This study aims to expand the application of handheld near-infrared spectroscopy combined with machine learning in quantitative analysis of Epimedium. After establishing and evaluating several promising models: Gradient Boosting Regression (GBR), Extra Trees (ET), Extreme Gradient Boosting (eXGB), Support Vector Regression (SVR), Random Forest (RF), Partial Least Squares Regression (PLSR), we selected PLSR and SVR for further optimization due to their excellent performance in terms of R2 and RMSE. Employing the NIPPY library for data preprocessing and a genetic algorithm for feature selection, models for predicting the contents of four active components in Epimedium were successfully established. The GA-SVR hybrid model delivered the following predictive performance for the four target components: icariin (R2:0.9556, RMSE:0.0031, Avg. MAPE:2.07%), epimedin A (R2:.9586, RMSE:0.0004, Avg. MAPE:3.60%), epimedin B (R2:0.9432, RMSE:0.0003, Avg. MAPE:3.58%) and epimedin C (R2:0.9232, RMSE:0.0182, Avg. MAPE:2.33%). These findings establish a novel paradigm for high-throughput analysis and quality assessment of food and traditional Chinese medicine active ingredients.

Isoflavones from legumes, particularly genistein, are highly valued as functional food ingredients. However, the poor water solubility and low bioavailability hinder its application, which can be improved through glycosylation. The glycosyltransferase GT2419 from Bacillus subtilis ATCC 6633 catalyzes the 7-OH and 4'-OH glycosylation of genistein, but its poor regioselectivity hampers the efficient synthesis of genistein 7-O-glucoside. Here, a semirational design strategy was employed to obtain the F231V mutant, which significantly increased the conversion rate of genistein 7-O-glucoside from 21.1% to 72.2% and improved the 7-O regioselectivity to 73.8%. Kinetic analysis revealed that the F231V mutant exhibited enhanced affinity and catalytic efficiency toward genistein. Computational analysis revealed that the F231V mutant enlarged the substrate-binding pocket and disrupted intermolecular hydrophobic interactions, thereby altering substrate orientation, and enhancing the regioselective formation of genistein 7-O-glucoside. This study provides an efficient biosynthetic method for genistein 7-O-glucoside, with promising applications in the food industry.

Severe burns can cause secondary pulmonary inflammation driven by endotoxin-related inflammatory activation and oxidative stress. Here, we integrated untargeted LC-MS, network pharmacology, transcriptomics, molecular docking, molecular dynamics simulation and target-engagement assays to identify and validate a candidate active constituent from Ganoderma lucidum. LC-MS identified 660 compounds, with terpenoids as a predominant class. Network pharmacology prioritized inflammatory signaling pathways related to PI3K-Akt, MAPK and NF-κB, and integration with RNA-seq identified 92 transcriptomics-supported candidate targets. Ganoderic acid L showed favorable predicted binding to PIK3CA and was selected for experimental validation. In an LPS-induced inflammatory lung injury mouse model, ganoderic acid L alleviated histopathological injury, pulmonary edema and bronchoalveolar lavage fluid protein leakage; reduced TNF-α, IL-6, IL-1β, MPO and MDA levels; and restored SOD and GSH-Px activities. RT-qPCR and western blotting indicated suppression of inflammatory gene expression and modulation of Akt/NF-κB/MAPK signaling. Surface plasmon resonance showed micromolar-level interaction with PI3K, and CETSA provided supportive evidence for potential target engagement. These findings identify ganoderic acid L as a candidate anti-inflammatory triterpenoid for LPS-induced lung injury, while causal PI3K engagement requires further validation.

