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Based on the hypoxia-inducible factor-1α(HIF-1α)/aryl hydrocarbon receptor nuclear translocator(ARNT) signaling pathway, this study aims to investigate the effects and potential mechanisms of artesunate in enhancing the effect of leflunomide on inhibiting bone destruction in rheumatoid arthritis(RA). By using in vitro osteoclast differentiation model induced by receptor activator of nuclear factor-κB ligand(RANKL) and a collagen-induced arthritis(CIA) mouse model, the intervention effect of combination treatment on bone destruction in RA was systematically evaluated at the cellular, histological, and molecular levels. Key mechanisms were analyzed and screened via transcriptomic sequencing, and relevant mechanisms were further validated through in vitro and in vivo experiments. The results show that artesunate can significantly enhance leflunomide's inhibitory effect on osteoclast formation, further reducing actin ring formation and bone resorption activity. In CIA mice, the combination treatment more effectively decreases the number of tartrate-resistant acid phosphatase(TRAP)-positive multinucleated osteoclasts in the joints compared with leflunomide alone, suggesting that artesunate potentiates leflunomide's ability to inhibit osteoclast formation and differentiation, thereby alleviating bone destruction in RA. Transcriptomic analysis indicates that artesunate may enhance leflunomide's inhibitory effect on bone destruction in RA by modulating the HIF-1α/ARNT signaling pathway. Further validation finds the nuclear co-localization of HIF-1α and ARNT during osteoclast differentiation. The combination treatment markedly suppresses HIF-1α/ARNT signaling and the expression of molecules related to downstream osteoclasts, while upregulating antioxidant-related proteins. The critical role of this signaling pathway in the combination treatment for inhibiting osteoclast differentiation is further supported by using an HIF-1α inhibitor. In summary, this study elucidated the critical role of the HIF-1α/ARNT signaling pathway in artesunate-enhanced leflunomide-mediated amelioration of bone destruction in RA, providing new experimental evidence and theoretical support for understanding the molecular mechanisms of bone destruction in RA and guiding combination treatment with leflunomide.

Understanding the heterogeneity of mammary epithelial cells and their contributions to postnatal development and regeneration is of great importance. Recent advances have defined the intrinsic complexity of the mammary epithelium, its stromal interactions, and the ontogeny of adult stem and progenitor cells. These insights have not only refined our understanding of normal mammary gland biology but also highlighted how specific epithelial subsets serve as cells of origin and targets of transformation in breast cancer. The close parallels between mammary stem cells and tumor-initiating cells, particularly their shared plasticity and self-renewal, highlight stemness as a central node linking developmental programs to malignant transformation. Both populations are shaped by microenvironmental cues and specialized niche components that regulate their maintenance, fate, and susceptibility to oncogenic reprogramming. Here, we review recent advances in mammary epithelial biology, emphasizing the key signaling pathways and stromal interactions that govern mammary stem cell behavior during development and tumorigenesis. We further discuss how developmental programs and microenvironmental cues that sustain mammary morphogenesis are subverted during cancer progression, with particular attention to pregnancy-associated transient susceptibility and aging-associated cancer risk. Overall, this Review offers a framework for understanding how normal mammary epithelial programs are redirected during malignant progression.

Chronic obstructive pulmonary disease(COPD) is a systemic disease characterized by persistent airflow limitation and airway inflammation, with a consistently high incidence and a lack of effective clinical interventions to reverse disease progression. Based on the core TCM pathogenesis theory of "deficiency, phlegm, stasis, and toxin", this paper systematically explores the critical role of the "gut lung axis-neutrophil extracellular traps(NETs)" pathway in the occurrence and development of COPD, as well as its intrinsic association with TCM pathogenesis. Specifically, dysbiosis of the intestinal microecology and dysregulation of pulmonary NETs persist throughout the disease course. Among these, "deficiency of healthy Qi" serves as the initiating factor; disruption of the gut microbiota and deficiency of metabolic products lead to decreased pulmonary defensive function and immunometabolic imbalance. Furthermore, impairment of the intestinal barrier results in the translocation of lipopolysaccharide(LPS) into the bloodstream. As an "endogenous toxin", LPS activates pulmonary neutrophils, triggers oxidative stress, and promotes the formation of NETs. Moreover, the explosive release of pulmonary NETs represents the microscopic entity of the pathological products of "phlegm, stasis, and toxin". The highly viscous DNA backbone of NETs contributes to the formation of "tenacious phlegm" that obstructs the airways; NETs-induced immunothrombosis leads to "stasis obstructing lung collaterals"; and the cytotoxic proteins carried by NETs act as "virulent toxins" that directly damage the lung parenchyma. These pathological processes are the microscopic manifestations of the core pathogenesis of COPD, namely "deficiency leading to excess", "intertwining of phlegm and stasis", and "toxin damaging lung collaterals". In this context, spleen Qi deficiency drives dysregulation of the gut-lung microecology; the influx of endogenous toxins induces NETs activation; and the accumulation of phlegm, stasis, and toxin aggravates lung tissue remodeling. Accordingly, this paper proposes the application of the "consolidating the foundation and clearing the source" method to reconstruct intestinal microecology and block the origin of endogenous toxins, as well as the "removing toxins and dredging collaterals" method to target NETs regulation for resolving phlegm and removing stasis. Through dual "gut-lung" targeting to restore systemic homeostasis, this study provides a theoretical basis and new insights for the integrated TCM and western medicine prevention and treatment of COPD.

