This study aimed to quantitatively investigate the role of marginal chordae of the mitral valve during static closure by analyzing the force within these chordae and its potential correlation with the transvalvular pressure. Porcine (n = 10) and human (n = 4) mitral valves were mounted in a saddle-shaped annulus clamp, while an in vitro system was utilized to replicate a physiological systolic blood pressure by vacuum pressure applied to the atrial side of the valve. A Millar pressure catheter and force transducers (“C-gauges”) were used to acquire the data. Strut chordal force was recorded simultaneously with the marginal chordal force, to serve as a verification of the employed equipment. The marginal chordal force in porcine and human averaged to 0.012 N ± 0.008 N and 0.006 N ± 0.008 N respectively at peak systolic pressure of 120 mmHg. Additionally, a simplified finite element study was conducted and supports low marginal forces relative to intermediary chords. This low force calls into question the traditionally-described role of the marginal chordae in preventing mitral prolapse at closure. The chords do, however, still show signs of assisting with the leaflets’ positioning. The relationship between marginal chordal forces and transvalvular pressure varies significantly across experiments, further obfuscating their role in normal valvular function.
Neuroinflammation is a key driver and an important therapeutic target of neurodegeneration. Dysregulated nuclear factor-κB (NF-κB) signaling plays a central role in neuroinflammation and depends on M1 and K63 ubiquitination of NF-κB essential modulator (NEMO). Ivangustin (IVA) is a sesquiterpene lactone isolated from Inula japonica Thunb., a traditional Chinese medicine has been used to treat inflammatory diseases. But its anti-neuroinflammatory effects and molecular targets remain unclear. This study aimed to evaluate the anti-neuroinflammatory effects of IVA and elucidate its direct functional target and underlying mechanism. Integrated in vitro, in vivo, and in silico/bioinformatics approaches were employed. The anti-neuroinflammatory activity of IVA was evaluated in LPS-stimulated BV2 cells, primary microglia, and an LPS-induced acute cognitive impairment mouse model. Mechanistic studies were performed using pull-down, liquid chromatography-tandem mass spectrometry (LC–MS/MS), cellular thermal shift assay (CETSA), drug affinity responsive target stability (DARTS), gene knockdown, point mutation and bioinformatics analyses. In vitro, IVA significantly inhibited nitric oxide (NO) production and mRNA and protein expression of pro-inflammatory factors in LPS-stimulated microglia. Furthermore, IVA significantly suppressed inhibitor of NF-κB kinase α/β (IKKα/β) and inhibitory subunit of NF-κB alpha (IκBα) phosphorylation, IκBα degradation, and NF-κB p65 phosphorylation and nuclear translocation, thereby blocking the expression of NF-κB-targeted genes. Its anti-neuroinflammatory activity was attenuated by thiol donors, indicating the importance of its electrophilic lactone moiety. Furthermore, ubiquitin conjugating enzyme E2L3 (UBE2L3) was identified as the functional target of IVA in BV2 cells, whereas UBE2L3 knockdown markedly attenuated IVA-mediated anti-neuroinflammatory action. Bioinformatics analysis suggests UBE2L3 overexpression as a potential risk factor for neurodegenerative diseases. Mechanistically, IVA covalently bound to the catalytic cysteine 86 of UBE2L3, thereby inhibiting UBE2L3-mediated M1 and K63 ubiquitination of NEMO, which is crucial for NF-κB activation. In vivo, IVA ameliorated LPS-induced cognitive impairment and neuroinflammation in mice via inhibiting Iba1 expression and suppressing NF-κB activation. IVA exerts anti-neuroinflammatory effects by covalently targeting UBE2L3 and suppressing NF-κB signaling. These findings also identify UBE2L3 as a potential therapeutic target for neuroinflammation and support IVA as a promising lead compound for further preclinical investigation of neuroinflammatory disorders.