Sustainable microbial biocatalysis provides an eco-friendly route for modifying complex natural products and supports the valorization of renewable resources. In this study, the catalytic diversity of Bacillus megaterium CGMCC 1.1741 and Bacillus subtilis ATCC 6633 was explored for the biotransformation of Soyasapogenol B (a), an aglycone derived from soybean oil-crop byproducts. This work extends our previous fungal-mediated studies by revealing the distinct bacterial-catalyzed transformation that further expands the chemical space of this soybean-derived aglycone. Ten metabolites (a1-a10) were isolated and structurally characterized by 1D/2D NMR and HR-ESI-MS, reflecting a diverse range of catalytic reactions. These included C-3 oxidation, C-11 methoxylation, regioselective hydroxylation at C-7 and C-15, and site-specific glycosylation, potentially influencing pharmacological relevance such as enhanced solubility, and bioavailability. The findings not only highlight the multifunctional catalytic potential of Bacillus strains in tailoring Soyasapogenol B (a) but also underscores their value as sustainable microbial platforms for converting soybean oil-crop byproducts into structurally enriched metabolites.

Abstract Background: Brain metastasis (BM) is a leading cause of death in lung cancer patients, and the brain’s unique microenvironment is key to metastatic initiation and progression. However, the molecular mechanisms of tumor microenvironment crosstalk in lung cancer BM remain unclear. Lipocalin-2 (LCN2) is associated with inflammation and cancer, but its specific role in lung cancer BM is undefined. Methods: Single-cell sequencing analysis was conducted on BM specimens from patients with lung cancer BM, and data mining of public databases was performed to identify key molecules. Cell lines with stable LCN2 knockdown or overexpression were constructed using lentiviral transduction. Transcriptome sequencing and immunofluorescence assays were applied to verify the in vitro functions of LCN2, including its regulatory effects on downstream signaling pathways and cellular behaviors. For in vivo studies, murine models of lung cancer BM were established via brain orthotopic injection and intracardiac injection of tumor cells. Bioluminescence imaging and multiplex immunofluorescence staining was utilized to investigate the role of LCN2 in BM tumor initiation and progression, as well as its interaction with brain microenvironmental components. Results: LCN2 was upregulated in BM compared to primary tumors, correlating with shorter intracranial disease-free and overall survival and plasma LCN2 was higher in BM patients. LCN2 is dispensable for BBB transmigration but is essential for promoting tumor growth within the brain microenvironment. LCN2 promoted intracranial tumor growth and upregulates VEGF-A via the JAK2/STAT3 signaling pathway to facilitate angiogenesis in lung cancer BM. More importantly, LCN2 bound to SLC22A17 on astrocytes to activate the JAK2/STAT3 signaling pathway and induce CCL2 secretion, which recruited macrophages that secreted IL-1β to upregulate LCN2 in tumor cells via the IL-1R-NF-κB signaling axis. Inhibition of IL-1β-IL-1R and JAK2/STAT3 signaling by IL-1R inhibitor anakinra combined with STAT3 inhibitor SH4-54 an suppress LCN2-driven tumor progression. Conclusions: Tumor-derived LCN2 orchestrates a brain-specific metastatic program through dual mechanisms: a paracrine loop involving astrocyte activation and macrophage recruitment and a tumor-intrinsic angiogenic pathway via SLC22A17-JAK2-STAT3-VEGF-A signaling. These mechanistic insights suggest that LCN2 may serve as both a therapeutic target and a prognostic biomarker in lung cancer BM. Citation Format: Danming He, Yixiang Zhu, Jian Zhang, Wei Zhuang, Hua Bai, Jie Wang. Lipocalin-2 drives brain metastatic progression through reciprocal tumor-microenvironment interactions in lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 4093.

