Cancer is a highly heterogeneous malignant disease. Currently, precision medicine in oncology urgently requires experimental models that can faithfully simulate the individual tumor characteristics of patients. Traditional two-dimensional cell lines and animal models exhibit significant limitations in preserving tumor heterogeneity, microenvironment, and predicting treatment responses. Organoid technology, with its three-dimensional structure and high fidelity in recapitulating the histopathological and molecular features of primary tumors, has emerged as a transformative tool in cancer research. This review systematically outlines the development of organoid technology, highlighting the methodologies and applications for modeling various solid tumors, including lung cancer, breast cancer, gastric cancer, renal carcinoma, and colorectal cancer. It encompasses multiple aspects such as investigations into tumorigenesis mechanisms, drug screening, personalized therapy, and immunotherapy assessment. Furthermore, the current challenges and future directions of organoid technology are discussed, with the aim of promoting its broader application in cancer research and clinical translation.

As a core biological process for the repair of liver injury, liver regeneration represents a key breakthrough in the treatment of liver failure and end-stage liver disease. Its mechanistic complexity and therapeutic translation potential have long been research hotspots. In recent years, with the advancement of molecular biology and regenerative medicine technologies, significant progress has been made in liver regeneration research, including mechanistic elucidation, identification of regulatory targets, and development of therapeutic strategies. This article systematically reviews the molecular mechanisms and therapeutic strategies of liver regeneration, analyzes current research bottlenecks, and discusses future directions, aiming to provide a reference for scientific research and clinical applications in this field.

As significant bioactive constituents within natural products, alkaloids exhibit characteristics of extensive sources, diverse structures, multi-targeted and multi-level actions, and possess potent immunomodulatory effects. Although numerous studies have explored the pharmacological effects and mechanisms of alkaloids, a systematic review of their immunomodulatory activities and underlying mechanisms remains to be systematically reviewed, particularly an integrative analysis of the most recent research advances. Therefore, this paper collates recent research findings on alkaloids within the field of immunoregulation. It reviews the modulatory effects of alkaloids on the immune system, summarises the principal molecular mechanisms underlying their actions, and outlines their applications in the treatment of immune-related diseases. This review aims to provide a reference for subsequent mechanistic studies and the development of related pharmaceuticals.

Stroke is characterized by high incidence, high disability, high mortality, and high recurrence rates. Current clinical management relies primarily on injectable thrombolytic therapy. While a variety of oral adjuvant medications are available, their therapeutic efficacy and underlying mechanisms have not been systematically evaluated. Traditional Chinese medicine (TCM), as a multi-component and multi-target therapeutic option, has demonstrated considerable potential in the prevention and treatment of stroke. Emerging evidence indicates that TCM exerts modulatory effects on stroke-related pathological processes by regulating the gut microbiota, maintaining gastrointestinal homeostasis, and acting through the gut-brain axis. The conventional pharmacological paradigm holds that drugs must enter the systemic circulation and traverse the blood-brain barrier (BBB) to exert central effects. Despite their low oral bioavailability and limited systemic exposure, certain pharmacologically active constituents nonetheless exert significant neuroprotective effects, suggesting that their in vivo activities may be mediated by indirect gut-derived regulatory mechanisms rather than direct systemic actions. Based on these observations, we propose the concept of "intestinal retention components" (IRC), referring to a class of orally administered bioactive constituents that are poorly absorbed across the intestinal epithelium and predominantly retained within the intestinal lumen due to their physicochemical properties. Although these compounds exhibit minimal systemic exposure, they are capable of modulating the intestinal microenvironment, thereby influencing brain function and improving stroke outcomes. This review aims to systematically elucidate the potential mechanisms by which IRC indirectly exerts anti-stroke effects via the gut-brain axis, encompassing microbiota-mediated biotransformation, attenuation of gut microbial dysbiosis and its associated metabolic perturbations, restoration of intestinal barrier integrity, and regulation of the intestinal mucosal immune microenvironment. Given that numerous central nervous system disorders are also accompanied by intestinal microenvironmental dysregulation, the IRC-centered research framework targeting the gut microenvironment may provide a valuable reference for mechanistic elucidation and the identification of candidate drugs among orally active constituents in the context of broader neurological diseases.

The mixed lineage kinase domain-like protein (MLKL) is a "pseudokinase" that not only functions as a key effector molecule in the necroptosis pathway but also participates in the regulation of other programmed cell death pathways, thereby playing a crucial role in modulating various physiological processes. Dysregulation of MLKL activity is closely associated with the initiation and progression of multiple diseases, rendering it a highly promising therapeutic target. However, drug discovery targeting MLKL is still in its early stages, with no candidates having advanced to clinical trials. Thus, further in-depth and systematic investigations are urgently needed. This review summarizes the structural features, biological functions, and current progress in drug development targeting MLKL.

As a prominent class of nitrogen-containing heterocycles, 1,3,5-triazines are characterized by their rigid, symmetrical structures, ease of functionalization, exceptional biological activities, making them a privileged scaffold in medicinal chemistry. This paper provides a systematic review of the research progress on 1,3,5-triazine derivatives over the past five years (2021-2025), covering their structure-activity relationships and mechanisms of action. Results indicate that 1,3,5-triazine compounds exhibit promising antimicrobial activity against methicillin-resistant Staphylococcus aureus, drug-resistant Escherichia coli, and Candida albicans. Furthermore, certain 1,3,5-triazine compounds target PD-L1, PI3K/mTOR, or EGFR, thereby inducing apoptosis or cell cycle arrest and inhibiting tumor cell growth. In addition, these compounds also have anticonvulsant, neuroprotective, antimalarial, antihyperglycemic, and anti-inflammatory properties. However, several 1,3,5-triazine compounds lack clear mechanisms and possess unsatisfactory efficacy, requiring further in-depth research to develop new chemical entities.

Sesquiterpenoids are a class of terpenoids composed of 15 carbon atoms, with a basic framework formed by three C₅ isoprene units. They exhibit diverse skeletal structures and an exceptionally rich variety of types. These compounds are widely distributed in plants and serve as bioactive metabolites in diverse medicinal species, demonstrating various pharmacological activities such as anti-tumor, antibacterial, and anti-inflammatory. The elucidation of biosynthetic pathways, their heterologous reconstruction, and the sustainable production of bioactive sesquiterpenoids have emerged as research focal points. In recent years, substantial advancements have been made in the field of sesquiterpenoid biosynthesis. Based on an overview of their biosynthetic pathways, this review provides a comprehensive progress in synthetic biology research on sesquiterpenoids derived from medicinal plants. This review summarizes the status of the heterologous biosynthesis of representative plant-derived sesquiterpenoids, and discusses the major bottlenecks along with their corresponding optimization strategies and future perspectives.

