for treating HCC, their effectiveness is limited by significant side effects and the requirement for optimal liver function, restricting their broader use. Furthermore, these drugs cannot be used in combination, leading to suboptimal treatment outcomes. This study introduces a novel therapeutic approach by repurposing two existing drugs, niclosamide and disulfiram, as a combination therapy to treat HCC. Despite their known anticancer properties, these drugs face challenges such as high lipophilicity and a significant first-pass effect, resulting in low bioavailability. To overcome this limitation, we used chemically modified inulin fibers as oral carriers to enhance drug absorption and protect the drugs from the acidic gastric environment. We compared the efficacy of this system across two HCC cell lines-HepG2 (from a Caucasian patient) and Huh6 (from an Asian patient)-to explore how genetic and ethnic variations impact sensitivity to the proposed therapeutic approach. Our findings indicate that the proposed inulin oral delivery approach improves drug efficacy both in vitro and in vivo. Moreover, our comparative study underscores the importance of considering ethnic and genetic diversity in designing effective cancer therapies. Overall, the proposed oral delivery system can potentially reduce the need for invasive procedures, lower infection risks, and minimize patient discomfort by avoiding the need for intravenous administration, while improving treatment outcomes.
Metabolomics provides direct insights into cellular physiology, yet it faces greater analytical challenges compared to genomics and transcriptomics due to the chemical diversity, instability, and environmental sensitivity of metabolites. Conventional bulk metabolomics averages signals across cell populations, thereby masking cellular heterogeneity critical for understanding disease mechanisms. Recent breakthroughs in single-cell metabolomics (SCM), driven by advances in mass spectrometry, microfluidics, isotope tracing, and spatial omics, have enabled the detection of metabolic diversity at unprecedented resolution. SCM has uncovered cell-type-specific biomarkers, revealed metabolic reprogramming in cancer and immunity and revealed disease progression. These studies highlight SCM's transformative potential in biomarker discovery, clinical diagnostics, and precision medicine. Despite rapid progress, SCM remains limited by low metabolite abundance, instability during cell handling, lack of standardized quantification methods, and challenges in integrative multi-omics analysis. Future developments will require innovations to improve sensitivity and spatial resolution, establish cross-laboratory quality control frameworks, and apply artificial intelligence for data interpretation. With continued technological convergence, SCM is poised to evolve from a niche research tool into a cornerstone platform for biological and biomedical research.
Oral squamous cell carcinoma (OSCC) remains a major global health challenge, and early detection is essential for improving patient outcomes. To identify reliable salivary biomarkers, we performed a targeted metabolomics study using 13C2/12C2-dansylation, integrating candidate metabolites from published OSCC metabolomics datasets and systematically verifying them through LC-MRM-MS. A panel of 26 metabolites was quantified in saliva samples from 299 subjects, including healthy controls (n = 98), oral potentially malignant disorders (OPMD I, n = 50; OPMD II, n = 53), and OSCC patients (n = 98). Among these, cadaverine, N-acetylcadaverine, choline, glycine, and tryptophan were significantly dysregulated across disease groups. Cadaverine, N-acetylcadaverine and choline were consistently elevated in OSCC and showed progressive increases with disease stages, whereas glycine levels declined. A four-metabolite panel (cadaverine, N-acetylcadaverine, choline, glycine) demonstrated strong diagnostic performance in distinguishing OSCC from healthy controls (AUC = 0.91). Correlation analyses further revealed coordinated regulation among polyamine-related metabolites, suggesting a reprogramming of ornithine-polyamine metabolism in OSCC progression. In conclusion, we establish a verified salivary metabolite panel with high discriminatory power for OSCC detection. These findings highlight polyamine-associated metabolic alterations as a hallmark of malignant transformation in the oral cavity and support salivary metabolomics as a clinically accessible tool for non-invasive screening and risk stratification.
