Lung adenocarcinoma (LUAD) is a leading cause of cancer-related mortality worldwide. Although established biomarkers guide targeted therapy, their utility in prognostic stratification remains limited. In this study, we. ASPH mRNA expression across pan-cancers and in LUAD was analyzed using the TIMER and The Cancer Genome Atlas (TCGA) databases. Immunohistochemical (IHC) staining was performed on 120 clinical LUAD specimens to validate ASPH protein expression. Kaplan-Meier survival analysis was used to assess the association between ASPH expression and overall survival (OS) as well as progression-free survival (PFS). Univariate and multivariate Cox proportional hazards regression models were applied to evaluate the prognostic value of ASPH. Gene Set Enrichment Analysis (GSEA) and Pearson co-expression analysis were conducted to explore potential molecular mechanisms. ASPH was significantly upregulated in LUAD tissues at both mRNA and protein levels compared with corresponding normal lung tissues (all p < 0.001). High ASPH expression was closely correlated with aggressive clinicopathological features, including larger tumor size, T-stage parameters, lymph node/distant metastasis, and advanced clinical stage (all p < 0.05). Kaplan-Meier analysis showed that high ASPH expression was associated with poorer OS and PFS (both p < 0.001). Multivariate Cox regression confirmed ASPH as an independent prognostic factor for OS (TCGA cohort: HR = 1.01, 95

This study proposed an innovative low-temperature enzymatic catalytic approach for efficient N-acetylneuraminic acid (Neu5Ac) production. By integrating homology search, clustering distribution, solubility prediction, and structural-based bioinformatics calculations, two novel cold-adapted enzymes of N-acetylglucosamine -2-epimerase (PraAGE) and N-acetyl-d-neuraminic acid lyase (PshNAL) were identified from Pricia antarctica and Psychromonas hadalis respectively. Both enzymes exhibit high catalytic activity, stability, and substrate tolerance at low-temperature. Notably, PraAGE demonstrated activity without ATP and required only 20 μM ATP for optimum activity, while PshNAL showed a high equilibrium constant (Kc) at low-temperature, effectively driving the forward reaction toward Neu5Ac synthesis. Leveraging these findings, a two-step enzymatic system for Neu5Ac production at low-temperature was established. After two rounds of fed-batch reactions, the highest Neu5Ac yield of 639.15 mM (197.67 g/L) with the molar conversion rate of 79.89% was achieved at 40 h. The low-temperature strategy lays a robust foundation for large-scale industrial Neu5Ac production.

Lung adenocarcinoma (LUAD) is the leading cause of cancer-related death worldwide. Despite advances in surgery, targeted therapy, and immunotherapy, the 5-year survival rate of advanced LUAD remains below 20%, indicating an urgent need for reliable molecular biomarkers for early detection and prognosis. In this study, the authors hypothesized that three consistently upregulated genes could act as effective diagnostic and prognostic biomarkers for LUAD. The authors analyzed transcriptomic data from two independent cohorts, TCGA-LUAD (535 tumors, 59 normal samples) and GSE115002 (52 tumors, 52 matched normal samples), to screen differentially expressed genes. Three core genes-B3GNT3, FERMT1, and SPP1-were consistently overexpressed in LUAD tumors in both datasets. These genes showed excellent diagnostic performance, with AUC values above 0.95 in TCGA-LUAD and high accuracy in GSE115002. Survival analysis showed that high expression of each gene was significantly associated with shorter overall and disease-free survival, and multivariate Cox regression verified their independent prognostic value. Functional enrichment analysis indicated that these three genes participate in epithelial-mesenchymal transition, extracellular matrix remodeling, and immune suppression, all of which are closely related to LUAD invasion and metastasis. The authors further constructed a prognostic nomogram combining the three genes and TNM stage, achieving a concordance index of 0.743 and demonstrating good predictive performance. These findings confirm that B3GNT3, FERMT1, and SPP1 are promising diagnostic and prognostic biomarkers for LUAD, supporting the clinical application in risk stratification and management.

