To enhance the accuracy of early prostate cancer diagnosis, we developed an electrochemical sensor based on synthesized mesoporous carbon nanospheres functionalized with ferrocenecarboxylic acid and encapsulated hemoglobin (HMCNs-FCA-Hb). This sensor enables the simultaneous detection of prostate-specific antigen (PSA) and sarcosine (SAR) as dual biomarkers. Unlike traditional electrochemical detection methods that frequently necessitate multiple electrode modifications, our sensor operates without the need for such modifications, thereby significantly simplifying the detection process. Under alkaline conditions, HMCNs-FCA-Hb can release ferrocenecarboxylic acid to generate an electrical signal for PSA detection. The linear range for PSA detection is from 0.001 to 30 ng/mL, with a detection limit of 0.11 pg mL-1 (S/N = 3). Additionally, HMCNs-FCA-Hb with excellent peroxidase-like activity allows for indirect detection of SAR with the linear range of 0.01-25 μM and a detection limit of 0.003 μM (S/N = 3). Specifically, we have integrated a micro electrochemical workstation and mobile smart devices to achieve portable and wireless detection of PSA and SAR in clinical serum samples with satisfactory results. The results can be visually and promptly displayed, highlighting the sensor's potential for clinical application in the early diagnosis of prostate cancer.

Prostate cancer (PCa) is the second most commonly occurring cancer among men worldwide. Although the clinical management of PCa has significantly improved, a number of limitations have been identified in both early diagnosis and therapeutic treatment. Because multiple studies show that prostate-specific antigen (PSA) screening frequently results in overdiagnosis and overtreatment, the use of PSA alone as a diagnostic marker for PCa screening has been controversial. For individuals with locally advanced or metastatic PCa, androgen deprivation therapy (ADT) is the standard initially successful treatment; nonetheless, the majority of patients will eventually develop lethal metastatic castration-resistant prostate cancer (CRPC). Alternative treatment options, including chemo-, immuno-,or radio-therapy, can only prolong the survival of CRPC patients for several months with the most developing resistance. Considering this background, there is an urgent need to discuss about selective prostate-specific biomarkers that can predict clinically relevant PCa diagnosis and to develop biomarker-driven treatments to counteract CRPC. This review addresses several PCa-specific biomarkers that will assist physicians in determining which patients are at risk of having high-grade PCa, focusing on the clinical relevance of these biomarker-based tests among PCa patients. Secondly, this review highlights the effective use of these markers as drug targets to develop precision medicine or targeted therapies to counteract CRPC. Altogether, translating this biomarker-based research into the clinic will pave the way for the effective execution of personalized therapies for the benefit of healthcare providers, the biopharmaceutical industry, and patients.

Metabolomics, a burgeoning field within systems biology, focuses on the comprehensive study of small molecules present in biological systems. Mass spectrometry (MS) has emerged as a powerful tool for metabolomic analysis due to its high sensitivity, resolution, and ability to characterize a wide range of metabolites thus offering deep insights into the metabolic profiles of living systems.

This review provides an overview of the methodologies, workflows, strategies, data analysis techniques, and applications associated with mass spectrometry-based metabolomics.

We discuss workflows, key strategies, experimental procedures, data analysis techniques, and diverse applications of metabolomics in various research domains. Nuances of sample preparation, metabolite extraction, separation using chromatographic techniques, mass spectrometry analysis, and data processing are elaborated. Moreover, standards, quality controls, metabolite annotation, software for statistical and pathway analysis are also covered. In conclusion, this review aims to facilitate the understanding and adoption of mass spectrometry-based metabolomics by newcomers and researchers alike by providing a foundational understanding and insights into the current state and future directions of this dynamic field.

Equally with other indigenous green leafy vegetables, Solunum nigrum L. has been widely consumed by the VhaVenda tribe found in the Limpopo Province of South Africa since ancient times as a source of food diversification due to its higher-quality nutritional value, sustainability, food security, and medicinal benefits. It is mostly cultivated from seeds in seedling trays and transplanted in the open field, and at the maturity stage, marketing and distribution are mainly conducting through informal markets (i.e., street vendors). However, recently, it can be found in selected supermarkets and commercial grocery stores in South Africa. The leaves and young shoots of S. nigrum are cooked solely and/or as a supplementary vegetable with Brassica rapa L. subsp. chinensis (Chinese cabbage), Spinacia oleracea L. (spinach), Amaranthus graecizans L. (green amaranth), Solanum lycopersicum L. (tomato), and/or cooking oil for flavor.

Contrary to other green leafy vegetables, few studies have been conducted on the metabolites released by S. nigrum and the influence of growing conditions on the metabolites thereof.

A 1H-nuclear magnetic resonance tool was used to identify the untargeted metabolites released by S. nigrum, and spectra were phase-corrected and binned with MestReNova and statistically analyzed with SIMCA 18.0.2.

The findings showed that a total of 12 metabolites were detected between the growing conditions. Eleven similar metabolites, such as glycocholate, chlorogenate (human health benefits), caffeine for its bitter taste, choline, 3-Chlorotyrosine (antidiabetic, blood pressure), etc., and a few vital soluble sugars, were detected in S. nigrum samples grown in the open field and greenhouse-cultivated. Glucose was exclusively detected in the S. nigrum grown under greenhouse conditions.

Despite clinical suspicion of prostate cancer (PCa), 20% to 25% of patients exhibit a tumor-negative biopsy result.

This work aimed to assess the serum metabolic profile of clinically significant (cs) compared to clinically insignificant (ci) PCa or benign (Be) patients.

A total of 1078 serum samples were analyzed. Nuclear magnetic resonance (NMR) spectroscopy was used to quantify 73 metabolites; random forest was used for the model algorithm.

We identified a 22-metabolite panel, which discriminated csPCa (International Society of Urological Pathology [ISUP] 2-5, n = 328) from ciPCa (ISUP 1, n = 101) and Be patients (negative biopsy, n = 649) with a higher performance when combined with the standard clinical parameters age, prostate-specific antigen (PSA), and percentage free PSA (%fPSA) (area under the curve [AUC] 0.84) than the clinical parameters alone (AUC 0.73). Our study further revealed significant dysregulations of the urea cycle and the choline pathway along with changes in tricarboxylic acid cycle, cholesterol metabolism, and a significant increase of the inflammation marker glycoprotein acetyls B in csPCa patients. In particular, ornithine and dimethylglycine were the 2 most important features to discriminate csPCa from Be + ciPCa with significantly higher ornithine and lower dimethylglycine levels in patients with csPCa (ornithine: 63.7 ± 26.5 µmol/L, dimethylglycine: 12.6 ± 6.3 µmol/L; P < .001) compared to Be + ciPCa patients (ornithine: 50.3 ± 31.6 µmol/L, dimethylglycine: 14.9 ± 7.7 µmol/L).

