Lysine-specific demethylase 1 (LSD1) is a member of the flavin adenine dinucleotide (FAD)-dependent monoamine oxidase family and is highly expressed in various tumor cells. LSD1 plays a crucial role in tumor progression by participating in multiple processes such as tumor cell proliferation, invasion, differentiation, and immune cell killing. Studies have shown that LSD1 exerts its oncogenic functions not only by demethylating histone and non-histone substrates but also through demethylase activity-independent protein-protein interactions, thereby diversifying its mechanisms of action. With the increasing depth of research into the tumor-promoting mechanisms of LSD1, targeting LSD1 for cancer therapy has become a major focus in the field. This review summarizes the oncogenic functions of LSD1 through its interactions with different proteins and provides an overview of LSD1 inhibitors that have entered clinical stages. A comprehensive understanding of the roles of LSD1 in tumors will offer theoretical support for developing more effective and specific LSD1 inhibitors, thereby guiding future directions for enhanced anti-tumor therapies targeting LSD1.

Hyperuricemia is a metabolic disease characterized by abnormally elevated level of serum uric acid, which is the key pathological basis of gout. Oral medications reduce blood uric acid level by inhibiting uric acid production and/or promoting uric acid excretion. Xanthine oxidoreductase is the rate-limiting enzyme for uric acid synthesis, by inhibiting its activity, the production of uric acid will be reduced. Urate transporter 1 (URAT1), one of the important proteins mediating uric acid reabsorption in kidney, is also a main target of anti-hyperuricemic drugs. By inhibiting URAT1 activity, the uric acid excretion will be promoted, thereby reducing the blood uric acid. At present, three xanthine oxidoreductase inhibitors and two urate transporter 1 inhibitors have been launched, and multiple compounds are currently in clinical trials. The physiological functions and metabolic pathways of uric acid, etiology of hyperuricemia and its therapeutic targets are introduced in this review. We analyze the efficacy and safety of oral therapeutic agents, to provide insights for the development of new drugs and the rational clinical use of medications in the treatment of hyperuricemia.

Ferroptosis, a novel programmed cell death mechanism characterized by the iron-dependent accumulation of lipid peroxides, is precisely regulated by multiple signaling pathways, including intracellular iron metabolism, lipid peroxidation, and antioxidant systems. Recent studies have highlighted the dual role of ferroptosis in the fate determination of hepatic stellate cells (HSCs) and hepatocytes, indicating its significant involvement in the progression of hepatic fibrosis. This is corroborated by ongoing research that explores the molecular mechanisms of ferroptosis and its potential as a therapeutic target in various liver diseases, including liver fibrosis. This article elucidates the key molecular mechanisms of ferroptosis and its dual role in liver fibrosis, while systematically reviewing the latest progress in anti-fibrotic research using natural products and synthetic compounds targeting key ferroptosis molecules. The study aims to offer a solid theoretical basis and strategic guidance for the development of innovative anti-fibrotic drugs by regulating ferroptosis pathways.

Atherosclerosis (AS) is a common pathological basis for cardiovascular and cerebrovascular diseases. Chinese ethnomedicine, with its distinct theoretical frameworks and abundant resources, shows unique potential in managing AS. This review systematically synthesizes the pathogenesis of AS and corresponding therapeutic strategies from various Chinese ethnomedical systems, revealing that "physiological imbalance" serves as a common underlying mechanism. We detail the mechanisms of these ethnomedicines, including improving the composition of gut microbiota, regulating lipid metabolism, mitigating oxidative stress, suppressing inflammation and modulating vascular homeostasis. Available clinical evidence is critically evaluated. Key challenges are discussed, such as the need for theoretical modernization, deeper mechanistic studies, and more rigorous clinical validation. Promising future research directions are also outlined.

