A systematic chemical characterization of Bidens pilosa var. radiata was performed using ultra-high performance liquid chromatography coupled with Orbitrap high-resolution mass spectrometry(UHPLC-Orbitrap-MS). A quantitative method based on ultra-performance liquid chromatography coupled with photodiode array detector(UPLC-PDA) was established for phenolic acids and flavonoids, and the impacts of growth stage, geographical environment, and medicinal parts on chemical constituents of B. pilosa var. radiata were comprehensively investigated. This study revealed for the first time its multi-dimensional dynamic variation patterns across harvest time, geographical origin, and medicinal parts. A total of 48 chemical constituents were identified, predominantly flavonoids and phenolic acids. The established UPLC-PDA method achieved accurate quantification of 10 major compounds, including neochlorogenic acid, chlorogenic acid, rutin, and hyperoside. Multivariate analysis demonstrated that the active ingredients of B. pilosa var. radiata showed significant time-dependent accumulation, with August identified as the optimal harvesting period for samples from Guangxi. The quality of B. pilosa var. radiata varied markedly across regions, with Guangxi specimens exhibiting balanced and higher constituent levels and thus being qualified as premium medicinal references. Climatic conditions significantly influenced component accumulation, with high temperature, humidity, and moderate sunlight in Guangxi favoring flavonoid and phenolic acid biosynthesis. Distinct tissue-specific distribution patterns of phenolic acids and flavonoids were observed, with content levels following the order of leaves > flowers > stems. Furthermore, principal component analysis(PCA) was employed for unsupervised pattern recognition of 14 batches of Bidens medicinal materials to characterize their distribution patterns, while supervised orthogonal partial least squares-discriminant analysis(OPLS-DA) was applied to differentiate samples from different botanical origins. Isochlorogenic acid A, chlorogenic acid, and isochlorogenic acid B were identified as potential differential components. By establishing a chemometric quality evaluation system based on multi-component analysis, this study systematically elucidated the dynamic variation patterns of chemical constituents in B. pilosa var. radiata, providing a scientific foundation for rational quality standards development, medicinal part optimization, origin suitability assessment, and germplasm resource utilization.

Fuganlin oral liquid (FOL) has been clinically employed for the treatment of pediatric qi deficiency colds, manifesting symptoms such as fever, cough, asthma, and sore throat. However, the chemical composition and bioactive components of FOL have not been clearly elucidated. In this study, a comprehensive qualitative analysis of FOL was conducted utilizing ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry in conjunction with network pharmacology. A total of 124 chemical components were tentatively characterized, comprising flavonoids, phenolic acids, saponins, coumarins, and others. Among these, 43 compounds were unequivocally identified by comparison with authentic reference standards. Furthermore, network pharmacology analysis revealed that the ingredients of FOL exhibited anti-inflammatory properties and demonstrated potential efficacy in relieving cough and asthma associated with respiratory tract infections. Collectively, this study provides the first comprehensive characterization of the chemical composition in FOL and explores the potential pharmacological mechanisms of its bioactive constituents, thereby providing scientific support for quality control standards and clinical applications.

Endothelial dysfunction, chronic inflammation, immune dysregulation, oxidative stress, mitochondrial dysfunction, and metabolic disturbances collectively contribute to cardiovascular diseases (CVDs) associated with blood stasis patterns. Xuefu Zhuyu Decoction (XFZYD) is widely used clinically for the management of CVDs. Based on serum-exposed prototype profiling in rats, two pharmacology-driven core component sets of XFZYD were defined as the core set for the promotion of blood circulation and the elimination of blood stasis (CPBEB; HSYA, GRo, FA, β-ECD, AMY, ALB, PF) and the core set for the regulation of qi and the relief of pain (CRQRP; LIQ, NR, NAR, ROF, HSD, NHP, LTG, NRG, ISL, FNT, NOB, PD, SSa). CPBEB primarily targets vascular pathology by regulating endothelial dysfunction with dyslipidemia-driven arterial lipid deposition. Mechanistically, CPBEB is associated with improved endothelial function, reduced plaque instability, attenuated chronic inflammation and oxidative stress, normalized lipid and bile acid metabolism, and decreased thrombosis. CRQRP primarily modulates vascular tone and systemic energy metabolism. These effects are linked to enhanced AMPK/SIRT1-driven antioxidant defenses and mitochondrial homeostasis, increased NO/cGMP signaling, coordinated crosstalk among the TLR4/NF-κB, JAK/STAT, NLRP3, and PPAR pathways, and remodeling of the gut microbiota-immune network. In summary, this review integrates modern analytical approaches with network pharmacology and the literature evidence to clarify the material basis underlying XFZYD's therapeutic effects in CVDs, thereby supporting the modernization and internationalization of traditional Chinese medicine.

