Adverse childhood experiences (ACEs) are a high risk factor for depression and anxiety in adulthood and are associated with the alterations in the hypothalamic‒pituitary‒adrenal (HPA) axis and monoamine neurotransmitters. This study aimed to elucidate the behavioral, neuroendocrine, and neural circuit mechanisms underlying the impact of ACEs effect on adult second-hit stress. We employed a "two-hit" mouse model that combines maternal separation (MS) with restraint stress (RS). To investigate the potential pathological effects of ACEs on depressive- and anxiety-like behaviors in second-hit model mice. The mice were assessed through the behaviors test of sucrose preference test (SPT), tail suspension test (TST), open field test (OFT) and elevated zero maze (EZM). We analyzed the neural circuits in mice by immunohistochemical (IHC) technique, and detected the neuroendocrine changes of the HPA axis and monoamine neurotransmitters by ELISA, flow cytometry (FCM), and high performance liquid chromatography-electrochemical detection (HPLC-ECD). Mice in the RS and MS + RS groups exhibited depressive- and anxiety-like behaviors in the SPT, TST, OFT and EZM. Serum levels of GR, CORT, ACTH and CRH levels were decreased in the RS and MS + RS groups. Limbic‑system‑related neural circuits were activated in short- and long-term stress, with the expression of c-Fos decreasing and that of FosB increasing. Moreover, the concentrations of 5‑HT, dopamine (DA), norepinephrine (NE), and epinephrine (E) were lowered in the prefrontal cortex, amygdala, hippocampus, and hypothalamus. Maternal separation exerts a time-dependent dual effect on adult mice's response to restraint stress: initially enhancing resilience, but later increasing susceptibility to depressive- and anxiety-like behaviors. These effects are likely mediated by neuroendocrine regulation of neural circuits involving HPA axis hormones and monoamine neurotransmitters. This study provides a novel two-hit mouse model for exploring how ACEs shape adult respond to stress.

Plant-derived exosome-like nanoparticles (PELNs) are lipid bilayer nanostructures containing endogenous lipids, proteins, nucleic acids, and phytochemicals, which have attracted increasing interest as bioinspired carriers for cancer therapy. This review evaluates how plant source, isolation, purification procedures, vesicle composition, cargo-loading strategy, and administration route shape the quality and tumor-delivery performance of PELNs. The available evidence indicates that plant source and processing are major determinants of particle size, purity, surface charge, cargo profile, and biological activity. Ultracentrifugation remains widely used but is limited by contaminant co-isolation and poor scalability, whereas density-gradient purification and size-exclusion chromatography improve purity, and ultrafiltration and tangential flow filtration offer greater potential for large-scale manufacturing. Passive incubation generally preserves vesicle integrity and is most suitable for hydrophobic small molecules, whereas electroporation, sonication, and extrusion can increase cargo loading but may cause aggregation, membrane remodeling, or loss of endogenous components. Preclinical studies suggest that PELNs can exert intrinsic antitumor effects, modulate the tumor microenvironment, improve chemotherapeutic delivery, and help overcome drug resistance. However, evidence for in vivo tumor-targeting remains less robust than evidence for cellular uptake, and direct comparisons with established nanocarriers remain scarce. Clinical translation will require standardized nomenclature and characterization, reproducible manufacturing, quantitative loading and release assays, route-specific biodistribution studies, and repeated-dose safety evaluation. These findings provide a framework for the rational development of PELNs as reproducible tumor-oriented nanocarriers.