Toxic Chinese medicines constitute a significant component of the traditional Chinese medicine system. Taking aristolochic acid-containing herbs as an example, their clinical application and industrial development face severe challenges due to safety incidents caused by aristolochic acid-containing components. The Pharmacopoeia of the People's Republic of China has deleted relevant varieties based on drug safety considerations, which, while mitigating medication risks, also results in the waste of medicinal resources. Existing methods such as processing and combination therapy also have limitations in reducing the toxicity of these herbs, including limited efficacy and difficulties in standardizing processes, making it challenging to balance toxicity control with the preservation of pharmacological effects. Based on this, this paper systematically reviews the deletion of herbal materials and proprietary Chinese medicine formulations from the last five editions of the Chinese Pharmacopoeia, summarizing the medicinal value and controversies surrounding herbs derived from the Aristolochia genus, the toxic mechanisms of aristolochic acid, and existing detoxification methods along with their limitations. Addressing these limitations, this paper proposes application strategies for gene editing, enzyme engineering, and fermentation engineering in reducing the toxicity of Aristolochia and other Chinese medicinal herbs, from the perspective of modern biotechnology, and explores new pathways for the precise regulation of the synthesis and degradation of toxic components. Compared with traditional detoxification methods, the use of modern biotechnology holds promise for precisely regulating the synthesis or degradation of toxic components, thereby reducing toxicity while preserving the overall therapeutic efficacy of the herbal materials, with the aim of achieving "detoxification while preserving efficacy". Future efforts should be intensified to verify the botanical sources of toxic Chinese herbal medicines, elucidate the biosynthetic pathways of toxic components, and investigate their toxic mechanisms. Additionally, the development and utilization of toxic Chinese herbal resources should be advanced to promote the high-quality development of traditional Chinese medicine through inheritance, innovation, modernization, and internationalization.

Pharmacokinetic studies of traditional Chinese medicine polysaccharides (TCMPs) are essential for elucidating the material basis of their pharmacological effects. A central challenge in this field lies in the paradox between their low oral bioavailability and significant systemic efficacy. Research has demonstrated that TCMPs exhibit low plasma concentrations following oral administration, and their therapeutic effects are largely independent of direct prototype absorption, instead being mediated through indirect mechanisms such as gut microbiota-derived metabolites and immunomodulation. In terms of distribution, TCMPs selectively target organs rich in mononuclear phagocytes, such as the liver and spleen, where they engage pattern recognition receptors to initiate immune regulation. Their metabolism involves a synergistic action between host enzymes and gut microbiota, with fecal excretion serving as the primary elimination route. A groundbreaking study by Ding et al. has recently provided direct evidence that specific structural types of TCMPs, such as 1, 3- and 1, 4-linked glucans, can be absorbed as intact macromolecules via a clathrin/dynamin-1/Rab5-mediated endocytic pathway, offering a mechanistic explanation for their systemic efficacy. Current research continues to face challenges, including insufficient sensitivity of analytical methods and the complexity of metabolic networks. Future integration of highly sensitive analytical techniques, multi-omics approaches, and artificial intelligence will drive the field toward more precise and systematic investigation, facilitating the clinical translation of TCMPs.

In recent years, conjugated drugs have become a hotspot in drug research and development due to their precise target binding. Radionuclide drug conjugates (RDC) are an emerging class of targeted conjugated drugs that combine diagnostic and therapeutic functions, representing a new drug type with immense development potential. Due to structural differences, RDC drugs exhibit distinct in vivo processes — absorption, distribution, metabolism, excretion, and toxicity (ADMET) — as well as unique bioanalytical methods compared to small molecule drugs. Starting from the analysis of the physicochemical properties of RDC, this article analyzed the pharmacokinetic characteristics and its application in the PK model selection of RDC and summarized the bioanalysis methods of such drugs, aiming to promote further development of RDC.

Metastasis is the leading cause of cancer deaths. It involves intricate interactions between circulating tumour cell (CTC) and blood components such as platelets, neutrophils and macrophages. This review first summarizes how CTC interact with these cells to promote their survival, immune evasion and seeding in distant organs. We then focus on recent advances in nanodelivery systems designed to counteract these processes — including the targeted elimination of single CTC, disruption of CTC homotypic clusters, blocking of CTC-platelet/neutrophil aggregates, and inhibition of neutrophil extracellular traps (NETs). Various strategies, such as ligand-mediated targeting, biomimetic cell membrane coating, and stimuli-responsive release, are critically discussed. Finally, we highlight key challenges and future directions for translating these systems into clinical anti-metastatic therapies.

Natural plant essential oils, as secondary metabolites of plants, possess hydrophobicity and volatility, and exhibit a wide range of biological activities, including antibacterial, antioxidant, anti-inflammatory, anti-aging, and antitumor effects. They are considered potential alternatives to chemically synthesized drugs. However, their poor stability and low water solubility limit clinical applications. Nanotechnology, through the construction of delivery systems such as liposomes, nanoemulsions, nanogels, and polymeric nanoparticles, significantly improves the stability, bioavailability, and targeting ability of essential oils, thereby enhancing their pharmacological activity and reducing toxic side effects. This review systematically summarizes the chemical composition, biological activity mechanisms, and recent advances in nanoformulations of natural plant essential oils, with a particular focus on the application strategies and effects of nanodelivery systems in improving essential oil performance. Furthermore, it highlights their translational potential and future development directions in modern medicine.

Ginsenoside Rg3 possesses a variety of pharmacological activities. However, ginsenoside Rg3 has poor water solubility, which limits its full exertion of pharmacological effects. Existing delivery systems can not only improve the solubility of ginsenoside Rg3 but also leverage the unique advantages of each delivery system, enabling ginsenoside Rg3 to exert its pharmacological effects more effectively. This review focuses on the impact of ginsenoside Rg3 delivery systems, summarizes and compares the pharmacodynamics and pharmacokinetics of ginsenoside Rg3, and highlights the advantages of various delivery systems, provides a basis for the selection of ginsenoside Rg3 delivery systems, and serves as a cornerstone for the full development of ginsenoside Rg3.

