Objectives: This study aimed to systematically investigate the dose-dependent effects of 60Co-gamma radiation on the entire growth cycle of Astragalus membranaceus, using a dose gradient ranging from 50 to 400 Gy. The specific objectives were to evaluate its impacts on seed germination, seedling physiology, agronomic traits of one-yearold plants, and the accumulation of medicinal components. Methods: A radiation dose gradient of 50-400 Gy was applied to Astragalus membranaceus seeds. Assessments included seed germination and seedling emergence rates, physiological and biochemical markers (e.g., antioxidant enzyme activities and secondary metabolites), agronomic traits, and medicinal compound accumulation. The median lethal dose (LD50) was calculated, and a multi-trait weighted index (I) was used to screen superior germplasm lines. Results: Low-dose radiation (50-100 Gy) significantly improved seed germination and emergence rates, while high doses (>= 200 Gy) were inhibitory, with an LD50 of 201.95 Gy. At 50 Gy, antioxidant defense mechanisms were activated, indicated by increased CAT and APX activities, and flavonoid content rose by 48.2 %. Doses >= 300 Gy caused severe damage, including a 57.8 % reduction in thylakoid density. In one-year-old plants, 50 Gy synergistically improved biomass and active compound content. Doses of 100-300 Gy induced transgressive segregation: one line (300 Gy-2) showed a root dry weight 4.63 times that of the control. Using a multi-trait index, seven superior lines were selected. Line 100 Gy-8 exhibited a 1.54-fold increase in total flavonoids (93.03 mg/g), while line 400 Gy-12 showed a threefold enhancement. Conclusion: The results indicate that 50 Gy is an optimal dose for simultaneously improving yield and quality in Astragalus membranaceus, whereas the 100-300 Gy range serves as a promising mutagenic window for generating high-flavonoid or high-biomass variants. This study provides theoretical support and core germplasm resources for targeted breeding strategies.

Corn silk, the stigma of the female flower of maize (Zea mays L.), is a medicinal and food-based homologous natural product. Previous studies have highlighted the lipid-modulating and hepatoprotective properties of corn silk; however, its effects on fatty liver disease remain unclear. C57BL/6J mice were fed an HFD and orally administered CSE. Hepatic histopathology, systemic inflammatory markers, and metabolic profiles were assessed. Gut microbiota and metabolomic analyses were integrated to identify the key regulatory axis. CSE administration significantly improved hepatic steatosis, reduced circulating lipid levels, and suppressed systemic and hepatic inflammation. Microbiota analysis revealed that CSE restructured the intestinal ecology by fostering a “competing guild” dominated by Akkermansia, which was negatively correlated with several pathobionts. Metabolomic profiling indicated that CSE significantly modulated pentose-glucuronate interconversion and primary bile acid biosynthesis. Notably, the enrichment of taurine and activation of the bile acid-FXR signaling axis were identified as pivotal mechanisms for enhancing fatty acid β-oxidation and restoring redox balance via the glutathione pathway. Our findings demonstrate that CSE effectively ameliorates FLD by modulating the gut microbiota-metabolite-liver axis. These results position CSE as a promising prebiotic and functional food candidate for the prevention and treatment of metabolic liver diseases.

Background Growth-regulating factors (GRFs) are transcription factors that exist only in plants. They are mainly responsible for regulating plant growth and development and play an important role in responding to abiotic stress. Astragalus membranaceus (A. membranaceus) is recognized as a significant medicinal plant, and its growth is often restricted by abiotic stresses such as cold. Although the GRF gene families have been recognized in diverse plant species, systematic studies on them in A. membranaceus are still lacking. Result Based on the whole-genome data of A. membranaceus, we identified 10 GRF gene family members (AmeGRF1-AmeGRF9). Then, it analyzed their properties, phylogenetic evolution, conserved motifs, cis-acting elements, Synteny, and functional enrichment. Promoter analysis revealed that multiple AmeGRF members contain low-temperature response elements. Further qRT-PCR analysis revealed that AmeGRF4 exhibited marked induction in response to cold stress, indicating its potential role in the cold adaptation of A. membranaceus. Conclusions This work sheds light on the biological functions and abiotic stress regulatory mechanisms mediated by GRF genes in A. membranaceus.

