Due to continuous application as a flavoring agent in the pesticide, pharmaceutical, and food industries, methyl eugenol (ME) persists in the environment and causes deleterious impacts including cytotoxicity, genotoxicity, and liver damage. This study utilized a comprehensive approach, integrating toxicokinetics, metabolomics, and gut microbiota analysis, to explore the mechanisms behind ME-induced hepatotoxicity in mice. The study observed significant rises in ALT and AST levels, along with significant weight loss, indicating severe liver damage. Toxicokinetic data showed delayed Tmax and plasma accumulation after 28 days of repeated ME exposure at doses of 20 mg/kg, 40 mg/kg, and 60 mg/kg. The metabolomic analysis pinpointed four critical pathways—TCA cycle; alanine, aspartate, and glutamate metabolism; arginine biosynthesis; and tyrosine metabolism—linked to 20 potential biomarkers. Gut microbiota analysis revealed that extended ME exposure led to microbial imbalance, particularly altering the populations of Akkermansia, Prevotella, and Ruminococcus, which are key to amino acid metabolism and the TCA cycle, thus contributing to hepatotoxicity. However, the causal relationship between changes in gut microbiota and liver metabolite levels still requires further in-depth research. This study underscores the significant role of liver metabolites and gut microbiota in ME-induced liver damage.

Background/Objectives: Epilepsy is a common chronic and recurrent neurological disorder that poses a threat to human health, and Acorus tatarinowii Schott (ATS), a traditional Chinese medicine, is used to treat it. This study aimed to determine its effects on plasma metabolites. Moreover, the possible mechanism of its intervention in epilepsy was preliminarily explored, combined with network pharmacology. Methods: An epileptic model of rats was established using pentylenetetrazol. The potential targets and pathways of ATS were predicted by network pharmacology. Ultra Performance Liquid Chromatography–Quadrupole–Time of Flight Mass Spectrometrynce Liquid Chromatography–Quadrupole–Time of Flight Mass Spectrometryance Liquid Chromatography–Quadrupole–Time of Flight Mass Spectrometry and statistical analyses were used to profile plasma metabolites and identify ATS’s effects on epilepsy. Results: Kyoto Encyclopedia of Genes and Genomes enrichment analysis revealed that ATS was involved in regulating multiple signaling pathways, mainly including the neuroactive ligand–receptor interaction and GABAerGamma-aminobutyrate transaminaseAminobutyrate Transaminaseapse signaling pathway. ATS treatment restored 19 metabolites in epiGamma-aminobutyrate transaminaseminobutyrate Transaminase rats, affecting lysine, histidine, and purine metabolism. GABA-T was found as a new key target for treating epilepsy with ATS. The IC₅₀ of ATS for inhibiting GABA-T activity was 57.9 μg/mL. Through metabolomic analysis, we detected changes in the levels of certain metabolites related to the GABAergic system. These metabolite changes can be correlated with the targets and pathways predicted by network pharmacology. One of the limitations of this study is that the correlation analysis between altered metabolites and seizure severity remains unfinished, which restricts a more in-depth exploration of the underlying biological mechanisms. In the future, our research will focus on conducting a more in-depth exploration of the correlation analysis between altered metabolites and seizure severity. Conclusions: These results improved our understanding of epilepsy and ATS treatment, potentially leading to better therapies. The identification of key metabolites and their associated pathways in this study offers potential novel therapeutic targets for epilepsy. By modulating these metabolites, future therapies could be designed to better manage the disorder. Moreover, the insights from network pharmacology can guide the development of more effective antiepileptic drugs, paving the way for improved clinical outcomes for patients.

Ulcerative colitis (UC) is a subtype of inflammatory bowel disease (IBD) that has been associated with overconsumption of calories and lipids, compared to the healthy population, and summer temperatures have been reported to be closely related to the prevalence of UC. To evaluate the effects of dietary and lifestyle factors on UC, a combination of 2.0% dextran sulfate sodium (DSS), a high-fat/high-sugar diet, and exposure to high temperature and humidity was used to construct mouse models of UC. Changes in body weight, disease activity index (DAI) scores, histopathological analysis, serum lipid levels, serum diamine oxidase (DAO), and D-Lactate (D-LA) levels, as well as the expression of inflammatory cytokines and tight junction proteins in colonic tissue, were all assessed to study the impacts of the high-fat/high-sugar diet and high-temperature/high-humidity exposure on the progression of UC. The symptoms observed in the UC mouse model induced by 2.0% DSS alone were similar to those seen in patients with UC, while the high-fat and high-sugar diet, along with humid and hot exposure, exacerbated DSS-induced UC in the mice. This included more severe histopathological damage to the colon tissue, increased expression of pro-inflammatory cytokines (IL-6, IL-17A, and IL-1β), and a more significantly compromised intestinal barrier, characterized by the destruction of ZO-1 and elevated levels of DAO and D-LA. Additionally, the high-fat/high-sugar diet and high-temperature/high-humidity exposure led to further disturbances in glucose and lipid metabolism in the mice, which were not observed in those treated with DSS alone. This study is the first to investigate the effects of a high-fat/high-sugar diet and high-temperature/high-humidity exposure on the progression of UC.

