Xiao-Qiang Li

Introduction: Japanese Encephalitis (JE) remains a serious health threat with limited treatment options. This study aims to construct comprehensive libraries of high-frequency compounds from Chinese herbs and to screen active compounds against JE by integrating bioinformatics, network pharmacology, and experimental validation. Methods: A natural compound library was constructed through data mining and frequency analysis of Traditional Chinese medicine (TCM) prescriptions for the treatment of JE. ADMET prediction was performed to select compounds with Blood-Brain Barrier (BBB) permeability and to assess their potential toxicity. The compound-JE intersection targets were used to establish a PPI Network and to perform GO and KEGG enrichment analysis. Core targets were identified based on the PPI network by Cytoscape software. Molecular docking was performed with Discovery Studio software. Finally, in vitro experiments were carried out further to screen and validate the anti-inflammatory and antiviral effects of the active compounds in Neuro2a or BV2 cell models infected with Japanese Encephalitis Virus (JEV). Results: Seven of the most commonly used herbs and 16 compounds were identified. Six compounds with BBB permeability and good druggability were screened. Network pharmacology revealed that these six compounds mainly targeted five core targets to exert anti-neuroinflammatory activity. In addition, molecular docking results suggested that JEV proteins were the major targets for these six compounds for antiviral activity. In JEV-induced cell models, Caryophyllene oxide, Qingdainone, and Isoliquiritigenin reduced inflammatory factors by regulating the PTGS2/NF-κB pathway in BV2 cells and exhibited the highest antiviral activity through JEV proteins in Neuro2a cells Discussion: This study establishes an integrative strategy bridging traditional medicine with modern pharmacology for anti-JEV drug discovery. The identification of Caryophyllene oxide, Qingdainone, and Isoliquiritigenin as dual-function agents, exerting both antiviral and anti-inflammatory effects, highlights the therapeutic potential of multi-target compounds against JE. Notably, this dual mechanism offers advantages over conventional singletarget therapies by simultaneously inhibiting viral replication and modulating host inflammation. Collectively, this work provides a framework for identifying multi-target anti-JEV agents from natural products, laying a foundation for the translational development of Caryophyllene oxide, Qingdainone, and Isoliquiritigenin. Conclusion: An integrative pipeline combining TCM-based compound library construction, bioinformatics, network pharmacology, and experimental validation has been established to identify novel anti-JE agents. Using this strategy, Caryophyllene oxide, Qingdainone, and Isoliquiritigenin were selected as candidates possessing a unique dual antiviral and anti-inflammatory mechanism for the treatment of JE.

Immunotherapy has demonstrated potential in treating various malignant tumors, but its efficacy in pancreatic cancer (PC) remains limited, possibly due to the dense stromal components and immunosuppressive microenvironment of PC. Focal adhesion kinase (FAK), a non-receptor tyrosine kinase, plays a crucial role in the tumor microenvironment and intracellular signaling pathways. However, the specific role of FAK in the development and progression of PC, as well as its regulatory mechanisms on the tumor immune microenvironment (TIM), are still not fully understood. In this study, we analyzed single-cell sequencing datasets and clinical specimens to evaluate the role of FAK in the immune response of PC. We verified the impact of FAK alterations on CD8⁺ T cell infiltration using a co-culture system of patient-derived organoids (PDO) and immune cells. Additionally, mouse PC models and dual humanized models are established to investigate the in vivo function of FAK and the potential of its inhibitors for immunotherapy. Our results demonstrate that FAK is associated with the immunosuppressive microenvironment in PC. Inhibiting FAK enhances CD8⁺ T cell infiltration by promoting CXCL10 secretion in PC. Moreover, FAK inhibitors exhibit a synergistic anti-tumor effect when combined with immune checkpoint inhibitors. This study explores the potential of FAK as a therapeutic target, particularly its role in modulating TIM, thereby providing new research directions for the treatment of PC.