Bile acids play a central role in maintaining cholesterol homeostasis and regulating lipid metabolism, exhibiting extensive medicinal value. However, conventional extraction, separation, and chemical synthesis methods are unable to meet the demands of current industrial production, rendering biocatalysis a highly promising alternative for bile acid manufacturing. Cytochrome P450 (CYP450) enzymes, which catalyze the hydroxylation of inert C(sp³)-H bonds, serve as the core enzymes in the bile acid biosynthetic pathway. Through strategies such as enzyme design, redox partner engineering, and optimization of catalytic systems, the catalytic efficiency of CYP450s has been significantly enhanced, enabling the efficient biosynthesis of various bile acids. This review summarizes the characteristics and synthetic strategies of bile acids, as well as the optimized application of CYP450 hydroxylases, providing a reference for the green production of bile acids and related drug research.

Papain-like protease (PL^pro or PLP) is a key non-structural protein in the replication process of coronaviruses. It is responsible for the cleavage and processing of viral polyproteins, and has deubiquitination and deISGylation activities, which can inhibit the host's innate immune response. Due to the high conservation of PL^pro among various coronaviruses such as SARS-CoV-2, SARS-CoV, and MERS-CoV, it has become an important target for the development of broad-spectrum coronavirus inhibitors. Since the outbreak of the COVID-19 pandemic, research on PL^pro has advanced rapidly, and multiple classes of candidate molecules with clear structures and definite mechanisms have been identified, including non-covalent inhibitors (e.g., GRL0617 and its derivatives), covalent inhibitors (e.g., vinyl ester compounds), and inhibitors derived from natural products. This article reviews the research progress of PL^pro-targeted inhibitors, covering their structural characteristics, activity data, mechanism of action, and discovery strategies, aiming to provide theoretical support and design ideas for the development of broad-spectrum anti-coronavirus drugs.

Toll-like receptors (TLR), as important pattern recognition receptors (PRR), are widely distributed on the endosomal membrane surface of immune cells such as macrophages and dendritic cells. Among them, TLR7 has attracted much attention due to its specific recognition of the molecular pattern of viral single-stranded RNA (ssRNA). After TLR7 recognizes single-stranded ribonucleic acid, it activates a series of signal cascade reactions, initiating a high level of systemic adaptive immune response to kill the virus. In recent years, artificially synthesized TLR7 agonists have been used in the clinical treatment of diseases such as HPV infection and basal cell carcinoma. They activate the immune microenvironment by mimicking the structure of viral RNA, significantly enhancing the function of antigen-presenting cells and promoting Th1-type immune response. This paper summarizes the research progress of TLR7 agonists in the past decade and their application in various diseases, aiming to provide references for relevant research and development.

Targeted anticancer agents have become a cornerstone of cancer therapy, specifically acting on targets such as enzymes, receptors, and proteins to inhibit the growth and proliferation of cancer cells. In addition to enhancing therapeutic efficacy, these agents significantly reduce adverse effects and improve patients' quality of life. Carboranes, a class of boron cluster compounds with unique structural features in medicinal chemistry, hold considerable potential for optimizing the performance of targeted anticancer drugs. Characterized by three-dimensional aromaticity, high hydrophobicity, and substantial steric bulk, carboranes have been incorporated into the molecular scaffolds of known targeted anticancer agents by numerous researchers. Their distinct physicochemical properties are exploited to modulate the biological behavior of parent molecules, thereby strengthening drug-target interactions, improving target selectivity, and enhancing metabolic stability. Furthermore, due to their exceptionally high boron content, carboranes serve as efficient boron carriers in boron neutron capture therapy. This review summarizes the latest research progress on the application of carboranes in the design of targeted anticancer drugs, aiming to provide valuable insights and references for the development of carborane-based targeted anticancer agents.