This study discovered a 22-metabolite panel to discriminate patients with csPCa from Be + ciPCa patients when combined with age, PSA, and %fPSA. It may therefore be used as a supportive biomarker to reduce the number of unnecessary biopsies and also to identify novel therapeutic targets in the future.

Hypertension is a highly prevalent medical condition that occurs when blood pressure is too high, which greatly increases the risk of developing other cardiovascular diseases and is generally associated with higher rates of morbidity and mortality. Due to the silent/asymptomatic nature of hypertension, although the methods currently available to diagnose it are easy, they generally do not allow for an early diagnosis and an efficient prognosis to avoid irreversible damage in the medium or long term. In fact, an early diagnosis of hypertension would be crucial to decrease hypertension-associated mortality. Since metabolomics using NMR can provide a global measurement of various serum metabolites, it is very suitable for detecting novel biomarkers. We therefore analyzed serum metabolomic profiles among normotensive and hypertensive elderly individuals by NMR and identified new potential biomarkers for hypertension and associated diseases. We found higher levels of acetate, formate, and glycerol, and lower levels of glutamine, glycine, and sarcosine in individuals with hypertension. Therefore, these metabolites could be used for early diagnosis of hypertension to avoid comorbidities derived from hypertension and associated mortality.

Moyamoya disease (MMD) is a rare cerebrovascular disease in humans. Although early revascularization can improve symptoms, it cannot reverse the progression of the disease. The current diagnosis still relies on traditional a Digital Subtraction Angiography (DSA) examination, which is invasive and expensive, leading to delayed diagnosis and affecting treatment timing and patient prognosis. The ability to diagnose MMD early and develop personalized treatment plans can significantly improve the prognosis of patients. Here, we have introduced the research on MMD biomarkers. By integrating proteomics and metabolomics data, we have successfully identified over 1700 features from more than 60 serum samples collected at the onset of symptoms in MMD patients. We use multiple computational strategies to interpret complex information in serum, providing a comprehensive perspective for early diagnosis of MMD. Diagnostic ability of our biomarker is significantly better than previous studies, especially when used in combination. In the study of molecular mechanisms, we found that the ferroptosis pathway was significant disruption in MMD patients, which was also confirmed by transcriptomics data. Finally, we validated the metabolites and proteins associated with ferroptosis pathways, as well as the biomarkers screened by machine learning, using another independent MMD cohort. Our research provides important clues for the diagnosis of MMD, and this assay can identify MMD early, thereby promoting stronger monitoring and intervention.

Ferroptosis, a regulated cell death driven by iron-dependent lipid peroxidation, is associated with chemoresistance in lung adenocarcinoma (LUAD). This study aims to investigate the role of sarcosine in ferroptosis and its underlying mechanisms.

An RSL3-induced ferroptosis model was used to screen a library of 889 human endogenous metabolites and metabolomic profiling was harnessed to identify metabolites associated with ferroptosis. Cell viability, lipid-reactive oxygen species (ROS), ferrous iron, malondialdehyde (MDA), and mitochondrial integrity were assessed to evaluate sarcosine's effects on ferroptosis. Metabolic fate was studied using 15N-labeled sarcosine. Next, we used untargeted metabolomic profiling and next-generation sequencing to dissect metabolic and transcriptomic changes upon sarcosine supplementation. The effects of sarcosine on ferroptosis and chemotherapy were further validated in patient-derived organoids (PDOs), xenograft models, and LUAD tissues.

Sarcosine emerged as a potent ferroptosis inducer in the metabolic library screening, which was further confirmed via cell viability, lipid-ROS, ferrous iron, and MDA measurements. Metabolic flux analysis showed limited conversion of sarcosine to other metabolites in LUAD cells, while untargeted metabolomic profiling and seahorse assays indicated a metabolic shift from glycolysis to oxidative phosphorylation. Sarcosine enhanced pyruvate dehydrogenase activity to generate more ROS by interacting with PDK4, reducing PDHA1 phosphorylation. As a co-activator of N-methyl-D-aspartate receptor (NMDAR), sarcosine also exerted its pro-ferroptosis effect via regulating ferrous export through the NMDAR/MXD3/SLC40A1 axis. Given the significance of ferroptosis in chemotherapy, we validated that sarcosine enhanced the sensitization of cisplatin by promoting ferroptosis in LUAD cells, PDOs, and xenograft models.

Sarcosine promotes ferroptosis and enhances chemosensitivity, suggesting its potential as a therapeutic agent in treating LUAD.

Clear cell renal cell carcinoma (ccRCC) is fundamentally a metabolic disease. To investigate the underlying metabolite changes in the occurrence of ccRCC, we analyzed untargeted metabolomics of 15 ccRCC samples and paired adjacent non-malignant renal tissues by UHPLC-MS/MS analyses. In this study, 511 differential metabolites were screened, of which the top ten up-regulated metabolites in ccRCC were histamine, 1-methylnicotinamide, L-kynurenine, cortisol, tetrahydrocorticosterone, calcitriol, D-erythrose 4-phosphate, citric acid, sedoheptulose 1,7-bisphosphate, and UDP-alpha-D-galactose, and the top down-regulated metabolites were D-cysteine, acetylcholine, pantothenic acid, cytosine, UMP, biocytin, dUMP, 5-phosphoribosyl 1-pyrophosphate, cytidine-5'-monophosphate, and 16α-hydroxyestrone. KEGG pathways enrichment analysis further demonstrated several highlighted pathways: steroid hormone biosynthesis, pyrimidine metabolism, and vitamin digestion and absorption. Our study reveals metabolic patterns of ccRCC and provides insight into the potential biomarker panel to diagnose ccRCC.

The development of simple, inexpensive, deployable clinical diagnostics could have a global impact on public health by making measurements of patient health status more widely accessible to patients regardless of socioeconomic status. Here, we report a novel biosensor for sarcosine using a colorimetric readout created by a hybrid catalyst system using copper nanocubes and the enzyme sarcosine oxidase. The enzyme catalyzes the reaction of sarcosine to generate H2O2, which the copper nanocubes then use as a substrate to create free radicals that convert colorless 3,3',5,5'-tetramethylbenzidine (TMB) to its blue, oxidized form. The sensor showed good substrate affinity for Cu nanocubes and yielded a wide linear response range (0-140 μM) for sarcosine detection, with high selectivity against various interfering species. The limit of detection and limit of quantification were found to be 1.43 μM and 4.7 μM, respectively. We showed that the biosensor maintains function in a complex serum sample matrix, suggesting potential utility in clinical applications. Finally, we demonstrated a prototype based on light emitting diodes (LEDs) and a light-dependent resistor (LDR) for unambiguous visual interpretation using an inexpensive microcontroller potentially suitable for use outside of traditional clinical or analytical laboratories.