As a natural green catalyst, laccase demonstrates significant potential in industries such as environmental remediation and food safety. However, the inherent limitations of the native free enzyme—including its catalytic performance, stability, and reusability—hinder its practical adoption. This review focuses on the key challenges associated with the real-world application of laccase and systematically summarizes recent advances in innovative strategies for enhancing its performance, as well as breakthroughs in developing alternative materials. Unlike previous reviews, this work provides, for the first time, an integrated and systematic overview of both performance optimization of native laccase—through source screening, protein engineering, and immobilization techniques—and the development of emerging laccase-mimicking nanozymes, thereby offering a novel perspective from “enzyme optimization” to “enzyme replacement”. The article provides a critical analysis of the mechanisms, advantages, and limitations of various strategies, along with an assessment of their practical potential. This evaluation is complemented by an outline of promising future research directions and key technologies for advancing laccase development. This review aims to provide a comprehensive, in-depth, and forward-looking resource for researchers in the field, facilitating the rational design and application of next-generation biocatalysts.

Medicinal herbs contain natural products (NPs) possessing rich scaffolds valuable for drug discovery, particularly in oncology. While most NP-derived cancer therapeutics directly kill tumor cells, emerging opportunities lie in modulating antitumor immunity. However, target-annotated NPs for cancer immunotherapy remain scarce. Herein we established a multiplexed platform combining virtual screening, affinity selection-mass spectrometry, and metabolomics profiling to identify bioactive NPs targeting the adenosine 2A receptor (A2AR), a master regulator of tumor immunosuppression. Screening the crude extract of a medicinal herb and isolating the active constituent resulted in the discovery of a novel dual antagonist for A2AR/A2BR with preferential activity on A2AR. This compound, ER-15, adopts a unique binding mode as revealed by structural modeling, MD simulations, mutagenesis, and SAR analysis. Functionally, ER-15 reversed adenosine-mediated immunosuppression and augmented the immune checkpoint inhibitor therapy in both the animal model and patient-derived tumor organoids, supporting its therapeutic potential in anti-PD-1-resistant tumors. Therefore, our strategy is expected to overcome traditional NP discovery bottlenecks, enabling efficient identification of target-annotated novel leads for drug development.