IntroductionThe geo-authenticity of medicinal plants, exemplified by Rehmannia glutinosa, is largely attributed to environmentally driven variation in bioactive compounds; yet, the underlying systemic molecular mechanisms remain elusive. MethodsThis study employed an integrated approach combining targeted quantitative analysis, transcriptomics, and metabolomics to dissect how geographical origin shapes the medicinal quality of Rehmannia glutinosa, using two cultivars ('Wen 85-5' and 'Jin Jiu') sourced from two distinct production regions (Henan and Hebei).ResultsTargeted quantification confirmed origin-specific accumulation patterns of key bioactive compounds, most notably a significantly higher acteoside content in roots from the Henan origin. Multi-omics profiling revealed a conserved core molecular response to geographical origin, involving 894 common differentially expressed genes and 443 common differentially abundant metabolites enriched in hormone signaling, primary metabolism, and specialized biosynthesis pathways. While the response amplitude was genotype-dependent (stronger in 'Wen 85-5'), its fundamental architecture was consistent. Crucially, we identified a coordinated upregulation of the entire acteoside biosynthetic network in Henan-sourced roots. This was evidenced by the concerted induction of key structural genes (PAL, C4H, 4CL, TyDC, UGT) across both phenylpropanoid and tyrosine-derived branches, coupled with elevated levels of pathway intermediates.ConclusionThis study elucidates that the geo-authenticity of Rehmannia glutinosa arises from an origin-triggered, systemic reconfiguration of interconnected transcriptional and metabolic networks. The identified core regulatory network provides a mechanistic framework for understanding quality formation and paves the way for molecular-assisted cultivation and breeding strategies.

Pseudostellaria heterophylla is an important medicinal herb whose seed propagation is hindered by physiological dormancy. We investigated cold stratification-induced dormancy release via integrative physiological, anatomical, and transcriptomic analyses. Optimal dormancy release was achieved after 55 days of cold stratification at 0-1 degrees C with 16% moisture, yielding 86.67% germination. Anatomically, embryo volume progressively increased while endosperm decreased, accompanied by asynchronous cotyledon elongation and vascular tissue formation. Physiologically, three distinct phases were identified. Soluble protein peaked at 20 days. Starch showed transient increases at 20 and 45 days before declining after 45 days. Soluble sugars accumulated only after 45 days. Endogenous hormone analysis revealed that ABA decreased while GA3 and IAA increased from 20 days onward. Respiratory metabolism shifted toward the TCA cycle (MDH increased) with no change in G6PDH. Transcriptomic analysis identified 52,582 unigenes and 11,425-15,212 differentially expressed genes across stratification stages. WGCNA revealed a blue module (5234 genes) strongly correlated with germination traits and enriched in starch/sucrose metabolism, glycolysis, phenylpropanoid biosynthesis, and photosynthesis-related pathways. p-coumaric acid, a phenylpropanoid intermediate, progressively declined during stratification, and exogenous application confirmed its dose-dependent germination inhibition, identifying it as a candidate endogenous germination inhibitor. These findings suggest that cold stratification is associated with a temporally structured, multi-stage reprogramming of embryo development, hormone signaling, energy metabolism, and secondary metabolite clearance, with the blue module integrating these processes. This study provides a theoretical foundation for developing evidence-based seed priming technologies for medicinal plants.

Coreopsis tinctoria Nutt. (CtN) has a widely used in traditional medicine and food applications in China. As a newly authorized food ingredient in China, CtN provides a safe, natural, and multitarget therapeutic strategy for skin aging intervention, offering unique advantages for the development of 'beauty-from-within' functional foods. However, there is currently a lack of its effects on skin glycation aging. The alcohol extract of Chinese C. tinctoria Nutt (CtNE) was analyzed using UHPLC-LTQ-Orbitrap-MS/MS-(MS2) and MS3 technology. A methylglyoxal (MGO)-induced glycation aging model was established in HaCaT cells, followed by intervention with the CtNE. A total of 28 components were identified from CtNE, which demonstrated the ability to improve MGO-induced cell proliferation inhibition. CtNE increased the number of cells in the S phase, promoted cell migration, reduced reactive oxygen species (ROS) and malondialdehyde (MDA) levels, elevated the antioxidant superoxide dismutase (SOD) level, decreased β-galactosidase activity, and mitigated the nuclear accumulation of K9 trimethylated histone H3 (K9M-H3). Furthermore, CtNE downregulated P21 expression and phosphorylation of P53, while upregulating the expression of P-PI3K,P-AKT,P-mTOR,ZO-1 and Occludin, and inhibiting P-P53 expression. The constituents of CtNE exhibit the potential to ameliorate skin glycation-induced aging.