Locoweeds, including Oxytropis and Astragalus species, are globally recognized as plants containing swainsonine (SW), a neurotoxic alkaloid that induces neurological dysfunction and growth inhibition in livestock. SW is produced by endophytic fungi in plants; the pyrroline-5-carboxylate reductase (P5CR) gene is critical in the fungal SW biosynthetic pathway. In this study, a P5CR gene knockout mutant (ΔP5CR) was constructed from the endophytic fungus Alternaria oxytropis OW7.8 isolated from Oxytropis glabra. Compared to the wild-type strain (A. oxytropis OW7.8), the SW content in the ΔP5CR mycelia was significantly reduced, indicating that the P5CR gene plays a crucial role in promoting SW biosynthesis. Compared to the wild-type strain A. oxytropis OW7.8, the ΔP5CR mutant exhibited distinct morphological alterations in both colony and mycelial structures. The transcriptomic analysis of A. oxytropis OW7.8 and ΔP5CR revealed the downregulation of six genes associated with SW biosynthesis. Metabolomic profiling further demonstrated altered levels of six metabolites linked to SW synthesis. These findings provide foundational insights into the molecular mechanisms and metabolic pathways underlying SW biosynthesis in fungi. They hold significant value for future strategies to control SW in Oxytropis glabra and contribute positively to the protection and sustainable development of grassland ecosystems.

The dysregulation of polyamines in tumors has made polyamine metabolism an appealing target for cancer therapy. Gene mutations drive the reprogramming of polyamine metabolism in tumors, presenting promising opportunities for clinical treatment. The proposed strategies involve inhibiting polyamine biosynthesis while also targeting the polyamine transport system as antitumor approaches. A growing number of drugs aimed at polyamine biosynthesis and transport systems are undergoing clinical trials. Polyamine metabolism plays a role in regulating cancer signaling pathways, suggesting potential combination therapies for cancer treatment. Furthermore, supplemental polyamine substances have demonstrated antitumor activity, indicating that combining polyamines with downstream targets or immunotherapy could offer significant clinical benefits. These discoveries open new avenues for leveraging polyamine metabolism in anticancer therapy.

Tobacco is an important economic crop and a model plant for molecular biology research. It exists in various cultivars and is processed using different curing methods. Fatty acids play a crucial role in the quality and flavor of tobacco leaves. However, there is limited information on the fatty acid composition across different cultivars, developmental stages, and curing methods. This study employed targeted metabolomics and transcriptomics to investigate the fatty acids and related pathway genes in tobacco leaves from different cultivars, developmental stages, and curing methods.

This study focused on four tobacco cultivars: K326, Basma, Samsun, and Cuba1, and investigated fatty acid differences in the leaves at four developmental stages (seedling, transplanting, budding, and topping) under two curing methods (air-curing and flue-curing). K326 was used as the main cultivar for comparison with the other three. The analysis included short-chain fatty acids (C2-C6), free fatty acids (C8-C24), and gene expression differences. The fatty acid metabolic profile of different tissue types in K326 at the budding stage was also examined. The results showed significant differences in fatty acid content among the different tissues of K326 at the budding stage, with the highest levels of short-chain fatty acids found in flower buds and upper leaves. At the seedling stage, there were marked variations in short-chain fatty acid content across different periods. Three key genes Nta01g31980, Nta08g22780, and Nta23g11140 were identified as major differential genes in fatty acid-related pathways in K326 compared to the other three cultivars during this stage. Regarding the four cultivars, the total short-chain fatty acid content at the budding stage was ranked as Basma > Samsun > Cuba1 > K326 before topping, but the order was reversed after topping. At the budding stage, 35 fatty acid pathway-related genes showed similar expression levels in Basma and Samsun, differing from K326 and Cuba1. Among the two curing methods, air-curing resulted in higher short-chain fatty acid content than flue-curing. Under air-curing, Samsun and Basma showed more downregulation of differential fatty acids compared to K326, while the opposite was observed under flue-curing.

This study expands our understanding of fatty acids in tobacco across different cultivars and developmental stages, providing a molecular basis for the study of fatty acids and genes related to their biosynthesis and metabolism.

Epithelial-mesenchymal transition (EMT) plays an irreplaceable role in the development of silicosis. However, molecular mechanisms of EMT induced by silica exposure still remain to be addressed. Herein, metabolic profiles of human alveolar type II epithelial cells (A549 cells) exposed directly to silica were characterized using non-targeted metabolomic approaches. A total of 84 differential metabolites (DMs) were identified in silica-treated A549 cells undergoing EMT, which were mainly enriched in metabolisms of amino acids (e.g., glutamate, alanine, aspartate), purine metabolism, glycolysis, etc. The number of DMs identified in the A549 cells obviously increased with the elevated exposure concentration of silica. Remarkably, glutamine catabolism was significantly promoted in the silica-treated A549 cells, and the levels of related metabolites (e.g., succinate) and enzymes (e.g., α-ketoglutarate (α-KG) dehydrogenase) were substantially up-regulated, with a preference to α-KG pathway. Supplementation of glutamine into the cell culture could substantially enhance the expression levels of both EMT-related markers and Snail (zinc finger transcription factor). Our results suggest that the EMT of human alveolar epithelial cells directly induced by silica can be essential to the development of silicosis.

Prostate cancer (PCa) remains a global health burden, with limited reliable biomarkers beyond prostate-specific antigen (PSA). Statins have been associated with survival benefits in advanced Pca, potentially by modulating cholesterol metabolism and tumor biology. However, the causal mechanisms are not well understood. A distinct three-lipid signature (3LS) has previously been proposed as a prognostic biomarker for PCa.

This study investigates the effects of atorvastatin intervention on PCa tissue markers, long-term clinical outcomes, and the prognostic value of the 3LS derived from prostate tissue lipidome.