This study aims to investigate the protective effect of psoralen against hydrogen peroxide(H_2O_2)-induced oxidative stress injury in mouse cranial apical osteoblasts(MC3T3-E1) and explore its molecular mechanism in promoting osteogenic differentiation through PTEN-induced putative kinase 1(PINK1)/Parkin-mediated mitophagy regulation. An oxidative injury model in MC3T3-E1 cells was established with 200 μmol·L~(-1) H_2O_2. The experiment was conducted with four groups: normal, model, psoralen, and inhibitor groups. Cell viability and proliferation were assessed by the CCK-8 assay and EdU incorporation assay, respectively. Apoptosis was analyzed by Annexin V-FITC/PI double-staining flow cytometry. Mitochondrial membrane potential was evaluated with the JC-1 probe, and reactive oxygen species(ROS) levels were measured by the DCFH-DA probe. Early osteogenic differentiation markers and late mineralized nodule formation were observed by alkaline phosphatase(ALP) staining and alizarin S staining, respectively. Transmission electron microscopy was employed to examine mitochondrial ultrastructure changes. Immunofluorescence staining and Western blot were employed to determine the expression levels of PINK1, Parkin, p62, Runt-related transcription factor 2(Runx2), and Osterix. Molecular docking and 100 ns molecular dynamics simulations were conducted to validate the binding mode and stability of psoralen with PINK1. The results showed that psoralen ameliorated H_2O_2-induced cell damage in a concentration-dependent manner, with the optimal concentration being 80 μmol·L~(-1). Psoralen significantly promoted cell proliferation, upregulated the expression of Runx2, Osterix, PINK1, and Parkin, enhanced mitochondrial membrane potential, increased mitophagy levels and osteogenic differentiation capacity, and enlarged mineralized nodule area. Simultaneously, it inhibited apoptosis, reduced intracellular ROS content, and downregulated the expression of autophagy substrate p62. Molecular docking results showed that the binding energy between psoralen and PINK1 was-7.024 kcal·mol~(-1), and 100 ns molecular dynamics simulations further confirmed the structural stability of the formed complex with persistent hydrogen bond interactions. In conclusion, psoralen directly targets and activates the PINK1/Parkin pathway to mediate mitophagy, clear damaged mitochondria, improve the oxidative stress microenvironment, promote the expression of osteogenic marker proteins and mineralization, thereby enhancing osteogenic differentiation of MC3T3-E1 cells, providing a new target and candidate drug for the prevention and treatment of osteoporosis.

Acute lung injury(ALI) and acute respiratory distress syndrome(ARDS) are severe respiratory diseases characterized by uncontrolled inflammatory responses and disruption of the alveolar-capillary barrier. This type of disease is often triggered by multiple factors such as infection, sepsis, trauma, or inhalation injury, and the pathological process involves multiple sections including inflammatory cascade reactions, oxidative stress imbalance, abnormal activation of immune cells, and increased pulmonary microvascular permeability. Owing to their complex pathogenesis and high mortality, available clinical therapeutic approaches remain limited. Therefore, the development of safe and effective therapeutic agents to attenuate ALI has become a major focus of current research field. As a TCM, Rheum palmatum has a long history of use in clearing heat and toxins, purging the bowels, and regulating systemic inflammatory responses. In recent years, increasing attention has been paid to its therapeutic effect in ALI. RESULTS:: show that R. palmatum and its major bioactive anthraquinones, such as emodin and rhein, exert protective effects through multi-target and multi-pathway mechanisms. These mechanisms mainly include inhibition of inflammatory signaling pathways such as nuclear factor-κB(NF-κB), mitogen-activated protein kinase(MAPK), and NOD-like receptor protein 3(NLRP3) inflammasome, attenuation of oxidative stress response, activation of the nuclear factor erythroid 2-related factor 2(Nrf2)-mediated antioxidant defense system, regulation of programmed death in immune cells including macrophages, improvement of immunometabolic status, modulation of macrophage polarization and the gut microbiota as well as the gut-lung axis, and so on. In addition, R. palmatum has been shown to preserve the integrity of the alveolar-capillary barrier and to suppress excessive neutrophil activation and the overformation of neutrophil extracellular traps(NETs), thereby reducing inflammation amplification effect and the risk of immune microthrombi. This paper summarized the action mechanisms and research progress of R. palmatum in the treatment of ALI, aiming to elucidate the unique advantages of R. palmatum anthraquinones in intervening in the complex pathological processes of ALI through systemic regulatory networks and to provide a solid theoretical basis for the research and development of innovative R. palmatum-based medicines and their clinical combination medication.