Background:Cistanche deserticola (CD), a functional plant with homology of medicine and food, is used for reinforcing kidney to strengthen yang and loosening bowel to relieve constipation. It is ordinarily processed with rice wine-steamed, which is known as wine-steamed CD (W-CD) to enhance effects in clinical practice. Nevertheless, timely processing of CD is an effective means to ensure quality; the processing techniques also played a crucial role in influencing the quality of CD and its products, which require further investigation. This study aimed to explore suitable drying methods for the efficient production of CDs and W-CDs. Methods:Herein, the fresh CD is collected and both CD and W-CD are prepared, which all drying mainly included forced-air drying (FAD, at 40, 60, and 80 °C), far-infrared air drying (FID, at 40, 60, and 80 °C), vacuum microwave drying (VMD, at 50, 55, and 60 °C), vacuum freeze drying (VDF), sun-drying (SD) respectively. Furthermore, drying kinetics were employed to analyze drying characteristics, establishing Weibull function models for different processing methods of CD and W-CD. Combining intelligent sensory technologies (E-nose, E-tongue, color difference meter) with texture analyzers, and employing scanning electron microscopy, the trait characteristics and microstructural features were investigated to examine the effects of different drying methods on CD and W-CD. The components content of Echinacoside, Cistanoside A, Tubuloside A, Verbascoside, Isoverbascoside, 2'-Acetylverbascoside, and total polysaccharides are analyzed by high-performance liquid chromatography (HPLC) and ultraviolet-visible spectrophotometry (UV), and the total extracts are also measured. Above those are combined with chemometrics to obtain important factors analysis to differentiate samples of quality. Results:The Weibull model of drying dynamics is established successfully for CD and W-CD drying processing. The microstructure, rehydration rate (RR, %), and porosity (%) of CD are significantly influenced by rice wine-steamed processing, as are the sweetness (ANS) and content of phenylethyl glycoside, which are also increased. The best drying condition for CD is FAD60-80 °C, and W-CD is FID 40 °C. Conclusion:Our study, which is comprehensive in comparing the quality of CD and W-CD across different drying processes based on "color-odor-taste-component content," revealed that improving quality can enhance the production of fresh CD. Besides, intelligent sensory technology can provide a foundation for future quality control of CD and W-CD.

The ligand-binding pocket of the androgen receptor (AR) is the targeting site of all clinically used AR antagonists. However, various drug-resistant mutations emerged in the pocket. We previously reported a new targeting site at the dimer interface of AR (dimer interface pocket) and identified a novel antagonist M17-B15 that failed in oral administration. In this study, the head part of M17-B15 was substituted with divergent structures. Potent antagonist Z10 with benzo[b]oxepine was first identified. Subsequent structural optimization on the 2-oxopropyl moiety of Z10 generated the more powerful Y5 (IC50 = 0.04 μM). Out of the ordinary, Y5 demonstrated dual mechanisms of action, antagonized AR by disrupting AR dimerization, and induced AR degradation via the ubiquitin-proteasome pathway. Furthermore, Y5 exhibited excellent activity against variant drug-resistant AR mutants comparable to recently approved darolutamide. Furthermore, Y5 effectively suppressed the tumor growth of the LNCaP xenograft via oral administration, providing a potential novel therapeutic for drug-resistant prostate cancer.

Mycotoxins, toxic fungal secondary metabolites, exhibit a diverse array of toxicological effects, including hepatotoxicity, carcinogenicity, estrogenicity, immunotoxicity, and neurotoxicity. These toxins cause severe contamination in food, feed, and traditional Chinese medicines (TCMs), threatening global food security and imposing substantial economic burdens. Among over 400 distinct mycotoxins identified to date, aflatoxin B1 (AFB1), ochratoxin A (OTA), and zearalenone (ZEN) stand out for their pervasive contamination and grave toxicities. Upon absorption, these toxins undergo biotransformation into reactive metabolites that exert multifaceted toxicities via mechanisms such as carcinogenesis, estrogenic effects, oxidative stress, inflammation, and abnormal apoptosis, collectively threatening human and livestock health. The application of natural and engineered enterosorbents suppresses intestinal absorption and subsequent bioactivation of mycotoxins, while dietary small-molecule bioactive compounds neutralize post-absorption toxicity via biotransformation intervention and cytoprotective reinforcement, collectively preventing the onset and progression of related diseases. This paper reviews the biosynthetic routes of three representative mycotoxins (AFB1, OTA, and ZEN), along with their biotransformation and underlying pathogenic mechanisms. Furthermore, nutritional intervention approaches targeting the underlying mechanisms to ameliorate mycotoxin-induced damage are discussed. This review not only provides valuable insights for future research on mycotoxin toxicity, but also establishes a theoretical foundation for utilizing dietary strategies to counteract mycotoxin-induced physical damage.