Ultraviolet-B (UV-B) radiation induces excessive reactive oxygen species (ROS) in plants, impairing stress tolerance and secondary metabolism. Flavonoids were recognized as key antioxidants in Astragalus membranaceus , which help mitigate UV-B damage. However, how antioxidant enzymes interact with flavonoid metabolism under UV-B remains unclear. Using whole-genome data, we identified 14 AmSOD genes classified into Cu/Zn-, Fe-, and Mn-SOD subfamilies. These genes share conserved structures but show divergent expression patterns. Under UV-B, AmCSD10 (Cu/Zn-SOD) was strongly and continuously upregulated, matching increased SOD activity. Transcriptomic, enzyme activity, and metabolomic data were integrated across time points (UV3h-UV18h), revealing 64 flavonoids that shift from stability to accumulation at later stages. Correlation analysis linked 29 flavonoids to six AmSOD genes. AmCSD10 showed the strongest associations with five upregulated flavonoids, contained formononetin, glabranin and aloeresin A. Co-expression networks suggest AmCSD10 coordinates antioxidant defense and flavonoid biosynthesis via ERF - and WRKY -type transcription factors. This study identifies an AmCSD10 -centered module linking ROS detoxification and flavonoid metabolism, highlighting its role in redox control and medicinal compounds accumulation. The findings reveal mechanistic links between stress adaptation and quality formation in Astragalus .

Background The GRAS transcription factor family plays essential roles in plant development and adaptation to environmental stress. However, its genomic organization and regulatory mechanisms remain largely unknown in medicinal plants. Result Through genome-wide analysis, 39 IiGRAS genes were identified in I. indigotica . Phylogenetic, structural, and synteny analyses indicated that family expansion was driven by tandem and segmental duplications under strong purifying selection. Promoter analysis revealed stress-responsive cis-elements related to abscisic acid, jasmonic acid, and drought signaling. Integrated transcriptomic and metabolomic analyses further showed that IiSCL genes are co-expressed with transcription factors such as AP2, bZIP, WRKY, and HLH, and correlate with metabolites including trehalose-6-phosphate, lysophosphatidylcholine (LPC 18:1), and N-lactoyl-phenylalanine. These associations suggest IiSCL genes participate in coordinated transcriptional and metabolic regulation, contributing to osmotic balance, membrane stability, and oxidative defense under stress. Conclusion This study establishes a molecular framework for understanding SCL-mediated regulatory networks in I. indigotica and identifies potential genetic targets for improving stress tolerance, thereby supporting the sustainable cultivation of medicinal plants.

