Objective To study the chemical constituents fromdried whole plant of alpine herb Gentiana nanobella, and to conduct preliminary screening for antimalarial activity of the isolated constituents. Methods Ethyl acetate and n-butanol fractions of 75% ethanol extract of the whole plant of G. nanobella were isolated and purified by repeated column chromatography padded with silica gel, RP-18 and Sephadex LH-20, respectively. The structures of the isolated compounds were identified based on their physicochemical properties and spectral data. The in vitro inhibitory activity of the testing compound against human Plasmodium falciparum 3D7 strain was achieved with the SYBR Green I method. Results A total of 11 phenol compounds were obtained and identified as 2,3-dihydroxy benzoic acid methyl ester-3-O-β-L-rhamnosyl-(1''→2')-b-D-glucopyranoside (1), 2,3-dihydroxy benzoic acid-3-O-β-L-rhamnosyl-(1''→2')-b-D-glucopyranoside (2), 2,3-dihydroxy benzoic acid methyl ester-3-O-β-D-glucopyranosyl-(1''→6')-β-D-glucopyranoside (3), benzyl β-D-xylopyranosyl-(1''→6')-β-D-glucopyranoside (4), apigenin 5-O-β-D-glucopyranoside (5), salcolin B (6), apigenin-7-O-β-D-glucopyranoside (7), benzoic acid (8), p-hydroxybenzoic acid (9), 3-hydroxy-2-methoxybenzoic acid (10), and isovanillic acid (11). Compound 6 exhibited an in vitro antimalarial activity against Plasmodium falciparum 3D7 strain with IC₅₀ 18.50 μmol/L and inhibition rate of 98.48% at a concentration of 50 μmol/L. Conclusion Compounds 1 and 2 are new compounds, named gentinanoside A and gentinanoside B. Compounds 1—11 were isolated from G. nanobella for the first time. G. nanobella is abundant in phenolic compounds, and some compounds had certain antimalarial activity.

Traditional Chinese medicine（TCM） and ethnic medicine, as integral components of traditional medicine, exhibit diverse therapeutic effects, complex chemical compositions, intricate physiological processes, and multifaceted mechanisms of action. This makes elucidating the pharmacological basis and mechanisms of action of TCM and ethnic medicine a key research direction in the modernization of TCM. However, challenges such as incomplete pharmacological evaluation models, low correlation with clinical efficacy, and low screening efficiency and throughput remain core obstacles. Artificial intelligence-driven organoid and organ-on-achip technologies offer an innovative research paradigm for the screening of bioactive compounds and the discovery of target mechanisms in TCM and ethnic medicines. By constructing highly biomimetic human organoids/organ-on-a-chip models that precisely simulate the microenvironment and pathological states of human organs, combined with artificial intelligence technologies for in-depth analysis of high-throughput pharmacodynamic data, it is possible to elucidate the synergistic effects of multi-component, multi-target interactions. Systematically screening groups of active components in TCM, quantifying their metabolic kinetic processes, and revealing potential target points and molecular pathways significantly enhances the precision and efficiency of bioactive substance identification. The deep integration strategy of organoids/organ-on-a-chip technology with artificial intelligence provides new technical support for accelerating the modernization of TCM and ethnic medicine research and promoting innovative drug development.