Essential oil is a key quality marker of Nardostachys jatamansi DC. (NJ), and is one of the main application forms of NJ alone or in a prescription with efficacies such as sedative and antiarrhythmic activities. Herein, GC/EI-MS and UPC2-QTOF-MS were simultaneously applied to characterize the chemical profile of the essential oil of N. jatamansi (EONJ), and a UPC2-PDA method was further established to determine the contents of the major sesquiterpenones in EONJ. As a result, a total of 81 and 35 constituents were tentatively identified by GC/EI-MS and UPC2-QTOF-MS, respectively. Among them, there were three major sesquiterpenones, 1,8,9,10-tetradehydroaristolan-2-one, aristolone, and kanshone H were accurately identified from EONJ. Then, the modifying agent, column temperature, backpressure, flow rate, wavelength, and injection volume were optimized for a UPC2-PDA method, whose linearity, precision, repeatability, stability, and recovery were further evaluated to make sure it was accurate and feasible for quantitative determination of the 1,8,9,10-tetradehydroaristolan-2-one, aristolone, and kanshone H in 38 batches of EONJ. Their contents fell into the ranges of 0.0804-0.4876 %, 0.0201-0.0870 %, and 0.0083-0.0444 %, respectively. This study supplies a simple, rapid, and environmental-friendly UPC2-PDA method dedicated to the content determination of sesquiterpenone isomers in EONJ, with a new reference for global quality control of N. jatamansi.

Amomi Fructus, a mature fruit from a ginger family plant, has various species, resulting in inconsistent sourcing and quality. Most studies distinguish species by volatile compounds, yet research shows it also contains flavonoids with notable pharmacological effects. Solely focusing on volatile compounds could lead to considerable resource waste. This study aims to establish flavonoid markers in Amomi Fructus to distinguish its species, assess quality, and promote efficient resource use. Utilizing natural deep eutectic solvents (NADES) and response surface methodology (RSM), an optimal extraction system (choline chloride-ethylene glycol) yielded 41.38 mg RE/g total flavonoids. LC-MS analysis of 18 Amomi Fructus batches identified 26 flavonoids, quantified 19, and highlighted three key markers—epicatechin, procyanidin B2, and procyanidin B4—that effectively differentiate Amomum villosum Lour. (AMV) from Amomum villosum Lour. var. xanthioides T.L. Wu et Senjen (AMVX). Finally, flow cytometry confirmed these markers' antioxidant activity, effectively reducing H₂O₂-induced oxidative damage in GES-1 cells.

Semen Sojae Praeparatum (SSP) is traditional dual-purpose product with both medicinal and edible applications. In this study, electronic nose, comprehensive two-dimensional gas chromatography-mass spectrometry, and ultra-performance liquid chromatography were employed to characterize the fermentation-associated attributes underlying "distinct aroma and umami-rich taste" and to analyze the dynamic isoflavones changes. Results revealed that moisture content and reducing sugars initially increased and subsequently decreased, while pH declined during pre-fermentation and remained stable thereafter. Key flavor contributors were identified as acids, aldehydes, alcohols, esters, and pyrazines, specifically 2-furanmethanol, furfural, 2,5-dimethylpyrazine, isovaleric acid, benzaldehyde, and acetic acid, which collectively defined the characteristic aroma profile. Free amino acids, metabolized from proteins by protease, accumulated mainly during pre-fermentation. Isoflavone aglycones gradually increased due to the deglycosylation of glycosides in the pre-fermentation phase, whereas both aglycones and glycosides remained stable during post-fermentation. These findings establish a scientific foundation for understanding the fermentation mechanisms of SSP.