The ESTO1 trial randomized 158 statin-naïve PCa patients to receive high-dose atorvastatin (80 mg daily) or placebo before prostatectomy. Long term outcomes were assessed for 102 patients through medical records review. Prostate tissue samples were pathologically characterized, and lipidome quantified. Cox regression models were used to analyse clinical outcomes between the groups. The 3LS score was calculated by identifying the constituent lipids from the prostate lipidome.

Higher intraprostatic atorvastatin lactone concentrations were associated with reduced Ki67 expression and PSA levels. After a median follow-up of seven years, no significant differences were observed in biochemical recurrence, overall mortality, or initiation of hormonal therapy. However, the atorvastatin arm had a lower risk of major acute cardiovascular events (HR 0.11, 95% CI 0.01-1.01). The intraprostatic 3LS correlated with higher baseline tumor aggressiveness but did not predict subsequent outcomes.

Higher atorvastatin lactone concentrations in the prostate tissue were linked to improved pathological variables. Pre-surgery statin intervention reduced MACE risk but no impact on other clinical outcomes was observed. The 3LS from prostate tissue does not seem to be prognostic marker in localized Pca.

Despite significant progress in nanozyme research and the advancement of analytical techniques, the inherent lack of specificity for target analytes often limits their utility in analysis. Integrating specific recognition capabilities into inorganic nanomaterials, independent of biological catalysts or adaptors, represents a crucial breakthrough in the field. Detecting Sarcosine (Sar) in human urine has recently emerged as a non-invasive biomarker for prostate cancer (PCa), presenting a valuable diagnostic tool. This study introduces a novel method for embedding molecular imprinting sites directly onto the surface of a Zn/Ce-based zeolitic imidazolate framework (Zn/Ce-ZIF) nanozyme, facilitating the development of a highly specific colorimetric assay for precise Sar measurement. By utilizing the lanthanide metal cerium as the catalytic element and ZIF-8 as the structural scaffold, we synthesized spherical Zn/Ce-ZIF nanozymes with exceptional oxidase-like catalytic efficiency. The efficiency of molecular imprinting experiments and the ability of molecularly imprinted polymers (MIPs) to identify target molecules were significantly enhanced by using theortical calculations to screen suitable functional monomers. The molecularly imprinted nanozyme (Zn/Ce-ZIF@MIP) initiates a colorimetric oxidation reaction of 3,3',5,5'-tetramethylbenzidine (TMB), wherein the presence of Sar facilitates selective recognition and capture by the MIP shell, modulating the colorimetric response by hindering TMB's access to the catalytic site. An intelligent color extraction detection device has been developed for the rapid perception of Sar. This colorimetric sensing platform has been validated through the detection of Sar in simulated urine samples. Overall, the application of surface molecular imprinting enhances the functionality of nanozymes in analytical fields.

Background: Pancreatic cancer is the most lethal of all human cancers. The disease has no obvious symptoms in its early stages and in the majority of cases, the cancer goes undetected until it has advanced to the point that surgery is no longer a viable option or until it has metastasized to other organs. The absence of reliable and sensitive biomarkers for the early detection of pancreatic cancer contributes to the poor ability to detect the disease before it progresses to an untreatable stage. Objectives: Here, an orthotopic xenograft mouse model of pancreatic cancer was investigated to determine if urinary metabolic biomarkers could be identified and used to detect the early formation of pancreatic tumors. Methods: The orthotopic xenograft mouse model of pancreatic cancer was established by injecting human MiaPaCa-2 cells, derived from a male patient aged 65 years with pancreatic adenocarcinoma, into the pancreata of severe combined immunodeficient mice. Orthotopic pancreatic tumors, allowed to grow for eight weeks, were successfully established in the pancreata in 15 out of 20 mice. At the time of sacrifice, tumors were excised and histologically analyzed and the masses and volumes recorded. Urine samples were collected prior to injection, at one-week post injection, and every two weeks afterwards for eight weeks. Results: NMR-based metabolic profiling of the urine samples indicated that 31 metabolites changed significantly over the course of tumor initiation and growth. Longitudinal metabolic profiling analysis indicated an initial increase in activity of the metabolic pathways involved in energy production and/or cell synthesis by cancer cells as required to support tumor growth that was followed by a diminished difference between control and orthotopic mice associated with tumor senescence as the tumors reached 7-8 weeks post injection. Conclusions: The results indicate that NMR-based urinary metabolic profiling may be able to detect the earliest stages of pancreatic tumor initiation and growth, highlighting the potential for translation to human clinical studies.

Metabolomics has emerged as a transformative tool in precision oncology, with substantial potential for advancing biomarker discovery, monitoring treatment responses, and aiding drug development. Integrating artificial intelligence (AI) into metabolomics optimizes data acquisition and analysis, facilitating the interpretation of complex metabolic networks and enabling more effective multiomics integration. In this opinion, we explore recent advances in the application of metabolomics within precision oncology, emphasizing the unique advantages that AI-driven metabolomics offers. We propose that AI not only complements but also amplifies the potential of current platforms, accelerating research progress and ultimately improving patient outcomes. Finally, we discuss the opportunities and challenges involved in translating AI-driven metabolomics into clinical practice for precision oncology.

Metabolomics is a valuable tool to assess glyphosate exposure and its potential impact on human health. However, few studies have used metabolomics to evaluate human exposure to glyphosate or glyphosate-based herbicides (GBHs). In this study, an untargeted and targeted metabolomics approach was applied to human skin fibroblasts exposed to the GBH Roundup (GLYP-R). Cytotoxicity, cell death, and oxidative stress assays were performed to evaluate potential damage caused by GLYP-R in fibroblasts. The herbicide showed a cytotoxic effect at concentrations above 100.0 mg L-1, with IC50 = 164.2 ± 8.7 mg L-1, inducing significant reactive oxygen species (ROS) production and necrosis. A GC×GC/Q-TOFMS method using derivatization with propyl chloroformate/propanol was developed for untargeted analysis, allowing the identification of 400 metabolites of different classes in the samples. The most significant compounds in the discrimination and classification of the samples were fatty acids and amino acids (AA). Based on the relevance of AA in untargeted analysis, a targeted analysis of 21 AA was performed using the same validated GC×GC method. Metabolomic analyses allowed the construction of two biomarker models with performance evaluated by receiver operating characteristic (ROC) curves: an untargeted model formed by four metabolites (methylcysteine, N-acetyl-l-methionine, methyl stearate, and linoleic acid) and a targeted model formed by three AA (l-glutamic acid, l-cysteine, and γ-aminobutyric acid). This study is the first to report the use of metabolomics to evaluate human skin cells exposed to GLYP-R, contributing to the toxicological research on glyphosate.