Coxsackievirus B3 is a major pathogen responsible for viral myocarditis and is associated with various severe diseases, including dilated cardiomyopathy and pancreatitis. However, no vaccines or specific drugs have been approved for CVB3 infection to date. High content screening technology, which integrates confocal microscopy with multidimensional intelligent image analysis, offers significant advantages for drug screening. In this study, utilizing an EGFP-CVB3 reporter virus, we successfully established a stable and reliable quantitative high-content imaging system for screening anti-CVB3 drugs. This system enables the simultaneous acquisition of multi-parametric data at the single-cell level, including infection rate, cell morphology, and organelle status. The assay was validated using known positive compounds. In conclusion, this system facilitates high-throughput screening of candidate compounds against CVB3 within a single viral life cycle, providing a rapid and sensitive technological tool to accelerate the discovery of anti-CVB3 drugs.

Tumor necrosis factor receptor-associated factor 2 (TRAF2) is a central signaling adaptor that integrates multiple oncogenic pathways and represents potential target for cancer therapy. In this study, leveraging the extensive protein-protein interaction network of TRAF2, 100 "undruggable" key tumor-associated proteins were selected to screen for TRAF2 interactors. It was found that cytokine receptor-like factor 2 (CRLF2) is a novel interacting protein of TRAF2, and the liquidambaric acid (LDA), which can directly target TRAF2, can inhibit the TRAF2/CRLF2 interaction. Binding site studies revealed that CRLF2 binds to the TRAF domain of TRAF2 and can be blocked by LDA. Functional studies of CRLF2 indicated that TRAF2 is a key protein in the CRLF2-STAT3 (signal transducer and activator of transcription 3) signaling pathway, and LDA can regulate CRLF2-related STAT3 phosphorylation. In summary, this study reveals the TRAF2-mediated tumor protein interaction network and elucidates a new mechanism by which LDA exerts antitumor effects by inhibiting TRAF2/CRLF2 interaction to regulate the STAT3 signaling pathway. It also provides new theoretical guidance and potential strategies for the development of anti-tumor drugs targeting undruggable targets.

Salidroside (SAL), a principal active constituent of the medicinal plant Rhodiola crenulata, exerts diverse pharmacological activities including antioxidation, anti-inflammation, and anti-aging. However, its protective efficacy against liver injury and the underlying molecular mechanisms remain incompletely elucidated. The low oral bioavailability of SAL implies potential interactions with the gut microbiota. Therefore, this study focused on the metabolic transformation of SAL mediated by the gut microbiota. The results demonstrated that SAL is metabolized to tyrosol (TYR) in the gut microbiota. Virtual molecular docking and pure enzyme metabolic experiments further validated the critical role of β-glucosidase (β-G) in this transformation process. In vitro cellular experiments have demonstrated that SAL and its primary metabolite TYR exert a synergistic hepatoprotective effect. Based on network pharmacology screening, hub targets including NF-κB1, ESR1, BCL2, MMP9, and TLR4 were revealed. Furthermore, SAL intervention significantly increased the levels of short-chain fatty acids (SCFAs) by the gut microbiota in an in vitro culture system. Untargeted metabolomics analysis suggested that SAL may reshape the metabolic profile of the gut microbiota by modulating pathways including the urea cycle and tryptophan metabolism. This study provides a novel perspective for a deeper understanding of the hepatoprotective mechanism of SAL and offers valuable scientific evidence for future clinical translational research on natural medicines. The experimental protocol was approved by the Laboratory's Institutional Animal Care and Use Committee of the Chinese Academy of Medical Sciences and Peking Union Medical College (Approval No.: 00009927).

This study systematically investigated the effects of Xihuang Pill on exosomes (Exo) secretion and how Exo pretreated with Xihuang Pill affect intercellular communication, thereby inhibiting the activity of triple-negative breast cancer cells. First, the inhibitory affected of Xihuang Pill on the activity and migration of 4T1 and MDA-MB-231 cells were verified using MTT, scratch assays, and crystal violet assays. Then, Exo and Exo secreted by cells pretreated with Xihuang Pill (XHP-Exo) were obtained by ultracentrifugation. The Exo were characterized by transmission electron microscopy, potential, particle size, and marker proteins. The inhibitory effects of XHP-Exo on the activity and migration of 4T1 and MDA-MB-231 cells were detected using crystal violet and scratch assays. The expression of programmed death ligand 1 (PD-L1), glutathione peroxidase 4 (GPX4), reactive oxygen species (ROS), and reduced glutathione (GSH) in cells was detected by Western blot and immunofluorescence. The chemical components of Xihuang Pill and XHP-Exo were analyzed by ultra-high performance liquid chromatography-quadrupole-electrostatic field orbital trap mass spectrometry (UPLC-Q-Exactive-MS/MS). The results showed that Xihuang Pill could significantly inhibit the proliferation and migration of breast cancer cells, downregulate kidney and brain expressed adaptor protein (KIBRA) expression, and effectively reduce the secretion of exosomes from breast cancer cells. Furthermore, XHP-Exo induced ferroptosis in recipient tumor cells, characterized by increased intracellular ROS levels and decreased GSH content, accompanied by downregulation of PD-L1 expression. Component analysis revealed 19 common active chemical components in both Xihuang Pill and XHP-Exo. In summary, Xihuang Pill inhibits the secretion of exosomes from breast cancer cells, and component-containing exosomes derived from it induce ferroptosis in recipient cells and downregulate PD-L1 expression.

