Background. Triple-negative breast cancer (TNBC) is a subtype of breast cancer with limited therapeutic options. Eupalinolide O (EO) was reported to inhibit tumor growth. This study is aimed at exploring the role of EO on TNBC both in vivo and in vitro. Methods. In in vitro experiments, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and clonogenic assay were conducted to measure the impact of EO on TNBC cell growth at different concentrations and time points. Flow cytometry was conducted to evaluate cell apoptosis. Mitochondrial membrane potential (MMP) loss, caspase-3 activity, and reactive oxygen species (ROS) generation were assessed. The expressions of apoptosis-related mRNAs and Akt/p38 MAPK signaling pathway-related proteins were measured. In in vivo experiments, by injecting TNBC cells into the nude mice to induce xenograft tumor, mice were treated with EO for 20 days. Then, in vivo bioluminescence imaging system was utilized to monitor the growth and distribution of TNBC cells. Tumor volume and weight were also recorded. Hematoxylin-eosin (HE) staining and ELISA assay were applied to observe tumor tissue morphology and ROS levels. Furthermore, western blotting was conducted to observe the expression of apoptosis-related proteins and Akt/p38 MAPK signaling pathway-associated proteins. Results. EO inhibited the cell viability and proliferation of TNBC cells but not normal epithelial cells. Furthermore, EO induced apoptosis, decreased MMP, and elevated caspase-3 activity and ROS content in TNBC cells. Meanwhile, the expression of apoptosis-related mRNAs and Akt/p38 MAPK pathway-related proteins was regulated by EO treatment. Besides, in vivo experiments demonstrated EO not only suppressed tumor growth, Ki67 expression, ROS generation, and Akt phosphorylation but also upregulated caspase-3 expression and p-38 phosphorylation. Conclusion. EO may induce cell apoptosis in TNBC via regulating ROS generation and Akt/p38 MAPK pathway, indicating EO may be a candidate drug for TNBC.

This review provides a comprehensive analysis of the molecular innovations driving vaccine adjuvant research, moving from empirical formulations to rational designs. After outlining clinical benchmarks, we systematically categorize and evaluate emerging experimental adjuvant classes, including cytokine-based agonists, advanced particulate delivery systems, emulsion-based platforms, and nucleotide-based strategies. Notably, we focus on natural product-derived adjuvants, highlighting how their structure-activity relationships (SAR) drive potent immunomodulatory activities. We further dissect the molecular mechanisms underpinning these adjuvants, detailing how they target specific pattern recognition receptors (PRRs) to orchestrate dendritic cell maturation, antigen cross-presentation, and Th1/Th2 polarization. Crucially, we discuss the rational selection of adjuvants based on specific clinical targets and antigen modalities. Finally, we propose a forward-looking perspective on developing multi-modal strategies that synergize molecular immunostimulants with synthetic delivery vectors.

Background: Plant-derived extracellular vesicles (PDEV) are emerging as natural nanomedicines for various diseases. Scutellaria baicalensis (S. baicalensis) is a traditional Chinese herb long used to treat intestinal inflammatory bowel disorders (IBD), with its therapeutic effects attributed to bioactive flavonoids such as baicalin and wogonin. However, whether SEV contribute to its anti-inflammatory activity remains unexplored. The assembled multi-component nature of SEV, which carry flavonoids, lipids, proteins, and miRNAs, suggests a potential to exert therapeutic effects against IBD through mechanisms distinct from isolated compounds, with potential advantages in bioavailability and multi-target engagement. Methods: We demonstrated that SEV exert potent antioxidant and anti-inflammatory effects in LPS-stimulated RAW264.7 macrophages and Caco-2 intestinal epithelial cells. Moreover, we assessed the therapeutic effects of SEV on dextran sulfate sodium (DSS)-induced IBD in a murine model. Results: In inflamed RAW264.7, SEV modulated the NF-κB/NLRP3 signaling axis to exert anti-inflammatory effects. They scavenged reactive oxygen species (ROS), restored mitochondrial membrane potential, upregulated the anti-inflammatory cytokine IL-10, and suppressed the pro-inflammatory cytokines TNF-α, IL-6, and IL-1β. In Caco-2 intestinal epithelial cells, SEV also repaired intestinal barrier function by restoring expression of the tight junction proteins Zonula Occludens-1 (ZO-1), Claudin-1, and Occludin (OCLN), alongside reduced TNF-α levels. In vivo, SEV accumulated at colonic inflammatory loci to effectively alleviate IBD, as evidenced by improved body weight and increased colon length. This protective effect was mediated through inhibition of the NF-κB/NLRP3 signaling axis in colon tissues, which subsequently restored intestinal barrier integrity by increasing goblet cell numbers, upregulating OCLN proteins, and enhancing Mucin2 (MUC2) secretion, while simultaneously rebalancing inflammatory cytokines through suppression of TNF-α/IL-1β and promotion of IL-10 production. Conclusion: SEV have the potential to protect the colon against DSS-induced colitis by inhibiting the NF-κB/NLRP3 signaling pathway, providing a promising therapeutic candidate for IBD. Infographic on Scutellaria baicalensis extraction and its effects on inflammatory bowel disease.The process begins with grinding the plant, followed by centrifugation, filtering and ultracentrifugation to obtain SEV. Sucrose gradient centrifugation is used to separate SEV. The SEV is then shown to be administered to a model of IBD, where it restores the inflammatory colon microenvironment. The restoration is indicated by increased levels of ZO-1, Claudin-1 and OCLN. SEV intake affects the inflammatory response by reducing ROS and increasing MMP. It also impacts the NF-kB/NLRP3 pathway, reducing TNF-alpha, IL-6 and IL-1beta, while increasing IL-10. The cells involved in this process include RAW264.7 and CaCO2, which intake SEV, leading to these changes.

Objective: The role of Nuclear factor E2-related factor 2 (NRF2) in cells is a double-edged sword, as it can maintain the stability of the inner environment for normal cells by resisting oxidation, but can also confer drug adaptability to cancer cells. Recent reports suggest that inhibiting NRF2 can weaken the drug resistance of cells, making it a potential target for cancer therapy. Our research on CXX-3, a norditerpenoid isolated from Podocarpus nakaii, demonstrated its ability to combat cancer cells. Methods: We observe for the first time that an active ingredient from P. nakaii induces cancer cell death accompanied by the accumulation of reactive oxygen species (ROS) and arrests the cell cycle in the S phase. The ROS inhibitor NAC(N-acetyl-l-cysteine) can partially rescue cell death. Additionally, we identified the NRF2/KEAP1 (Kelch-like ECH-associated protein 1, KEAP1) complex as the critical site conquered by CXX-3. Dysfunction of this complex ruined the antioxidation ability of glioma and colon carcinoma. Interestingly, CXX-3 seems to disable the KEAP1/NRF2 complex through a separate mechanism. Results: In conclusion, CXX-3 can disrupt the antioxidation ability of cancer cells, leading to the accumulation of ROS, thereby arresting the cell cycle, and inducing cell death. Conclusion: As such, CXX-3 holds promise as a prospective compound for cancer therapy.