Citri reticulatae pericarpium (CRP), a traditional Chinese medicine and food homologous material, is rich in bioactive flavonoids with multiple pharmacological effects. However, traditional organic solvent extraction methods for flavonoids suffer from low efficiency, high cost, and environmental pollution. In this study, ultrasound-assisted deep eutectic solvent extraction (UAE-DES) was developed as an efficient and green alternative for flavonoid extraction from CRP. The 41 DESs were screened, and choline chloride-acetamide (mole ratio 1:3) with 30% water content was identified as the optimal solvent. The extraction performance was improved by 92% compared to 70% acidified methanol. Response surface methodology (RSM) based on BoxBehnken Design (BBD) was further used to optimize the extraction process, with the optimal conditions determined as follows: solid-liquid ratio 1:28 (g/mL), extraction temperature 30 degrees C, extraction time 61 min, and ultrasonic power 210 W. Under these conditions, the maximum hesperidin content (the most abundant flavonoid in CRP) reached 27.46 +/- 0.33 mg/g. Fourier transform-infrared spectrometry (FTIR) verified the formation of strong hydrogen bonds between choline chloride and acetamide, which enhanced flavonoid solubility. Scanning electron microscopy (SEM) observations revealed that UAE-DES treatment caused more severe disruption of CRP cell structures compared to water and 70% methanol extraction, facilitating flavonoid release. Molecular dynamics (MD) simulations further elucidated the extraction mechanism at the molecular level: The multicomponent (chloride ion, acetamide, choline cation) in Des synergizes to keep flavonoids monodisperse, increasing solvent accessibility and mass transfer efficiency. In conclusion, UAE-DES is a promising green extraction technology for CRP flavonoids, and the insights into the molecular interaction mechanism provide a theoretical basis for the design and application of DES in plant active ingredient extraction.

ETHNOPHARMACOLOGICAL RELEVANCE:Cyclocarya paliurus (Batal.) Iljinsk, a millennia-old traditional Chinese herb, is prized for its ability to clear the lungs and nourish the liver. Additionally, it is employed in traditional Chinese medicine practice for heat clearance and detoxification, addressing conditions such as lung diseases. AIM OF THE STUDY:This study aimed to evaluate beneficial effects of polysaccharides (CPP) from C. paliurus on chronic obstructive pulmonary disease (COPD) and its potential mechanisms. MATERIALS AND METHODS:The chemical characterization of the isolated and purified CPP was conducted using fourier-transform infrared spectroscopy, ultraviolet spectroscopy and scanning electron microscopy. COPD was induced in male BALB/c mice by intranasal infusion of LPS and exposure to cigarette smoke for 28 days. Lung tissues were then collected for subsequent histopathological and molecular analyses. The mechanism of CPP against COPD was investigated through transcriptomic data mining and Western blot analysis. Additionally, acute toxicity of CPP was assessed in mice following a single oral dose of 15 g/kg. RESULTS:Experimental evidence established that CPP consists of six monosaccharides: fucose, arabinose, rhamnose, galactose, glucose, and xylose. CPP treatment significantly reduced the levels of PCO2 and HCO3- in the blood of COPD mice, concurrently alleviating pulmonary inflammation. Mechanistic investigations have revealed that CPP exerts its anti-inflammatory effect by modulating the AhR/NF-κB pathway. In addition, CPP demonstrated safety at doses exceeding 100 times the effective level. CONCLUSION:The results suggest that CPP holds promise as a potential therapeutic agent for the intervention of COPD. These findings provide a theoretical basis for the development of the ethnic medicinal herb Cyclocarya paliurus.