As a phytotherapeutic agent with historical applications in epilepsy management, Acorus tatarinowii Schott (ATS) remains pharmacologically enigmatic, particularly regarding its pathophysiological mechanisms. This knowledge gap significantly hinders the clinical application of ATS-based treatments. To explore the potential of ATS in combating epileptogenesis, we utilized a pentylenetetrazole (PTZ)-induced chronic epilepsy rat model. Brain metabolomic analysis was performed by ultra-performance liquid chromatography coupled with mass spectrometry (UPLC/MS). Principal component analysis (PCA) and orthogonal projections to latent structures-discriminant analysis (OPLS-DA) were performed for screening differential metabolites. Gut microbiota composition was analyzed through 16S rRNA gene sequencing and examined using Spearman correlation analysis. The results show that oral ATS (50 mg/kg) significantly improved the seizure latency and pathology of rats with epilepsy. Ascorbate and aldarate metabolism, glycerophospholipid metabolism, arachidonic acid metabolism, and intestinal flora were crucial for ATS’s ability to counteract epilepsy. The therapeutic effects of ATS against epilepsy were investigated with brain metabolomics and gut microbiota analysis, providing the basis for further comprehensive research.

Cinnamomum migao, a plant in the Lauraceae, is a rare and endangered species endemic to China (Yan et al. 2022). In February 2025, symptoms of leaf spot were observed on C. migao plants growing in Pingtang County, Guizhou Province, China (25°29'55″N, 106°40'29″E). Disease incidence reached 100% (n=200). Foliage exhibited ash-white necrotic lesions that were elliptical or irregular in shape, surrounded by a brown halo at the margin. Six symptomatic leaf samples (5 × 5 mm) were surface-sterilized with 75% ethanol for 30 s, followed by 2.5% NaClO for 1 min. After air-drying, the tissues were placed on potato dextrose agar (PDA) and incubated at 28°C for 7 days. Hyphal tips from emerging colonies were transferred to fresh PDA plates to obtain pure cultures. A total of 16 purified isolates were obtained, 12 of which shared similar morphological characteristics. One representative isolate (designated ZYX) was selected for further study. Colonies on PDA were villous, with dense aerial mycelia, initially white and turning grayish over time; the reverse side was white with a yellow gradient and a red concentric ring. Conidia were ellipsoidal, aseptate, and 3.7 - 6.4 x 1.4 - 4.0 μm (average 5.04 × 2.61 μm; n= 60). Morphological characteristics were consistent with those described for Epicoccum latusicollum, a fungal pathogen causing leaf spot on tobacco in China (Guo et al. 2021). For molecular identification, the internal transcribed spacer (ITS) region was amplified with primers ITS1/ITS4 (White et al. 1990), the large subunit ribosomal RNA gene (LSU) with LROR/LR5 (Vilgalys and Hester 1990), the beta-tubulin gene (TUB2) with Btb2Fd/Btub4Rd (Woudenberg et al. 2009), and the RNA polymerase II second largest subunit gene (RPB2) with RPB2-5F2/fRPB2-7cR (Liu et al. 1999). Resulting sequences were deposited in GenBank with accession numbers PV946034 (ITS), PV946035 (LSU), PV976759 (TUB2), and PV976758 (RPB2). BLASTn analysis showed isolate ZYX shared 99% to 100% sequence identity with the E. latusicollum type strain CGMCC 3.18346, ITS (KY742101; 478/479 bp), LSU (KY742255, 859/863 bp), TUB2 (KY742343, 333/333 bp), and RPB2 (KY742174, 595/596 bp) sequences in the GenBank database. Furthermore, the amplified sequences were used to construct a phylogenetic tree in PhyloSuite 1.2.3 which showed that ZYX clustered with E. latusicollum. Thus, both morphological and molecular data support the identification of the isolate as E. latusicollum. To fulfill Koch's postulates, pathogenicity tests were conducted using conidial suspensions (1×106 conidia/mL) sprayed onto three healthy C. migao plants. Three additional plants were treated with sterile distilled water as controls, and plants were maintained at 28°C with 85% relative humidity and a 12-hour photoperiod. After 7 days, the inoculated plants developed symptoms identical to those observed in the field, while the control plants remained asymptomatic. The pathogen was successfully re-isolated from symptomatic leaves and confirmed based on morphological features and DNA sequences, fulfilling Koch's postulates. No pathogens were isolated from the control plants. E. latusicollum has been reported to cause disease in various medicinal plants (Yan et al. 2024, Wang et al. 2023). To our knowledge, this is the first report of E. latusicollum causing leaf spot on C. migao in China. This report contributes to the development of targeted and efficient disease management strategies for growers.