Objective To identify members of the cycloartenol synthase (CAS) gene family (PgCAS) using the Panax ginseng transcriptome database, and analyze the expression patterns of PgCAS04-5 in overexpressed and RNAi-positive hair root materials), as well as explore the function of this gene in ginsenoside biosynthesis. Methods Bioinformatics approaches were adopted to characterize the chromosomal localization, conserved motifs, phylogenetic relationships and expression profiles, and the correlations with the gene expression of key saponin synthesis enzymes and the content of monomer saponin of PgCASs. The protein structure and subcellular localization of PgCAS04-5 were predicted. Meanwhile, the expression levels of key enzyme genes and variations in ginsenoside contents were detected in overexpression and RNAi-positive hairy root lines. Results A total of 20 PgCAS transcripts were identified from the P. ginseng transcriptome database. Chromosomal mapping showed that PgCAS genes were unevenly distributed on five chromosomes of P. ginseng. Phylogenetic tree analysis divided the PgCAS family into three subfamilies, and most PgCAS members belonged to subfamily Ⅱ and subfamily Ⅲ. The PgCAS genes displayed obvious spatiotemporal specificity in expression across different cultivars, growth years and tissues. Additionally, the PgCAS family formed an interaction network with key genes involved in ginsenoside biosynthesis, and their content was significantly correlated with the accumulation of monomeric ginsenosides. Subcellular localization prediction demonstrated that PgCAS04-5 was localized to chloroplasts. Moreover, the overexpression of this gene was inversely regulated with the expression of four genes including PgCAS04-5, PgDS-1, PgSE2-4 and PgCYP137 in RNAi positive materials, with significant differences. Conclusion A total of 20 members of the PgCAS gene family were identified in this study, and functional differentiation exists among PgCAS family members. PgCAS04-5 positively regulates the biosynthesis of ginsenosides Rg₂and Rf, and negatively modulates the synthesis of ginsenosides Rb₂and Rd. The results provide a valuable reference for further research on the PgCAS gene fami

Objective To prepare paeoniflorin-glycyrrhiza protein self-assembled nanoparticles (Pae-GP/SAN) with good bioavailability and investigate its intestinal absorption mechanism. Methods In this study, glycyrrhiza protein (GP) was used as the carrier to prepare Pae-GP/SAN by ultrasonic dispersion method. The formulation and preparation process were optimized using average particle size, polydispersity index (PDI), encapsulation efficiency, and drug loading as evaluation indicators, and the prepared nanoparticles were characterized. A rat in situ single-pass intestinal perfusion model was established to investigate and compare the intestinal absorption behaviors of paeoniflorin solution (Pae/Sol), paeoniflorin-glycyrrhiza protein physical mixture (Pae-GP/PM), and Pae-GP/SAN. Additionally, an aggregation-caused quenching (ACQ) fluorescent probe was used to observe the intestinal absorption of GP/SAN by confocal laser scanning microscopy (CLSM), aiming to initially clarify its penetration-enhancing mechanism. Results The optimized Pae-GP/SAN had an average particle size of (178.2 ± 6.3) nm, a PDI of 0.152 1 ± 0.011 2, a ζ potential of (−14.91 ± 1.13) mV, an encapsulation efficiency of (36.45 ± 2.32)%, and a drug loading of (21.70 ± 1.30)%, with a uniform spherical micromorphology. Intestinal perfusion experiments indicated that the absorption efficiency of Pae-GP/SAN in the ileum was superior to that in the jejunum (P < 0.05, 0.01). The formation of nanoparticles significantly promoted the absorption of paeoniflorin. Moreover, the absorption parameters of high mass concentration Pae-GP/SAN were significantly higher than those of low mass concentration Pae-GP/SAN (P < 0.001), while mass concentration had no significant effect on the absorption of Pae/Sol and Pae-GP/PM. Transporter inhibitor intervention experiments showed that indomethacin and reserpine had no significant effect on the absorption of the three formulations. Verapamil significantly increased the absorption of Pae/Sol and Pae-GP/PM (P < 0.01, 0.001) but had no significant effect on Pae-GP/SAN. CLSM observations confirmed that GP/SAN can be absorbed by the intestine in the form of intact nanoparticles. Conclusion Pae-GP/SAN can be endocytosed and absorbed by the intestine as intact nanoparticles, effectively evading the barrier effect of P-gp efflux protein, thereby significantly improving the oral bioavailability of Pae.