Objective To develop the determination methods for the content of vanillic acid, isovanillic acid, rhamnetin, luteolin, quercetin, diosmetin, oleanolic acid, ursolic acid, anemarrhenasaponin A2, gracillin, prosapogenin A, dioscin, protogracillin, β-sitosterol, stigmasterol, total ash and acid insoluble ash in Smilax riparia, and evaluate the quality of S. riparia by combining multivariate statistical analysis and entropy weight TOPSIS model. Methods HPLC was launched on an Agilent Symmetry C₁₈ chromatographic column（250 mm×4.6 mm,5 μm） by gradient elution with a mobile phase of acetonitrile-0.3% phosphoric acid at a flow rate of 1.0 mL/min, detection wavelength were 280 nm（for detecting vanillic acid and isovanillic acid）, 345 nm（for detecting rhamnetin, luteolin, quercetin, diosmetin） and 210 nm（for detecting oleanolic acid, ursolic acid, anemarrhenasaponin A2, gracillin, prosapogenin A, dioscin, protogracillin, β-sitosterol, stigmasterol）, column temperature was 30 ℃, and an injection volume was 10 μL. The contents of total ash and acid insoluble ash were detected according to the method of Chinese Pharmacopoeia 2025 edition. Based on the content measurement value, the quality of 18 batches of S. riparia samples was comprehensively evaluated by using the multivariate statistical analysis and entropy weight TOPS model. Results A total of 15 chemical components had good linear relationships in their respective mass concentration ranges, and the average recovery rates were 96.99%—100.14%. The contents of vanillic acid, isovanillic acid, rhamnetin, luteolin, quercetin, diosmetin, oleanolic acid, ursolic acid, anemarrhenasaponin A2, gracillin, prosapogenin A, dioscin, protogracillin, β-sitosterol, stigmasterol, total ash and acid insoluble ashin in 18 batches of S. riparia were 0.338—1.213, 0.074—0.200, 0.197—0.360, 0.423—0.676, 0.620—1.814, 0.496—1.007, 0.192—0.410, 0.095—0.187, 1.360—2.281, 0.379—0.651, 0.711—1.221, 2.120 —3.657, 0.965—1.652, 0.113—0.275, 0.063—0.123 mg/g, 3.6%—9.5%, 1.4%—3.5%, respectively. Multivariate statistical analysis classified 18 batches of S. riparia into three categories, principal component analysis yielded two principal components, with a cumulative variance contribution rate of 88.149%. A total of eight components such as dioscin, quercetin, luteolin, anemarrhenasaponin A2, oleanolic acid, vanillic acid, protogracillin and β-sitosterol were screened by VIP analysis, which could be used as differential markers of S. riparia in different producing areas. The results of entropy weight TOPSIS method showed that the quality of 18 batches of S. riparia was significantly different, and the comprehensive score was 0.230 6—0.706 2. The quality of samples numbered S16 and S15 was ranked in the front. Conclusion The quality evaluation model based on chemometrics and entropy weight TOPSIS is convenient and effective, and can be used for the quality evaluation of S. riparia.

Objective To investigate the chemical constituents and their myocardial cell protective activity from the fruits of Daemonorops draco. Methods Compounds were isolated and purified using a combination of silica gel, RP-C₁₈, Sephadex LH-20 and semi-preparative HPLC. Their structures were elucidated through comprehensive spectroscopic analysis, UV, IR, HRESIMS and NMR. The protective effects of all isolates on the H9c2 myocardial cells injury induced by oxygen-glucose deprivation (OGD) were evaluated. Results Three pairs of new flavans were purified and elucidated as (2S)-2-ethoxy-7-hydroxy-5-methoxy-6-methylflavan (1a), (2R)-2-ethoxy-7-hydroxy-5-methoxy-6-methylflavan (1b), (2S,3S)-2-ethoxy-3,7-dihydroxy-5-methoxy-6-methylflavan (2a), (2R,3R)-2-ethoxy-3,7-dihydroxy-5-methoxy-6-methylflavan (2b), (2R,3S)-2-ethoxy-3,7-dihydroxy-5-methoxy-6-methylflavan (3a) and (2S,3R)-2-ethoxy-3,7-dihydroxy-5-methoxy-6-methylflavan (3b). Compounds 1b and 2b demonstrated protective effects against OGD-induced H9c2 cell injury, which increased cell viability by (6.36 ±1.26)% and (10.78 ±0.76)% at a concentration of 20 μmol/L, respectively. Conclusion Compounds 1a/1b, 2a/2b, and 3a/3b are three pairs of enantiomers, and they are all new compounds, named as dracoropin I₁, dracoropin I₂, dracoropin J₁, dracoropin J₂, dracoropin K₁, dracoropin K₂. Among them, compounds 1b and 2b exhibited protective effects on myocardial cells.