This study aimed to investigate the mechanism of the traditional Chinese medicinal formula Qijing Mingmu (QJMM) in the treatment of conjunctivochalasis (CCH) using network pharmacology and cellular experiments. Effective compounds and action targets of QJMM components were screened using the TCMSP and TCM-ID databases. Disease targets for CCH were obtained from the DisGeNET database, with gene names converted from the UniProt database. The Venny database yielded disease-drug intersection targets, which were input into the STRING database to generate protein-protein interaction (PPI) networks, followed by gene ontology (GO) and KEGG pathway analyses. Cytosccape software was employed to construct target networks for core components and therapeutic effects of QJMM, facilitating molecular docking. Conjunctival tissue was obtained from CCH and normal control groups (NC) (ethics approval number: PTEC-A-2024-30-1), subjected to immunohistochemical staining, and cultured as primary conjunctival fibroblasts. Fibroblasts were divided into NC, CCH, CCH+QJMM, and CCH+ nuclear factor κappa-B (NF-κB) inhibitor pyrrolidine dithiocarbamate (PDTC) groups. Immunofluorescence staining, cytokine assays, Western blot (WB), and quantitative reverse transcription PCR (qRT-PCR) were performed on each group. Results showed that 176 active compounds were screened from QJMM, yielding 293 target molecules in total. CCH identified 27 targets, with 12 overlapping targets identified via Venn diagram analysis. The PPI network diagram indicated tumor necrosis factor (TNF), matrix metalloproteinase (MMP)-2, MMP-9, and interleukin (IL)-6 as key targets. GO analysis yielded 91 enriched entries, while KEGG analysis identified 45 related entries and 110 signaling pathways, revealing the NF-κB signaling pathway. Molecular docking revealed that the core components of CCH exhibited strong binding affinity with the key target NF-κB, indicating potent biological activity. Subsequent immunohistochemical staining demonstrated significantly higher Toll-like receptor 4 (TLR4) and NF-κB expression in conjunctival tissues of the CCH group compared to the control group (P < 0.05). Conjunctival fibroblast experiments confirmed that NF-κB and its upstream factors TLR4 and myeloid differentiation primary response 88 (MYD88) exhibited higher protein and mRNA expression in the CCH group than in the NC group. Furthermore, expression of downstream inflammatory cytokines IL-6, IL-1β, and TNF-α was significantly elevated compared to the NC group (P < 0.05). Following administration of QJMM and PDTC, the expression of NF-κB and its upstream factors TLR4 and MYD88 protein, mRNA and immunofluorescence intensity decreased compared to the CCH group (P < 0.05). In conclusion, QJMM exerts multi-targeted, multi-pathway regulatory effects in treating CCH. Both network pharmacology and experimental studies confirm NF-κB plays a pivotal role in CCH pathogenesis and progression. QJMM effectively treats CCH by inhibiting the NF-κB signaling pathway.

This study aims to investigate the pharmacological effects of procyanidin B3 (Pro B3) on non-alcoholic fatty liver disease (NAFLD) and its molecular mechanism underlying the regulation of fatty acid oxidation (FAO) in hepatocytes during NAFLD, thereby providing experimental evidence for the clinical application of Pro B3 in NAFLD. Mice were fed a high-fat diet (HFD) for 12 weeks to induce NAFLD and received oral gavage administration for 12 weeks. The pharmacological effect of Pro B3 in improving NAFLD was assessed through serum biochemical indexes, liver pathological changes and lipid droplet formation in the liver using Hematoxylin-eosin (H & E) staining and oil red O staining. This study protocol was approved by the Experimental Animal Ethics Committee of Gannan Medical University (approval No.: 2025443). In the in vitro experiments, the HepG2 cells were induced with a mixture of palmitic acid and oleic acid (2∶1) to create a cellular model of NAFLD and evaluate the cytotoxicity and cell viability of ProB3 on HepG2 cells. The expressions of FAO-associated genes such as acyl-CoA synthetase long chain family member 5 (ACSL5), carnitine palmitoyltransferase 1 (CPT1), acetyl-CoA carboxylase (ACC), and GATA binding protein 3 (GATA3) were detected by immunohistochemical, immunofluorescence, qRT-PCR and Western blot. Mechanistically, the JASPAR database and dual-luciferase reporter gene assays were used to explore the transcription factors and binding sites of ACSL5. By silencing GATA3 using siRNA, the effects of Pro B3 on the expression of ACSL5 and lipid deposition were examined. The results demonstrated that Pro B3 significantly improves the NAFLD-related indicators in mice, reduces serum lipid levels and lipid droplet accumulation in HepG2 cells, and up-regulates the expression of FAO-related genes, such as ACSL5, CPT1, ACC. Furthermore, GATA3 might be a transcription factor of ACSL5, and the transcriptional regulatory site might be "ACATAG". Moreover, Pro B3 significantly upregulated the expression of GATA3 mRNA and protein levels, while silencing GATA3 significantly inhibited the expression of ACSL5 mRNA and protein levels, as well as weakened the inhibitory effect of Pro B3 on lipid accumulation in HepG2 cells. In conclusion, Pro B3 may be an effective lead compound for treating NAFLD by promoting FAO in hepatocytes through regulation of the GATA3/ACSL5 pathway, thereby ameliorating NAFLD.

Exploring cellular senescence inducers is one of the approaches to screen anti-tumor active lead compounds. Whether glaucocalyxin B induces cellular senescence remains to be elucidated. This study aims to investigate the effect and mechanism of glaucocalyxin B on the senescence of lung adenocarcinoma A549 cells. 2 and 5 μmol·L^-1 of glaucocalyxin B induced senescence in A549 cells. Glaucocalyxin B significantly increased the levels of cyclin-dependent kinase inhibitor 1A (p21) and the proportion of β-galactosidase staining-positive cells in A549 cells. Glaucocalyxin B had no significant effect on the levels of peroxiredoxin Ⅰ (Prdx Ⅰ) protein in A549 cells but significantly inhibited its enzymatic activity. Knockdown of Prdx Ⅰ induced senescence in A549 cells. Glaucocalyxin B inhibited the enzyme activity of Prdx Ⅰ, leading to the increase of reactive oxygen species (ROS), and the ROS scavenger N-acetylcysteine attenuated glaucocalyxin B-induced the senescence of A549 cells. Either glaucocalyxin B or Prdx Ⅰ knockdown significantly upregulated the levels of tumor protein p53 in A549 cells, while knockdown of p53 significantly inhibited glaucocalyxin B-induced the senescence of A549 cells. In conclusion, glaucocalyxin B can induce senescence in lung adenocarcinoma cells by inhibiting the Prdx Ⅰ/ROS/p53 pathway, which might be one of its anti-tumor mechanisms.