Traditional Chinese medicine, with its rich history and profound influence on healthcare, is deeply rooted in medicinal plants, which serve as the foundation for Chinese herbal medicines (CHMs). However, as demand rises and wild resources become increasingly scarce, herbal medicines face significant challenges, including inconsistent production, variable quality, and safety concerns. AI technologies, such as bionic sensors and advanced spectroscopic techniques, offer precise, objective, and efficient solutions to address these issues. This review explores the application of these AI-driven detection technologies across various aspects of Chinese medicinal agriculture (CMA), covering field management (plant health assessment, pest and disease management, and species identification), and quality control (detection of adulteration and admixture, source identification, detection of hazardous substances, and physicochemical component analysis). This paper highlights the significant benefits of AI technologies in driving CMA modernization, including improved standardization, reduced human error, and increased operational efficiency. However, it also discusses the challenges related to complex-terrain monitoring, data fusion, model optimization, equipment limitations, and domain-specific complexities in CHM quality evaluation. This review further proposes key opportunities in AI-enabled complex-terrain monitoring, multi-omics breeding for Dao-di herbs, nanomaterial-enhanced pharmacology, and large language models-powered decision support. These opportunities could overcome existing challenges while harnessing AI's potential to transform CMA into an AI-driven ecosystem for standardized, precise, and consistent production of therapeutic-grade CHMs.

Ultraviolet (UV) exposure accelerates skin aging and increases the risk of skin-related diseases. Amentoflavone (AMF), the major compound isolated from Selaginella tamariscina, exhibits potent antioxidant and anti-inflammatory activities. This study aimed to investigate the therapeutic effects and mechanisms of S. tamariscina extract (STE) and AMF on UVB-induced skin photoaging. In vitro and in vivo photoaging models were established to evaluate the protective effects of STE and AMF. The therapeutic target of AMF was identified using network pharmacology, bioinformatic analysis, and molecular docking. In vitro, STE significantly reduced UVB-induced oxidative stress, inflammation, and apoptosis. In vivo, both STE and AMF effectively mitigated UVB-induced skin injury. Mechanistically, AMF directly interacted with AMP-activated protein kinase (AMPK), thereby promoting autophagy and protecting cells from UVB-induced damage. In conclusion, STE and its active compound AMF alleviate UVB-induced photoaging via activation of the AMPK signaling pathway, supporting their potential use in skin photoaging therapy.

Pseudostellaria heterophylla (Miq.) Pax (P. heterophylla) was a valued traditional Chinese herbal medicine. Previous studies have shown that P. heterophylla TuMV spreads during the vegetative propagation cycle using tuberous roots as carriers. However, the transmission mechanism of TuMV in P. heterophylla and its effects on host growth remain to be elucidated. In this study, virus-free P. heterophylla culture seedlings were infected with control, TuMV-ZR, and TuMV-ZR-EGFP, thereby resulting in the initial infection cycle of IF1 (TIF1, TEIF1) and control NIF1, and used these roots to propagate the subsequent infection cycle IF2 (TIF2, TEIF2) and control NIF2. The transmission of TuMV-ZR seedlings was tracked by EGFP signal, and their yield, quality, and resistance were analyzed simultaneously in the critical growth period of the plants. The results indicated that TuMV-ZR accumulated in the tuberous roots of IF1 plants, subsequently migrated to IF2 during seedling growth, and was re-stored in IF2 tuberous roots, thereby forming a simple virus transmission cycle. Meanwhile, the tuberous roots of IF1 and IF2 P. heterophylla showed lower fresh weight, dry weight, soluble sugar, and saponin levels compared to NIF1 and NIF2, respectively. TuMV caused a significant reduction in chlorophyll synthesis in IF1 and IF2 P. heterophylla, resulting in impairment to their photosynthetic organs and efficiency. The measurement of stress resistance in IF1 and IF2 P. heterophylla revealed that continuous viral infection disrupted antioxidant enzyme activity, increased the content of MDA, enhanced the activity of PAL, and elevated the levels of intracellular osmotic substances in both propagation cycles. The findings indicated that the accumulation of the TuMV-ZR virus during two successive vegetative propagation cycles induced physiological stress, impaired photosynthesis, and caused progressive yield and quality decline with each cycle. This study systematically examined the impact of TuMV-ZR persistence during vegetative propagation on key physiological and biochemical indices in P. heterophylla, providing critical data to clarify vegetative-propagation-mediated germplasm degradation.