Traditional Chinese Medicine (TCM) clinical practice depends on both codified theoretical knowledge and practitioner-specific experience, which differ in their data sources, reasoning patterns, and scope of generalization. We developed DFGLM-TCM, a modular large language model-based service system that separately models these two knowledge types through task-oriented components and coordinates them within a unified multi-task architecture. The knowledge-oriented component integrated a curated 22-GB TCM corpus containing approximately 3 million structured entries, a manually validated knowledge graph with more than 200,000 entities, and retrieval-augmented generation for literature retrieval and general TCM question answering. The experience-oriented component was trained using approximately 5000 authentic outpatient records from a senior TCM practitioner and 2000 expert-reviewed augmented cases to provide practitioner-specific diagnostic and prescription references. Through standardized interfaces and role-based access, the system supports knowledge retrieval, question answering, structured consultation, and prescription reference for clinicians, patients, and students. In an expert-rated evaluation of 100 TCM knowledge questions, DFGLM-TCM achieved the highest descriptive mean score among the evaluated models (4.48 ± 0.76). In 454 independent pulmonary-nodule cases, the experience-oriented component achieved a prescription-consistency score of 8.95 ± 0.63, exceeding that of the model additionally trained with general TCM knowledge (7.52 ± 0.44). A one-month assessment involving 35 physicians at three primary-care institutions suggested favorable short-term usability and acceptance. These findings highlight the value of separating general TCM knowledge from practitioner-specific experience while coordinating task-oriented training and multi-task services within a unified system, and support further evaluation of DFGLM-TCM as an auxiliary reference tool in broader clinical settings.
Traumatic brain injury (TBI) is associated with high morbidity, disability, and long-term neurological sequelae. Mitochondrial dysfunction is a central component of secondary injury after TBI, contributing to impaired energy metabolism, oxidative stress, calcium dysregulation, neuroinflammation, and neuronal apoptosis. This review summarizes current evidence regarding acupuncture-mediated regulation of mitochondrial homeostasis after TBI, focusing on structural homeostasis, quantitative homeostasis, and functional/metabolic homeostasis. We distinguish direct evidence from TBI models, indirect evidence from related brain injury models, and hypothesis-generating mechanisms. Particular attention is given to mitochondrial dynamics, mitochondrial biogenesis, mitophagy, energy metabolism, oxidative stress, calcium signaling, mitochondrial membrane potential, apoptosis, intercellular mitochondrial transfer, and putative upstream neural and humoral pathways. Current preclinical evidence suggests that acupuncture, particularly electroacupuncture, may influence mitochondrial homeostasis after TBI through multidimensional and context-dependent mechanisms. However, direct TBI-specific evidence remains limited, and several proposed mechanisms require further validation before acupuncture-mediated mitochondrial regulation can be considered a clinically established therapeutic strategy.
Aortic stenosis (AS) develops from calcific aortic valve disease (CAVD), which narrows the aortic valve opening as leaflet stiffness increases due to calcium deposition. This study extends the Reverse Calcification Technique (RCT) by incorporating a time dimension to develop a quantitative parametric model for patient-specific prediction of CAVD progression from sequential CT scans. Seventeen pre-transcatheter aortic valve replacement (TAVR) patients underwent sequential CT scans (1.2–6.5 years); baseline aortic valve calcification (AVC) volumes: ≈ 250 to ≈ 1,600 mm3 (cohort mean ≈ 730 mm3 at the first scan and ≈ 920 mm3 at the follow-up scan). A parametric model was developed using two approaches: forward prediction (mild to severe stages) and backward reconstruction (from severe to moderate stages). 34 test cases were assessed through alternating calibration and verification, with performance evaluated using Bland–Altman analysis, paired t-tests, and relative error calculations. For scan intervals under 3 years, forward prediction achieved a mean absolute error of 77 mm3 (7.0