Herpes simplex virus type 1 (HSV-1) is a neurotropic virus that establishes latent infection in the trigeminal ganglia following primary infection. Its reactivation can be triggered by both internal and external factors, including heat stress, emotional stress, excessive consumption of warm-natured foods, and the administration of tonic and warming traditional Chinese medicines (TCMs). The fire-heat symptoms in the upper facial region induced by HSV-1 reactivation are consistent with the "Shanghuo" syndrome described in TCM. In this study, HSV-1 infection models established by corneal scarification and intranasal inoculation were systematically compared. Dynamic changes throughout the entire "primary infection-latency-reactivation" cycle were monitored using multiple parameters, including mortality, body weight, ocular manifestations, viral glycoprotein B (gB) mRNA expression, and restless behavior. This study aimed to provide a basis for model selection in investigating the biological mechanisms underlying TCM "Shanghuo" and for screening anti-HSV-1 therapeutics. The results showed that during the primary infection phase (0-14 d), mice in both groups exhibited body weight loss and ocular surface inflammation, with an earlier onset (3 d vs 5 d) and more severe ocular damage observed in the corneal scarification group. During the latency phase (28 days post-infection), latency-associated transcript (LAT) was detected in the trigeminal ganglia in both groups, with a higher latency rate in the corneal scarification group (100% vs 87.5%). Following heat stress-induced reactivation, the intranasal inoculation group exhibited a significantly higher reactivation rate (85.7% vs 66.6%), more pronounced restless behavior, and elevated levels of gB mRNA expression and viral titers in brain tissue compared with the corneal scarification group. In conclusion, both corneal scarification and intranasal inoculation successfully establish HSV-1 models encompassing the full cycle of primary infection, latency, and reactivation. Dynamic monitoring of infection stages can be achieved by detecting gB mRNA in eye wash fluid. The corneal scarification model, characterized by a high latency rate and prominent ocular manifestations, is suitable for studies of viral keratitis and related therapeutic evaluation. In contrast, the intranasal inoculation model, with a higher reactivation rate and marked restless behavior, exhibits similarities to the restless manifestations of the TCM "Shanghuo" heat pattern, suggesting its potential utility in evaluating the efficacy of heat-clearing TCMs. Animal experiments conducted in this study were approved by the Ethics Committee of Jinan University (approval No. IACUC-20260311-06).

It is well established that baicalin induces the polarization of macrophages toward the M1 phenotype via glycolytic pathways; however, the underlying regulatory mechanisms remain unclear. This study aimed to identify the key upstream targets and regulated transcription factors involved in baicalin's regulation of glycolytic metabolism. Using the RAW264.7 cell model, RNA-seq and ATAC-seq technologies were employed, combined with Western blot, qRT-PCR, siRNA, and dual-luciferase reporter assays for detection and validation. The results showed that baicalin, at concentration of 100 μmol·L^-1, promoted RAW264.7 cell M1 polarization accompanied by high expression of Trem1; co-treatment with the Trem1 inhibitor LP17 reversed baicalin-mediated RAW264.7 cell polarization and the activation of downstream glycolytic signaling pathways. Among the transcription factors associated with the Trem1 promoter region, Fos was shown to be involved in the regulation of RAW264.7 cell polarization by baicalin. This study reveals that baicalin promotes downstream glycolytic metabolism by regulating the transcription factor Fos to activate Trem1, thereby driving macrophage polarization toward the M1 phenotype, providing a new metabolic mechanism to support its therapeutic potential in tumor immunoregulation and inflammation-related diseases.

To rapidly screen natural antibacterial active components targeting lipopolysaccharide (LPS) through the detection of traditional Chinese medicine (TCM) monomers via plasmonic near-field sensing. In this study, a plasmonic near-field biosensor based on gold nanoislands (AuNIs) and LPS (LPS@AuNIs biosensor) was constructed, and the LPS antagonistic activity of natural products derived from TCM was evaluated by the change in phase response. The limit of detection (LOD) and limit of quantification (LOQ) of the LPS@AuNIs biosensor for polymyxin B (PB), which specifically binds to LPS, were 6.34 ± 0.002 374 and 61.7 ± 0.001 735 nmol·L^-1, respectively. Detection results of 14 TCM monomers showed that the LPS@AuNIs biosensor exhibited excellent concentration-dependent responses to both 1, 2, 3, 4, 6-O-pentagalloylglucose (β-PGG) and geniposide (GP). Comparative analysis revealed that β-PGG displayed potent binding activity, with a dissociation constant (k_a) superior to that of PB. Further verification via minimum inhibitory concentration (MIC) and displacement assays confirmed that β-PGG possessed strong binding affinity comparable to PB. The developed biosensor enables specific binding between LPS and analytes, featuring high sensitivity and detection efficiency for high-precision quantitative analysis. This study innovatively integrates nanophotonics with natural medicine research, establishing an integrated system of "drug screening-target validation" and providing a high-efficiency technical platform for the rapid and effective screening of natural antibacterial active components.

Polypyrimidine tract-binding protein 1 (PTBP1) is a critical regulatory target in cancers and neurological disorders. However, the development of its small-molecule inhibitors has been hampered by the lack of traditional druggable pockets on its RNA-binding domains. This study employed a computer-aided drug design (CADD) strategy focusing on a dynamic binding pocket mediated by the flexible linker3 within the PTBP1 RBD3/4 domain. A commercial library containing approximately 1.6 million compounds was subjected to structure-based virtual screening. Following initial filtering based on lead-likeness rules and subsequent molecular docking, 14 candidate compounds were synthesized. Their binding activities were experimentally validated using prokaryotically expressed and purified PTBP1-RBD3/4 recombinant protein via bio-layer interferometry (BLI). Six compounds with micromolar affinity were identified, among which C17414 exhibited the highest binding affinity (K_D = 1.6 µmol·L^-1). Molecular docking analysis revealed that C17414 efficiently occupies the target pocket through multiple interactions, including hydrogen bonds, salt bridges, π-π stacking and hydrophobic interactions. Systematic ADMET (absorption, distribution, metabolism, excretion and toxicity) property predictions indicated that C17414 and C7485 possess a favorable balance of lipophilicity, polar surface area and membrane permeability, suggesting promising drug-likeness and potential for central nervous system drug development. This work not only discovers a series of high-affinity PTBP1 inhibitor hits and validates the feasibility of targeting its dynamic pocket but also lays a solid foundation for developing novel therapies against PTBP1.