Bladder cancer remains one of the most common malignancies of the urinary tract. Although the treatment landscape has expanded rapidly in recent years, gemcitabine still occupies a central position in intravesical treatment for non-muscle-invasive bladder cancer, in perioperative systemic therapy for muscle-invasive disease, and in platinum-based first-line regimens for advanced urothelial carcinoma. Yet the long-term benefit of gemcitabine is frequently curtailed by primary non-response or acquired resistance. In practice, this problem is often recognized only after radiographic progression or clear clinical deterioration has occurred. This review summarizes recent progress in bladder cancer therapy and translational research, with a particular emphasis on the biological basis and hierarchical evolution of gemcitabine resistance. We establish a 3-stage operational model of resistance, distinguishing: (1) early pharmacologic resistance driven by impaired drug uptake/activation or enhanced inactivation; (2) intermediate resistance driven by enhanced DNA damage repair, replication stress tolerance, and pro-survival autophagy signaling; and (3) late adaptive resistance driven by epithelial-mesenchymal transition (EMT), stemness maintenance, metabolic reprogramming, non-coding RNA-mediated epigenetic regulation, inflammatory microenvironmental remodeling, and extracellular vesicle-based intercellular transmission. These layers function as an interactive network, with sequential emergence under treatment pressure and parallel activation in context-dependent clinical settings. We stratify key mechanistic nodes (including the HYAL4-V1/CD44/JAK2-STAT3/CDA axis, AKR1C3, AP1M2-RAD54B, PRPF19-DDB1, AKT/mTOR signaling, Beclin-1-dependent autophagy, the MINCR/ZEB1/PHGDH axis, and IL-6-associated inflammatory states) by their clinical evidence quality and translational readiness, explicitly distinguishing preclinical discovery from clinically validated findings. Critically, most mechanistic findings remain at the preclinical or retrospective validation stage, with no markers yet approved for routine clinical use. Future work must prioritize longitudinal paired clinical samples, standardized analytic assays for dynamic biomarkers, and the integration of functional models (organoids, microfluidic systems), multi-omics technologies (single-cell sequencing, spatial transcriptomics), and liquid-biopsy approaches to translate mechanistic discoveries into clinically actionable predictive tools and therapeutic strategies.

The northeastern region of China experiences a distinctly cold climate influenced by the Siberian High during the winter months, thus resulting in severe cold weather conditions. Snow cover is prevalent and can persist for several months. This prolonged exposure to low temperatures necessitates specific adaptations in terms of agriculture and plant life, particularly for perennial herbs. Saposhnikovia divaricate (Turcz.) Schischk (SD) is a widely distributed perennial herb in the northeastern and northern provinces of China. However, there is limited documentation on the molecular mechanism through which this plant adapts to cold stress. Therefore, we elucidated the SD response to cold stress by transcriptome and metabolome analysis. Cold stress induced chlorosis and wilting in plants, thus leading to added function of antioxidant enzymes and higher levels of malondialdehyde, proline, soluble sugars. Notably, the differentially expressed genes (DEGs) were primarily related with sugar metabolism, ROS sweep, flavonoid and terpenoid biosynthesis, plant hormone signalling pathways, lipid metabolism, and transcription factors. Additionally, the differentially expressed metabolites (DEMs) mainly included lipids, flavonoids, terpenoid compounds, sugar-related metabolites, alkaloids and other metabolites. Furthermore, integrated analysis revealed coexpression patterns between carbohydrate metabolism-related genes and genes reference flavonoid and terpenoid biosynthesis, along with their corresponding metabolites. Finally, the qPCR results revealed notable over-expression levels of stress-related genes, including those participated in plant hormone signalling pathways (PP2C and AUX), flavonoid biosynthesis (CH3), antioxidant enzymes (AOX and CAT), and sugar-related metabolite metabolism (TPS, SPS, and SS). In conclusion, our findings suggest that cold stress strongly affects plant hormone signalling pathways, ROS scavenging mechanisms, unsaturated fatty acid synthesis and flavonoid and terpenoid biosynthesis in SD. These discoveries provide valuable insights into the impact of cold climates on herbaceous plants.

Hyperuricemia (HUA) is a globally prevalent metabolic disease characterized by excessive production or insufficient excretion of uric acid in the serum. Although several drugs are available for the treatment of HUA, they have been associated with undesirable side effects. Therefore, this study aims to evaluate the therapeutic effects of sunflower head extract (KHE) on HUA in a mouse model and explore its potential mechanisms. All mice were randomly divided into three groups: Normal control (NC, 0.5% CMC-Na), HUA model (MD, yeast extract paste 20 g/kg), and KHE treatment group (KHE, 1 g/kg). Biochemical indicators, the oxidative stress state, and metabolomics were analyzed. KHE reduced the levels of 5-aminoimidazole ribonucleotide, xanthine, hypoxanthine, and uric acid in the serum of mice with HUA but increased the levels of adenine and taurine. KHE decreased the activities of superoxide dismutase (SOD) enzymes, the hepatic hydrogen peroxide (H2O2) and malondialdehyde (MDA) levels, and the serum levels of betaine aldehyde and beta-D-glucosamine. KHE improved oxidative stress levels and mitigated potential damage to the kidneys and joints caused by urate deposition. These findings provide comprehensive evidence supporting the anti-HUA effects and underlying mechanisms of KHE in HUA mice.