BACKGROUND AND AIM:Hepatic ischemia-reperfusion (IR) injury represents a critical clinical challenge characterized by excessive inflammation. HMGB1, known as a proinflammatory mediator released after liver IR injury, has been reported to worsen the damage and inflammation via a positive feedback loop. Herein, shikonin has been explored to alleviate hepatic IR injury through direct interaction with HMGB1. METHODS:The ligation on the hilum of the right liver lobe lasted for 90 min and then removed to induce hepatic IR injury in SD rats. Serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were determined. At the cellular level, HMGB1-induced NO release from RAW264.7 was assessed to test the inhibitory effect of shikonin. The surface plasmon resonance (SPR), saturation transfer difference nuclear magnetic resonance (STD-NMR), and molecular dynamics simulation were employed to study the interaction between shikonin and HMGB1. RESULTS:After administration of shikonin, the degree of liver IR damage in rats was attenuated significantly in a dose-dependent manner. In vitro, shikonin can effectively inhibit HMGB1 activation. Furthermore, mechanistic studies indicated that shikonin could directly bind to HMGB1. The results of 1H STD-NMR confirmed that shikonin interacted with HMGB1, and multiple protons in shikonin participated in the binding process. Notably, shikonin exhibits a selective binding affinity toward the I122 and L129-bearing region of HMGB1, which were supported by molecular dynamics simulation. CONCLUSION:In general, our results suggest that shikonin is a promising natural candidate that directly targets HMGB1 to exert hepatoprotection for the development of clinical therapeutic anti-IR agents.

Drug-resistant Klebsiella pneumoniae (K. pneumoniae) represents a formidable clinical challenge. Targeting its quorum sensing (QS) system has emerged as a promising anti-virulence strategy to combat antibiotic resistance. In this study, eighteen pentacyclic triterpenes (PTs) were screened for their in vitro activity against K. pneumoniae. Among them, madecassic acid (PT-5) exhibited a negligible bactericidal effect but significantly suppressed key virulence traits in a highly virulent, multidrug-resistant K. pneumoniae strain (KP31). These suppressed traits included biofilm formation, AI-2 production, capsular polysaccharide synthesis, and swarming motility, identifying PT-5 as a potent quorum sensing inhibitor with anti-virulence properties. Moreover, PT-5 demonstrated a synergistic effect at a subinhibitory concentration (32 μg/mL) when combined with conventional antibiotics, enhancing the bactericidal activity of kanamycin by 31 %, streptomycin by 29 %, azithromycin by 24 %, and chloramphenicol by 24 %. In a Galleria mellonella larval infection model, PT-5 in combination with kanamycin markedly boosted larval survival to 60 %, compared with 20 % for kanamycin monotherapy. Quantitative PCR analysis further revealed that PT-5 significantly downregulated the expression of key QS-related genes (luxS, sdiA, lsrK) and major virulence-associated genes (wbbM, wzm, pgaA, mrkA) in KP31. In summary, PT-5 acts as an effective QS inhibitor that suppresses virulence and enhances antibiotic efficacy, representing a promising combinational therapy against drug-resistant K. pneumoniae.

To address the challenges of regioselective modification of steroidal saponins, a tandem microbial biotransformation strategy was developed using Bacillus megaterium CGMCC 1.1741 and Bacillus subtilis ATCC 6633. Three Δ5-steroids—diosgenin (1), pennogenin (2), and 25(R,S)-ruscogenin (3) were employed as substrates. In the first stage, B. megaterium facilitated highly regioselective hydroxylation at the C-7 position to yield metabolites 1a–3a. Subsequently, these intermediates were successfully glycosylated at the newly introduced C-7 hydroxyl group by B. subtilis, resulting in three novel steroidal glycosides 1b–3b. The structures of all metabolites were rigorously elucidated using HR-ESI-MS and 1D/2D NMR spectroscopy. Notably, while B. subtilis alone failed to glycosylate diosgenin directly, this tandem approach bypassed the catalytic limitation through preliminary enzymatic “priming”. This study provides an efficient, green, and highly selective enzymatic route for the structural diversification of steroidal saponins.