Nuclear receptor-binding SET domain protein 3 (NSD3) plays an important role in the initiation and progression of lung squamous cell carcinoma (LUSC), whereas small-molecule inhibitors targeting NSD3 remain limited. This study targeted the NSD3-PWWP1 domain and identified 37 potential NSD3-binding compounds through Glide-based hierarchical virtual screening followed by drug-likeness filtering. Surface plasmon resonance (SPR) analysis showed that compounds 2, 4, 20 and 27 could directly bind to NSD3, among which compound 2 exhibited the strongest equilibrium dissociation constant (K_D = 0.95 μmol·L^-1), approaching that of the positive control BI9321 (0.46 μmol·L^-1). Molecular docking analysis indicated that all four candidate compounds could occupy the NSD3-PWWP1 binding pocket and form stable interactions with key residues. At 30 μmol·L^-1, the four candidate compounds showed different degrees of inhibitory activity against three NSD3-high LUSC cell lines. Notably, compound 2 displayed a half maximal inhibitory concentration (IC₅₀) of 4.08 ± 1.01 μmol·L^-1 in HCC95 cells, which was superior to that of BI9321 (17.62 ± 3.11 μmol·L^-1). Meanwhile, these candidate compounds did not show obvious cytotoxicity toward HEK293T cells and exhibited favorable ADMET profiles and physicochemical property distributions. In conclusion, several potential NSD3 inhibitors were identified in this study. Among them, compound 2, a novel coumarin-based scaffold distinct from BI9321, represents a promising lead for further optimization as an anti-LUSC agent.

In this study ten new compounds Sd-01-Sd-10 were synthesized via acylation from sitagliptin and various fatty acyl chlorides. Animal experiments approved by the Institutional Animal Care and Use Committee of Jilin Provincial Institute of Pharmaceutical Research (approval No. 2025001). The target compounds were evaluated for their in vivo hypoglycemic activity using a type 2 diabetic mouse model, with subsequent structure-activity relationship analysis and further assessment of the most active compounds via an oral glucose tolerance test. Meanwhile, a maximum feasible dose test was conducted in healthy mice for preliminary safety evaluation. The results indicated that this series of compounds exhibited promising hypoglycemic activity and a high level of safety, which was supported by the observation of no abnormal behavior or mortality in healthy mice after a single oral dose of 2 000 mg·kg^-1. Among them, Sd-06 showed activity comparable to sitagliptin, whereas Sd-05 was more potent. Notably, Sd-05 at the lower dose of 30 mg·kg^-1 effectively improved glucose tolerance in normal mice, surpassing the efficacy of sitagliptin at 50 mg·kg^-1, highlighting its value for further development.

Three new eudesmane sesquiterpenoids (oxyphylleudnes K-M) were separated and purified from the ethyl acetate fraction of Alpinia oxyphylla Fructus, by using various column chromatography methods such as MCI Gel CHP-20, YMC GEL ODS-A, and silica gel columns, combined with preparative thin-layer and semi-preparative liquid chromatography methods. Their structures were identified by modern spectroscopic techniques (1D and 2D NMR, UV, IR and MS) and with computational NMR and ECD methods. Among them, there are two nor-eudesmane sesquiterpenes (1, 2) and one seco-eudesmane sesquiterpene (3).

Anti-renal fibrosis activity screening was performed on the extracts of Liquidambar formosana via the activity-tracing approach, which led to the identification of the bioactive fractions. Subsequent separation and purification afforded seven abietane-type diterpenoids, including one new diterpenoid, liquidformosane A (1) and six known ones. The planar structure of the new compound was established by nuclear magnetic resonance spectroscopy (NMR), and its stereoconfiguration was confirmed by quantum chemical calculations for simulation and prediction of electronic circular dichroism (ECD) spectra. The Western blot method was used to evaluate the in vitro anti-renal fibrosis activity of compounds 1-7, and the results showed that compounds 5-7 all exhibited significant anti-renal fibrosis activity.

The chemical constituents of Stellera chamaejasme L. were separated by comprehensive column chromatography on MCI, ODS C-18, Toyopearl HW-40C, and silica gels. Structures of the isolated compounds were determined by NMR (nuclear magnetic resonance), MS (mass spectrometry), IR (infrared spectroscopy), and quantum chemical calculations. Nine compounds were isolated from the chloroform fraction 95% ethanol extract of the titled plant and identified as chamaejaslactone A (1), stelleranoid A (2), stelleranoid D (3), stelleranoid E (4), wikstronone A (5), wikstronone C (6), chamaejasmone D (7), daphnauranol A (8), and holosericin B (9), respectively. Among them, compound 1 was characterized as the first example of a sesquiterpenoid with an unprecedented 7,8-seco-guaiane skeleton. In the in vitro bioassay, compound 1 exhibited moderate inhibitory activity against Escherichia coli-originated β-glucuronidase with an IC₅₀ value of 51.79 ± 3.21 μmol·L^-1.

The ethanol extract of Pteris multifida was isolated and purified using various column chromatographic methods, including silica gel, Sephdex LH-20 and semi-preparative HPLC. Based on the physicochemical properties of the isolated compounds, their structures were elucidated by spectroscopic data such as MS, NMR, and CD. From the ethyl acetate fraction of P. multifida,18 terpenoids were isolated, including 14 sesquiterpenoids possessing a 1H-inden-1-one nucleus (1-14) and four nortriterpenoids with a megastigmane skeleton (15-18). They were identified as (2S,3S)-2,5,7-trimethyl-3-hydroxyl-6-carboxyl-indan-1-one (ptemulic A) (1), (2S,3S)-2,7-dimethyl-5,6-dihydroxymethyl-indan-1-one (ptemulinol A) (2), dehydropterosin B (3), 6-(1,2-dihydroxyethyl)-2,5,7-trimethyl-indan-1-one (4), pterosin D (5), (3R)-pterosin (6), pterosin D 3-O-β-D-glucopyranoside (7), (3R)-pteroside W (8), (3R)-pteroside D (9), 14-acetyl-pterosin A (10), pterosin A (11), 11-acetyl-pterosin A (12), pterosin A2 (13), (2S,3S)-pterosin C 3-O-β-D-glucopyranoside (14), dearabinosyl pneumonanthoside (15), grasshopper ketone (16), vomifoliol (17), roseoside Ⅰ (18). Compounds 1 and 2 were new compounds, while compounds 3, 4, 10-13, 16 and 18 were isolated from this plant for the first time. New compounds 1 and 2 showed anti-inflammatory activity by inhibiting the production of nitric oxide (NO), with IC₅₀ values of 36.7 and 61.2 μmol·L^-1, respectively.