IntroductionGlyphosate (GLP) is one of the most widely used herbicides in the world. However, its underlying effects on the liver remain unclear. This study aims to investigate the toxic effects and the gut microbiome- and serum metabolite-related mechanisms of GLP on the liver in mice.Methods16S rDNA sequencing and UPLC-Q-TOF-MS/MS were used to investigate the mechanisms of GLP toxicity in mice administered with 0, 50, 250 and 500 mg/kg/day GLP for 30 days.ResultsGLP induced hepatocyte edema and ballooning as well as inflammatory cell infiltration. Exposure to GLP resulted in increased levels of serum ALT, TBIL, DBIL, and GLU. Microbiota analysis at the phylum level demonstrated that the proportions of Patescibacteria decreased in the GLP-treated group. The genus-level analysis identified 11 different genera, with eight decreased and three increased in the GLP-exposed group. Metabolomics analysis of serum showed 42 differential metabolites between the GLP and control groups. The metabolic pathway enrichment analysis revealed that the pentose phosphate pathway (PPP) and pyrimidine metabolism were significantly activated. Spearman analysis showed that the changes in the differential metabolites of the PPP and pyrimidine metabolism and gut microbiota were strongly associated with the biochemical index.DiscussionIn conclusion, GLP exposure induces hepatic injury through alterations in the gut microbiome and metabolic pathways, particularly by activating the pentose phosphate pathway and pyrimidine metabolism.

Background:Stomach adenocarcinoma (STAD) is the most common gastrointestinal cancer. A clear diagnosis and molecular targeted therapy have important implications for prolonging survival of patients. RAD51 is the central catalyst of homologous recombination that plays important role in maintaining genomic integrity. However, the clinical significance of RAD51 expression in STAD patients remains unclear. This study aimed to assess the association of RAD51 expression with clinicopathological characteristics and patient outcomes. Methods:In this study, RAD51 mRNA expression in STAD patients was assessed using the UALCAN and GEPIA databases. The diagnostic value of RAD51 was evaluated by analyzing the ROC curve (data from the The Cancer Genome Atlas (TCGA) database). The protein expression level of RAD51 in STAD patients and its relationship with clinicopathological characteristics and prognosis were evaluated by immunohistochemistry. Co-expression analysis of RAD51 in STAD was performed by Coexpedia and Gene Expression Profiling Interactive Analysis (GEPIA) databases. The associations of RAD51 and its co-expression genes with immune infiltrates were analyzed in TIMER database. Results:Our bioinformatic analysis revealed that RAD51 demonstrates elevated expression in STAD. The ROC curve analysis yielded an AUC value of 0.9366 (95% CI [0.9075-0.9658]), confirming its potential as a biomarker for STAD. Immunohistochemical assessments validated the up-regulation of RAD51 in STAD, highlighting its significant correlation with TNM stage and T stage, but not with age, sex, grade, N stage, M stage, or P53 expression. Patients exhibiting high RAD51 expression exhibited significantly reduced overall survival. Multivariate analysis identified RAD51 expression may serve as an independent prognostic biomarker of poor prognosis in patients with STAD. Additionally, our bioinformatic analysis identified eight RAD51 co-expression genes (AURKA, CKS1B, NUSAP1, PFDN4, CCNE1, CDCA4, KIF4A, and MCM10) in STAD. Moreover, we discovered that RAD51 and its main co-expressed genes were significantly negatively associated with most or all immune cell infiltration. Conclusions:RAD51 overexpression was related to disease progression and poor prognosis, as well as infiltration of immune cells in gastric cancer.