Yiqi Huoxue Jiedu Formula (YHJF) is a traditional Chinese medicine formula that has been used as an adjunctive therapy for sepsis for nearly two decades. Previous clinical studies showed that YHJF improves Sequential Organ Failure Assessment (SOFA) scores and modulates gut microbiota in elderly patients with pneumonia-associated sepsis. However, the mechanism by which YHJF protects against sepsis-associated acute lung injury (SALI) remains unclear. A murine SALI model was established by cecal ligation and puncture (CLP). Therapeutic effects were evaluated by histopathology, micro-CT, pulmonary function assessment, and ELISA. Mechanistic studies included proteomic analysis of lung tissues and LPS-stimulated MH-S macrophages, pharmacological modulation with Mdivi-1 and urolithin A (UA), macrophage-epithelial co-culture, HPLC fingerprinting, UPLC-HRMS, and molecular docking. YHJF significantly improved 7-day survival and ameliorated lung injury, pulmonary edema, respiratory dysfunction, and systemic inflammation in mice with CLP-induced SALI. Proteomic profiling and subsequent functional assays suggested that enhanced mitophagy in macrophages represents a central protective mechanism. In vivo, YHJF increased autophagosome formation and PINK1/Parkin co-localization in BALF-derived alveolar macrophages. In vitro, YHJF restored mitochondrial homeostasis by activating PINK1/Parkin-dependent mitophagy in macrophages. This was accompanied by reduced cytoplasmic mtDNA leakage, downregulated cGAS expression, and suppression of the STING–TBK1–IRF3 pathway and subsequent type I interferon responses. Pharmacological inhibition of mitophagy with Mdivi-1 abolished these protective effects of YHJF, whereas activation with UA augmented them, demonstrating that mitophagy is necessary for YHJF-mediated protection. In a macrophage–epithelial co-culture system, YHJF-treated macrophages alleviated LPS-induced apoptosis in MLE-12 alveolar epithelial cells. Furthermore, chemical analysis integrated with molecular docking identified aloe-emodin, rhein, and genistein as candidate bioactive constituents of YHJF that likely contribute to its regulation of macrophage mitophagy. YHJF protects against SALI by restoring macrophage mitophagy and suppressing mtDNA-STING-mediated inflammatory signalling. These findings support YHJF as a potential therapeutic strategy for sepsis-associated lung injury.
Cardiovascular health depends critically on the integrity of the vascular extracellular matrix (ECM) and the behavior of vascular smooth muscle cells (VSMCs), both of which can be adversely affected in vascular diseases. This study quantitatively evaluates the therapeutic potential of conditioned medium (CM) derived from bone marrow (BM-MSCs) and adipose-derived stem cells (ADSCs) in an elastase-injured human aortic smooth muscle cell (HASMC) model. We systematically varied seeding densities (2000, 5000, and 10,000 cells/cm2) and serum conditions to optimize the SC-SMC secretome for vascular repair. Our results indicate that neither BM-SMC nor AD-SMC CM significantly enhanced lysyl oxidase (LOX) activity. In fact, serum-supplemented BM-SMC CM significantly suppressed LOX activity at seeding densities of 2000 cells/cm2 (p = 0.0378) and 10,000 cells/cm2 (p = 0.0080) compared to injured untreated controls. High-density AD-SMC CM (10,000 cells/cm2) also resulted in a significant decrease in elastin levels (p < 0.05). In addition, serum presence was critical for maintaining the reparative phenotype. Serum-free (SF) conditions for both cell sources led to widespread, statistically significant reductions (p < 0.0001) in key repair and inflammatory biomarkers, including PDGF-AA, Leptin, Lipocalin-2, Osteopontin, RBP4, MMP-1, and IL-6. IL-11 emerged as a primary discriminatory biomarker, showing significant differences between BM and AD treatments at high seeding densities, with both sources causing a significant decrease (p < 0.0001) compared to injured untreated controls. These findings demonstrate that seeding density and serum conditions are critical variables that quantitatively modulate the efficacy of SC-SMC-CM. The study highlights that BM-SMC-derived CM offers a more stable platform for elastin maintenance under serum-free conditions, providing a foundation for developing tailored, cell-free regenerative therapies for cardiovascular disease.