Anoectochilus roxburghii (Wall.) Lindl. (A. roxburghii) is an increasingly popular medicinal herb. Arbuscular mycorrhiza (AM) fungi, known for their symbiotic relationships with plant roots, enhance nutrient uptake and disease resistance in host plants. However, their specific regulatory mechanisms in A. roxburghii are not fully understood. In this study, Fujian A. roxburghii was inoculated with the AM fungus Glomus intraradices, and successful root colonization was observed. Following AM fungal colonization, there was a significant upregulation of photosynthesis-related genes in the stems, accompanied by improved canopy phenotypes and root architecture. Consequently, AM-inoculated plants exhibited increased fresh and dry biomass, as well as elevated levels of polysaccharides and flavonoids. Additionally, the incidence of Fusarium oxysporum-induced stalk rot was reduced in AM-inoculated plants. Analysis of defense-related enzymes indicated that AM-inoculated plants exhibited a rapid and robust response to pathogen infection, mitigating oxidative stress. Transcriptomic analysis revealed significant upregulation of genes associated "Fatty acid degradation", "MAPK signaling pathway-plant", and "Plant-pathogen interaction", suggesting their involvement in enhanced disease resistance. A regulatory network centered on ACX1 and calmodulin, involving multiple transcription factors such as WRKY, bHLH, ERF, NAC, and HSF, was implicated in defense responses. These findings demonstrated the beneficial effects of AM fungi on yield, quality, and disease resistance in A. roxburghii, providing a theoretical foundation for its cultivation and genetic improvement.

The balance between growth and defense in response to nearby or canopy shading in heliotropic plants has been deeply understood. However, the adaptive traits developed by shade-tolerant plants through long-term evolution remain unclear. In this study, the typical shade-tolerant medicinal plant Anoectochilus roxburghii was used as the experimental material.(1) Different planting densities were set, including 8 cm(row spacing) × 8 cm(plant spacing), 6 cm × 6 cm, 4 cm × 4 cm, and 2 cm × 2 cm, to monitor the individual plant responses to nearby shading.(2) Different light environments, including blue light∶red light=3∶2(B3R2), blue light∶red light∶far-red light=3∶2∶1(B3R2FR1), blue light∶red light∶far-red light=3∶2∶2(B3R2FR2), and blue light∶red light∶far-red light=3∶2∶4(B3R2FR4), were set to monitor the morphological and physiological changes in plants in response to actual shading conditions. The results showed that:(1) Moderate increases in planting density helped optimize morphological traits such as stem diameter and leaf area. This not only slightly increased biomass but also significantly improved SOD activity in both leaves and stems, as well as lignin content in stems, thereby enhancing the plant's defense capabilities.(2) Increasing the far-red light in the light environment negatively regulated the plant height of A. roxburghii, which was contrary to the typical shade-avoidance response observed in heliotropic plants. However, it significantly enhanced SOD and POD activity in both stems and leaves, as well as lignin content in stems. Furthermore, it reduced the incidence and disease index of stalk rot, effectively defending against biotic stress. Therefore, the shade-tolerant plant A. roxburghii has specific adaptive strategies for shading conditions. Reasonable dense planting or light environment modulation can synergistically improve yield, medicinal quality, and resistance of A. roxburghii. This study provides a theoretical foundation and technical support for optimizing the regional deployment and cultivation strategies of ecological planting for Chinese medicinal materials.