WRKY transcription factors (TFs) are important transcriptional regulators in plants, with their members widely involved in plant growth and development as well as responses to abiotic stresses. However, researches on WRKY genes in the medicinal plant A. membranaceus are scarce. Specifically, the roles of AmWRKYs in cold stress adaptation and their regulatory effects on flavonoid biosynthesis, which determines both medicinal quality and stress resistance, remain largely unexplored. Given its high economic value and extreme sensitivity to cold in its main cultivation regions, identifying key regulators of its cold tolerance is crucial for genetic improvement. In this study, 94 AmWRKY were identified based on genome analysis, distributed across 8 chromosomes. AmWRKYs are structurally conserved, all carrying the core conserved domain "WRKYGQK" and classified into 6 subgroups. Cis-acting elements responsive to plant growth and development, abiotic stress, and hormone responses were identified in the promoter regions. Additionally, the transcriptome and metabolome data under cold stress were analyzed, and a co-expression and metabolite association network of AmWRKY genes was constructed. Sixteen AmWRKY transcription factors showed dynamic expression under cold stress, among which AmWRKY22/24/44/65 were continuously upregulated, indicating their core roles in cold adaptation. Co-expression network analysis revealed the synergistic effects of AmWRKY with AP2/ERF, MYB, and NAC transcription factors, forming a regulatory module integrating hormone signaling, antioxidant pathways, and circadian rhythm regulation. Metabolomics analysis indicated that AmWRKY24/44 expression was positively correlated with the upregulation of key flavonoid biosynthesis genes (AmCHS, AmFLS) and the accumulation of nine cold-responsive flavonoids. These findings suggest a new regulatory pathway of AmWRKY24/44 → flavonoid biosynthesis → cold resistance, linking secondary metabolism with environmental adaptation. This study reveals a novel regulatory pathway—“AmWRKY24/44” → flavonoid biosynthesis → cold resistance—in A. membranaceus, providing deeper mechanistic insights into how WRKY transcription factors modulate secondary metabolism under cold stress. These findings offer a valuable theoretical foundation for genetic improvement of cold tolerance in this medicinally important species.

Deep eutectic solvents (DESs) are receiving increased attention from researchers in many areas of science and technology. During the actual operation, the high viscosity of DESs, especially for solid-phase sample application, can be diminished by combination of water in the solvent system, heating, and consuming a large volume of DESs. In addition, plant matrix properties of Schisandra chinensis Baill fruits should be considered including sugars-rich and full of stone cells, which also dissolve into the solvent and to reduce extraction capacity. In this study, an ultrasonic probe-based system with a direct delivery of ultrasound to sample was developed to help the matrix materials become hydrated and swelled, which increases the diffusion and flow of solvent into the dry matrix and increases mass transfer, and also minish ultrasonic energy loss. Some parameters that potentially influence circulating and pulsating ultrasonic probe extraction (CPUPE) were methodically examined and improved using the Plackett-Burman, Box-Behnken design, and the optimum practical parameters were as follows: water content of DES, 20.2%; liquid-to-solid ratio, 11.0 mL/g; total ultrasonic treatment time, 24.7 min; ultrasonic setting temperature, 323 K; ultrasonic irradiation power, 90 W; and circulating rate, 25 mL/min. CPUPE provided a accessibility to use substituted green solvents of DESs (choline chloride-lactic acid) by improving their extraction performance. The high sample extraction capacity and circulation function of the suggested CPUPE technique makes it a quick and effective method for complex substrate plant samples. Under the circulating system, the osmotic pressure difference of the solvent is significantly increased. The CPUPE make the kind method operation simple, fast, can be apply in the on-line production analysis.