Sarcosine plays a key role in screening for early prostate cancer. However, the several already reported peroxidase mimics immobilized with sarcosine oxidase (SOX) utilized to detect uriary sarcosine still have some limitations such as complex synthesis process, using of expensive heavy metals to mimic enzyme activity and long color development time. Herein, an inexpensive peroxidase-like 2D Fe/Co-MOF nanosheet was prepared by a simple solvent modulation method. The resultant 2D Fe/Co-MOF nanosheets have strong peroxidase activity, with its Vmax value for H2O2 of 15.3 × 10-8 M/s, being 1.76 times that of HRP. Then, using the 2D Fe/Co-MOF as a peroxidase model for anchoring natural SOX to construct 2D Fe/Co-MOF/SOX , which can act as a cascade reactor for detection of sarcosine. Considering the above properties, a platform for the detection of sarcosine was built based on a colorimetric method. Because of presence of the high ratio of Fe2+ caused by the electron transfer from Co2+ to Fe3+, large specific surface area and plentiful active sites, 2D Fe/Co-MOF/SOX with TMB (3,3',5,5'-tetramethylbenzidine) colorimetric reagent could have fast color development and can be applied conveniently and fastly in an early screening tool for prostate cancer patients. The sarcosine could be quantified by peroxidase activity with a detection range of 1-400 μM and a limit of detection (LOD) of 0.324 μM. More importantly, the average sarcosine concentration of 21.367 μM and 1.871 μM was detected in patient's and normal urine (n = 5), respectively, which showed an excellent screening effect and a great potential in early prostate cancer.

Cancer due to its heterogeneous nature and large prevalence has tremendous socioeconomic impacts on populations across the world. Therefore, it is crucial to discover effective panels of biomarkers for diagnosing cancer at an early stage. Cancer leads to alterations in cell growth and differentiation at the molecular level, some of which are very unique. Therefore, comprehending these alterations can aid in a better understanding of the disease pathology and identification of the biomolecules that can serve as effective biomarkers for cancer diagnosis. Metabolites, among other biomolecules of interest, play a key role in the pathophysiology of cancer whose levels are significantly altered while 'reprogramming the energy metabolism', a cellular condition favored in cancer cells which is one of the hallmarks of cancer. Metabolomics, an emerging omics technology has tremendous potential to contribute towards the goal of investigating cancer metabolites or the metabolic alterations during the development of cancer. Diverse metabolites can be screened in a variety of biofluids, and tumor tissues sampled from cancer patients against healthy controls to capture the altered metabolism. In this review, we provide an overview of different metabolomics approaches employed in cancer research and the potential of metabolites as biomarkers for cancer diagnosis. In addition, we discuss the challenges associated with metabolomics-driven cancer research and gaze upon the prospects of this emerging field.

Cognitive impairment associated with schizophrenia (CIAS) and related deficits in learning (plasticity) are among the leading causes of disability in schizophrenia. Despite this, there are no Food and Drug Administration-approved treatments for CIAS, and the development of treatments has been limited by numerous phase 2/3 failures of compounds that showed initial promise in small-scale studies. NMDA-type glutamate receptors (NMDARs) have been proposed to play an important role in schizophrenia; moreover, the NMDAR has a well-characterized role in cognition, learning, and neuroplasticity. We review previously published clinical trials in CIAS that focused on NMDAR modulator treatments, focusing on published and recent developments of the use of novel NMDAR-modulating treatments for CIAS both alone and combined with plasticity/learning paradigms to enhance learning. We use this discussion of previous studies to highlight the importance of incorporating pharmacodynamic target engagement biomarkers early in treatment development, which can help predict which compounds will succeed or fail in phase 3. A range of direct and indirect NMDAR modulators are covered, including D-serine, D-cycloserine, memantine, and glycine and first-generation glycine transport inhibitors (e.g., sarcosine and bitopertin), as well as recent positive studies of iclepertin, a novel glycine transport inhibitor, and luvadaxistat, a D-amino acid oxidase inhibitor that increases brain D-serine levels, and indirect noninvasive brain stimulation NMDAR-modulating treatments. Several examples of successful use of pharmacodynamic target engagement biomarkers for dose/drug discovery are emphasized, including the mismatch negativity, auditory steady state, and time-frequency event-related potential approaches.

How mitochondria reconcile roles in functionally divergent cell death pathways of apoptosis and NLRP3 inflammasome-mediated pyroptosis remains elusive, as is their precise role in NLRP3 activation and the evolutionarily conserved physiological function of NLRP3. Here, we have shown that when cells were challenged simultaneously, apoptosis was inhibited and NLRP3 activation prevailed. Apoptosis inhibition by structurally diverse NLRP3 activators, including nigericin, imiquimod, extracellular ATP, particles, and viruses, was not a consequence of inflammasome activation but rather of their effects on mitochondria. NLRP3 activators turned out as oxidative phosphorylation (OXPHOS) inhibitors, which we found to disrupt mitochondrial cristae architecture, leading to trapping of cytochrome c. Although this effect was alone not sufficient for NLRP3 activation, OXPHOS inhibitors became triggers of NLRP3 when combined with resiquimod or Yoda-1, suggesting that NLRP3 activation requires two simultaneous cellular signals, one of mitochondrial origin. Therefore, OXPHOS and apoptosis inhibition by NLRP3 activators provide stringency in cell death decisions.

Cancer is one of the leading causes of death globally, and is ranked second in the United States. Early detection is crucial for more effective treatment and a higher chance of survival rates, reducing burdens on individuals and societies. Genitourinary cancers, in particular, face significant challenges in early detection. Finding new and cost-effective diagnostic methods is of clinical need. Metabolomic-based approaches, notably volatile organic compound (VOC) analysis, have shown promise in detecting cancer. VOCs are small organic metabolites involved in biological processes and disease development. They can be detected in urine, breath, and blood samples, making them potential candidates for sensitive and non-invasive alternatives for early cancer detection. However, developing robust VOC detection methods remains a hurdle. This review outlines the current landscape of major genitourinary cancers (kidney, prostate, bladder, and testicular), including epidemiology, risk factors, and current diagnostic tools. Furthermore, it explores the applications of using VOCs as cancer biomarkers, various analytical techniques, and comparisons of extraction and detection methods across different biospecimens. The potential use of VOCs in detection, monitoring disease progression, and treatment responses in the field of genitourinary oncology is examined.