Objective To explore the differences in the in vivo drug metabolism of raw and processed Dahuang (Rhei Radix et Rhizoma) in a rat model of middle cerebral artery occlusion and reperfusion (MCAO/R). Methods The MCAO/R rat model was established by the suture method. The successfully modeled rats were randomly divided into model group, raw Rhei Radix et Rhizoma (2.50 g/kg) group and processed Rhei Radix et Rhizoma group, with six rats in each group. Rats were administered continuously for 7 d. 12 h before the last administration, the rats were fasted, and 1 h after administration, blood and brain tissues were collected. Ultra-high performance liquid chromatography-quadrupole electrostatic field orbitrap linear ion trap mass spectrometry (UPLC-Orbitrap Fusion Lumos Tribrid-MS) was used to characterize the prototypes and metabolites of raw and processed Rhei Radix et Rhizoma in plasma and brain tissues of MCAO/R rats. Multivariate statistical analysis was used to screen the differential components, and desorption electrospray ionization mass spectrometry imaging (DESI-MSI) technique was combined to visualize the in situ spatial distribution of the differential components of raw and processed Rhei Radix et Rhizoma in brain tissue. Results A total of 109 prototype components and metabolites were identified in MCAO/R rats. According to variable important in projection (VIP) > 1 and P < 0.05, 23 and eight differential components of raw and processed Rhei Radix et Rhizoma were screened from the plasma and brain tissues, respectively. In the plasma of processed Rhei Radix et Rhizoma group, anthraquinone glycosides were decreased, while in the brain tissue, aglycones and metabolites (such as gallic acid, M31) were increased, and were mainly distributed in hippocampus, midbrain and cortex of brain tissue. The common differential components gallic acid and metabolite M31 were significantly increased in the processed Rhei Radix et Rhizoma group and were specifically enriched in the striatum. Conclusion This study characterized and spatially located the differential components of raw and processed Rhei Radix et Rhizoma in MCAO/R rats and found that processed Rhei Radix et Rhizoma increased the exposure levels of active aglycones and metabolites in the brain, enhancing its protective effect on brain tissue, providing a scientific basis for the “different treatment for raw and processed” processing theory.

Objective To prepare an amino acid-based deep eutectic gel dressing and study in vitro performance and the in vivo therapeutic effect on oral ulcer. Methods Deep eutectic solvents (DESs) were screened using lysine, proline, arginine, and glycine as hydrogen bond donors, and lactic acid, citric acid, glycerol, mannitol, and malic acid as acceptors, with curcumin solubility identifying the optimal system. Gel dressings were prepared by optimizing the formulation process based on the composite matrix of gelatin methacryloyl/chitosan methacryloyl (GelMA/CSMA) in combination with 3D printing technology. Its mechanical properties, adhesiveness, swelling, and biocompatibility were evaluated, and in vivo efficacy was verified in an SD rat oral ulcer model. Results The optimal DES system (proline-lactic acid, 1:12) increased curcumin solubility over 700-fold (0.789 mg/mL) compared to its solubility in water. The optimal printing formulation and process parameters were as follows: 1% CSMA, 5% GelMA, 0.1% lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP), 0.02% tartrazine, 20% DES content, 100 μm layer height, and a layer-by-layer exposure time of 25 s. The obtained amino acid-based gel dressing had excellent mechanical properties, with a 2.5 N adhesion force at 25 min and an approximately 500% swelling ratio at 6 h. In vitro biocompatibility showed that the survival rate of L929 cells was all above 90%, and the DES system had no significant cytotoxicity. It also showed high in vitro biocompatibility (L929 viability > 90%, non-cytotoxic). In vivo, the gel achieved localized, sustained curcumin release in SD oral ulcer model rats, reducing ulcer area by 40% with decreased neutrophil density and inflammatory infiltration. Conclusion Successfully prepared an amino acid-based DES gel dressing loaded with curcumin, which exhibits excellent wound adaptability and precise drug release behavior, offering an efficient and safe therapeutic strategy for oral ulcers.