Abstract Diabetic wounds represent one of the most intractable complications of diabetes, arising from a highly complex and self-perpetuating pathological microenvironment. This milieu is characterized by chronic inflammation, oxidative stress, impaired angiogenesis, peripheral neuropathy, immune dysregulation, recurrent infection, and marked dysfunction of resident cells and the extracellular matrix (ECM). These intertwined abnormalities not only hinder wound closure but also significantly increase the risk of chronic infection, amputation, and disability, thereby imposing substantial clinical and socioeconomic burdens. Conventional therapies, which typically target isolated aspects of wound pathology, often fail to disrupt the vicious cycle of non-healing, underscoring the urgent need for integrated, multi-target therapeutic strategies. Chinese herbal medicine (CHM) encapsulates a holistic therapeutic paradigm that is intrinsically aligned with the multi-factorial nature of diabetic wound management. CHM offers a diverse array of bioactive ingredients, including polysaccharides, saponins, flavonoids, alkaloids, and phenolics, that collectively exert anti-inflammatory, antioxidant, pro-angiogenic, neuroprotective, immunomodulatory, antimicrobial, and pro-proliferative effects. Moreover, many of these ingredients exhibit pleiotropic effects, enabling concomitant modulation of multiple pathological pathways to address the inherent complexity of diabetic wounds. However, the clinical translation of raw CHM is frequently hindered by poor bioavailability and rapid degradation at the wound site. Recent advances in CHM-based innovative delivery platforms have provided a transformative framework to overcome these limitations. This review highlights the emergence of CHM-loaded hydrogels and CHM-derived plant exosome-like nanovesicles (PELNs) as next-generation interventions. CHM-loaded hydrogels act as biomimetic scaffolds that facilitate controlled release and adaptively respond to microenvironmental cues (e.g., pH and reactive oxygen species (ROS)). Simultaneously, CHM-derived exosome-like nanovesicles serve as natural, biocompatible nanocarriers that enhance cellular uptake and molecular signaling, promoting regenerative healing through intercellular communication pathways. In this review, we systematically summarize the pathological features of diabetic wounds and synthesize the modern pharmacological mechanisms of representative CHM ingredients. We further evaluate the synergistic integration of CHM with advanced biomaterials and nanotechnology, emphasizing their translational potential from preclinical models to clinical evidence. By bridging traditional wisdom with cutting-edge bioengineering, these platforms offer a robust evidence-based framework for the modernized management of diabetic wounds.
The NOTCH1 gene and its signaling pathway play a critical role in the oncogenesis and progression of various carcinomas through complex cellular and molecular mechanisms. This integrative literature review examines 21 selected studies across multiple carcinoma types, including head and neck, prostate, penis, breast, and hepatocellular carcinomas, to elucidate the functional impact of notch1 alterations on tumor behavior and clinical outcomes. NOTCH1 functions as a context-dependent regulator, acting either as an oncogene or tumor suppressor according to cellular environment and molecular context. Aberrations in notch1 signaling influence cell proliferation, differentiation, apoptosis, and angiogenesis, thereby contributing to cancer initiation and progression. Despite some variability in findings, the majority of studies indicate that notch1-related molecular changes have significant implications for prognosis and potential therapeutic targeting. This review highlights the importance of understanding notch1 signaling pathways in the cellular and molecular biology of carcinomas, aiming to pave the way for novel diagnostic and treatment strategies.