This study aims to explore the effect and mechanism of a mitochondrion-targeted derivative of ergosterol peroxide(Mito-EP) on breast cancer. The methyl thiazolyl tetrazolium(MTT) assay was employed to examine the proliferation of MDA-MB-231 cells treated with different concentrations(0, 0.075, 0.15, 0.3, 0.6, 1.2, and 2.4 μmol·L~(-1)) of Mito-EP. Cells were grouped for treatment with water(blank control), low, medium, and high concentrations(0.15, 0.3, and 0.6 μmol·L~(-1)) of Mito-EP, and ergosterol peroxide(EP)(0.6 μmol·L~(-1)). After the cells were treated for 48 h, flow cytometry was employed to examine the apoptosis rate, reactive oxygen species(ROS) level, mitochondrial membrane potential, and cell cycle distribution, and the apoptosis, ROS, and mitochondrial membrane potential were observed by laser confocal microscopy. A mouse model bearing subcutaneous xenograft tumor was established by injecting 4T1 cell suspension and used to study the inhibitory effect of Mito-EP on breast cancer. Western blot was employed to determine the protein levels of B-cell lymphoma 2(Bcl-2), Bcl-2-associated X protein(Bax), cytochrome C(Cyt C), cleaved caspase-7, and cleaved caspase-9 in cells and the tumor tissue. The results showed that Mito-EP reduced the proliferation rate of MDA-MB-231 cells in a concentration-dependent manner. Compared with the blank control group, EP(0.6 μmol·L~(-1)) caused slight changes in the apoptosis rate, ROS level, and mitochondrial membrane potential. However, Mito-EP increased the apoptosis rate, elevated the ROS level, decreased mitochondrial membrane potential, up-regulated the protein levels of Bax, Cyt C, cleaved caspase-7, and cleaved caspase-9, and down-regulated the protein level of Bcl-2(all P<0.05). Moreover, Mito-EP reduced the tumor volume and weight. In summary, Mito-EP may promote apoptosis in breast cancer cells by activating the mitochondrial apoptosis pathway.

In order to evaluate the feeding safety of Astragalus flower, acute oral toxicity tests, Ames mutagenicity tests, mammalian erythrocyte micronucleus tests, mouse spermatogonial chromosomal aberration tests, and a 90-day subacute toxicity test were conducted on Astragalus flower in accordance with national food safety standards. The results showed that there were no obvious signs of toxicity or mortality in rats and mice 14 days after gastric administration of Astragalus flower. The maximum tolerated doses (LD50) for the acute oral toxicity test in rats and mice were greater than 20.00 g/kg BW. Three genetic toxicity tests were carried out using Astragalus flower extract, and the results indicated that the Astragalus flower extract did not induce reverse mutations in the tested bacterial strains. It had no significant impact on the micronucleus cell rate in mouse erythrocytes and showed no significant effects on the rate of chromosomal abnormal cells, autosomal univalent, and sex chromosome univalent. The 90-day oral toxicity test revealed that there were no significant changes in the weight gain, total food intake, and food utilization in the rats fed with different doses of Astragalus flower. Blood indicators and organ weights in rats showed no apparent toxicological changes. Anatomical and histological observations also did not reveal any significant pathological changes, and the no-observable-adverse-effect level was determined to be 1.20 g/kg BW. The study suggests that Astragalus flower has a high safety profile, exhibiting no mutagenicity and subacute toxicity at the tested doses.

To explore the corn silk’s effect and possible mechanism on patients with type 2 diabetes mellitus (T2DM) by untargeted metabolomics. Newly diagnosed patients with T2DM admitted to the endocrinology department of the author’s hospital from March 2020 to September 2021 were chosen and then allocated to either the intervention or the control group (NC) randomly. Patients in the intervention group were administered corn silk in the same way as the patients in the NC were given a placebo. A hypoglycemic effect was observed, and an untargeted metabolomics study was done on patients of both groups. Compared with the NC, the glycosylated hemoglobin and fasting blood glucose of patients in the intervention group significantly decreased after 3 months of treatment (P < .05), identified using tandem mass spectrometry, and analyzed by orthogonal partial least squares-discriminant analysis. A total of 73 differential metabolites were screened under the conditions of variable important in projection value >1.0 and P < .05. Differential metabolites are mainly enriched in signaling pathways such as oxidative phosphorylation, purine metabolism, and endocrine resistance. Through untargeted metabolomic analysis, it is found that corn silk water extract may reduce blood glucose in patients with T2DM through multiple pathways, including oxidative phosphorylation and purine metabolism.