Liver cancer is the malignancy with the highest mortality rate among digestive system cancers worldwide. Flavonoid plant extracts have shown significant safety and extensive function, especially anti-tumor activity. In this study, the therapeutic effects of a combination of baicalein and luteolin were evaluated in vitro. Cell proliferation, migration, and apoptosis were performed respectively through MTT assay, clone formation, wound healing, transwell, and JC-1 staining. Protein levels of EGFR, p-ERK, NFκB, and cleaved-caspase3 via western blotting and the interaction with key proteins EGFR and NFκB by molecular docking were determined in order to explore the underlying mechanism. In addition, a mouse heterotopic transplant tumor model was established to assess the anticancer activity in vivo. The experimental results showed that the combination of baicalein and luteolin exhibited more potent in suppressing cell proliferation and migration and inducing cell apoptosis when compared with baicalein or luteolin. The western blot and molecular docking studies demonstrated that the potential mechanism may relate to the inhibiting of the EGFR/NFκB signaling pathway. Moreover, the HepG2 cell xenograft model, hematoxylin and eosin, and immunohistochemical staining results also confirmed that the combination of baicalein and luteolin was more effective than a single compound in inhibiting tumor growth. In summary, the combination of baicalein and luteolin may resist the proliferation, migration, and apoptosis activity of HepG2 cells through the EGFR/NFκB signaling pathway, which provides new insights for further exploring plant extract treatment for liver cancer.

Chemical constituents of n-butanol fraction of ethanol extract from the leaves of Cyclocarya paliurus(Batalin)Iljinskaja were studied.Ten compounds were purified by silica gel,MCI,ODS,Sephadex LH-20 column chromatography and semi-preparative high-performance liquid chromatography.Based on the physicochemical properties and spectroscopic data,these compounds were identified as(4S)-4,8-dihydroxy-α-tetralone-5-[6'-(3",5"-dimethoxy-(E)-p-coumaroyl)]-β-D-glucopyranoside(1),(4S)-4,8-dihydroxy-α-tetralone-5-[6'-(E)-feruloyl]-β-D-glucopyranoside(2),(1S,3R,4S,5R)-4,5-di-O-caffeoyl quinic acid methyl ester(3),(1R,3R,4S,5R)-3,4-di-O-caffeoyl quinic acid methyl ester(4),(3R,5R)-3,4,5-tri-O-caffeoylquinic acid methyl ester(5),(1R,3R,4S,5R)-3-O-caffeoyl-5-O-p-coumaroyl quinic acid methyl ester(6),(1R,3R,4S,5R)-3-O-p-coumaroy-4-O-caffeoyl quinic acid methyl ester(7),(1R,3R,4S,5R)-3-O-caffeoyl-4-O-p-coumaroy quinic acid methyl ester(8),(3R,5R)-3,5-di-O-caffeoyl quinic acid methyl ester(9)and(1S,3R,4R,5R)-4-O-caffeoyl-5-O-feruloyl quinic acid methyl ester(10).Among them,compound 1 was a new compound,and compounds 3-10 were isolated from the genus C yclocarya for the first time.

Methicillin-resistant Staphylococcus aureus (MRSA) poses a significant clinical challenge owing to its antibiotic resistance and biofilm-forming ability. In this study, we conducted a screening of natural pentacyclic triterpenoids to identify lead compounds with high antibacterial activity. Senegenic acid 28-methyl ester (4) emerged as a potent antibacterial agent against MRSA and clinically isolated multidrug-resistant strains, with minimum inhibitory concentrations (MICs) of 8-64 μg/mL. We predicted the antibacterial mechanism of compound 4 using network pharmacology and validated by time-kill assays and membrane function tests. The results showed that compound 4 disrupted bacterial membranes and inhibited biofilm formation in a dose-dependent manner, reducing biomass and extracellular DNA levels. In vivo, compound 4 significantly improved survival in murine infection models, while demonstrating low cytotoxicity and minimal hemolytic activity. Furthermore, in-depth research on antibacterial mechanisms found that compound 4 downregulated key biofilm-associated genes (e.g., sarA and atl) in MRSA, which indicated by molecular docking and structural visualization analyses. Untargeted metabolomics revealed that compound 4 treatment induced widespread changes in metabolites and significant enrichment in amino acid, energy, and nucleotide metabolism pathways. Overall, these findings establish compound 4 as a dual-action agent with both anti-biofilm properties and regulate bacterial substance and energy metabolism against MRSA infections.