Colon cancer is one of the most prevalent cancers globally, characterized by the abnormal growth of cells in the intestines. Numerous studies have explored the effects of pomegranate-derived products, such as pomegranate seed oil (PSO), as anti-proliferative, anti-invasive, and pro-apoptotic agents against various cancer cell lines. Additionally, previous research has highlighted the anti-cancer properties of low-frequency electromagnetic fields (LF-EMF). In the present study, we investigated the combined effects of these two factors on the expression of the Caspase 3, Caspase 9, BAX, and Bcl2 genes. Human colon cancer cells of the HT29 line were sourced from the Pasteur Institute of Iran cell bank and maintained in a complete culture medium. The cells were categorized into four groups: control, PSO, EMF, and PSO+EMF. To assess cell viability and the type of cell death, MTT and Annexin V-FITC assays were employed. Changes in the expression levels of Caspase 3, Caspase 9, BAX, and Bcl2 were analyzed using Real-Time PCR. The results from the MTT and Annexin V-FITC assays indicated that both PSO and EMF reduced cell viability and promoted apoptosis in colon cancer cells. The Real-Time PCR results showed an upregulation of Caspase 3, Caspase 9, and BAX genes, along with a downregulation of Bcl2 expression in the treatment groups compared to the control group. This study demonstrated that the combination of PSO and EMF enhances apoptotic gene expression, thereby diminishing the proliferation and viability of cancer cells. Based on these findings, both PSO and LF-EMF exhibit cytotoxic effects on colon cancer cells, suggesting their potential as candidates for future research in the field of colon cancer.
Human topoisomerase 1B (hTopo1B) is a validated anticancer target due to its key role in relieving torsional stress during DNA replication and transcription, as well as its overexpression in rapidly proliferating tumor cells. Camptothecin (CPT) and its derivatives are principal hTopo1B-targeting agents but face challenges including lactone ring instability, dose-limiting toxicities, and acquired drug resistance. This research investigated seven protoberberine alkaloids from Coptis teeta alkaloids-berberine, coptisine, epiberberine, berberastine, jatrorrhizine, palmatine, and fetidine as potential CPT-like hTopo1B inhibitors using comprehensive computational methods. Density functional theory (DFT) calculations showed that six of the seven alkaloids had HOMO-LUMO gaps (2.608-3.015 eV) and electrophilicity indices similar to CPT, suggesting a capacity for charge transfer and DNA intercalation. Molecular docking of the hTopo1B-DNA binary complex (PDB:1A36) revealed that all compounds stabilized the covalent cleavage complex through π-π stacking and hydrogen bonds at the scissile site, with coptisine and epiberberine showing the strongest binding affinities (----10.32 and -10.53 kcal/mol, respectively). Molecular dynamics simulations over 250 ns confirmed the structural stability of the complexes, with low RMSD and RMSF values and minimal fluctuations in the radius of gyration. MM/GBSA and MM/PBSA binding free energy analyses consistently ranked epiberberine as the strongest binder (DG = -18.86 and -14.12 kcal/mol), followed by berberine and coptisine. Per-residue decomposition identified key contacts with the DNA bases DT118, DA17, and DA14, as well as with the protein residues GLU179 and GLY201. All protoberberine analogs showed drug-likeness in ADME profiling, good oral availability, and no PAINS alerts. These findings suggest that protoberberine alkaloids from C.teeta, especially epiberberine, coptisine, and berberine, are promising candidates for next-generation hTopo1B-targeted anticancer therapeutics. Experimental validation is required to confirm the proposed mechanism.
Human Serum Albumin (HSA) is abundant plasma protein in human blood. It plays a critical role in the transport and distribution of endogenous and exogenous compounds. Protein binding affects free fraction of a drug thus affecting pharmacokinetics and therapeutic efficacy. Glycation of HSA under diabetic conditions and the accumulation of uremic toxins in uraemia may alter drug-protein interactions. Therefore, the present study aimed to investigate the binding interactions of ribavirin and selected uremic toxins with binding interactions involving glycation-sensitive HSA residues using molecular docking analysis. Molecular docking results demonstrated that ribavirin bind to HSA with hydrogen bonding and hydrophobic interactions. Key glycation-prone residues, including ARG222, ARG218, and LYS195, were identified at the ribavirin binding site. Several of these residues were also involved in the binding of uremic toxins, indicating potential competition for ligand binding in diabetic and uremic conditions. The overlap of interaction sites suggests that glycation and toxin accumulation may influence the structural and biochemical properties of HSA, thereby affecting ribavirin binding affinity and distribution. In conclusion, this study highlights the important role of glycation-sensitive HSA residues in mediating ribavirin and uremic toxin interactions. These insights may support the future design and optimization of ribavirin-based therapeutics.