Triptolide, as a key active ingredient in the traditional Chinese medicine Tripterygium wilfordii, has shown significant potential in the treatment of kidney diseases. This article systematically reviews the research progress of its application in the treatment of kidney diseases, covering chemical properties, pharmacological and toxicological mechanisms, and targeted drug delivery strategies. In terms of pharmacological mechanisms, triptolide mainly exerts renal protection through multiple pathways such as protecting podocytes, inhibiting inflammation, and anti fibrosis. However, its significant toxicity is a key limiting factor for clinical applications, involving complex metabolic pathways, signaling pathways, and cellular processes. To enhance therapeutic efficacy and reduce toxicity, researchers have developed various targeted kidney delivery systems, such as delivery systems based on coupling technology, nanoparticles, polymer micelles, liposome modification, and derivative strategies. Although positive progress has been made in current research, in order to ensure safe clinical application, it is still necessary to further elucidate its pharmacological and toxicological mechanisms, and conduct long-term toxicological evaluations of existing strategies for enhancing efficacy and reducing toxicity. These works will contribute to the scientific evaluation of the clinical application prospects of triptolide and lay the foundation for its translational application in the treatment of kidney diseases.

Disruption of protein homeostasis is a central pathological mechanism in a variety of major diseases. However, traditional small-molecule inhibitors are often limited by issues such as undruggable targets and drug resistance, falling short of clinical needs. Targeted protein degradation (TPD) technologies offer a promising alternative by eliminating pathogenic proteins and thus overcoming key bottlenecks in conventional drug development. Among these, proteolysis-targeting chimera (PROTAC), which hijack the ubiquitin–proteasome system to degrade target proteins, have shown significant translational potential in the treatment of cancer and immune-related disorders. To further enhance the specificity and therapeutic index of PROTAC, bioorthogonal strategies have been increasingly explored in recent years for the development of next-generation degraders. These approaches not only reduce systemic toxicity but also enable spatiotemporal precision control, offering new avenues for the rational design of PROTAC.

This review focuses on recent advances in the component structure theory and associated research technologies for exploring the material basis of traditional Chinese medicine (TCM) preparations. The study first outlines the conceptual evolution of TCM "components" across three levels: individual compounds, synergistic groups, and integrated systems. It then introduces the development of the "component structure" theory from its initial three-tier structural framework (version 1.0), through quantitative proportionality modeling (version 2.0), to the current integration of in vivo phase states and biopharmaceutical disposition (version 3.0), which together form a unified theoretical system of element structure, quantity ratio structure, and phase structure. Furthermore, the technological progress of the fundamental research of Chinese medicine under the guidance of this theory is systematically summarized, covering the techniques of fine analysis of small molecules and macromolecules, the analysis and optimization of the component-volume ratio relationship and the analysis of the component-phase structure. Finally, it emphasizes the need for future research to build multidimensional analytical systems, integrate artificial intelligence and big data technologies, and promote standardization, in order to support the modernization and innovation of TCM.

Macromolecular drugs, due to their high specificity and excellent biological activity, play a crucial role in disease treatment. The oral administration route, with its advantages of high patient compliance and safety, is considered an ideal approach for delivering macromolecular drugs. However, various biological barriers in the gastrointestinal tract significantly limit the oral bioavailability and targeted delivery efficiency of macromolecular drugs. Existing research has primarily focused on overcoming single barriers or designing single carriers, this paper systematically addresses the multiple biological barriers encountered during the targeted delivery of orally administered macromolecular drugs. It provides an overview of various nanocarriers suitable for the targeted oral delivery of macromolecular drugs and delves into the latest advances in organ-specific targeting strategies, offering new perspectives for the precision treatment of orally administered macromolecular drugs. Furthermore, it incorporates examples of applications involving macromolecular drugs such as peptides, proteins, and nucleic acids, providing theoretical insights and research directions for enhancing the oral delivery efficiency of macromolecular drugs.