Rapid and intelligent identification of prostate cancer (PCa) is critical for early diagnosis. Herein, a convenient, reliable, and intelligent strategy is proposed to screen PCa individuals through indirectly quantifying sarcosine (Sar), an early indicator of PCa, in clinical urine samples. Success is achieved by integrating sarcosine oxidase (SOX) as a specific recognition unit; nanozyme-assisted multicolor intelligent visualization platform as a signal reporter. With the Fe-MOFs and peroxidase, the synergetic action of SOX and response gold nanorods (Au NRs) is controlled etched to exhibit a multicolored signal. The sensor exhibits excellent linearity with Sar within 1-60 × 10-6 m, boasting a remarkable detection limit of 0.12 × 10-6 m. The RGB value of the display color can be directly extracted using a mobile phone camera. PCa diagnosis can be swiftly made (within 15 min) and directly by identifying two RGB colors (R < 175 or B > 135). The enzyme-assisted multicolor intelligent visualization platform is adept at detecting minute differences in Sar concentration in urine samples between PCa patients and healthy individuals. The concept of enzyme-assisted multicolor sensing can be further expanded by modifying the type of immobilized enzymes, providing a valuable guideline for the rational design of multiple probes to measure specific biomarkers in biological samples.

Tobacco black shank (TBS), caused by Phytophthora nicotianae, is a severe disease. Plant root exudates play a crucial role in mediating plant-pathogen interactions in the rhizosphere. However, the specific interaction between key secondary metabolites present in root exudates and the mechanisms of disease resistance remains poorly understood. This study conducted a comprehensive comparison via quasi-targeted metabolomic analysis on the root exudate metabolites from the tobacco cultivar Yunyan87 and K326, both before and after inoculation with P. nicotianae. The results showed that the root exudate metabolites changed after P. nicotianae inoculation, and the root exudate metabolites of different tobacco cultivar was significantly different. Furthermore, homovanillic acid, lauric acid, and isoliquiritigenin were identified as potential key compounds for TBS resistance based on their impact on the mycelium growth of the pathogens. The pot experiment showed that isoliquiritigenin reduced the incidence by 55.2%, while lauric acid reduced it by 45.8%. This suggests that isoliquiritigenin and lauric acid have potential applications in the management of TBS. In summary, this study revealed the possible resistance mechanisms of differential metabolites in resistance of commercial tobacco cultivar, and for the first time discovered the inhibitory effects of isoliquiritigenin and homovanillic acid on P. nictianae, and attempt to use plants secondary metabolites of for plant protection.

The effect of sexual dimorphism on the metabolism of patients with Parkinson's disease has not been clarified. A group of patients with Parkinson's disease and healthy controls were recruited, and their clinical characteristics and plasma were collected. Untargeted liquid chromatography-mass spectrometry-based plasma metabolomics profiling was performed. Differentially expressed metabolites between patients and healthy controls were respectively identified in the male and female participants and metabolite set enrichment analyses were further employed. A total of 75 patients with Parkinson's disease (37 males and 38 females) and 31 healthy controls (16 males and 15 females) were enrolled while no significant differences can be discovered in clinical characteristics. The constructed male-specific metabolic model from orthogonal partial least squares-discriminant analysis can't well recognize female patients and the female-specific model also can't accurately identify male patients. There were 55 differentially expressed metabolites in the male participants, and fatty acids and conjugates and eicosanoids were the significantly enriched metabolite sets. Meanwhile, 86 metabolites were differentially expressed in the female participants while fatty acids and conjugates and glycerophosphocholines were enriched. Only 17 metabolites were simultaneously changed in both male and female patients. Significant sex differences of lipid metabolism were found in patients with Parkinson's disease.

The transition period is a crucial stage in the reproductive cycle for dams and is linked closely with postpartum recovery, reproduction performance, and health. The confronting problem in the yak industry is that transition yaks under a conventional grazing feeding regime endure nutritional deficiency since this period is in late winter and early spring of the Qinghai-Tibet Plateau with the lack of grass on natural pasture. Therefore, this study aimed to investigate the effects of perinatal nutritional supplementation and early weaning on serum biochemistry, reproductive performance, and metabolomics in transition yaks. Eighteen healthy yaks in late pregnancy (233.9 ± 18.3 kg, 2-4 parity) were randomly assigned to three groups: conventional grazing feeding (GF, n = 6), additional nutrition supplementation (SF, n = 6), and additional nutrition supplementation with early weaning (SW, n = 6). Yaks in the GF, SF, and SW groups were free grazing on the same pasture in the daytime from -30 to 90 d relative to parturition. Yaks in SF and SW groups received total mixed ration supplementation in the barn during the night throughout the trial. Calves in the SW group were early weaned and separated from the dam at 60 d postpartum. Maternal body weight was measured at -30 and 90 d, and serum samples were collected to analyze serum biochemistry, hormones, and metabolomics at -15, 30, and 90 d relative to calving. In the SF and SW groups, yaks showed significantly higher body weight gain, serum glucose, globulin, and total protein concentrations. Lipid transportation molecules apolipoprotein B100 and very low-density lipoprotein of SF and SW yaks were significantly increased along with the decreased lipid mobilization products non-esterified fatty acid and β-hydroxybutyric acid when compared to GF yaks at -15 and 30 d. At 90 d, serum non-esterified fatty acid and β-hydroxybutyric acid levels were significantly lower in SW yaks than in SF ones, while apolipoprotein B100 and very low-density lipoprotein levels were significantly higher in SW yaks than in GF yaks. The serum levels of metabolic regulatory hormones, including insulin, leptin, and insulin-like growth factor I were significantly increased, and glucagon was significantly reduced in the SF and SW groups than in the GF group at -15 and 30 d. Among serum reproductive hormones, SF and SW yaks had significantly higher estradiol and progesterone concentrations than GF ones at -15 and 30 d. Follicle-stimulating and luteinizing hormone levels were increased in SW group than in SF and GF ones at 90 d. The calving rates in the following year were 0% (GF), 16.7% (SF), and 83.3% (SW), respectively. The serum metabolomics analysis revealed 848 metabolites in positive mode and 350 in negative mode. With the perinatal nutritional supplementation, the lipid and energy metabolism of transition yaks were improved, meanwhile, lipid mobilization and estrogen production-related pathways were down-regulated. These data suggest that perinatal nutrition supplementation reduces body weight loss, improves glucose and lipid metabolic adaptation to the transition period, and improves yaks' reproductive performance. Additionally, the combination of early weaning and nutritional supplementation results in lower lipid mobilization and up-regulation of lipid transportation and reproductive hormone secretion, which may further contribute to postpartum recovery and acceleration of the reproductive cycle.