Objective To explore the mechanism of tubeimoside II (TUB-II) in inhibiting the growth and metastasis of colorectal cancer. Methods Human colorectal cancer cell lines RKO and SW620 were used as research subjects, the effect of TUB-II on viability of RKO and SW620 cells was detected. The cell proliferation ability was detected by colony formation assay. The cell migration ability was detected by scratch assay. Western blotting was used to detect the effect of TUB-II on expressions of lethal (2) giant larvae homolog 2 (LLGL2) and epithelial-mesenchymal transition (EMT)-related proteins in RKO and SW620 cells. Co-immunoprecipitation was used to detect the effect of TUB-II on ubiquitination of LLGL2. Subcutaneous tumor model and colorectal cancer lung metastasis model in vivo were constructed to investigate the effect of TUB-II on tumor growth and lung metastasis. Results TUB-II significantly inhibited the proliferation and metastasis of colorectal cancer both in vitro and in vivo (P < 0.05, 0.01, 0.001). TUB-II significantly upregulated the expression of LLGL2 (P < 0.05, 0.01, 0.001) and inhibited the activation of phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) pathway (P < 0.05, 0.01, 0.001), thereby suppressing tumor cell proliferation and metastasis. At the same time, TUB-II inhibited the interaction between murine double minute 2 (MDM2) and LLGL2 to suppress the ubiquitination of LLGL2, thereby stabilizing its expression and exerting an anti-colorectal cancer effect. Conclusion TUB-II inhibits the occurrence and development of colorectal cancer, and its mechanism may be related to the inhibition of LLGL2 ubiquitination and PI3K/Akt signaling pathway.

Postmenopausal osteoporosis (PMOP) results from a combination of multiple pathological mechanisms, including osteogenic impairment and osteoclast activation triggered by estrogen deficiency-induced chronic low-grade inflammation and mitochondrial dysfunction, as well as systemic inflammation and metabolic dysregulation caused by gut microbiota imbalance. Faced with this intricate and multidimensional pathological network, conventional single-target therapeutic strategies exhibit obvious limitations. Traditional Chinese medicine (TCM), with its theoretical advantages of “holistic view” and “syndrome differentiation-based treatment,” demonstrates potential for systematic intervention through multi-target and multi-pathway coordinated regulation. Current studies indicate that various TCM formulas and active components can effectively restore the homeostasis of the bone-immune microenvironment and balance between gut microbiota and bone metabolism by comprehensively modulating these multidimensional pathological networks. Based on this, this review systematically summarizes the multidimensional mechanisms of PMOP, highlighting the multi-target regulatory effects of TCM within this pathological network, thereby providing theoretical support and research ideas for in-depth elucidation of PMOP pathogenesis and for advancing integrated prevention and treatment strategies combining TCM and western medicine.