Aberrant activation of the phosphoinositide 3-kinase (PI3K)-AKT signaling pathway has been identified in numerous cancer types, contributing to uncontrolled cellular proliferation, inhibition of apoptosis, enhanced metastatic potential, and the development of resistance to chemotherapy and radiotherapy. Consequently, targeting the PI3K-AKT pathway has emerged as a promising therapeutic strategy. Current approaches to inhibit this pathway include small-molecule inhibitors, natural compounds, siRNAs, and miRNAs. However, these therapeutic approaches are often limited in clinical application due to prominent adverse effects, poor biological stability, low solubility, or lack of targeting ability. The integration of nanotechnology into drug delivery systems offers a compelling solution by improving drug stability and solubility, minimizing off-target accumulation, and enhancing tumor-specific targeting. Nanocarrier-based delivery of agents targeting the PI3K-AKT pathway holds the potential to improve therapeutic efficacy, mitigate drug-induced toxicity, sensitize tumors to conventional therapies, and ultimately enhance patient outcomes. This review provides a comprehensive overview of the PI3K-AKT signaling pathway and its oncogenic role, provides a detailed review of the latest progress of nano-based drug delivery system in cancer treatment based on pathways, and discusses future directions, aiming to inform and guide ongoing research in this field.

As a fundamental tool for enhancing ultrasound imaging contrast, ultrasound contrast agents (UCAs) have made significant advances in medical diagnosis and therapeutics in recent years. Their development has processed through four generations, with each generation achieving breakthroughs in core gas composition, shell materials, stability, and clinical applicability. The imaging mechanism of UCAs is based on the interaction between microbubbles and ultrasound waves, producing enhanced backscatter to generate clearer images. The in vivo behavior of microbubbles is influenced by various physiological factors. Both the core gas and shell materials are ultimately metabolized or safely eliminated from the body. Common preparation methods include ultrasonic cavitation, high-shear processing, mechanical agitation and so on. In modern medical applications, UCAs are widely employed in the diagnostic imaging of the heart, liver and tumors. Moreover, through targeted ligand modification and drug-loading strategies, they offer potential for theranostic application, i.e., integrating diagnosis and treatment. UCAs can also provide unique therapeutic benefit through gas-mediated therapies. This review systematically presents the development history of UCAs, elaborating on their imaging mechanisms, in vivo behavior, preparation techniques, physicochemical characteristics, and emerging applications at diagnostic-therapeutic interface.

Microalgae, as a class of photosynthetic microorganisms, possess excellent biocompatibility and metabolic diversity. They are capable of synthesizing various natural products with antioxidant, immunomodulatory, and anticancer activities, while also demonstrating great potential as drug delivery vehicles. Their inherent properties — such as self-propelled motility, photosynthetic oxygen generation, and biodegradability — facilitate enhanced accumulation of therapeutics within tumor tissues and improve treatment efficacy. However, natural microalgae still face limitations in targeting ability and drug loading capacity, necessitating engineering strategies to meet the complex demands of cancer therapy. This review summarizes representative microalgal species, their structural features, and biomedical applications. It further provides a systematic overview of current engineering approaches, including physical interactions, chemical modifications, biomembrane coatings, and genetic engineering. Particular attention is given to their emerging roles in multimodal tumor therapies, such as photoresponsive, magnetically guided, and immuno-combined treatment platforms. Finally, future development directions and prospects for clinical translation are discussed.

Nanocarriers, have been extensively explored as a novel drug delivery system, demonstrating significant advantages in improving drug solubility, stability, and targeting capability, and have thus become a research focus in the field. However, their clinical application is substantially limited by a poor capacity to traverse cellular membranes and overcome complex physiological barriers. Cell-penetrating peptides (CPPs), characterized by high membrane penetration efficiency, low toxicity, and favorable biocompatibility, can effectively facilitate the translocation of nanocarriers across biological membrane barriers via direct penetration or endocytosis, thereby significantly enhancing drug delivery efficiency. Concurrently, homing peptides (HPs) enable precise accumulation of nanocarriers at pathological sites by recognizing tissue-specific receptors, while reducing non-specific penetration into healthy tissues. As two crucial classes of functional peptides, CPPs and HPs have been widely utilized to modify nanocarriers, enhancing their capacity to overcome physiological barriers and improving targeted accumulation at diseased sites through distinct mechanisms. This review summarizes the classification and transport mechanisms of CPPs and HPs, as well as advances in their application for modifying nanocarriers to overcome physiological barriers, aiming to provide a reference for future research and applications in this field.