Enzyme-based portable amperometric biosensors are precise and low-cost medical devices used for rapid cancer biomarker screening. Sarcosine (Sar) is an ideal biomarker for prostate cancer (PCa). Because human serum and urine contain complex interfering substances that can directly oxidize at the electrode surface, rapid Sar screening biosensors are relatively challenging and have rarely been reported. Therefore, highly sensitive and selective amperometric biosensors that enable real-time measurements within <1.0 min are needed. To achieve this, a chitosan-polyaniline polymer nanocomposite (CS-PANI NC), a carrier for dispersing mesoporous carbon (MC), was synthesized and modified on a screen-printed carbon electrode (SPCE) to detect hydrogen peroxide (H2O2). The sarcosine oxidase (SOx) enzyme-immobilized CS-PANI-MC-2 ternary NCs were referred to as supramolecular architectures (SMAs). The excellent electron transfer ability of the SMA-modified SPCE (SMA/SPCE) sensor enabled highly sensitive H2O2 detection for immediate trace Sar biomarker detection. Therefore, the system included an SMA/SPCE coupled to a portable potentiostat linked to a smartphone for data acquisition. The high catalytic activity, porous architecture, and sufficient biocompatibility of CS-PANI-MC ternary NCs enabled bioactivity retention and immobilized SOx stability. The fabricated biosensor exhibited a detection limit of 0.077 μM and sensitivity of 8.09 μA mM-1 cm-2 toward Sar, demonstrating great potential for use in rapid PCa screening.

Salt stress poses a significant challenge to plant growth and restricts agricultural development. To delve into the intricate mechanisms involved in soybean's response to salt stress and find targets to improve the salt resistance of soybean, this study integrated transcriptomic, proteomic, and metabolomic analyses to explore the regulatory networks involved in soybean salt tolerance. Transcriptomic analysis revealed significant changes in transcription factors, hormone-related groups, and calcium ion signaling. Notably, the biosynthetic pathways of cutin, suberine, and wax biosynthesis play an important role in this process. Proteomic results indicated salt-induced DNA methylation and the enrichment of phosphopyruvate hydrase post-salt stress, as well as its interaction with enzymes from various metabolic pathways. Metabolomic data unveiled the synthesis of various metabolites, including lipids and flavonoids, in soybean following salt stress. Furthermore, the integrated multiomics results highlighted the activation of multiple metabolic pathways in soybean in response to salt stress, with six pathways standing out prominently: stilbenoid, diarylheptanoid, and gingerol biosynthesis; carotenoid biosynthesis; carbon fixation in photosynthetic organisms; alanine, aspartate, and glutamate metabolism; thiamine metabolism; and pyruvate metabolism. These findings not only offer valuable insights into leveraging multiomics profiling techniques for uncovering salt tolerance mechanisms but also identify candidate genes for soybean improvement.

The Xihuang Pill (XHP), a venerated traditional Chinese medicine, has demonstrated significant anti-cancer capabilities. Despite its proven efficacy, the scarcity of comprehensive pharmacological studies limits the widespread application of XHP. This research endeavor seeks to demystify the therapeutic underpinnings of XHP, particularly in the realm of colorectal cancer (CRC) therapy.

In this study, mice harboring CT26 tumors were divided into four groups, each administered with either XHP monotherapy, 5-fluorouracil (5-FU), or a combination of both. The tumor growth trajectory was closely monitored to evaluate the effectiveness of these anti-neoplastic interventions. Advanced techniques, including 16S-rDNA gene sequencing and ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS), were harnessed to scrutinize the gut microbiota and serum metabolite profiles. Immunohistochemical assays were employed to gauge the expression levels of CD4, CD8, and Foxp3, thereby providing insights into the dynamics of tumor-infiltrating lymphocytes within the tumor microenvironment.

Our findings indicate that XHP effectively suppresses the initiation and progression of colorectal tumors. The combinatorial therapy of XHP with 5-FU exhibited an enhanced inhibitory effect on tumor growth. Metabolic profiling revealed that XHP induced notable metabolic shifts, particularly impacting pathways such as steroid hormone synthesis, arachidonic acid metabolism, purine biosynthesis, and renin secretion. Notably, 17α-ethinyl estradiol and α-ergocryptine were identified as serum metabolites with the most substantial increase following XHP administration. Analysis of the gut microbiome suggested that XHP promoted the expansion of specific bacterial taxa, including Lachnospiraceae_NK4A136_group, Clostridiales, Desulfovibrionaceae, and Anaerotignum_sp., while suppressing the proliferation of others such as Ligilactobacilus, Lactobacillus_taiwanensis, and Candidatus_saccharimonas. Immunohistochemical staining indicated an upregulation of CD4 and CD8 post-XHP treatment.

This study delineates a potential mechanism by which XHP inhibits CRC tumorigenesis through modulating the gut microbiota, serum metabolites, and reshaping the tumor immune microenvironment in a murine CRC model. These findings contribute to a more profound understanding and potentially broaden the clinical utility of XHP in oncology.

Antlers are hallmark organ of deer, exhibiting a relatively high growth rate among mammals, and requiring large amounts of nutrients to meet its development. The rumen microbiota plays key roles in nutrient metabolism. However, changes in the microbiota and metabolome in the rumen during antler growth are largely unknown. We investigated rumen microbiota (liquid, solid, ventral epithelium, and dorsal epithelium) and metabolic profiles of sika deer at the early (EG), metaphase (MG) and fast growth (FG) stages. Our data showed greater concentrations of acetate and propionate in the rumens of sika deer from the MG and FG groups than in those of the EG group. However, microbial diversity decreased during antler growth, and was negatively correlated with short-chain fatty acid (SCFA) levels. Prevotella, Ruminococcus, Schaedlerella and Stenotrophomonas were the dominant bacteria in the liquid, solid, ventral epithelium, and dorsal epithelium fractions. The proportions of Stomatobaculum, Succiniclasticum, Comamonas and Anaerotruncus increased significantly in the liquid or dorsal epithelium fractions. Untargeted metabolomics analysis revealed that the metabolites also changed significantly, revealing 237 significantly different metabolites, among which the concentrations of γ-aminobutyrate and creatine increased during antler growth. Arginine and proline metabolism and alanine, aspartate and glutamate metabolism were enhanced. The co-occurrence network results showed that the associations between the rumen microbiota and metabolites different among the three groups. Our results revealed that the different rumen ecological niches were characterized by distinct microbiota compositions, and the production of SCFAs and the metabolism of specific amino acids were significantly changed during antler growth.