Coronary heart disease (CHD) is a global cardiovascular disease with high morbidity and mortality, and its complex pathological mechanism poses great challenges to clinical prevention and treatment. Paeoniflorin (PA), a monoterpene glycoside active ingredient from Ranunculaceae plants, has shown potential in cardiovascular protection, but its specific anti-CHD molecular targets and systematic regulatory networks remain unclear. In this study, a mouse model of CHD was established, and a multi-omics strategy combining label-free quantitative proteomics and metabolomics was adopted to explore the mechanism of PA in treating CHD. The results showed that PA significantly improved cardiac function, alleviated myocardial pathological injury and fibrosis, and regulated lipid metabolism in CHD model mice, with the high-dose group showing the optimal effect. Proteomic analysis identified 51 key differentially expressed proteins (DEPs) reversed by PA, which were mainly enriched in complement and coagulation cascades, and neutrophil extracellular trap formation pathways, with the C3-Cfd-C3aR signaling axis as the core hub. Further verification confirmed that PA could downregulate the expression of C3, Cfd, C3aR, and their downstream molecule BTK, thereby inhibiting myocardial inflammatory response and cardiomyocyte apoptosis. In addition, PA downregulated the expression of platelet activation markers ITGA2B/ITGB3. Metabolomic analysis revealed that PA reversed 57 abnormal metabolites in CHD mice, which were enriched in GABAergic synapse, retrograde endocannabinoid signaling and other pathways. Molecular docking confirmed that PA could stably bind to C3, Cfd, C3aR, BTK, and ITGA2B/ITGB3 with strong binding activity. In conclusion, PA exerts anti-CHD effects through a multi-target and multi-pathway synergism, mainly by targeting the C3-Cfd-C3aR axis to inhibit inflammation, apoptosis and platelet activation, and regulating metabolic disorders. This study provides experimental evidence and theoretical support for the clinical application of PA as a multi-target therapeutic drug for CHD.

This study aimed to systematically elucidate the antihyperlipidemic mechanism of paeoniflorin, and we adopted an integrated multi-omics strategy to screen the key molecular targets and regulatory pathways involved in its action, followed by experimental validation to verify the potential regulatory effects of paeoniflorin on the screened targets and metabolic processes. Rats with high-fat diet-induced hyperlipidemia received paeoniflorin treatment. Liver histopathology was evaluated using hematoxylin–eosin and Oil Red O staining. Serum levels of total cholesterol, triglycerides, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, total bile acids, activated partial thromboplastin time, prothrombin time, thrombin time, and fibrinogen were measured using a biochemical analyzer. Integrated multi-omics analyses were performed to investigate paeoniflorin’s lipid-lowering mechanism. Critical pathways and targets identified were validated using Western blotting. Paeoniflorin alleviated pathological liver damage in hyperlipidemic rats and improved blood lipid levels, coagulation function, and liver function markers. Multi-omics analyses verified that paeoniflorin downregulated the expression of TREM-1, TLR4, NF-κB, TNF-α, and IL-1β, thereby alleviating hepatic inflammation. Paeoniflorin also upregulated the expression of low-density lipoprotein receptors (LDLR), liver X receptor alpha (LXRα), and ATP-binding cassette subfamily G member 1 (ABCG1), while downregulating proprotein convertase subtilisin/kexin type 9 (PCSK9) expression, contributing to balanced cholesterol metabolism. Paeoniflorin normalized glycerophospholipid and branched-chain amino acid metabolism, which correlated with reduced inflammation and improved cholesterol metabolism. Paeoniflorin ameliorates hyperlipidemia through multitarget mechanisms, potentially by suppressing the TREM-1-TLR4-NF-κB signaling pathway to reduce inflammation and by regulating cholesterol metabolism via the PCSK9-LDLR and LXRα-ABCG1 pathways.