Flavonoids are a major component of Artemisia argyi and play a crucial role in its pharmacological properties. However, the molecular mechanisms underlying flavonoid biosynthesis in A. argyi remain unclear. To address this, transcriptome and quantitative metabolome analyses were conducted across five developmental stages of A. argyi. In total, 85 flavonoid compounds were identified across these different stages. Differentially expressed candidate genes and metabolites involved in flavonoid biosynthesis were also identified. Differentially accumulated flavonoid metabolites (DFMs) were observed among the 10 comparison groups, with 29 DFMs identified from the five developmental stages of A. argyi leaves. The biosynthesis process identified 38 differentially expressed genes (DEGs) from seven gene families. Furthermore, 26 DEGs exhibited a significant correlation with the levels of seven active flavonoid metabolites, as revealed by weighted gene co-expression network analysis. These DEGs included eight HCT genes, six CHI genes, two CHS genes, three CCoAOMT genes, two F3'H genes, two C4H genes, two CYP98A genes, and one F3H gene. Based on preliminary analysis, HCT1 may be associated with accumulating hispidulin and jaceosidin. This study investigated the relationship between differential gene expression and flavonoid accumulation using an integrated transcriptomic and metabolomic approach, providing valuable insights into the mechanisms of flavonoid biosynthesis and quality formation in A. argyi.

This study, conducted as part of a multicenter phase III clinical trial, aimed to assess the utility of circulating tumor DNA (ctDNA)-based minimal residual disease (MRD) in comparing the efficacy of short-course and long-course chemoradiotherapy (CRT) for locally advanced rectal cancer (LARC). A total of 244 plasma samples from 79 LARC patients undergoing neoadjuvant therapy (NAT) before surgery were collected at various time points. Targeted deep sequencing using a novel MRD panel was performed. During NAT, ctDNA levels declined significantly. Baseline ctDNA-MRD status did not correlate significantly with treatment response. Notably, compared to long-course radiotherapy, microsatellite instability increased significantly after short-course radiotherapy (shortRT). Additionally, ctDNA negativity or lower levels were significantly associated with pathological complete response (pCR). Clearance of ctDNA and MRD after shortRT correlated significantly with pCR. A predictive model based on ctDNA-MRD, combined with carcinoembryonic antigen (CEA), outperformed models using only MRD or only CEA in predicting pCR/non-pCR. These findings provide insights into NAT for LARC and highlight ctDNA-based MRD assessment's potential in tailoring treatment strategies, emphasizing the need for personalized approaches.

BACKGROUND:Rehmannia glutinosa, a herb of significant medicinal and food value, is highly susceptible to pathogen infections. Fusarium oxysporum f.sp. R. glutinosa (FORg), a pathogenic strain isolated from R. glutinosa, induces root rot disease, leading to severe yield losses. Currently, resistant cultivars and effective control strategies for R. glutinosa root rot remain very limited. This study aims to investigate the feasibility and potential of SIGS technology in preventing and controlling R. glutinosa root rot caused by FORg. RESULTS:This study demonstrates that FORg hyphae can capably take up dsRNA, confirming their suitability for SIGS-based disease control. dsRNA targeting SGE1 and CYP51, synthesized in vivo and extracted through low-cost ethanol method, significantly suppressed the expression of virulence-associated genes in both hyphae and conidia. Root disc assays further validated that exogenous dsRNA effectively attenuated FORg pathogenicity. Combined applications of SGE1 and CYP51S dsRNAs synergistically reduced disease severity compared to single-target treatments. Additionally, dsRNA encapsulated in layered double hydroxide (LDH) prolonged functional durability, with sustained plant protection observed for 15 days post-spraying, outperforming unencapsulated dsRNA. CONCLUSION:This finding establishes the feasibility of SIGS for controlling FORg-induced root rot in R. glutinosa and highlights the potential of nanomaterial encapsulation in addressing persistent fungal epidemics in agricultural systems. © 2025 Society of Chemical Industry.

Ligand fishing with immobilized enzymes offers a promising approach for screening natural active compounds in complex extracts. Key challenges in enzyme immobilization include maintaining structural integrity and enhancing loading capacity. This study employed hydrogen-bonded organic frameworks (HOFs), eco-friendly porous materials synthesized via hydrogen bonding, to immobilize elastase (ELA) through self-assembly for affinity screening of ELA inhibitors from Coreopsis tinctoria Nutt. The successful fabrication of ELA@HOF was validated using techniques such as SEM, TEM, FT-IR, XRD, XPS, TGA, and BET. HOFs significantly improved enzyme loading capacity (524.4 mg/g), immobilization rate (85.7%), specific activity (25.7 U/mg) and stability (notable acid resistance) compared to conventional methods. ELA@HOF demonstrated remarkable repeatability for ligand fishing which was reusable for 12 cycles. Seven inhibitors of ELA were extracted from extracts of C. tinctoria, which were identified by UPLC-MS combined with comparison with authentic samples as isookanin, taxifolin, marein, 7,3',5'-trihydroxyflavanone, okanin, eriodictyol, and sulfuretin. The ELA inhibitory activity and enzymatic kinetic study were further investigated, revealing a significant level of inhibition. Molecular docking technique was used to simulate the interaction between the ligand and ELA. These findings suggest the great potential of hydrogen-bonded organic frameworks for the rapid screening of active natural compounds.