This study investigates the impact of thermal decomposition on the ion mobility spectrum of L-alanine using ion mobility spectrometry (IMS) and computational methods. By employing a post-injection delay system, we examined the evolution of ion peaks corresponding to thermal decomposition products and their interaction with protonated alanine. Experimental results revealed that the observed ion mobility spectra predominantly feature protonated isomers and adduct ions. Computational analysis using Density Functional Theory (DFT) predicted the thermodynamically favored structures and stabilities of these products. Findings indicate that protonation at the nitrogen site in alanine is more stable than at the oxygen site, and observed peaks correspond to protonated isomers and adducts formed with ammonium ions. Further investigations showed that thermal decomposition of alanine generates ammonia, contributing to the formation of new adduct ions. This research provides new insights into the behavior of amino acids under thermal conditions with implications for analytical chemistry and biochemistry.

Prostate cancer is one of the most commonly diagnosed cancers in men and is a major cause of cancer-related deaths worldwide. Among the molecular processes that contribute to this disease, the weight of metabolism has been placed under the limelight in recent years. Tumours exhibit metabolic adaptations to comply with their biosynthetic needs. However, metabolites also play an important role in supporting cell survival in challenging environments or remodelling the tumour microenvironment, thus being recognized as a hallmark in cancer. Prostate cancer is uniquely driven by androgen receptor signalling, and this knowledge has also influenced the paths of cancer metabolism research. This review provides a comprehensive perspective on the metabolic adaptations that support prostate cancer progression beyond androgen signalling, with a particular focus on tumour cell intrinsic and extrinsic pathways.

Although immune checkpoint inhibitors (ICIs) have revolutionized immuno-oncology with effective clinical responses, only 30 to 40 % of patients respond to ICIs, highlighting the need for reliable biomarkers to predict and enhance therapeutic outcomes. This study investigated how amino acid, glycolysis, and bile acid metabolism affect ICI efficacy in non-small cell lung cancer (NSCLC) patients. Through targeted metabolomic profiling and machine learning analysis, we identified amino acid metabolism as a key factor, with histidine (His) linked to favorable outcomes and homocysteine (HCys), phenylalanine (Phe), and sarcosine (Sar) linked to poor outcomes. Importantly, the His/HCys+Phe+Sar ratio emerges as a robust biomarker. Furthermore, we emphasize the role of glycolysis-related metabolites, particularly lactate. Elevated lactate levels post-immunotherapy treatment correlate with poorer outcomes, underscoring lactate as a potential indicator of treatment efficacy. Moreover, specific bile acids, glycochenodeoxycholic acid (GCDCA) and taurolithocholic acid (TLCA), are associated with better survival and therapeutic response. Particularly, TLCA enhances T cell activation and anti-tumor immunity, suggesting its utility as a predictive biomarker and therapeutic agent. We also suggest a connection between gut microbiota and TLCA levels, with the Eubacterium genus modulating this relationship. Therefore, modulating specific metabolic pathways-particularly amino acid, glycolysis, and bile acid metabolism-could predict and enhance the efficacy of ICI therapy in NSCLC patients, with potential implications for personalized treatment strategies in immuno-oncology. ONE SENTENCE SUMMARY: Our study identifies metabolic biomarkers and pathways that could predict and enhance the outcomes of immune checkpoint inhibitor therapy in NSCLC patients.

Biosensors based on carbon nanotube field-effect transistors (CNT-FETs) have shown great potential in biomarker detection due to their high sensitivity because of appreciable semiconducting electrical properties. However, background signal interferences in complex mediums may results in low signal-to-noise ratio, which may impose challenges for precise biomarker detection in physiological fluids. In this work, we develop an enzymatic CNT-FET, with scalable production at wafer scale, for detection of trace sarcosine that is a biopsy-correlated biomarker of prostate cancer. Enzymatic cascade rectors are constructed on the CNT to improve the reaction efficiency, thereby, enhancing the signal transduction. As such, a limit of detection as low as 105 zM is achieved in buffer solution. Owing to the enhanced reaction efficiency, the testing of clinical serum samples yields significant signal difference to discriminate the prostate cancer (PCa) samples from the benign prostatic hyperplasia (BPH) samples (P = 1.07 × 10-5), demonstrating immense potential in practical applications.

Long-term use of rhubarb (RH) commonly leads to diarrhea, which can be alleviated by steaming with wine. However, the specific mechanism by which wine steaming alleviates RH-induced diarrhea remains unknown.

This study aims to reveal the underlying mechanisms of wine steaming in alleviating RH-induced diarrhea by examining small intestinal flora and serum metabolomics.

Major anthraquinone and anthrone components were detected using ultra-performance liquid chromatography-mass spectrometry (UPLC-MS). Eighty-four ICR mice were randomly divided into control, RH, and RH steamed with wine (PRH) groups and were administered RH and PRH (1, 4, and 8 g/kg, i.g). for 14 consecutive days. Histopathological analysis was performed using hematoxylin-eosin staining. Levels of inflammatory factors and tight junction proteins, zonula occludens-1 (ZO-1) and occludin, in the small intestine were measured. The small intestine content was analyzed using 16S rRNA sequencing, and UPLC-MS was used to analyze endogenous metabolites.

Levels of major anthraquinone and anthrone components decreased in PRH. Both RH and PRH groups showed varying degrees of loose stools and increased fecal water rates; the RH group exhibited more severe effects. Compared with the control group, RH caused small intestine injuries, increased levels of inflammatory cytokines, downregulated the expression of ZO-1 and occludin, and induced gut microbiota (GM) imbalance. The relative abundance of Lactobacillus decreased, while the relative abundance of Shigella and Streptococcus increased. However, PRH had a milder impact than RH. The glycerophospholipid metabolic pathway was involved in this effect. The levels of inflammatory cytokines and potential metabolites (sn-glycero-3-phosphoethanolamine) were positively correlated with Streptococcus infection, while the levels of ZO-1 and occludin were negatively correlated with Streptococcus infection. GM imbalance and abnormal glycerophospholipid metabolism contributed to impaired intestinal barrier function and inflammatory factor release, which may underlie RH-induced diarrhea, though PRH had a weaker effect.

PRH alleviated RH-induced diarrhea by recovering GM balance, reducing ZO-1 and occludin expression, and decreasing the release of inflammatory factors. This mechanism may be linked to the reduced anthraquinone content. This study is the first to explore the mechanism of wine steaming in alleviating RH-induced diarrhea through small intestinal flora and serum metabolomics. It provides data to support the broader clinical use of RH and its safer application.