BackgroundDysbiosis of the gut microbiota (GM) has been linked to inflammatory bowel disease (IBD), yet its associated molecular mechanisms remain poorly defined. Identifying causal host genes mediating GM-IBD interactions is therefore of great importance.ObjectiveTo identify GM-associated causal genes for IBD and to prioritize key targets and cell types underlying GM-host crosstalk.MethodsWe integrated GWAS datasets of GM, UC, and CD using a two-sample Mendelian randomization (MR) framework with IVW as the primary estimator. Causal SNPs were mapped to genes for enrichment analyses. Candidate genes were refined by intersecting MR-derived genes with bulk RNA-seq DEGs (training: GSE87473, validation: GSE75214) and prioritized using nested cross-validated machine-learning models. Single-cell RNA-seq (GSE116222) was used to localize key genes to specific cell types. The functional role of TNIK was validated in IL-10-/- IBD mice via AAV9-mediated overexpression. Immunohistochemical staining of Ki67 and Cleaved caspase 3 was conducted to evaluate epithelial proliferation and apoptosis in colonic tissues.ResultsMR analysis identified 307 and 360 GM-associated causal genes for UC and CD, respectively. TNIK (TRAF2 and NCK-interacting kinase) was highlighted as a key candidate gene. Seven TNIK-associated immune cell subsets showed altered infiltration in UC. Single-cell transcriptomics revealed TNIK dysregulation in colonocytes, goblet cells. T/NK cells in UC. TNIK overexpression in IL-10-/- mice reduced disease severity and downregulated IL-1β, IL-6, and TNF-α. Immunohistochemistry confirmed that TNIK overexpression enhanced Ki67 expression and reduced Cleaved caspase 3 expression.ConclusionBy integrating MR with transcriptomics and single-cell seq results, we identified TNIK as a potential GM-associated host kinase linking dysbiosis to epithelial and immune dysfunction in IBD. TNIK emerges as a promising node for IBD prognosis through barrier maintenance and immune regulation.

ETHNOPHARMACOLOGIC RELEVANCE:The rapid increase in cardiovascular and cerebrovascular diseases (CCVDs) is a significant threat to human health. Traditional Chinese medicine (TCM) offers unique therapeutic advantages. Xuefu Zhuyu Formula (XFZYF), a classic TCM prescription, has been widely used in clinical practice to treat CCVDs and other related conditions. AIM OF THE STUDY:The study aimed to comprehensively elucidate the bioactive components and mechanisms of XFZYF and serve as a reference for future research. The current development, structural characteristics, blood- and intestine-migratory components, quality control measures, pharmacokinetics, pharmacological mechanisms, and clinical applications of XFZYF were systematically summarized. MATERIALS AND METHODS:A comprehensive literature search was conducted up to 2024 in PubMed, Web of Science, and the China National Knowledge Infrastructure (CNKI) database using the keywords: "Xuefu Zhuyu", "cardiovascular disease", "cerebrovascular disease", "chemical constituents", "migratory components", "quality control", and "pharmacological properties". RESULTS:Eight distinct dosage forms of XFZYF have been developed for clinical application. XFZYF consists of 11 medicinal ingredients, and various compounds have been identified or preliminarily characterized. These compounds are broadly classified into phenolic acids, flavonoids, triterpenoid saponins, monoterpene glycosides, spermidines, and phthalides. Clinically, XFZYF is widely applied in both internal medicine and surgical settings. CONCLUSION:XFZYF may exert protective effects against the onset and progression of CCVDs by modulating amino acid metabolism, non-coding RNAs, inflammatory responses, synaptic plasticity, chemokines, oxidative stress, lipid metabolism, mitochondrial function, platelet aggregation, angiogenesis, and gut microbiota. Additionally, perspectives on current limitations and directions for future research were discussed. 1) Establishment of a comprehensive quality control standard for XFZYF. 2) Enhancement of the clinical safety assessment of XFZYF, especially in combination therapies. 3) Advancement of the systematic study of the interactions and compatibility of migratory components. 4) Promotion of AI-driven strategies in the standardization and modernization of TCM.

Aptamer-based biosensors have emerged as an important and promising technology for applications in food safety, environmental monitoring, and pharmaceutical analysis. Obtained via Systematic evolution of ligands by exponential enrichment (SELEX) screening, these recognition elements exhibit antibody-comparable affinity and specificity, alongside superior chemical stability, easy synthesis, and broad target adaptability. Substantial advances in the field have been marked by the systematic development of food contaminant-specific aptamers, elucidation of their binding mechanisms, and construction of versatile biosensing platforms. The integration of these aptamers with conventional electrochemical and optical sensors has substantially enhanced detection sensitivity and lowered detection limits, particularly for trace-level analytes in complex food matrices. Furthermore, the integration of aptamer technology with novel nanomaterials has facilitated the development of high-performance detection platforms for a wide range of food contaminants, including heavy metals, antibiotics, foodborne pathogens, mycotoxins, pesticides, and food additives. This review systematically summarizes recent advances in SELEX techniques for aptamer screening, highlights the application of aptamer-based biosensors in detecting these contaminants, and discusses current challenges and future prospects in the field of food safety, which establishes a comprehensive framework to advance aptamer-based biosensing technologies for rapid detection and early warning in food safety monitoring.