Objective To explore the mechanism of morin, an active ingredient of Sangzhi (Mori Ramulus), against gastric cancer based on network pharmacology, molecular docking and in vitro experiments. Methods Potential targets of Mori Ramulus and gastric cancer-related targets were screened by network pharmacology, and the intersection of targets was obtained. Protein-protein interaction (PPI) network topological analysis, gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis, and GEO dataset validation were used to screen core targets. Molecular docking and molecular dynamics simulation were applied to verify the binding ability of morin (the active ingredient of Mori Ramulus) to phosphatidylinositol 3-kinase regulatory subunit 1 (PIK3R1) and the stability of the formed complex. In in vitro experiments, human gastric adenocarcinoma AGS cells were treated with 100, 200, 300, 400 μmol/L morin alone or combined with phosphatidylinositol 3-kinase (PI3K) agonist 740Y-P. Cell proliferation ability was detected by CCK-8 assay and plate clone formation assay. Cell apoptosis rate and cell cycle distribution were detected by flow cytometry. Expressions of PI3K/protein kinase B (Akt) pathway, apoptosis and cycle related proteins were detected by Western blotting. Results A total of 178 potential targets of Mori Ramulus, 13 100 gastric cancer-related targets, and 159 intersecting targets were identified. Enrichment analysis showed that the key pathway was the PI3K/Akt pathway. Morin had strong binding affinity with PIK3R1, was significantly highly expressed in gastric cancer tissues, and the constructed morin-PIK3R1 complex maintained excellent stability. The in vitro experiment results showed that morin could inhibit AGS cells proliferation in a dose-dependent manner, induce G₀/G₁ phase arrest, and promote cell apoptosis (P < 0.05, 0.01). Meanwhile, morin downregulated the expressions of PI3K/Akt pathway, B-cell lymphoma-2 (Bcl-2), cyclin D1 (CCND1), cyclin-dependent kinase 4 (CDK4) and cyclin-dependent kinase 6 (CDK6) protein (P < 0.05, 0.01), and upregulated the expressions of Bcl-2 associated X protein (Bax) and p21 (P < 0.01). Combined treatment with 740Y-P significantly reversed the effects of morin on proliferation, apoptosis and cell cycle (P < 0.05, 0.01). Conclusion Morin can exert anti-gastric cancer activity by targeting and stably binding to PIK3R1, inhibiting the activation of PI3K/Akt pathway, inducing cell cycle arrest, promoting apoptosis, and inhibiting proliferation in gastric cancer cells.

Objective To investigate the chemical compositions of the ethyl acetate (EtOAc) fraction of Sophora flavescens and evaluate their anti-liver fibrosis activity. Methods The EtOAc extraction of S. flavescens was separated and purified by various column chromatography methods, including silica gel, polyamide, Sephadex LH-20 and C₁₈reversed-phase column chromatography. The structures of the isolated compounds were elucidated based on modern spectroscopic techniques and physical and chemical identification method. Transforming growth factor beta-1 (TGF-β1)-induced LX-2 cells were used as an in vitro model of hepatic fibrosis, and the anti-fibrotic activities of the compounds were evaluated using the CCK-8 assay. Additionally, cell wound scratch assay, real-time quantitative polymerase chain reaction (RT-qPCR), Western blotting, and flow cytometry assays were employed to investigate the anti-liver fibrosis activity and mechanism of compound 14 (kushenol T). Results A total of 28 compounds were isolated from the EtOAc of S. flavescens, and they were identified as follows: ethyl 2,4-dihydroxybenzoate (1), trifohrhizin (2), 2,4- dihydroxybenzoic acid (3), kuraridin (4), kushenol N (5), kurarinol A (6), kushenol C (7), 8-isopentenyl-5-methoxy-7,2′,4′-trihydroxy flavanonol (8), 2′-hydroxy-isoxanthohumol (9), kurarinone (10), methyl 2,4-dihydroxybenzoate (11), sophoflavescenol (12), umbeliferone (13), kushenol T (14), sophophenoside B (15), 6′′-β-D-xylosegenistin (16), 3′,4′-methylenedioxyisoflavone-7-O-β-D-apiofuranosyl-(l→6)-β-D-glucopyranoside (17), ambocin (18), kushenol O (19), 4′-hydroxy-3′-methoxyisoflavone-7-O-β-D-xylopyranosyl-(l→6)-β-D-glucopyranoside (20), 3′-hydroxy-4′-methoxyisoflavone-7-O-β-D-apiofuranosyl-(1→6)-β-D-glucopyranoside (21), genistein (22), isoxanthohumol (23), 8-isopentenyl-5-methoxy-7,4′-dihydroxyflavanonol (24), 2-(2¢,4¢-dihydroxyphenyl)-5,6-methylenedioxy- benzofuran (25), calycosin (26), daidzein (27), narirutin (28). Bioactivity screening revealed that compounds 9, 11—14, 17, 20, 23—25 and 28 exhibited significant inhibitory effects on LX-2 cells. Among these, 14 demonstrated the most potent activity, markedly downregulating the expression of hepatic fibrosis markers, such as α-smooth muscle actin (α-SMA), fibronectin (FN) and Collagen I, at both the gene and protein levels. Furthermore, 14 promoted cell apoptosis and suppressed TLR4 in the Toll-like receptors 4 (TLR4)/nuclear factor kappa-B (NF-κB) signaling pathway, myeloid differentiation primary response 88 (MyD88), and TGF-β-activated kinase 1 (TGF-β-activated kinase 1), tumor necrosis factor α (TNF-α), and interleukin-1β (IL-1β). Conclusion Compounds 18, 25 and 28 were isolated from this species for the first time. Compounds 9, 11—14, 17, 20, 23—25 and 28 significantly inhibited LX-2 cell proliferation. Among them, compound 14 had the most significant activity and exerts an anti-liver fibrosis effect through the TLR4/NF-κB signaling pathway.