Morus alba L., a common traditional Chinese medicine (TCM) with a centuries-old medicinal history, owned various medicinal parts like Mori folium, Mori ramulus, Mori cortex and Mori fructus. Different medical parts exhibit distinct modern pharmacological effects. Mori folium exhibited analgesic, anti-inflammatory, hypoglycemic action and lipid-regulation effects. Mori ramulus owned anti-bacterial, anti-asthmatic and diuretic activities. Mori cortex showed counteraction action of pain, inflammatory, bacterial, and platelet aggregation. Mori fructus could decompose fat, lower blood lipids and prevent vascular sclerosis. The main chemical components in Morus alba L. covered flavonoids, phenolic compounds, alkaloids, and amino acids. This article comprehensively analyzed the recent literature related to chemical components and pharmacological actions of M. alba L., summarizing 198 of ingredients and described the modern activities of different extracts and the bioactive constituents in the four parts from M. alba L. These results fully demonstrated the medicinal value of M. alba L., provided valuable references for further comprehensive development, and layed the foundation for the utilization of M. alba L.

Background Medicine and food homological (MFH) products exhibit enhanced safety and tolerability, minimizing notable side effects, making them pivotal for prolonged use in cardiovascular diseases. This study aims to identify functional compounds in MFH based on cardiac remodeling-related target, employing reliable, comprehensive, and high-throughput methods.Methods By bioinformatics and in vivo verifications, we initially investigated the key target in the progression of cardiac remodeling. Subsequently, we performed molecular docking among medical homology compound database (MHCD), and then performed drug-likeness evaluations to recognize functional component based on disease-related target. Pharmacological verifications and data mining including cardiac and medullary transcriptomics, neurotransmitter metabolomics, resting-state functional magnetic resonance imaging (rs-fMRI), and correlationship analysis were utilized to define the benefical effects of MFH functional components, as well as its in-depth mechanims.Results The critical roles of oxidative stress and the key target of NRF2 in cardiac remodeling were discovered, and β-ecdysterone was screened as the most promising NRF2 enhancer in MHCD. Dose-dependent efficacy of β-ecdysterone in countering oxidative stress and ameliorating cardiac remodeling were then verfied by in vivo and ex vivo experiments. By data mining, the crosstalk mechanism between cardiac remodeling and neuromodulation was identified, and further unveiled Slc41a3 as a potential key factor influenced by β-ecdysterone. Additionally, β-ecdysterone mitigated increases in norepinephrine (NE) and its metabolites DHPG in the sympathetic nerve center hypothalamic paraventricular (PVN), as indicated by rs-fMRI. Cardiac and medullary transcriptomes revealed central-peripheral regulation signaling pathways during cardiac remodeling with the involvement of core gene of Dhx37 .Conclusions Our study identified β-ecdysterone as a natural MFH functional compound countering cardiac remodeling by targeting NRF2 elevation. It elucidates crosstalk between cardiac remodeling and neuromodulation, facilitating precise drug screening and mechanistic insights, providing substantial evidence for β-ecdysterone application and molecular mechanisms in cardiovascular diseases.### Competing Interest StatementThe authors have declared no competing interest.