Gardenia blue (GB), a natural active blue pigment, can be synthesized from iridoid glycosides and primary amino-containing compounds in vivo and in vitro. However, limited studies have reported about GB synthetic intermediates, leading to unawareness of the formation mechanism of GB. Here, we reported that GB could be detected in rats′ feces, whose formation was simulated by in vitro reaction of amino acid-rich feces extract from rats and genipin (GP) metabolized from geniposide (GE) through β-glucosidase. Firstly, we proved that 14 amino acids (AAs) detected in rats′ feces containing primary amino groups can react with GP to produce GB. Then, taking histidine (His) as example, we characterized the key intermediates during the reaction process, including basic units genihistidine A (GH-A) and dimers. As an active basic unit, GH-A underwent covalent polymerization to form dimer, which then self-assembled into supramolecular spherical nanoparticles through multiple noncovalent interactions, including hydrogen bonds, π-π interactions, and van der Waals interactions. Our study revealed the formation mechanism of GB and provided insights into the bioavailable form of GE from traditional Chinese medicine in vivo.

To decipher subtle chemical variation of both non-volatile and volatile components during bran-roasting of nutmeg (Myristica fragrans), we innovatively established a "three-source comparison" approach (raw nutmeg, RN; bran-roasted nutmeg in lab, BN-L; bran-roasted nutmeg from market, BN-M) and developed chemometrics/machine learning (ML) models to identify processing-related markers. Untargeted high-resolution liquid chromatography-mass spectrometry-based metabolomic analysis of the RN and BN-L samples unveiled 57 differential metabolites, and five thereof (involving increased 5-hydroxymaltol, maltol, dipterine, fragransin B2, and decreased adipic acid) were selected as processing-associated markers. These five markers achieved 90 % accuracy in identifying the BN-M samples in the external validation, much better than that gained by seven potential volatile markers recorded by gas chromatography-mass spectrometry. Conclusively, integration of chemometrics and ML reveals the subtle chemical changes during the processing of nutmeg, and it is the first study reporting robust markers that are diagnostic to discriminate between the raw nutmeg and processed products.

Chebulae Fructus (CF) is known as one of the richest sources of hydrolyzable tannins (HTs). In this study, ultra-performance liquid chromatography coupled with a photodiode array detector method was established for simultaneous determination of the 12 common phenolcarboxylic and tannic constituents (PTCs). Using this method, quantitative analysis was accomplished in CF and other four adulterants, including Terminaliae Belliricae Fructus, Phyllanthi Fructus, Chebulae Fructus Immaturus, and Canarii Fructus. Based on a quantitative analysis of the focused compounds, discrimination of CF and other four adulterants was successfully accomplished by hierarchical cluster analysis and principal component analysis. Additionally, the total contents of the 12 compounds that we focused on in this study were unveiled as 148.86 mg/g, 96.14 mg/g, and 18.64 mg/g in exocarp, mesocarp, and endocarp and seed of CF, respectively, and PTCs were witnessed to be the most abundant in the exocarp of CF. Noticeably, the HTs (chebulagic acid, chebulanin acid, chebulinic acid, and punicalagin) were observed to be ultimately degraded to chebulic acid, gallic acid, and ellagic acid during sunlight-drying of the fresh fruits. As a result, our study indicated that CF and its adulterants could be distinguished by the observed 12 PTCs, which were mainly distributed in the exocarp of the fruits. The HTs were prone to degrade into the three simple phenolcarboxylic acids during drying or processing, allowing us to obtain a more comprehensive understanding of the PTCs, with great significance in the improved quality of CF and related products.