Plant-derived nanovesicles are nano-sized extracellular vesicles that encapsulate nucleic acids, proteins, and lipids, and exert antioxidant, anti-inflammatory, antitumor, and tissue-regenerative effects, showing great potential for disease intervention. Schisandra chinensis (Turcz.) Baill., a traditional medicinal plant, has been extensively studied by our group and others for its hepatoprotective constituents and mechanisms of action. Excessive acetaminophen (APAP) intake is the leading cause of drug-induced liver injury in clinical practice, yet effective and low-toxicity preventive or therapeutic strategies are still lacking. To investigate the protective effect and mechanism of Schisandra chinensis-derived nanovesicles (SSDNs) against APAP-induced acute liver injury in mice, SSDNs were first isolated from Schisandra chinensis by gradient ultrahigh-speed centrifugation. Transmission electron microscopy and nanoparticle tracking analysis revealed an exosome-like bilayer membrane, uniform morphology, and a mean diameter of 205.4 nm. In vitro CCK-8 assays showed that SSDNs exhibited no cytotoxicity. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis identified schisandrol A as the major lignan component of SSDNs. Furthermore, we evaluated the protective effect of SSDNs on APAP-induced acute liver injury in mice. All experimental protocols were approved by the Animal Ethics Committee of Southern Medical University (approval No. SMUL202408016). C57BL/6 mice were pretreated with SSDNs via tail vein injection for 4 days and then given a single intraperitoneal injection of APAP (400 mg·kg^-1). Pretreatment with SSDNs significantly lowered serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels, reduced hepatic necrosis, and restored lobular architecture. Concurrently, SSDNs elevated the hepatic glutathione (GSH) level, decreased protein expression of APAP-metabolizing enzymes cytochrome P450 (CYP) 2E1 and CYP3A11, and upregulated proliferating cell nuclear antigen (PCNA) and cell-cycle proteins [cyclin-dependent kinase 4 (CDK4), cyclin A1 (CCNA1), cyclin D1 (CCND1), and cyclin E1 (CCNE1)]. In conclusion, SSDNs mitigate APAP-induced acute liver injury in mice by preserving hepatic GSH, suppressing CYP450 enzymes involved in APAP metabolic activation, and promoting hepatocyte proliferation and repair. These findings offer a novel strategy for the modern application of Schisandra chinensis and the clinical prevention of APAP-induced hepatotoxicity.

Based on the traditional Chinese medicine theory that "excessive Qi transforms into fire", this study investigates whether overdose of the Qi-tonifying herb Astragalus membranaceus (AM) induces "Shanghuo" and analyzes the related components and underlying mechanisms. Using a mouse model of "Shanghuo" with latent herpes simplex virus type 1 (HSV-1) infection, we systematically observed the general state and behavioral changes of the mice, and detected the expression of virus-related genes and proteins as well as lipid peroxidation-related indicators. The results showed that after intervention with AM and its components, the mice exhibited various "Shanghuo" symptoms, including eye inflammation, increased thirst and water intake, dry stool, and hyperactivity. These symptoms were accompanied by reactivation of latent HSV-1 in the brain, upregulation of lipid peroxidation-related proteins arachidonate 15-lipoxy-genase (ALOX15) and 4-hydroxynonenal (4-HNE), downregulation of cluster of differentiation 117 (CD117; c-Kit) protein expression in intestinal tissue, and elevated levels of the lipid peroxidation product 4-HNE. Further component analysis revealed that AM polysaccharides had a particularly significant impact on behavioral indicators, while low-content AM isoflavones and saponins also reactivated latent HSV-1. In conclusion, various components of AM may synergistically reactivate latent HSV-1 and induce "Shanghuo" potentially through enhancing lipid peroxidation in the body.This experiment has been approved by the Animal Experimental Ethics Committee of Jinan University (IACUC-20250417-04).