Rehmannia glutinosa is an important medicinal herb; but its long-term cultivation often leads to continuous cropping problems. The underlying cause can be attributed to the accumulation of and alterations in root exudates; which interact with soil-borne pathogens; particularly Fusarium oxysporum; triggering disease outbreaks that severely affect its yield and quality. It is therefore crucial to elucidate the mechanisms by which root exudates induce F. oxysporum CCS043 outbreaks. In this study; the genome of F. oxysporum CCS043 from R. glutinosa’s rhizosphere microbiota was sequenced and assembled de novo; resulting in a 47.67 Mb genome comprising 16,423 protein-coding genes. Evolutionary analysis suggests that different F. oxysporum strains may adapt to the host rhizosphere microecosystem by acquiring varying numbers of specific genes while maintaining a constant number of core genes.The allelopathic effects of ferulic acid; verbascoside; and catalpol on F. oxysporum CCS043 were examined at the physiological and transcriptomic levels. The application of ferulic acid was observed to primarily facilitate the proliferation and growth of F. oxysporum CCS043; whereas verbascoside notably enhanced the biosynthesis of infection-related enzymes such as pectinase and cellulase. Catalpol demonstrated a moderate level of allelopathic effects in comparison to the other two. Furthermore; 10 effectors were identified by combining the genomic data. Meanwhile; it was found that among the effector-protein-coding genes; ChiC; VRDA; csn; and chitinase exhibited upregulated expression across all treatments. The expression patterns of these key genes were validated using qRT-PCR. Transient overexpression of the two effector-encoding genes in detached R. glutinosa leaves provided further confirmation that ChiC (GME8876_g) and csn (GME9251_g) are key effector proteins responsible for the induction of hypersensitive reactions in R. glutinosa leaf cells. This study provides a preliminary indication that the use of allelochemicals by F. oxysporum CCS043 can promote its own growth and proliferation and enhance infection activity. This finding offers a solid theoretical basis and data support for elucidating the fundamental causes of fungal disease outbreaks in continuous cropping of R. glutinosa and for formulating effective mitigation strategies.

ObjectiveThis study aims to develop a model for long non-coding RNA (lncRNA) associated with cuproptosis and assess the efficacy of immunotherapy and chemotherapy in children with Wilms tumor (WT) based on individualized risk scores.MethodsData was obtained from the online database. Cox proportional hazards analysis and LASSO Cox regression were employed to generate cuproptosis-related lncRNA signatures. Patients were classified into high- and low-risk groups and clinical outcomes were further analysed. Tumor mutation burden and immunoinfiltration were calculated and potential immunotherapy response was evaluated. The sensitivity of immunotherapy and chemotherapy was ultimately analyzed based on individual risk scores associated with cuproptosis.ResultsA eight cuproptosis-related lncRNAs signature was established and high-risk group showed a worse prognosis than the low-risk group. This model showed a good diagnostic performance. Low-risk group displayed an elevated tumor immune dysfunction and was more sensitive to 13 drugs.ConclusionThe current study introduces a novel approach for predicting clinical prognosis and determining the appropriate therapy for patients with WT.