Objective: This case aims to look into and analyze the prophylactic and therapeutic effects of Tanyu tongzhi decoction (TYTZD) in atherosclerosis (AS), and to explore its possible regulating mechanism, including that of bile acid metabolism, using the integrated methodology in combination with network pharmacology and experimental validation. Methods: ApoE ^-/- mice were fed with a high-fat diet and administered trimethylamine N-oxide (TMAO) for 12 weeks to establish an atherosclerosis model. At the end of the eighth week, the murine group was divided into 4 groups: high-fat diet (HFD), HFD plus trimethylamine N-oxide (TMAO), Tanyu Tongzhi decoction (TYTZD), and TMAO inhibitor (DMB), one of which was a regular chow diet group as a control. Following a 4-week intervention period, TMAO concentrations across all experimental groups were quantified using HPLC-MS methodology. TYTZD’s therapeutic benefits on lipid profiles and atherosclerotic lesion development in murine models were assessed via serum biochemical tests, HE staining, and Oil Red O staining techniques. The TCMSP and ETCM databases facilitated the prediction of molecular targets for TYTZD bioactive constituents. AS-related disease targets were retrieved from the GEO database repositories. Network pharmacology approaches enabled the identification of differentially expressed genes linking “TYTZD-AS-bile acid metabolism” pathways. Enrichment analyses combined with PPI network construction revealed critical regulatory targets. TYTZD’s influence on bile acid metabolic processes in atherosclerotic mice was examined through targeted metabolomic profiling of serum and fecal samples. Finally, we validated the expression of proteins in core signaling pathways by western blotting. Results: In vivo experiments demonstrated that a high-fat diet combined with TMAO intervention significantly promoted AS progression in ApoE ^-/- mice, manifested primarily as a significant increase in aortic plaque burden, markedly elevated serum lipid levels, and disordered serum bile acid metabolism. Both DMB and TYTD treatments effectively reduced the in vivo TMAO levels, regulated blood lipids, alleviated aortic lipid deposition, and inhibited the formation of AS plaques. Network pharmacology analysis indicated that the potential target of TYTZD for dealing with AS was mainly enriched by cholesterol conversion and primary bile acid synthesis pathways. CYP7A1 and CYP27A1 were identified as key genes through which TYTZD regulates bile acid metabolism. Biochemical and metabolomic analyses demonstrated that DMB and TYTZD treatments substantially elevated total circulating bile acids, augmented bile acid reserves, and restored homeostatic balance in serum and fecal bile acid profiles. Specifically, both interventions decreased blood concentrations of CA, CDCA, DCA, and LCA while concurrently increasing TUDCA levels in both serum and feces. Immunoblot analysis indicated that DMB and TYTZD suppressed hepatic FXR along with its negative regulator SHP, enhanced liver CYP7A1 expression, and reduced ileal FXR and FGF15 protein abundance. Collectively, these data indicate that TYTZD counteracts TMAO-mediated inhibition of CYP7A1, the rate-limiting enzyme governing bile acid biosynthesis, thus facilitating cholesterol catabolism into bile acids. Conclusion: TYTZD prevents the events of atherosclerosis progression by regulating the bile acid metabolism. It does this by controlling liver FXR-SHP and intestinal FXR-FGF15 signaling pathways together. Thus, bile acid synthesis facilitated by CYP7A1 increases.

Background: Coronary heart disease (CHD) has a high global disease burden. According to traditional Chinese medicine theory, the main syndrome type of CHD is the syndrome of intermingled phlegm and blood stasis (SI-GPBS). Tanyu Tongzhi Decoction (TYTZD) exerts clear cardioprotective effects on CHD patients with SI-GPBS, while its specific regulatory mechanism remains unclear. Methods: Clinical serum proteomics and network pharmacology were used to screen key targets and pathways for CHD with SI-GPBS. An APOE^−/− mouse model of CHD complicated with SI-GPBS was established and treated with TYTZD. Transcriptomics, proteomics and WGCNA were combined to screen core genes, with Western blotting, immunofluorescence, co-localization analysis and Carstairs staining for target verification and observation of coronary microthrombosis and endothelial injury. Results: A total of 754 differentially expressed proteins were identified in CHD patients with SI-GPBS, significantly enriched in the platelet activation pathway, with ITGA2B as the upregulated core hub protein. Network pharmacology found 94 active ingredients and 144 therapeutic targets of TYTZD for CHD with SI-GPBS, and key components bound well with ITGA2B. In APOE^−/− mice with SI-GPBS, TYTZD improved cardiac function, reduced blood lipids, myocardial enzymes, aortic lipid deposition and myocardial damage, downregulated ITGA2B, F2RL2, FGA and FGB, inhibited integrin αIIbβ3 signaling, restrained endothelial activation and reduced coronary microthrombosis. Conclusions: TYTZD treats CHD with SI-GPBS mainly by inhibiting platelet activation, improving endothelial dysfunction, and reducing coronary microthrombosis. This study provides experimental basis for TYTZD’s clinical application in CHD with SI-GPBS and new ideas for TCM syndrome–disease combination research.