Social isolation is an environmental stressor that can adversely affect behavioral function and neuroplasticity-related pathways. Brain-derived neurotrophic factor (BDNF) is one of the key molecules implicated in these changes. Lithium has been reported to influence neuroplasticity-related signaling and may modulate BDNF expression. In this study, we investigated the effects of lithium carbonate on behavioral alterations induced by chronic post-weaning social isolation in male rats and assessed BDNF mRNA expression in the prefrontal cortex. Rats were subjected to social isolation from postnatal day (PND) 21 to PND 70. Lithium carbonate (20 mg/kg, i.p.) was administered once daily during PND 64-70. Behavioral performance was evaluated using the open field test, hot plate test, marble burying test, and forced swim test. Compared with controls, socially isolated rats showed hyperlocomotion, increased rearing, lower pain threshold, altered immobility behavior in the forced swim test, obsessive-compulsive-like behavior, and reduced prefrontal BDNF mRNA expression. Lithium treatment attenuated several isolation-induced behavioral alterations and was accompanied by a partial reversal of reduced prefrontal BDNF mRNA expression toward control levels. Interpretation of molecular findings should be considered preliminary because gene expression analysis was conducted on a limited number of biological samples (n = 3 per group) and only BDNF mRNA, not BDNF protein, was measured. In conclusion, lithium carbonate improved several behavioral abnormalities associated with chronic social isolation and was associated with changes in prefrontal BDNF mRNA expression in male rats. Further studies are needed to clarify the molecular pathways and causal mechanisms underlying these effects.
Crohn's disease (CD) is a chronic inflammatory bowel disorder driven by complex interactions between genetic susceptibility, gut microbiota, and immune dysregulation, typically associated with Th1 and Th17 responses. However, emerging evidence highlights that type 2 allergic mechanisms also significantly contribute to CD pathogenesis. This review examines the cellular and molecular basis of allergic pathways in CD, focusing on IgE-mediated hypersensitivity, mast cell and eosinophil effector functions, and Th2 cytokines (IL-4, IL-5, IL-13). Key epithelial-derived alarmins-namely IL-33, TSLP, and IL-25-are discussed as critical upstream mediators linking mucosal barrier injury to type 2 immune activation in the CD gut. Elevated serum IgE, intestinal mast cell infiltration, and tissue eosinophilia have consistently been documented in CD patients. Mechanistically, FcεRI signaling on mast cells triggers degranulation and release of histamine, proteases, and pro-inflammatory cytokines such as TNF-α and IL-6, leading to increased epithelial permeability and visceral hypersensitivity. Moreover, crosstalk between Th2 and Th1/Th17 pathways further amplifies chronic intestinal inflammation. Understanding these allergic mechanisms opens novel therapeutic avenues, including anti-IgE (omalizumab), anti-IL-4/IL-13 (dupilumab), anti-IL-33 (itepekimab), anti-TSLP (tezepelumab), and mast cell stabilizers (ketotifen). This review integrates current molecular evidence on allergic reactions in CD and highlights their clinical and therapeutic relevance. Recognizing type 2 immune signatures in subsets of CD patients may enable more personalized treatment strategies.