Tilorone is an antiviral agent with interferon-inducing properties. The present study focused on evaluating the apoptosis-inducing effects of tilorone in triple-negative breast cancer (TNBC) using in silico and in vitro models. Molecular docking and dynamics were performed to investigate the interactions of cGAMP and tilorone with human STING. Time- and dose-dependent effects of tilorone on the proliferation of TNBC cells were measured by MTT assay. Colony and mammosphere formation assays were performed to test the effects of tilorone on cell proliferation and stemness. Transwell and wound healing assays were performed to test the antimetastatic potential of tilorone. IFNβ levels in cultured supernatants were measured by ELISA; loss of mitochondrial membrane potential (MMP) and nuclear changes were measured by JC-1 and DAPI staining, respectively. Apoptotic proteins were measured by Western blot technique and caspase-3 assay. The in silico molecular docking and dynamics simulation studies revealed that tilorone establishes similar binding with human STING as that of the natural ligand, cGAMP. In vitro experiments further demonstrated that tilorone significantly suppressed TNBC cell proliferation in a time- and dose-dependent manner. Tilorone markedly reduced the ability of TNBC cells to form colonies and mammospheres and cell migration, indicating its inhibitory effects on proliferation, stemness, and metastasis. Mechanistically, treatment with tilorone led to an increase in the level of IFN-β released in culture supernatants of TNBC cell lines. Additionally, tilorone modulated the expression of caspase-3 and PARP proteins with loss of MMP. Thus, tilorone exerts antiproliferative, antistemness, antimetastatic, and pro-apoptotic effects in TNBC, highlighting its potential as a promising therapeutic candidate in TNBC.

This study integrated network pharmacology, computational simulation, spatial transcriptomics, and animal experiments to systematically reveal the multi-level mechanism of the traditional Chinese medicine compound Zuopi Pills in treating atherosclerosis(AS) by inhibiting ferroptosis via the acyl-CoA synthetase long-chain family member 4(ACSL4)/lysophosphatidylcholine acyltransferase 3(LPCAT3)/arachidonate 15-lipoxygenase(ALOX15) signaling axis. Network pharmacology was used to screen for the intersecting targets between the active components of Zuopi Pills and AS, followed by protein-protein interaction(PPI) construction, Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analyses. Molecular docking and molecular dynamics simulations were conducted on the core components against key ferroptosis-related targets, glutathione peroxidase 4(GPX4), ACSL4, LPCAT3, and ALOX15, to assess binding stability. Spatial transcriptomics was utilized to directly observe, within the tissue spatial dimension, the distribution and expression patterns of key targets in atherosclerotic plaque regions of the aorta, thereby validating their association with pathological sites. An ApoE~(-/-)AS mouse model of AS induced by a high-fat diet was established. Mice were treated with different doses of Zuopi Pills to assess its effects on body weight, aortic pathology(hematoxylin-eosin staining), serum lipid profiles, inflammatory cytokines, and ferroptosis-related indices, as well as protein/mRNA expression in aortic tissue. Network pharmacology analysis identified 175 intersecting targets, with KEGG enrichment highlighting ferroptosis as a key pathway. Molecular docking showed favorable binding between core components and the targets, and molecular dynamics simulations confirmed the stability of these complexes. Spatial transcriptomics analysis visually demonstrated the specific high expression of GPX4, ACSL4, LPCAT3, and ALOX15 within the aortic plaque region, providing in situ evidence to support subsequent mechanistic studies. Animal experiments confirmed that Zuopi Pills significantly reduced body weight, improved serum lipid profiles, alleviated plaque formation, and inhibited inflammation in AS mice. Mechanistically, Zuopi Pills decreased Fe~(2+) and malondialdehyde(MDA) content while increasing superoxide dismutase(SOD) activity and glutathione(GSH) levels in aortic tissue. It downregulated the protein and mRNA expression of prostaglandin-endoperoxide synthase 2(PTGS2), transferrin receptor 1(TFR1), ACSL4, LPCAT3, and ALOX15, while concurrently upregulating GPX4 expression. In conclusion, Zuopi Pills exert a clear therapeutic effect on AS mice, and its mechanism is closely related to the regulation of the ACSL4/LPCAT3/ALOX15 signaling axis and the inhibition of ferroptosis in vascular cells.