Objective To screen high-efficiency phosphate-solubilizing bacteria (PSB) in the rhizosphere soil of Fritillaria taipaiensis and provide data support for the development of microbial fertilizers. Methods Rhizosphere soil samples were collected from ten production areas. Strains of inorganic phosphate-solubilizing bacteria (IPSB) and organic phosphate-solubilizing bacteria (OPSB) were isolated and purified. Their phosphate-solubilizing capacities were evaluated qualitatively and quantitatively, and the strains were identified based on physiological and biochemical characteristics as well as 16S rDNA sequencing. The effects of selected strains were further verified using pot experiments. Results A total of 42 IPSB and 31 OPSB strains were isolated. Quantitative analysis showed that among the IPSB strains, WP2-2 exhibited the strongest solubilizing capacity, increasing available phosphorus by 231.30 mg/L, followed by WP7-2 (211.05 mg/L). Among the OPSB strains, YP3-1 showed the highest phosphorus solubilization (28.57 mg/L), followed by YP5-1 (22.30 mg/L). Strain WP2-2 was identified as Serratia plymuthica, WP7-2 as Bacillus cereus, and both YP3-1 and YP5-1 were also identified as B. cereus. Pot experiments revealed that inoculation with these PSB strains altered the total phosphorus content in bulbs and fibrous roots, and also influenced the total alkaloid content in the bulbs. Conclusion The strains WP2-2, WP7-2, YP3-1, and YP5-1 show great potential for developing specialized microbial fertilizers for the cultivation of F. taipaiensis.

As a classic method of traditional Chinese medicine (TCM) processing, wine-processing has been employed for toxin reduction and preservation of medicinal properties, efficacy enhancement and medicinal property modification, or purposely guiding the therapeutic effects of herbal medicines. This review retrieved literature from PubMed, Web of Science, China national knowledge infrastructure (CNKI), and Wanfang databases using keywords including “wine-processing” “medicinal properties” and “efficacy-enhancing mechanisms”. Findings indicate that traditional wine-processing predominantly utilizes HuangJiu, with distinctive techniques such as wine rinsing, wine stir-frying and wine steaming remaining in current practice. Modern processing technology emphasizes optimization of critical parameters. Wine-processing facilitates component transformation through physical dissolution, enhanced cellular permeability, and synergistic enzymatic/non-enzymatic reactions. Mechanisms involving multi-target networks and microbiota-targeted metabolic regulation, enable directional modulation of herbal cold/hot properties alongside efficacy enhancement and toxicity reduction. Future research should prioritize establishing digitalized process quality control standards, conduct interdisciplinary investigations into wine-processing effects on efficacy-related component groups, and elucidate directional regulation mechanisms of medicinal properties. These advancements are anticipated to promote standardization and modernization of wine-processing techniques while scientifically interpreting the theoretical foundations of TCM processing.