Fufang E jiao Jiang (FFEJ), a Chinese patent medicine with the effects of tonifying Qi and nourishing blood, as well as nourishing Yin and moistening dryness, is clinically used as an adjuvant therapy for tumor patients and has been proven to possess certain anti-tumor activity. To identify the material basis for its anti-tumor efficacy, this study employed the cell counting kit-8 (CCK-8) assay to screen the anti-tumor active fractions of FFEJ on Lewis cells, and validated their activity through cell cloning, apoptosis, invasion, and migration assays. Ultra-high performance liquid chromatography coupled with quadrupole-Orbitrap high-resolution mass spectrometry (UHPLC-Q-Exactive-Orbitrap-MS), integrating network pharmacology analysis was used to systematically analyze the chemical components and potential action mechanisms of the active fractions. The results showed that the n-hexane (F2) and ethyl acetate (F3) extracts were the main anti-tumor active fractions of FFEJ. Both fractions dose-dependently inhibited Lewis cell colony formation, induced cell apoptosis, significantly reduced the horizontal and vertical migration abilities of Lewis cells, and suppressed their invasion and distant metastasis. F2 was dominated by primary metabolites such as organic acids and sugars, while F3 was rich in secondary metabolites including ginsenosides and iridoids. Network pharmacology analysis indicated that 60 key active components in F3 act on core targets such as proto-oncogene tyrosine-protein kinase Src (SRC), signal transducer and activator of transcription 3 (STAT3), and RAC-alpha serine/threonine-protein kinase (AKT1), regulating signaling pathways closely related to tumor apoptosis, migration, and invasion, including phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT), interleukin-17 (IL-17), tumor protein 53 (p53), and hypoxia-inducible factor-1 (HIF-1), thereby exerting synergistic anti-tumor effects. This study systematically elucidates the multi-component, multi-target, multi-pathway anti-tumor characteristics of FFEJ, identifies its active fractions and potential pharmacodynamic substances, and provides experimental evidence for the clinical application of FFEJ in adjuvant tumor therapy.

This study aims to analyze the components absorbed into the blood and brain after intragastric administration of Paeonia rockii pollen in rats, and to explore the pharmacodynamic material basis. The chemical constituents of Paeonia rockii pollen, as well as those in medicated serum and brain tissue, were investigated using ultra-performance liquid chromatography-electrospray ionization-triple quadrupole-linear ion trap mass spectrometry (UPLC-ESI-Q-TRAP-MS/MS) based on widely targeted metabolomics technology. Differential components were screened using principal component analysis and orthogonal partial least squares discriminant analysis, followed by functional annotation and pathway enrichment analysis, and the mass spectrometric fragmentation patterns of blood-brain barrier-penetrating components were deduced. A total of 340 components were identified from Paeonia rockii pollen, primarily including flavonoids, terpenoids, and phenolic acids. In the medicated serum, 38 prototype absorbed components such as methyl gallate, myricetin, trachelogenin, tyrosol, and albiflorin, along with two metabolites (inosine and hypoxanthine) were identified. In the brain tissue, 15 prototype components such as gallic acid, isorhamnetin, 4-hydroxycoumarin, resveratrol, and paeoniflorin were detected, and the plasma concentration was the highest after 2 h of intragastric administration. These components were mainly enriched in pathways such as longevity regulation, microbial metabolism in different environments, biosynthesis of secondary metabolites, flavonoid degradation, nicotinate and nicotinamide metabolism, and purine metabolism. The animal experiment protocol was approved by the Experimental Animal Ethics Review Committee of Lanzhou Institute of Food and Drug Control, Gansu Province (Approval No. 2023-010).

This study comprehensively evaluated the reproductive safety of Ramulus Mori (Sangzhi) alkaloids (SZ-A) in rats, analyzed the toxicokinetics of the main active ingredient 1-deoxynojirimycin (DNJ) during embryo-fetal development, and determined the no-observed-adverse-effect level. Reproductive toxicity assessment was conducted in three stages (this study was reviewed and approved by the Institutional Animal Care and Use Committee of Beijing Union-Genius Pharmaceutical Technology Development Co., Ltd., ethics numbers: SN22072-DS, SN21288-DS, SN22074-DS). In stage Ⅰ, rats were orally administered SZ-A at doses of 100, 200 and 500 mg·kg^-1 (calculated as total alkaloids) to evaluate its effects on fertility and early embryonic development, including sperm count and motility in males, as well as mating and pregnancy rates in females. In stage Ⅱ, pregnant rats were given SZ-A by gavage at 100, 200 and 500 mg·kg^-1 to examine its impact on maternal health, embryo-fetal development, and fetal growth, while DNJ concentrations in maternal plasma, placenta and fetus were measured. In stage Ⅲ, pregnant rats were orally administered SZ-A at doses of 250, 500 and 750 mg·kg^-1. Indicators were examined to evaluate perinatal safety, including maternal reproductive function; offspring growth and development, behavior and learning/memory function, reproductive function; and F2 embryonic development. The results showed that SZ-A had no adverse effects on fertility, early embryonic development in both male and female rats, and on embryo-fetal development in pregnant rats. The plasma exposure of DNJ in pregnant rats showed a linear relationship with the administered dose. However, only limited distribution of DNJ was observed in the placenta, and the drug was undetectable in whole fetal tissues. At all tested doses, SZ-A showed no reproductive toxicity. This was evidenced by normal sex ratio, growth and development, and reproductive function in the F1 generation; embryonic development of the F2 generation was also unaffected. The no-observed-adverse-effect level (NOAEL) of SZ-A over the entire reproductive cycle in rats was determined to be 500 mg·kg^-1, indicating favorable safety. This finding provides essential reproductive safety data to support the potential clinical application of SZ-A in special populations such as diabetic patients of childbearing age, gestational diabetes mellitus (GDM), and patients with polycystic ovary syndrome (PCOS).

As an important form of drug delivery, the transdermal drug delivery system has demonstrated unique advantages and potential in the treatment of various diseases such as chronic pain and skin diseases. However, in the prevention and treatment of skin photo-damage, it still faces issues like low transdermal efficiency and poor drug retention. Therefore, there is an urgent need to develop a highly efficient and safe drug delivery technology. In this study, solid lipid nanoparticles loaded with vitamin A palmitate (VAP-SLN) were designed and prepared to improve the encapsulation efficiency and stability of vitamin A palmitate (VAP), aiming to enhance the therapeutic effect of VAP on skin photodamage via transdermal drug delivery. The high-speed shearing and high-pressure homogenization method were used to prepare VAP-SLN. After prescription screening, it was determined that VAP-SLN was mainly composed of solid lipid glyceryl behenate and liquid lipid medium-chain triglycerides, phospholipids, Tween 80, alkyl polyglycosides and VAP. The prepared VAP-SLN had a particle size of approximately 200 nm, a spherical structure, and an encapsulation efficiency of 91.1%. Its phase transition temperature was 59.8 ℃. This characteristic endowed it with excellent thermal stability and significantly improved the stability of the encapsulated VAP. VAP-SLN exhibited the advantage of sustained-release characteristics and intradermal retention, which could significantly prolong the local drug action and enhance the transdermal drug delivery efficiency. At the cellular level, VAP-SLN exhibited good cellular uptake and retention ability in HaCaT cells, thereby enhancing the effect of VAP in alleviating cellular oxidative stress and inflammatory responses. In the mouse skin photodamage model, VAP-SLN could effectively improve the reduction of collagen fibers and the decrease in collagen volume in photodamaged skin, restore the epidermal thickness and morphology to a healthy level, and effectively prevent and treat ultraviolet-induced skin damage. The animal experiments were approved by the Institutional Animal Care and Use Committee of Yantai Institute of Materia Medica (Approval No.: 20250701-01). In conclusion, VAP-SLN significantly alleviates skin photodamage through antioxidant and anti-inflammatory mechanisms, providing a new strategy for the transdermal delivery of VAP and the prevention of skin photodamage.