Gouty arthritis (GA), a common inflammatory arthropathy characterized by monosodium urate crystal deposition in joints and soft tissues, currently lacks safe and effective therapeutic agents. Piper sintenense Hatusima, a plant used for both medicinal and dietary purposes, is widely utilized in ethnic minority regions of China and is believed to alleviate joint swelling and pain. This study aims to identify the active components of P. sintenense responsible for its anti-GA effects, and to investigate their composition and underlying mechanisms. Our results demonstrated that the alkaloid fraction (PAL) serves as the primary active component for treatment of GA in P. sintenense. In both rats and mice GA animal models, administration of PAL led to reduced serum levels of inflammatory factor tumor necrosis factor-α (TNF-α) and increased levels of anti-inflammatory factor transforming growth factor-β (TGF-β). PAL treatment also reduced serum uric acid and creatinine levels, alleviated pain-related gait abnormalities in GA model animals. Piperine, pipernonaline, and dehydropipernonaline were identified as the major constituents in PAL. Network pharmacology and experimental validation revealed that PAL can suppress reactive oxygen species (ROS) and malondialdehyde (MDA) levels, enhance glutathione (GSH) release in the GA model. Furthermore, Western blot analysis indicated that the PAL promotes Nrf2/HO-1 protein expression and inhibits the release of inflammatory cytokines, exerting anti-GA effects. These effects were reversed upon administration of the Nrf2-specific inhibitor ML385. These findings suggest that PAL are the principal bioactive constituents in P. sintenense responsible for its therapeutic effects against GA. PAL activate the Nrf2/HO-1 signaling pathway, which subsequently alleviates oxidative stress and contributes to the anti-GA effects.

Targeting bromodomain and extra-terminal (BET) proteins is a promising therapeutic strategy for acute myeloid leukemia (AML). However, clinical development of conventional BET inhibitors has been limited by dose-related toxicities and compensatory resistance. Proteolysis-targeting chimeras (PROTACs) offer a catalytic and often more selective alternative by degrading target proteins via the ubiquitin-proteasome system. In this work, we designed and synthesized a series of novel BET PROTACs based on the clinical inhibitor ABBV-075. Through systematic optimization of linker length/composition and E3 ligase ligands (CRBN or VHL), we identified two highly potent PROTACs, A10 (CRBN-based) and A12 (VHL-based). Both compounds effectively degraded BET proteins, inhibited cell proliferation, induced cell-cycle arrest, and promoted apoptosis in MV4-11 AML cells. In MV4-11 xenograft models, A10 (6 mg/kg) and A12 (2 mg/kg) demonstrated significant tumor growth inhibition (76.2% and 60.5%, respectively). A comprehensive drug-likeness assessment revealed that while A12 exhibited higher systemic exposure in pharmacokinetic studies, A10 displayed a markedly superior safety profile with minimal hepatorenal toxicity. The favorable efficacy-safety balance of A10 underscores its strong potential as a preclinical candidate for AML therapy. This study highlights how rational PROTAC optimization can yield degraders with enhanced therapeutic windows, providing a promising path forward for targeted protein degradation in AML.