Objective To investigate the antifungal secondary metabolites from the rice fermentation products of the Valeriana officinalis endophytic fungus Chaetomium elatum FH-3. Methods The one strain many compounds OSMAC (OSMAC) strategy and global natural products social molecular networking (GNPS) molecular networking technology were employed for targeted separation. Chromatographic and spectroscopic methods were used for structural identification. The antifungal activity of the compounds against Colletotrichum gloeosporioides was evaluated using the mycelial growth rate method. Results A total of 12 compounds were targeted and isolated from the ethyl acetate extract of Chaetomium elatum FH-3. These compounds were identified as 4′′-methoxy-asperianas A (1), (3S,4S)-4-hydroxy-6-methoxymellein (2a), (3R,4R)-4-hydroxy-6-methoxymellein (2b), cis-4,6-dihydroxymellein (3), cis-4-hydroxymellein (4), 6-methoxymellein (5), 6,8-dihydroxy-3-methyl-3,4-dihydroisocoumarin (6), 3-methyl-6-hydroxy-8-methoxy-3,4-dihydroisocoumarin (7), xenofuranone B (8), flavipesin B (9), p-hydroxybenzaldehyde (10), and p-hydroxybenzoic acid methyl ester (11). Antifungal testing showed that compounds 1—4 exhibited significantly stronger inhibitory activity against C. gloeosporioides than the positive control carbendazim, with half-maximal effective concentration (EC₅₀) values of 15.02, 25.31, 37.89, and 47.86 μg/mL, respectively. Conclusion Compounds 1 and 2a are new butenolide and dihydroisocoumarin derivatives, named butenolide A and dihydroisocoumarin C respectively, and compounds 8 and 9 are reported for the first time from this strain. Compounds 1—9 demonstrate significant inhibitory activity against C. gloeosporioides.

Objective Compare the differences in material basis and anti-insomnia and anti-anxiety efficacy between Suanzaoren Decoction (SZRD, 酸枣仁汤) and Suanzao Decoction (SZD, 酸枣汤) in the Jin Gui Yao Lue. Methods This study quantifies 20 chemical ingredients as evaluation indicators to establish the boiling methods of SZRD and SZD, based on ultra-high performance liquid chromatography-triple quadrupole linear ion trap mass spectrometry (UPLC-Qtrap-MS) technology. Moreover, the ultra-high performance liquid chromatography-orbitrap tandem mass spectrometry (UPLC-Orbitrap-MS/MS) metabolite identification method specific to the Chinese herbal medicine was used to identify and compare the chemical profile of the two decoctions. Lastly, the p-chlorophenylalanine (PCPA) induced insomnia model mice were used to evaluate and compare the efficacy of the two decoctions in anti-insomnia and anti-anxiety. Results The method of boiling Suanzaoren in ceramic-coated iron pot for 25 min and adding other herbs then continuing to boil for 40 min can match well with the original records in Jin Gui Yao Lue. When Suanzaoren and Suanzao fruit were both used with the amount of 2 litters in Han Dynasty, the contents of jujuboside A, jujuboside B, spinosin, 6‴-feruloylspinosin, magnoflorine, pachymic acid and ligustilide in SZRD were over 6-fold higher than those in SZD, whereas the contents of liquiritigenin, isoliquiritin, isoliquiritigenin and timosaponin AIII in SZD were over 2 times than those in SZRD. Among the top 100 compounds with the strongest mass spectrometric signals detected in the two decoctions, 42 compounds exhibited higher contents in SZD and 21 compounds were enriched in SZRD. Among the differential compounds, three isoquinoline alkaloids [(-)-isocorypalmine, cheilanthifoline and asimilobine], one pyrrolidine alkaloid (stachydrine) and one steroidal alkaloid (zhebeinine) were further identified. The relative abundances of cheilanthifoline and asimilobine in SZD were 5.2 times and 4.8 times higher than those in SZRD, respectively; meanwhile, the relative abundances of (-)-isocorypalmine, stachydrine and zhebeinine in SZD only reached 0.3, 0.4 and 0.4 times the corresponding values in SZRD. The anti-insomnia and anti-anxiety efficacies of the two decoctions in the PCPA induced insomnia mice are similar to each other, and no statistically significant intergroup difference in their pharmacological efficacy was observed.. Conclusion Under the original decoction method recorded in Jin Gui Yao Lue, SZRD and SZD show significant differences in material basis, yet their similar anti-insomnia and anti-anxiety effects under specific condition may result from the combined actions of multiple active components.