This study aims to elucidate the material basis and mechanism of action of the Aconitum pendulum Busch product processed with highland barley wine against rheumatoid arthritis(RA). UPLC-Q-TOF-MS/MS was employed to preliminarily identify 55 compounds from the product. Through network pharmacology, 19 potential active ingredients and 140 corresponding targets were screened out. The drug targets were intersected with 89 RA targets collected from the OMIM database to yield a set of potential intervention targets. The protein-protein interaction(PPI) network was constructed via STRING and topological analysis was conducted via Cytoscape, which identified eight core targets: PPARG, EGFR, SLC6A3, OPRM1, PRKCA, CYP2D6, HTR1A, and HMGCR. Gene Ontology(GO) functional and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analyses performed in the Metascape database revealed that these targets were significantly enriched in processes such as TRP channels, the phosphatidylinositol 3 kinase(PI3K)-protein kinase B(Akt) signaling pathway, and neutrophil extracellular traps(NETs) formation. An integrated "drug-ingredient-disease-target-pathway" network was constructed, which selected seven core ingredients(dehydrolucidusculine, linoleic acid, pyraconitine, 16-epipyroaconitine, delphinine, 1-monopalmitin, and songorine) for molecular docking with the eight core targets. The results demonstrated favorable binding activity between each ingredient and the targets. Animal experiments showed that the A. pendulum product processed with highland barley wine alleviated joint swelling and inflammatory pathological changes of synovial tissue in the rat model of adjuvant-induced arthritis(AA) in a dose-dependent manner. In terms of the serum inflammatory cytokine levels, it downregulated the expression of pro-inflammatory cytokines tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), interleukin-17A(IL-17A), and interleukin-6(IL-6). Furthermore, the product significantly reduced the phosphorylation levels of PI3K and its downstream signaling molecule Akt of knee synovial tissue in the rat model of AA. In conclusion, this study preliminarily reveals that the A. pendulum product processed with highland barley wine may exert anti-RA effects through multi-ingredient, multi-target, and multi-pathway mechanisms, particularly by regulating the PI3K-Akt signaling pathway and inflammatory responses.

This study focused on the protective effect of costunolide(COS) against sorafenib(SOR)-induced cardiomyocyte injury and its association with mitophagy. Cell viability was assessed using the CCK-8 assay. H9c2 cells were divided into six groups: control(ctrl) group, SOR group, SOR + low-dose COS(SOR+low-COS) group, SOR + high-dose COS(SOR+high-COS) group, SOR + high-COS + rapamycin group, and SOR + high-COS + adenosine monophosphate-activated protein kinase(AMPK) activator(AICAR) group, with each group receiving the respective treatments for 24 hours. Concurrently, male C57BL/6J mice were randomly assigned to six groups(10 per group): ctrl group, SOR group, SOR + low-COS group, SOR + high-COS group, SOR + high-COS + rapamycin group, and SOR + high-COS + AICAR group. Following the respective interventions, cardiac function and myocardial pathological changes were evaluated. The autophagosome formation labelled by microtubule-associated protein 1A/1B light chain 3(LC3) and the overall autophagic status, as well as the expression levels of proteins related to the AMPK/UNC-51-like autophagy activating kinase 1(ULK1)/FUN14 domain containing 1(FUNDC1) signaling pathway, were examined in both cellular and murine samples. The results demonstrated that 0-20 &#x3bc;mol&#xb7;L~(-1) SOR significantly reduced H9c2 cell viability in a dose-dependent manner(P<0.05). Conversely, 2.5-10 &#x3bc;mol&#xb7;L~(-1) COS dose-dependently and significantly increased the viability of SOR-treated H9c2 cells(P<0.05). Compared with the ctrl group, the SOR group exhibited notable changes in multiple indicators, including a significant increase in lactate dehydrogenase(LDH) leakage rate, LC3-related indices, and the expression levels of various autophagy-related proteins, as well as a significant decrease in mitochondrial membrane potential and p62 protein(p62) expression(P<0.05). Following COS administration, these indices were significantly reversed in the SOR+low-COS and SOR+high-COS groups(P<0.05). Further addition of rapamycin and AICAR resulted in changes opposite to those observed in the SOR+high-COS groups(P<0.05). Animal experiments exhibited a similar trend: the SOR group showed significant differences in multiple cardiac functional parameters and protein expression levels compared with the ctrl group, which were ameliorated by COS treatment. Upon further addition of rapamycin and AICAR, both cardiac functional indices and protein expression levels displayed trends opposite to those in the SOR+high-COS group(P<0.05). In conclusion, COS inhibits excessive mitophagy in cardiomyocytes and exerts a protective effect against SOR-induced cardiomyocyte injury, a mechanism potentially mediated through suppression of the AMPK/ULK1/FUNDC1 signaling pathway.

Postmenopausal osteoporosis (PMOP) arises from a bone remodeling imbalance in which osteoclast-mediated resorption exceeds osteoblast-driven formation. However, current therapies primarily target osteoclasts without restoring osteoblast function, underscoring the need for mechanism-driven dual-action approaches. Here, we identify neddylation as a post-translational modification that reciprocally regulates osteoclast and osteoblast differentiation. In bone tissue from patients with PMOP, NEDD8 and its activating enzyme NAE1 were significantly upregulated, indicating heightened pathway activity. Mechanistically, c-Cbl neddylates NFATc1 to promote osteoclastogenesis, whereas Rbx1 neddylates Runx2 to suppress osteoblast maturation. Pharmacological blockade with the NAE1 inhibitor MLN4924 counteracted both effects and, in ovariectomized mice, reduced osteoclast activity while preserving osteoblast function, preventing deterioration of bone mass and microarchitecture. In human PMOP bone tissues, elevated NEDD8 and NAE1 expression was associated with increased NFATc1 in osteoclast-lineage cells and reduced Runx2 in osteoblast-lineage cells, suggesting translational relevance. These findings reveal neddylation as a unifying mechanism that coordinates opposing effects on osteoclast and osteoblast master transcription factors and highlight its potential as a mechanism-driven dual-action therapeutic target.