Objective To prepare a self-assembled nanoparticles of saikosaponin D (SSD) and cannabidiol (CBD) and preliminarily investigate their anti-hepatoma mechanism in vitro. Methods The SSD-CBD self-assembled nanoparticles were constructed using the nano co-precipitation method at a mass ratio of 3∶1, and their assembly mechanisms were characterized by techniques including X-ray photoelectron spectroscopy (XPS), Fourier transform infrared spectroscopy (FTIR), and ¹H-¹H nuclear overhauser enhancement spectroscopy (¹H-¹H NOESY). Its physicochemical properties were evaluated by transmission electron microscopy (TEM) and dynamic light scattering (DLS). The antitumor activity, apoptosis induction, and preliminary mechanisms were systematically assessed by MTT assay, flow cytometry (Annexin V-FITC/PI staining), and cysteinyl aspartate specific proteinase (Caspase) activity assays. Results The assembly was self-assembled through hydrogen bonding and hydrophobic interactions, with SSD and CBD showing a synergistic effect against HepG2 cells (CI = 0.79). It exhibited pH-responsive drug release, specifically releasing drugs in the tumor microenvironment (pH 6.8) while remaining stable at normal physiological environment (pH 7.4). Mechanistic studies indicated that it primarily induced HepG2 cell death by activating the mitochondrial apoptotic pathway. Conclusion The constructed SSD-CBD self-assembled nanoparticles offer a carrier-free strategy that can simultaneously improve the delivery efficiency of CBD and achieve precise collaborative drug delivery, providing an experimental basis for the development of intelligent nanomedicine based on SSD.

Alzheimer’s disease (AD) is a neurodegenerative disorder characterized by progressive cognitive decline. Its pathological mechanisms are complex, involving multiple key links such as beta-amyloid protein (Aβ) deposition, excessive phosphorylation of Tau protein, neuroinflammation and synaptic dysfunction. Traditional Chinese medicine (TCM) categorizes AD under conditions like “dementia” and “forgetfulness”, with the core pathogenesis being “deficiency at the root and excess at the surface”, where kidney essence deficiency is the root cause and phlegm turbidity and blood stasis obscuring the orifices are the surface manifestations. As a distinctive TCM therapy, aromatic orifice-opening drugs, with their pungent, fragrant and penetrating nature that can reach and regulate meridians, follow the principle of “awakening the orifices and refreshing the mind, resolving phlegm and dispelling foulness”, demonstrating unique advantages in the prevention and treatment of AD. Representative drugs include Shichɑngpu (Acori Tatarinowii Rhizoma), Yuanzhi (Polygalae Radix), Shexiang (Moschus), Bingpiɑn (Borneolum Syntheticum), etc. Modern pharmacological research has revealed their multi-target intervention potential, including inhibiting Aβ generation and aggregation, reducing Tau protein phosphorylation, anti-neuroinflammation, anti-oxidative stress, and regulating neurotransmitters and promoting neurogenesis. Commonly used TCM formulas with the effect of aromatic orifice-opening such as Angong Niuhuang Pill (安宫牛黄丸), Suhexiang Pill (苏合香丸) and Changpu Yujin Decoction (菖蒲郁金汤) have shown good efficacy in improving patients’ cognitive function, memory, and daily living abilities through compatibility and synergy. This review systematically summarizes the research and application status of aromatic orifice-opening drugs in the prevention and treatment of AD, providing theoretical basis and practical references for the intervention of AD by aromatic orifice-opening TCMs.