Rheumatoid arthritis (RA) is a chronic systemic autoimmune disease that seriously affects the life quality of patients. As a patent medicine of Chinese traditional medicine, YuXueBi capsule (YXBC) is widely used for treating RA with significant effects. However, its active compounds and therapeutic mechanisms are not fully illuminated, encumbering the satisfactory clinical application. In this study, we developed a method for identifying the chemical compounds of YXBC and the absorbed compounds into blood of rats using ultra performance liquid chromatography/ion mobility-quadrupole time-of-flight mass spectrometry (UPLC/IM-QTOF-MS) combined with UNIFI analysis software. A total of 58 compounds in YXBC were unambiguously or tentatively identified, 16 compounds from which were found in serum of rats after administration of YXBC. By network pharmacology, these prototype compounds identified in serum were predicted to regulate 30 main pathways (including HIF-1 signaling pathway, neuroactive ligand-receptor interaction, IL-17 signaling pathway, and so on) through 146 targets, resulting in promoting blood circulation and removing blood stasis, analgesia, and anti-inflammatory activities. This study provides a scientific basis for the clinical efficacy of YXBC in the treatment of RA.

Febrile seizures (FS) are the most common type of seizures for children. As a commonly used representative cold formula for resuscitation, Zixue Powder (ZP) has shown great efficacy for the treatment of FS in clinic, while its active ingredients and underlying mechanism remain largely unclear. This study aimed to preliminarily elucidate the material basis of ZP and the potential mechanism for the treatment of FS through ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS), network pharmacology, and molecular docking. UPLC-Q-TOF-MS was firstly applied to characterize the ingredients in ZP, followed by network pharmacology to explore the potential bioactive ingredients and pathways of ZP against FS. Furthermore, molecular docking technique was employed to verify the binding affinity between the screened active ingredients and targets. As a result, 75 ingredients were identified, containing flavonoids, chromogenic ketones, triterpenes and their saponins, organic acids, etc. Through the current study, we focused on 13 potential active ingredients and 14 key potential anti-FS targets of ZP, such as IL6, STAT3, TNF, and MMP9. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analysis showed that inflammatory response, EGFR tyrosine kinase inhibitor resistance, AGE-RAGE signaling pathway in diabetic complications, and neuroactive ligand-receptor interaction were the main anti-FS signaling pathways. Licochalcones A and B, 26-deoxycimicifugoside, and hederagenin were screened as the main potential active ingredients by molecular docking. In conclusion, this study provides an effective in-depth investigation of the chemical composition, potential bioactive components, and possible anti-FS mechanism of ZP, which lays the foundation for pharmacodynamic studies and clinical applications of ZP.

Various enzymes secreted by microorganisms during fermentation of Massa Medicata Fermentata (MMF) play a crucial role in conversion of the chemical substances for better clinical application. However, enzyme-driven transformation of chemical compounds during MMF fermentation has not been fully unveiled. Our findings revealed that MMF fermented with a mixture of wheat bran (WB)-flour (2:1) was observed with high enzymatic activities, alongside distinguished appearance. The activities of protease, ester bond hydrolase for cCGA and EF, and glycoside hydrolase all increased with the extended fermentation time of MMF, reaching 14.84x10(-8)+/- 0.3436x10(-8), 0.05456x10(-8)+/- 0.0017x10(-8), 0.5715x10(-8)+/- 0.1285x10(-8), and 15.25x10(-8)+/- 0.01389x10(-8) Kat/g on the 8th day of fermentation. Moreover, protease has been proven to drive the conversion of WB proteins into amino acids by precolumn phenyl isothiocyanate derivatization. Besides, transformation patterns of ester bond hydrolase on substrate components were revealed, in which cryptochlorogenic acid was decomposed into caffeic acid and quinic acid, and ethyl ferulate was metabolized into ferulic acid. By glycoside hydrolase, amygdalin was transformed into prunasin and benzaldehyde. This study reveals the enzyme-mediated transformation patterns of the focused components during fermentation of MMF, laying foundation for further elucidation of its fermentation mechanism and elevation of quality control.