Postmenopausal osteoporosis(PMOP) is triggered by estrogen deficiency. Insufficient estrogen lead to bone resorption exceeding bone formation, thereby causing decreased bone mass. Studies have confirmed that the total flavonoids extract from Epimedii Folium(TFE) can ameliorate osteoporosis, while the bone nutrient consisting of chondroitin sulfate(CS) combined with glucosamine hydrochloride(GLcN) exerts therapeutic effects on bone metabolism-related diseases. Since osteoporosis is fundamentally characterized by metabolic disorders, the combination of TFE-bone nutritional supplement(TG) may exert synergistic multi-target regulatory effects, thereby offering a new approach for the prevention and control of PMOP. This study systematically explored the ameliorating effects and mechanisms of TG on PMOP through in vivo and in vitro experiments combined with metabolomics technology. In vivo, a rat model of OVX was established by bilateral ovariectomy. The pharmacodynamic effects of TG on PMOP were verified based on bone histopathological staining, bone mineral density, and physiological indicators. In addition, metabolomics technology was used to quantify urine metabolites to explore the potential biomarkers of PMOP regulated by TG and the associated metabolic networks. In vitro, MC3T3-E1 cells were treated with TFE and TG to screen safe drug doses. Subsequently, osteogenic induction was performed, and the effects and mechanisms of TG on the osteogenic differentiation of MC3T3-E1 cells were studied by ALP staining, ARS staining, and Western blot analysis. Pharmacodynamic results showed that TG reduced the generation of osteoclasts and significantly restored bone mineral density and physiological indicator levels. Metabolomic results indicated that TG alleviated bone loss mainly by intervening with multiple metabolic pathways, including nicotinate and nicotinamide metabolism, histidine metabolism, pyrimidine metabolism, tryptophan metabolism, alanine, aspartate and glutamate metabolism, purine metabolism, vitamin B6 metabolism, etc. In vitro experiments confirmed that TG promoted the proliferation of osteoblasts and osteogenic differentiation via the bone morphogenetic protein-2(BMP-2)/Sma-and Mad-related protein(Smad) signaling pathway. In summary, TG can effectively ameliorate PMOP through a multi-target synergistic effect. This study provides an experimental basis and theoretical support for the clinical application of TG.

This paper constructed a reactive oxygen species(ROS)-responsive hydrogel loaded with sodium tanshinone &#x2161;_A sulfonate(STS) and salvianolic acid B(SAB) and evaluated its therapeutic effect on hypertrophic scars. A hydrogel was prepared by using hyaluronic acid(HA) and polyvinyl alcohol(PVA) bridged by phenylboronic acid(PBA) as the matrix with two drugs physically encapsulated inside. Its structure, rheological properties, and drug release behavior were characterized. Scar models of rabbit ears were established and divided into a blank control group, a model group, an asiaticoside cream group, a regular drug-loaded hydrogel group, and a drug-loaded ROS-responsive hydrogel group. After 21 days of intervention, the net increased thickness of the car was detected. The tissue morphology was detected by using hematoxylin-eosin and Masson staining. The expressions of &#x3b1;-smooth muscle actin(&#x3b1;-SMA) and collagen-&#x2160;(COL-&#x2160;) were detected by immunohistochemistry, and the levels of transforming growth factor-&#x3b2;1(TGF-&#x3b2;1), tissue inhibitor of metalloproteinases-1(TIMP-1), matrix metallopeptidase-9(MMP-9), ROS, and 8-hydroxy-2'-deoxyguanosine(8-OHdG) were assessed. The results show that, compared to those in the model group, the net increased thickness of scar, TGF-&#x3b2;1, TIMP-1, ROS, 8-OHdG, &#x3b1;-SMA, and COL-&#x2160; in each treatment group were significantly reduced(P<0.05), while MMP-9 levels were increased(P<0.05), and the improvement effect in the drug-loaded ROS-responsive hydrogel group was better than that in the regular drug-loaded hydrogel group(P<0.05). These findings indicate that the hydrogel can achieve smart drug release and significantly inhibit scar formation, providing a new strategy for treatment.