Objective This study investigated the regulatory mechanism by which methyl jasmonate (MeJA) modulates triterpenoid saponin biosynthesis in Jixuecao (Centella asiatica). Methods Plants were treated with 200 μmol/L MeJA, and temporal changes in leaf asiaticoside and madecassoside contents were quantified by high-performance liquid chromatography (HPLC). Transcriptome data were analyzed using weighted gene co-expression network analysis (WGCNA) to identify modules significantly correlated with asiaticoside accumulation, followed by Mfuzz clustering to refine gene subclusters. Expression patterns of candidate key genes were validated by quantitative real-time PCR (qRT-PCR). Results MeJA treatment markedly promoted asiaticoside and madecassoside accumulation, increasing their contents by 18.28% and 14.58%, respectively. WGCNA revealed 33 co-expression modules, with the darkred module displaying a highly significant positive correlation with asiaticoside content (P < 0.01). Mfuzz clustering subdivided the 286 genes in this module into four subclusters; Notably, 239 genes in Clusters 1-3 were significantly enriched in the terpenoid backbone biosynthesis, sesquiterpenoid biosynthesis, and triterpenoid biosynthesis pathways. Twenty-eight structural genes directly involved in asiaticoside synthesis were identified, exhibiting expression profiles closely aligned with saponin accumulation. qRT-PCR confirmed significant MeJA-induced upregulation of 11 key structural genes associated with triterpenoid backbone synthesis and modification, consistent with the transcriptome findings. Conclusion MeJA enhances triterpenoid saponin biosynthesis in C. asiatica through coordinated upregulation of these 11 key structural genes within the darkred module. By integrating multi-omics profiling with experimental validation, this work systematically elucidates the molecular mechanisms underlying MeJA-induced triterpenoid saponin accumulation, offering valuable candidate genes for future functional characterization and genetic enhancement of C. asiatica.

Objective To explore the potential active components, targets and molecular mechanism of Tianma Gouteng Yin (天麻钩藤饮) in improving vascular endothelial function in hypertension based on network pharmacology, molecular docking and experimental verification, and to clarify whether it exerts effects via phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/endothelial nitric oxide synthase (eNOS) signaling pathway. Methods Active components and targets of Tianma Gouteng Yin were screened through HERB and SwissTargetPrediction databases. Hypertension-related targets were obtained from OMIM and GeneCards databases. A protein-protein interaction (PPI) network was constructed, and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis was performed. Molecular docking was used to verify the binding affinity between core components and key targets in PI3K/Akt/eNOS pathway. Human umbilical vein endothelial cells (HUVECs) injured by angiotensin Ⅱ (Ang-Ⅱ) were used as the cell model. CCK-8, qRT-PCR, Western blotting, reactive oxygen species (ROS) and nitric oxide (NO) assays were applied to evaluate the effects of Tianma Gouteng Yin on cell viability, oxidative stress, inflammatory response and PI3K/Akt/eNOS pathway. Results Network pharmacology identified 228 active components and 969 overlapping hypertension-related targets of Tianma Gouteng Yin. Core targets included serine/threonine kinase 1 (AKT1) and PI3K family members [phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA), phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit beta (PIK3CB), etc.]. KEGG enrichment indicated that PI3K/Akt pathway was the key pathway. Molecular docking confirmed that core components such as kaempferol and apigenin showed strong binding affinity to PI3K/Akt pathway targets. The results of cell experiment showed that 25-100 mg/mL Tianma Gouteng Yin had no cytotoxicity in normal HUVECs, and dose-dependently attenuated Ang-Ⅱ-induced reduction in cell viability (P < 0.001), significantly inhibited the expressions of angiotensin-Ⅱ type 1 receptor (AT1R) and NADPH oxidase 4 (NOX4), as well as ROS production (P < 0.001), upregulated the levels of antioxidant enzymes including superoxide dismutase 1 (SOD1), catalase (CAT) and glutathione peroxidase 1 (GPX1) (P < 0.05, 0.01, 0.001), reduced the expressions of inflammatory factors including interleukin-6 (IL-6), IL-1β, tumor necrosis factor-α (TNF-α), intercellular adhesion molecule 1 (ICAM1) and vascular cell adhesion molecule 1 (VCAM1) (P < 0.01, 0.001), increased NO production and suppressed endothelin-1 (ET-1) expression (P < 0.01, 0.001), upregulated the protein expressions of p-PI3K, p-Akt and p-eNOS (P < 0.001). After administering the PI3K specific inhibitor Wortmannin, the activation effect of Tianma Gouteng Yin on PI3K/Akt pathway was significantly weakened, indicating that its endothelial protective effect depends on PI3K/Akt/eNOS signaling pathway. Conclusion Tianma Gouteng Yin may activate PI3K/Akt pathway through multi-target synergistic effects of core components such as kaempferol and apigenin, improve oxidative stress and inflammatory response, regulate the balance of NO and ET-1, and protect endothelial function in hypertension.