As damage-associated molecular patterns (DAMPs), the high mobility group box 1 (HMGB1) mediates the transmission of intercellular damage, inflammatory signals and plays a key role in pathological processes such as aseptic inflammation, autoimmune diseases and cancer. Celastrol, a natural product extracted from Tripterygium wilfordii Hook.f, exerts a neuroprotective effect by binding to HMGB1 in cerebral ischemia-reperfusion injury. To explore the binding characteristics between celastrol and HMGB1, surface plasmon resonance (SPR), dynamic light scattering (DLS) and multi-spectral technology, including fluorescence spectroscopy and circular dichroism (CD) spectra, were applied. Molecular docking as well as molecular dynamic (MD) simulation were also performed to predict the binding poses of celastrol and HMGB1. The SPR results showed that the KD value of celastrol and HMGB1 was 5.57 × 10-5 M. In fluorescence spectroscopy, the binding of celastrol can dose-dependently quench the endogenous fluorescence of HMGB1, and the quenching type is static quenching. Moreover, celastrol can also reduce the content of α-helix and enhance the random coil content of HMGB1, which could increase its particle size. Molecular docking celastrol was engaged in interactions with the amino acids Lys95, Arg104 and Ala133, resulting in the formation of multiple hydrogen bonds within the length of 1.8-2.0 Å. The main forces involved were electrostatic interaction, hydrophobic interaction and hydrogen bonds. The MD simulation further showed that a stable complex was formed between HMGB1 and celastrol. The in vitro biological evaluation showed that celastrol could inhibit NO release in the HMGB1-induced RAW264.7 inflammatory cell model with an IC50 value of 0.89 μM. Celastrol could bind to HMGB1 and slightly change its secondary structure and spatial conformation, subsequently affecting its pro-inflammatory function.

As a plant with a rich history of medicinal and dietary use, Gynostemma pentaphyllum (Thunb.) Makino is predominantly characterized by the gypenosides. Contemporary research has demonstrated that these saponins possess significant hypolipidemic and hypoglycemic effects. This study focused on developing an LC-MS/MS approach to elucidate the pharmacokinetic profiles of 13 gypenosides in rat models. The separation was accomplished using a ZORBAX Eclipse Plus C18 column (2.1 mm × 100 mm, 1.8 µm). Detection was facilitated by employing positive electrospray ionization (ESI) in multiple reaction monitoring (MRM) mode. The analytical method was fully validated, including selectivity, linear response, measurement precision, analytical accuracy, extraction efficiency, matrix interference, and sample stability. The lower limit of quantification (LLOQ) for all compounds was 10 ng/mL, with each exhibiting a favorable linear relationship (r ≥ 0.9960). The precision, as measured by the relative standard deviation (RSD) for intra-day and inter-day variability, did not exceed 8.41 %. The accuracy, expressed as the relative error (RE), varied from -4.42-8.42 %. The recovery rate of the extraction process was between 92.02 % and 114.77 %, and no notable matrix effects were detected. The stability of the compounds was confirmed under four distinct storage conditions. Pharmacokinetic results indicated that all analytes displayed rapid absorption, prolonged retention, and slow elimination in rats, and exhibited nonlinear pharmacokinetics at low, medium, and high dosage levels. This study has established a quantification method for 13 gypenosides and successfully applied it to their pharmacokinetic studies in rats, providing a foundation for their further development and utilization.

This study investigates the potential for the microbial transformation of camelliagenin B, a saponin derived from Camellia oleifera seed cake meal, to develop novel metabolites. We employed three microbial strains, specifically Bacillus subtilis ATCC 6633, Bacillus megaterium CGMCC 1.1741, and Streptomyces griseus ATCC 13273, to biotransform camelliagenin B into its derivatives. The compounds were purified and separated using chromatographic techniques, such as high-performance liquid chromatography (HPLC). Structural identification was carried out using spectroscopic methods, including nuclear magnetic resonance (NMR) and mass spectrometry (MS). Ten bioactive compounds were obtained (1a-1j), of which nine were novel with multiple tailoring reactions, such as allyl oxidation, C-C double-bond rearrangement, hydroxylation, dehydrogenation, and glycosylation, observed in camelliagenin B analogs. The structures of these compounds were determined by 1D/2D NMR and HR-ESI-MS analysis. Therefore, this study showcases the capacity of microbial transformation as a sustainable and environmentally friendly method for generating bioactive compounds from C. oleifera seed cake meals. The individual chemicals can potentially facilitate the design of novel medicinal agents, functional foods, and natural preservatives.