This study investigated the therapeutic efficacy of Sijunzi Decoction on chronic atrophic gastritis(CAG) in rats and its potential mechanism of action in mitigating gastric mucosal injury through the "gut microbiota-hepcidin-ferroptosis" pathway. Specific pathogen-free(SPF) grade male Wistar rats were randomly divided into a normal group, a model group, a positive drug vatacoenayme group, and low-, medium-, and high-dose Sijunzi Decoction groups. A CAG model was established using a composite modeling method combining multiple pathogenic factors with irregular feeding. Gut microbiota composition and functional changes were analyzed by 16S rRNA high-throughput sequencing. Hepcidin expression in gastric tissues was assessed by immunofluorescence. Western blot was performed to measure the expression of ferroportin 1(FPN1) and ferroptosis-related proteins, including glutathione peroxidase 4(GPX4) and solute carrier family 7 member 11(SLC7A11). The levels of ferrous iron(Fe~(2+)), malondialdehyde(MDA), superoxide dismutase(SOD), and glutathione peroxidase(GSH-Px) were determined by colorimetric assay. Histopathological alterations in gastric tissue were observed via hematoxylin-eosin(HE) staining. The 16S rRNA sequencing results indicated a significant gut microbiota dysbiosis in the model group compared to the normal group. The model group exhibited significant enrichment of Prevotella, Allobaculum, Bacteroides, Enterococcus, and members of Enterobacteriaceae, whereas the normal group was relatively enriched with Turicibacteraceae/Turicibacter, Roseburia, and Veillonellaceae. Following pharmacological intervention, the vitaminazyme group showed enrichment of Clostridiaceae and Bifidobacterium. Distinct microbial signatures were observed across different doses of Sijunzi Decoction: the low-dose group was enriched with Bacteroides, Prevotella, and Veillonellaceae; the medium-dose group was enriched in Turicibacter and Actinobacteria; and the high-dose group was enriched with Sutterella, Allobaculum, and Blautia. Overall, the microbiota structure in all treatment groups shifted back towards that of the normal group compared to the model group. Functional prediction indicated that Sijunzi Decoction could upregulate metabolic pathways related to the degradation of aromatic compounds. In gastric tissues, the model group exhibited significant iron overload, oxidative stress, and ferroptosis activation, manifested as elevated Fe~(2+) and MDA levels, reduced GSH-Px and SOD activities, upregulated hepcidin expression, and markedly downregulated FPN1 and the key ferroptosis proteins GPX4 and SLC7A11. Sijunzi Decoction intervention effectively reversed these changes, restored iron homeostasis and redox balance, significantly upregulated the expression of FPN1, GPX4, and SLC7A11, and notably ameliorated gastric mucosal atrophy, glandular structural damage, and inflammatory cell infiltration in CAG rats. In summary, Sijunzi Decoction may improve gastric tissue injury and oxidative stress in CAG by remodeling the structure and function of the gut microbiota, downregulating hepcidin expression, promoting iron export, and inhibiting ferroptosis.

Understory ecological cultivation of Chinese medicinal herbs is a crucial component of China's national strategy for developing forest-based economies, yet it frequently faces challenges such as low yields and high production costs. This review synthesizes comparative studies on the active constituent content of nine medicinal herbs-Panax ginseng, P. quinquefolius, P. notoginseng, Polygonatum, Dendrobium, Paris polyphylla, Coptis chinensis, Bletilla striata, and Epimedium-between understory cultivation and field cultivation. The findings indicate that Dendrobium in understory cultivation systems generally exhibit higher levels of polysaccharides than greenhouse-grown counterparts. For C. chinensis, alkaloid content shows no significant difference between understory cultivation and shaded field cultivation. Extended growth periods enable P. ginseng and P. quinquefolius to achieve total and rare ginsenoside concentrations surpassing those of short-cycle cultivated counterparts. The quality of Polygonatum, P. notoginseng, P. polyphylla, and B. striata is influenced by forest stand type and shade intensity, with no clear patterns yet established. Flavonoid accumulation in Epimedium is sensitive to canopy closure, exhibiting an optimal light-intensity range. Moreover, understory cultivation enhances soil microbial diversity and reduces the abundance of certain phytopathogens, suggesting potential ecological pest/disease-suppression effects, although direct evidence regarding pest/disease incidence rates and agrochemical residues remains lacking. Overall, the impact of understory cultivation on medicinal herb quality is species-specific and environmentally contingent, with quality advantages likely arising from moderate abiotic stress, extended growth duration, and enhanced biodiversity. Future research should standardize experimental designs, quantify key ecological factors, and deepen mechanism understanding of forest-medicinal herb interactions to support high-quality industry development.

This study systematically investigated the mechanism of Scutellariae Radix against atherosclerosis(AS) through the integrated application of network pharmacology, molecular docking, and animal experiments. A total of 34 active ingredients of Scutellariae Radix were screened from the TCMSP database, and 65 potential therapeutic targets were identified by combining SwissTargetPrediction and GEO data analysis. Protein-protein interaction(PPI) network construction, molecular docking, and Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analyses indicated that Scutellariae Radix might exert the anti-AS effect by modulating inflammatory responses, negatively regulating pyroptosis, and influencing the NOD-like receptor(NLRP) signaling pathway. In vivo, AS was induced in ApoE~(-/-) mice by 16-week high-fat diet feeding, followed by intervention with Scutellariae Radix extract. The results showed that Scutellariae Radix significantly attenuated lipid deposition and plaque area in the aortic wall, decreased serum levels of total cholesterol(TC), low-density lipoprotein cholesterol(LDL-C), and triglycerides(TG), and increased the high-density lipoprotein cholesterol(HDL-C) level, thereby improving lipid metabolism disorders. Meanwhile, it markedly inhibited the expression of proinflammatory cytokines tumor necrosis factor-&#x3b1;(TNF-&#x3b1;), interleukin(IL)-1&#x3b2;, and IL-6. In addition, Scutellariae Radix significantly suppressed nuclear factor-&#x3ba;B(NF-&#x3ba;B) phosphorylation and downregulated the protein expression of NOD-like receptor pyrin domain-containing protein 3(NLRP3), cleaved caspase-1, and the N-terminal fragment of gasdermin D(N-GSDMD), which suggested that Scutellariae Radix effectively inhibited pyroptosis in arterial tissues. Collectively, these findings suggested that Scutellariae Radix ameliorated AS by suppressing pyroptosis in arterial tissues, attenuating inflammation, and improving lipid metabolism via regulating the NLRP3/caspase-1/GSDMD signaling pathway.