Oplopanax elatus Nakai is a deciduous shrub within the Araliaceae family and is classified as a first-class protected plant in Jilin Province. To elucidate the structural characteristics of its chloroplast genome and phylogenetic relationships, this study conducted chloroplast genome sequencing of one cultivated individual and five wild populations of O. elatus, followed by comparative chloroplast genomic and phylogenetic analyses with published congeneric species of Oplopanax and allied genera of Araliaceae deposited in GenBank. The results revealed that the chloroplast genomes of the six O. elatus accessions ranged from 156 108-156 193 bp in length, with a uniform GC content of 38.03%. A total of 129-133 genes were annotated, comprising 87-88 protein-coding genes, 6-8 rRNA genes, and 36-37 tRNA genes. The chloroplast genome of cultivated O. elatus contained one additional protein-coding gene compared to those of wild accessions. Using the chloroplast genome sequence of cultivated O. elatus as reference, sequence alignment against wild populations identified 36 single nucleotide polymorphism (SNP) loci and 10 insertion-deletion (InDel) loci. SNPs were distributed in both intergenic spacers and coding sequences (CDS), whereas all InDels were restricted to the large single-copy (LSC) region. Nucleotide diversity (Pi) analysis of the 13 chloroplast genomes detected two hypervariable regions, clpP1-exon3_clpP1-exon2 and ycf1, which are proposed as candidate DNA barcodes and molecular markers for O. elatus. Analysis of codon preference indicated that all 13 samples exhibited a preference for codons terminating in A or T. Discrepancies in codon numbers were observed between cultivated and wild O. elatus chloroplast genomes. A total of 545 simple sequence repeat (SSR) loci and 498 dispersed repeat sequences were identified within the 13 chloroplast genomes. Phylogenetic reconstruction based on branch support values and clustering patterns partitioned the eight Araliaceae genera into four clades. The genus Oplopanax formed a highly supported monophyletic lineage positioned centrally within the tree, diverging subsequently to Panax and constituting a sister group to the fourth major clade, with its closest affinity to Schefflera. This investigation advances the understanding of evolutionary dynamics within Oplopanax and allied genera, with the acquired genomic resources providing a foundation for DNA barcode screening and germplasm conservation of the genus.

Hyperforin is a key pharmacologically active constituent responsible for the antidepressant and other therapeutic effects of Hypericum perforatum. However, its content within medicinal materials varies significantly according to the harvest season and the plant organ, leading to inconsistencies in drug quality. To elucidate the dynamic patterns of hyperforin biosynthesis, this study systematically analyzed the accumulation of hyperforin and the expression levels of key biosynthetic genes in the roots, stems, leaves, and flowers of H. perforatum collected across different seasons. The results revealed that hyperforin accumulation exhibits distinct seasonal fluctuations and tissue specificity, following the order of flower > leaf > stem > root. Notably, the hyperforin content in leaves reached its peak in the second season. The expression patterns of key biosynthetic genes (e.g., HpPKS, HpPT1-4) were inconsistent with the accumulation trends of the final product in terms of both temporal (seasonal) and spatial (tissue) dynamics. Gene expression was predominantly active during the first season and was significantly high in roots, where hyperforin content was the lowest. These findings suggest that the regulation of hyperforin biosynthesis involves complex multi-level mechanisms, with an asynchronous regulatory relationship between gene expression and the accumulation of the final product. By dissecting the spatiotemporal characteristics of hyperforin biosynthesis from both the perspectives of metabolite accumulation and gene expression, this study not only provides a theoretical basis for further research into its regulatory mechanisms but also offers valuable insights for the quality control, metabolic regulation, variety breeding, and the development of science-based cultivation and harvesting strategies of H. perforatum.

The traditional Chinese medicinal material Mugua is derived from the dried fruit of Chaenomeles speciosa (Sweet) Nakai (Rosaceae). The material produced in Xuancheng, Anhui Province, is regarded as a geo-authentic medicinal material and is commonly referred to as "Xuan Mugua". C. speciosa is a medicinal and edible plant with a long history of use in traditional Chinese medicine. In this study, we sequenced and assembled the complete chloroplast genome of C. speciosa using the Illumina platform and performed comprehensive genome annotation. Repetitive sequences were analyzed with REPuter and MISA, codon usage bias was evaluated via relative synonymous codon usage (RSCU), and comparative analyses of inverted repeat (IR) boundaries and phylogenetic relationships were conducted among closely related Chaenomeles species. The chloroplast genome is 159 991 bp in length, exhibiting a typical quadripartite structure with a GC content of 36.5%, and encodes 126 genes, including 83 protein-coding genes, 35 tRNAs, and 8 rRNAs. Seventy-six simple sequence repeats (SSRs) and 55 long repeat sequences were identified, with SSRs primarily consisting of A/T mononucleotides and predominantly located in the large single copy (LSC) and intergenic regions. Codon usage analysis showed that 30 codons have a preference (RSCU > 1), mostly ending with A/U. Comparative analysis of IR boundaries indicated high conservation within the genus. Genome variation analysis revealed four highly variable regions in the LSC (trnR-atpA, petN-psbM, trnT-GGU-psbD, trnV-M). Phylogenetic analysis confirmed that Chaenomeles is monophyletic, with C. speciosa closely related to C. japonica and C. tibetica. This work provides the first detailed characterization of the C. speciosa chloroplast genome and phylogeny, and the identified SSRs and highly variable regions represent valuable resources for germplasm identification and evolutionary studies in Chaenomeles.