Sulfonamides are not only widely applied in clinics but also highly valued in animal husbandry. Recently, it has become common for sulfonamide residues to exceed the standard limits in food, which can affect human health. Current regulations limit these residues. Therefore, we constructed a new limit test method to rapidly determine the levels of sulfonamide residues. Six sulfonamides were detected using the latest method called TLC-SERS, namely, sulfamethasone (A), sulfamethazine (B), sulfadoxine (C), sulfamethoxydiazine (D), sulfamethoxazole (E), and sulfathiazole (F). The optimal conditions for SERS detection were investigated for these six drugs, and the separation effects of different TLC spreaders on them were compared. Then, we successfully established a separation system using dichloromethane–methanol–ammonia in a ratio of 5:1:0.25 (v/v/v), which provided good separation effects on the six drugs. The residues were preliminarily separated via TLC. A silver sol solution was added to the spot on the silica gel G plate at the corresponding specific shift values, and SERS detection was performed. The sample solution was placed on the spot under a 532 nm laser, and the SERS spectrum was collected and analyzed for the six sulfonamides. The results showed obvious variations in the SERS spectrum among the six sulfonamides, with the LODs being 12.5, 6.4, 6.3, 7.1, 18.8, and 6.2 ng/mL from A to F, respectively, and an RSD of <3.0%. Within 48 h, the SERS signal for each sulfonamide drug was kept stable, with an RSD of <3.0%. The detection results of 20 samples using the TLC-SERS method were consistent with those obtained by UPLC-MS/MS. The established TLC-SERS method is simple and fast, providing a useful reference for the rapid detection of residue limits in food.

ETHNOPHARMACOLOGICAL RELEVANCE:Trollius chinensis Bunge has a long history of use in China as traditional Chinese medicine and functional tea for the treatment of respiratory infections, such as pharyngitis, tonsillitis and bronchitis. Pharyngitis can impact the entire throat and adjacent lymphoid tissues, and may lead to significant systemic complications. However, the active components and mechanism of Trollius chinensis Bunge for treating acute pharyngitis remains unclear. AIM OF THE STUDY:Trollius chinensis Bunge is recognized in China both as a medicinal herb and a functional tea. Research into its properties aimed to establish its effectiveness against pharyngitis and to pinpoint the active components and mechanism. MATERIALS AND METHODS:A 70% ethanol extract from the herb was prepared, which was refined using chromatography through a column containing D101 macroporous resin and varying ethanol solutions. The efficacy of the initial and refined extracts was tested using a rat model of ammonia-induced acute pharyngitis. Pathological examination, HE staining and ELISA were applied to screen activity fraction. The compounds were isolated by silica gel, sephadex LH-20 column chromatography and semi-preparative HPLC chromatography from active fraction. All of the isolated compounds were assessed for anti-inflammatory activity by acting on LPS-induced RAW 264.7 macrophage cells in vitro. Cytotoxicity of compounds was detected by CCK-8 assay. The Griess reaction was applied to evaluate the inhibitory effects of isolated compounds on NO production in RAW 264.7 cells induced by LPS. TNF-α, IL-6, IL-1β and PGE2 levels in macrophage supernatant were detected by ELISA. Molecular docking and western blot analysis were applied to study the anti-inflammatory mechanism of active compound. RESULTS:The fraction extracted with 30% ethanol proved particularly effective, significantly reducing pharyngitis symptoms. This was evidenced by decreased levels of cytokines (TNF-α, IL-6, IL-1β and PGE2) and visible improvements in the pharyngeal tissue histology. In pursuit of pharyngitis treatments, 23 phenolic acids and 13 flavonoids were isolated from the 30% ethanol fraction and identified using spectral analysis. Of these, three were newly discovered compounds and eight were first-time isolates from the Trollius genus. These compounds were further investigated for their ability to suppress nitric oxide production in RAW 264.7 cells triggered by lipopolysaccharide. Compounds 3, 19, and 26 exhibited strong anti-inflammatory properties. HPLC analysis of the 30% ethanol fraction revealed that orientin was the predominant component, accounting for 44.4% of this fraction. Western blot analysis demonstrated that orientin reduced the expression levels of the protein p-p65 relative to p65, p-IκBα relative to IκBα and iNOS, indicating an anti-inflammatory effect potentially through the modulation of the NF-κB signaling pathway. CONCLUSION:The finding of this study provided strong support for the use of T. chinensis as a potential functional food for treating pharyngitis.