Soil salinization seriously threatens global food security with continuously expanding affected areas. Bacillus megaterium-based bioinoculant serves as an eco-friendly strategy for saline-alkali land remediation, whereas wild-type (WT) strains generally exhibit poor salt-alkali tolerance. To address this limitation, atmospheric and room temperature plasma (ARTP) mutagenesis combined with a microbial microdroplet culture (MMC) high-throughput screening system was adopted to screen high salt-tolerant B. megaterium mutant followed by multi-omics characterization. Using WT B. megaterium LD (CGMCC No.21828) as the original strain, a mutant library was constructed via optimized ARTP treatment, and the elite mutant SLD48 was isolated under dynamically increasing salt gradients. Compared with the wild type, SLD48 possesses a larger genome of 6,252,538&#xa0;bp and acquires 28 extra functional genes including spoIVCA and pJ_gene0003. Intracellular osmoprotectants (proline, glycine betaine) and antioxidant &#x3b1;-tocopherol were markedly accumulated in SLD48. moreover, exogenous supplementation of these substances in the culture medium can effectively alleviate the damage caused by salt stress to the cells. In pot trials with saline-alkali soil, the rhizosphere colonization of SLD48 reached 3.52&#x2009;&#xd7;&#x2009;10&#x2075; CFU/g (2.46-fold of LD), and maize shoot fresh weight increased by 40.5% after inoculation. Collectively, this work confirms that mutant SLD48 is a promising microbial resource for saline-alkali soil improvement and crop yield promotion.

This study aimed to investigate the effects of Zhenwu Decoction(ZWT) on cardiac fibrosis in mice with heart-kidney Yang deficiency-induced chronic heart failure(CHF). The research further explored the therapeutic mechanisms of ZWT in CHF treatment in the hope of providing novel insights for TCM approaches to managing heart-kidney Yang deficiency-induced heart failure. Sixty C57BL/6 mice were randomly divided into the following groups: the control group(normal untreated mice),the DOX group(doxorubicin-induced CHF model),the low-dose ZWT group(ZWT-L,4.7 g&#xb7;kg~(-1)),the medium-dose ZWT group(ZWT-M,9.3 g&#xb7;kg~(-1)),the high-dose ZWT group(ZWT-H,18.6 g&#xb7;kg~(-1)),and the dopamine group(DA,positive control). The CHF mouse model was established over a 4-week period, which was followed by an additional 4-week treatment regimen. After 8 weeks, the spontaneous locomotor activity of mice was assessed, and TCM syndrome scoring was performed. Mouse serum samples were collected and analyzed to measure the levels of cardiac-specific biomarkers including cardiac troponin &#x2160;(cTn-&#x2160;),brain natriuretic peptide(BNP), creatine kinase-MB isoenzyme(CK-MB), creatine kinase(CK),lactate dehydrogenase(LDH),triiodothyronine(T3),succinate dehydrogenase(SDH) and myocardial cyclic adenosine monophosphate(cAMP). Cardiac tissue was collected for pathological examination. Western blot and real-time quantitative polymerase chain reaction(RT-PCR) were used to detect the expression of myocardial cAMP-dependent protein kinase catalytic subunit(PKA C),protein kinase A(PKA),ryanodine receptor 2(RyR2),phospholamban(PLB),B-cell lymphoma-2(Bcl-2),caspase-3,cleaved caspase-3 and Bcl-2-associated X protein(Bax). The experimental results indicated that, compared with the control group, the DOX group exhibited significantly elevated TCM syndrome scores accompanied by decreased heart rate, reduced body weight, and increased cardiac index(P<0.05); serum levels of CK, CK-MB, BNP, cTn-&#x2160;, LDH and myocardial cAMP were significantly increased(P<0.05), while SDH and T3 levels were decreased(P<0.05); meanwhile, myocardial interstitial fibrosis was aggravated and myocardial hypertrophy appeared; the expression of PKA C, PKA, caspase-3, cleaved caspase-3, Bax and phosphorylation of RyR2 were increased(P<0.05), the expression of Bcl-2 and phosphorylation of PLB were decreased compared with the DOX group, the ZWT treated groups demonstrated reduced TCM syndrome scores, increased heart rate, improved body weight, and decreased cardiac index. Additionally, serum levels of CK, CK-MB, BNP, cTn-&#x2160;, LDH and myocardial cAMP were decreased(P<0.05), while SDH and T3 levels were increased(P<0.05); myocardial hypertrophy and fibrosis were also improved; the expression of PKA C, PKA, Bax, caspase-3, cleaved caspase-3 and phosphorylation of RyR2 was reduced(P<0.05), and the expression of Bcl-2 and phosphorylation of PLB were increased(P<0.05). These findings suggested that ZWT may exert therapeutic effects on CHF by regulating sarcoplasmic reticulum calcium-related proteins, reducing apoptosis, and improving cardiac function.