Emerging evidence indicates that mitochondrial dysfunction is not merely a consequence but a driving force in cancer progression. Unlike normal cells, cancer cells rewire their mitochondrial metabolism to support uncontrolled proliferation, a process that includes aerobic glycolysis, persistent reactive oxygen species production, and adaptation to hypoxic conditions. A common hallmark across many tumors is the suppression of the intrinsic apoptotic pathway, primarily achieved through an imbalance between anti-apoptotic and pro-apoptotic BCL-2 family proteins. This evasion of cell death not only facilitates tumor initiation and metastasis but also contributes to resistance against conventional therapies. Here, we provide an overview of major mitochondrial alterations in cancer, with a detailed focus on how the mitochondrial apoptotic machinery is disabled in malignant cells. We then discuss current therapeutic strategies designed to re-activate mitochondria-mediated apoptosis, including BH3 mimetics, direct activators of pro-apoptotic proteins like BAX, immune checkpoint inhibitors, CAR‑T cell therapy, and mitochondria-targeted nanomedicine. Finally, we address the limitations and safety concerns of existing pro-apoptotic drugs and propose future directions to develop more selective and effective cancer treatments. Understanding the molecular mechanisms that govern mitochondrial apoptosis may open new avenues for inducing tumor cell death while minimizing harm to normal tissues.
Telomeres are protected by the shelterin complex and consist of TTAGGG repeats. Their gradual erosion triggers replicative senescence unless counteracted by telomerase or the telomerase-independent Alternative Lengthening of Telomeres (ALT) pathway, utilized by 10-15% of cancers. Accumulating evidence indicates that the decision to activate ALT is intimately linked to the plasticity of telomeric chromatin. This review integrates recent data showing that both constitutive and facultative heterochromatin marks shape ALT activity. We summarize the dynamic regulation of telomeric chromatin and explore the contrasting evidence for two models: the 'open telomeric chromatin model,' where loss of constitutive heterochromatin (e.g., reduced H3K9me3) promotes ALT by increasing chromatin accessibility for homologous recombination (HR) factors, and the 'closed telomeric chromatin model,' where a specific gain of heterochromatic features (e.g., H3K9me3 or H3K27me3) facilitates ALT by creating a specialized phase-separated environment that promotes telomere clustering and break-induced replication (BIR). Resolving this paradox is crucial for understanding ALT initiation and for developing promising synthetic-lethal strategies against ALT-dependent cancers.
Osteoporosis is increasingly linked to metabolic dysregulation. Medicine–food homologous (MFH) materials contain diverse natural bioactives with reported osteoprotective effects, but their overall evidence landscape remains insufficiently integrated. To synthesize preclinical evidence on MFH-derived bioactives that improve osteoporosis-related phenotypes and to discuss their possible trans-organ actions under metabolic disturbance. PubMed, Web of Science, Scopus, and CNKI were searched from January 2000 to September 2025. Original in vivo or in vitro studies were included when a defined MFH-derived constituent or standardized single-material extract was tested in an osteoporosis-relevant model and reported both bone-related and metabolism-related outcomes. A total of 38 bioactive components from 24 MFH materials were identified and regrouped into seven higher-level natural-product categories, including flavonoids, phenolic and polyphenolic compounds, polysaccharides, saponins, terpenoids, proteins/peptides, and other specialized metabolites. Across studies, osteoprotective effects were frequently accompanied by parallel improvements in lipid metabolism, inflammatory status, oxidative stress, gut microbiota, or related metabolites. These findings suggest that MFH bioactives may act beyond bone-local signaling alone. In the discussion, this pattern was further interpreted through liver-bone and gut-bone. MFH-derived bioactives show potential to improve osteoporosis-related phenotypes and may exert broader systemic regulatory effects, although specific mediators and causal links still require validation.