Alhagi camelorum (AC) is an herbal medicine known for its anti-inflammatory and antioxidant properties. However, the mechanism by which AC affects skin aging remains unclear. In this study, a polysaccharide with antioxidant activity was extracted from AC seeds (ACSP). The structure of ACSP was characterized by molecular weight determination, Fourier transform infrared spectroscopy, monosaccharide composition analysis, scanning electron microscopy, and physicochemical analysis. The molecular weight of ACSP was 736,831 Da. ACSP, devoid of a triple-helical conformation, comprised mannose, rhamnose, galacturonic acid, glucose, galactose, and arabinose at a molar ratio of 39.65:0.85:0.59:11.57:40.25:6.60. The anti-skin-aging activity and potential mechanisms of action of ACSP were investigated in vitro. We observed that ACSP delayed the onset of senescence, promoted cell migration, decreased the expression of DNA damage markers and the production of reactive oxygen species, and increased the mitochondrial membrane potential in damaged cells. ACSP also significantly reduced the expression of the inflammatory mediators interleukin (IL)-6, IL-1β, and tumor necrosis factor-α in HaCaT cells. In addition, ACSP attenuated the expression of proteins within the IL-17 signaling pathway and suppressed the phosphorylation of JAK2/STAT3/NF-κB proteins, thereby exerting a senescence-delaying effect on MGO-induced HaCaT cells. In conclusion, we elucidated the potential of ACSP in delaying skin aging and offer a novel plant-derived adjuvant to delay aging.

Acteoside (ACT), a prominent compound of the hydroxytyrosol-type phenylethanol glycoside (HPG) class, is present in plants and holds significant potential for food and pharmaceutical applications. However, the limited production of ACT in plants restricts its broader utilization. Although the biosynthetic pathways of ACT are increasingly understood, its transport mechanisms within plants remain unclear. RgMATE6, a vacuolar-type Multidrug and Toxic Compound Extrusion (MATE) transporter identified in Rehmannia glutinosa (a plant known for ACT p roduction), was selected for investigation. This study aims to elucidate the role of RgMATE6 in ACT transport and its impact on ACT biosynthesis. Our study utilized a multidisciplinary approach, including in silico analysis to predict substrate specificity, quantitative real-time PCR (qRT-PCR) to quantify gene expression, HPLC to measure HPG levels, vacuolar membrane vesicle uptake assays to validate RgMATE6 transport activity in vitro, and genetic transformation in R. glutinosa to assess its functional roles in vivo. In silico analysis identified RgMATE6 as a phenolic compound transporter, and correlation analysis revealed a strong positive association between the HPG accumulation and RgMATE6 expression in R. glutinosa. Functional validation through vacuolar membrane vesicle uptake assays in Nicotiana benthamiana confirmed RgMATE6’s role as an HPG transporter, demonstrating a significant preference for ACT. Overexpression and repression experiments in R. glutinosa further demonstrated that RgMATE6 facilitates ACT import into vacuoles and enhances its production. Additionally, tissue-specific expression analysis revealed the coordinated expression patterns between RgMATE6 and six ACT biosynthetic genes in the transgenic plants. RgMATE6 facilitates the transport and accumulation of ACT within vacuoles, and its expression might synergize with ACT biosynthesis. These findings establish a framework for improving ACT and other HPG production through targeted manipulation of plant MATE transporters.

The obstacle of continuous cropping poses a key challenge limiting the sustainable and safe production of Chinese herbal medicines, with the dysregulation of plant immune responses being identified as the primary contributing factor. LRR-RLPs, as major members of plant pattern recognition receptors (PRRs), are extensively involved in mediating plant immune responses. However, these receptors remain unidentified in the medicinal plant Rehmannia glutinosa. In this study, we identified 14 RgRLP genes in R. glutinosa through bioinformatics approaches and systematically analyzed their differential expression patternas under potted cultivation versus controlled simulated continuous cropping conditions. Subsequently, we constructed overexpression systems for functional characterization of three pivotal RgRLP genes (RgRLP4, RgRLP10, and RgRLP12), which were identified as central regulators of continuous cropping stress responses. Notably, continuous cropping stress induced severe oxidative damage in root tip tissues and promoted rhizosphere proliferation of Fusarium oxysporum via exudate-mediated interactions, thereby exacerbating stress effects. Importantly, overexpression of RgRLP genes significantly alleviated continuous cropping-induced stress damage. Furthermore, transcriptomic analysis of transgenic lines revealed marked upregulation of three LRR-RLK genes (RgRLK19, RgRLK27, and RgRLK30), suggesting their potential roles in RgRLP-mediated stress signaling pathways. This study provides a good theoretical basis and technical support for in-depth study of the formation mechanism and reduction strategies of continuous cropping obstacles of R. glutinosa.