This study aimed to investigate the effects of loganin on the osteogenic differentiation of mouse embryonic osteoblast precursor cells(MC3T3-E1) under oxidative stress and to explore its underlying mechanisms. First, a network pharmacology approach was employed to identify common targets of loganin and osteoporosis(OP) using SwissTargetPrediction, OMM, and GeneCards databases. A total of 31 potential targets were identified, with signal transducer and activator of transcription 3(STAT3), interleukin-2(IL-2), matrix metalloproteinase-9(MMP-9), and caspase-3 being central in the protein-protein interaction(PPI) network. Further enrichment analysis based on Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) suggested that the JAK-STAT signaling pathway might be a key pathway through which loganin exerts its effects. In vitro experiments utilized hydrogen peroxide(H_2O_2, 200 &#x3bc;mol&#xb7;L~(-1)) to establish an oxidative stress injury model in MC3T3-E1 cells. The cells were divided into a control group, an H_2O_2 model group, and low-, medium-, and high-dose(50, 100, 200 &#x3bc;mol&#xb7;L~(-1)) loganin groups. The results showed that loganin significantly reversed the H_2O_2-induced decreases in cell viability, reduced intracellular reactive oxygen species(ROS) levels, enhanced alkaline phosphatase(ALP) activity, and inhibited apoptosis. Western blot analysis revealed that loganin upregulated the expression of osteogenic differentiation markers, including Runt-related transcription factor 2(Runx2), ALP, and osterix, while downregulating the expression of apoptosis markers such as cleaved caspase-3 and Bcl-2-associated X protein(Bax) and upregulating the anti-apoptotic protein B-cell lymphoma-2(Bcl-2). Furthermore, loganin inhibited the phosphorylation of the JAK2/STAT3 pathway. Notably, the promoting effect of loganin on osteogenic differentiation and its anti-apoptotic effect were significantly reversed upon treatment with a JAK2/STAT3 pathway agonist, further confirming the critical role of this pathway in its mechanism of action. In conclusion, this study suggests that loganin may alleviate oxidative stress by inhibiting the JAK2/STAT3 signaling pathway, thereby suppressing apoptosis and promoting osteogenic differentiation in MC3T3-E1 cells.

This study aimed to investigate the ameliorative effects and underlying mechanisms of the combined use of Platycodon grandiflorum and Sonchus oleraceus on pulmonary nodules in mice. C57BL/6J mice were randomly divided into blank group, model group, positive control group, P. grandiflorum extract group, S. oleraceus extract group, and combination group. The pulmonary nodule model was established by intratracheal instillation of 1&#xd7;10~9 CFU Cutibacterium acnes for three consecutive days, followed by seven consecutive days of drug administration. Serum amyloid A(SAA) levels were measured, pathological changes in lung tissue were observed, and changes in interleukin(IL)-6, tumor necrosis factor-&#x3b1;(TNF-&#x3b1;), IL-1&#x3b2;, glutathione peroxidase(GSH-PX), superoxide dismutase(SOD), total antioxidant capacity(T-AOC), and malondialdehyde(MDA) levels in lung tissue were evaluated. Additionally, macrophage M1/M2 polarization and activation of the Toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)/nuclear factor-&#x3ba;B p65 subunit(NF-&#x3ba;B p65) signaling pathway were analyzed. The results showed that all treatment groups ameliorated lung injury in mice with pulmonary nodules. Specifically, the combination group alleviated body weight loss, reduced SAA levels, downregulated the expression of IL-6, TNF-&#x3b1;, and IL-1&#x3b2;, enhanced the activities of GSH-PX, SOD, and T-AOC, decreased MDA accumulation, restored the balance of M1/M2 macrophage phenotypes, upregulated arginase-1(ARG1) levels, suppressed the mRNA expression of inducible nitric oxide synthase(iNOS), TLR4, MyD88, and NF-&#x3ba;B, increased the expression of inhibitor of nuclear factor-&#x3ba;B alpha(I&#x3ba;B&#x3b1;), and downregulated the p-p65/p65 ratio. Meanwhile, compared with the single-herb groups, the combination group demonstrated significant advantages in suppressing inflammatory cytokines and oxidative damage. In conclusion, the combined preparation of P. grandiflorum and S. oleraceus can alleviate pulmonary inflammation and oxidative stress injury and improve pulmonary nodules by inhibiting the NF-&#x3ba;B signaling pathway and suppressing macrophage M1 polarization.