Inflammatory bowel disease (IBD) is a relapsing inflammatory disorder of the gastrointestinal tract with increasing global incidence. Current therapies are often limited by side effects, loss of efficacy, and high cost, underscoring the need for safer and more effective alternatives, particularly multi-target agents derived from natural products. This study aimed to elucidate the protective mechanisms of Yiyi Fuzi Baijiang formula (YFB), a traditional Chinese medicine (TCM) formulation, against dextran sulfate sodium (DSS)-induced acute colitis, focusing on its systemic regulation of the gut barrier–microbiota–metabolism axis. We employed an integrated approach combining network pharmacology, UPLC-Q-TOF-MS/MS-based phytochemical analysis, in vivo evaluation in a DSS-induced colitis mouse model, 16S rRNA gene sequencing, and untargeted metabolomics to assess the effects of YFB and uncover its mechanisms of action. Network pharmacology predicted, and experiments confirmed, that core YFB components (e.g., quercetin, kaempferol) act via IL-17, TNF, and NF-κB pathways. YFB administration dose-dependently improved disease activity index, colon shortening, and histopathology in colitis mice. It restored intestinal barrier integrity by upregulating ZO-1, Occludin, and MUC2, while suppressing pro-inflammatory cytokines (TNF-α, IL-6, IL-1β, IL-17A) and NF-κB activation. Critically, YFB promoted epithelial repair by restoring the expression of intestinal stem cell marker LGR5 and progenitor cell marker SOX9, and by normalizing the aberrant increase in endocrine cell marker CHGA. YFB treatment was associated with reversal of DSS-induced gut microbiota dysbiosis, restoration of diversity, enrichment of beneficial bacteria (e.g., Lachnospiraceae), and suppression of opportunistic pathogens (e.g., Enterobacteriaceae). Untargeted metabolomics showed that YFB treatment was associated with modulation of DSS-altered fecal metabolites (e.g., fatty acids, bile acids) and pathways such as "microbial metabolism in diverse environments". YFB, when administered concomitantly with DSS, protects against DSS-induced colitis via the synergistic effects of its multi-component system. Its mechanism entails systemic regulation of the gut barrier–microbiota–metabolism axis, involving suppression of NF-κB-driven inflammation, promotion of intestinal epithelial repair (via LGR5/SOX9/CHGA modulation), restoration of the intestinal barrier, alterations in gut microbiota, and modulation of host–microbial co-metabolism. These findings provide a scientific basis for YFB's clinical application and highlight the value of TCM formulations in managing complex multi-factorial diseases.
Heart failure with preserved ejection fraction (HFpEF) has currently emerged as a predominant and challenging subtype of heart failure, with high morbidity and mortality. However, the efficacy of current therapeutic strategies for HFpEF remains unsatisfactory. The traditional Chinese medicine formulation Shenfu Qiangxin pill (SFQX) ameliorates clinical symptoms in patients with heart failure, but its precise mechanisms for HFpEF remain to be elucidated. This study aimed to investigate the therapeutic potential of SFQX for HFpEF and to elucidate the mechanisms underlying its effects. UPLC-Q-TOF-MS/MS analysis was performed to identify the major active ingredients of SFQX. The HFpEF mouse model was established using a high-fat diet and an Nω-Nitro-L-arginine methyl ester hydrochloride (L-NAME) to evaluate the therapeutic efficacy of SFQX. We employed an integrated approach combining single-cell RNA sequencing (scRNA-seq) with functional and molecular validation to characterize SFQX-induced changes in cardiac cellular composition, cell-state remodeling, and tissue-level phenotypes in HFpEF. We show that SFQX exerts therapeutic effects against HFpEF by coordinately modulating maladaptive cardiac cell subsets. Specifically, SFQX was associated with reprogramming of pathogenic immune cell polarization, normalization of fibroblast state heterogeneity, enhanced endothelial metabolic adaptability, and restoration of lymphatic endothelial homeostasis. These multicellular changes were accompanied by improved cardiac structure and function, reduced fibrosis and inflammation, enhanced lymphatic drainage capacity, and alleviated myocardial edema. Our findings highlight the ability of SFQX, as a multicomponent agent, to precisely regulate the highly heterogeneous pathology of HFpEF at a network level. This work not only establishes a mechanistic link between holistic principles of traditional medicine and modern biology but also provides a novel theoretical basis for SFQX's efficacy in multifactorial diseases.