Osimertinib, as a first-line therapeutic agent for non-small cell lung cancer (NSCLC), significantly prolongs patients' progression-free survival and overall survival. However, the development of acquired resistance to this agent remains an unavoidable clinical challenge. This study investigated platycodin D (PD) as a potential resistance modulator. Cell viability was assessed using the CCK-8 assay; cell proliferation was detected using the colony formation assay; cytotoxic effects were evaluated via lactate dehydrogenase (LDH) release; apoptosis was detected by PI/Hoechst double staining and flow cytometry; and the expression of apoptosis-related proteins and key components of the Akt/p70S6K pathway as well as phosphorylation levels were examined by Western blot. Results demonstrated that platycodin D suppressed the viability of osimertinib-acquired resistant NSCLC cells and exhibited synergistic antiproliferative effects when combined with osimertinib. The combination treatment significantly inhibited cell growth and induced apoptosis, with effects markedly stronger than either agent alone. Mechanistic studies revealed that the combined treatment more effectively suppressed the phosphorylation of Akt and p70S6K as compared to the single treatment group. In conclusion, this study reveals that platycodin D can enhance the cytotoxic effect of osimertinib against resistant NSCLC cells and reverse osimertinib resistance by inhibiting the Akt/p70S6K signaling pathway, thereby providing a promising strategy to overcome osimertinib resistance.

This study aims to investigate whether Xiaoxianxiong Decoction (XXXD) can exert a protective effect in atherosclerosis (AS) by activating growth arrest specific 6 (GAS6)/receptor tyrosine kinase AXL (AXL) and inhibiting endothelial cell ferroptosis. This experiment was approved by the Animal Experiment Ethics Committee of Shaanxi Academy of Traditional Chinese Medicine (approval number: 2022-10). At the animal level, the apolipoprotein E knockout (ApoE^-/-) mice were fed with a high-fat diet (HFD) to establish the AS model. At the cellular level, the endothelial cell injury model was established with oxidized low-density lipoprotein (ox-LDL), and the cell viability, lactate dehydrogenase (LDH), reactive oxygen species (ROS), Fe²⁺, and lipid peroxidation indicators malondialdehyde (MDA), superoxide dismutase (SOD), total glutathione peroxidase (GPx), and glutathione reductase (GR) were detected. Western blot was used to detect the expression of GAS6, AXL, glutathione peroxidase 4 (GPX4), and solute carrier family 7 member 11 (SLC7A11) proteins. The results showed that XXXD could reduce body weight, aortic plaque area and lipid accumulation in AS mice, improve dyslipidemia, increase lipid antioxidant capacity in aorta and endothelial cells (increase SOD, GPx and GR, inhibit MDA), reduce LDH, ROS, MDA and Fe²⁺ levels in endothelial cells, and up-regulate the expression of GAS6, AXL, GPX4, and SLC7A11 proteins to improve AS and endothelial cell injury. The above results confirmed that XXXD can alleviate AS by regulating ferroptosis of endothelial cells through the GAS6/AXL signaling pathway.

This study aims to explore the therapeutic effect and mechanism of action of Zhisou Huatan Wan on rats with chronic obstructive pulmonary disease (COPD) of phlegm-heat obstructing the lungs. Animal experiments were approved by the Experimental Animal Ethics Committee of Henan University of Chinese Medicine (approval number: IACUC-202506002). Sixty male SD rats were randomly divided into six groups: control group, model group, erythromycin group (100 mg·kg^-1), and low-dose (206.55 mg·kg^-1), medium-dose (413.1 mg·kg^-1), and high-dose Zhisou Huatan Wan (826.2 mg·kg^-1) group. The rats were constructed with COPD of phlegm-heat obstructing the lungs by using smoke inhalation combined with intratracheal injection of lipopolysaccharide (LPS, 1 mg·kg^-1) and cold air stimulation. The lung function, cough frequency, tracheal phenol red excretion, the degree of pulmonary tissue edema, arterial partial pressure of oxygen (PaO₂), arterial partial pressure of carbon dioxide (PaCO₂) of each group of rats were detected. H & E, Masson, and PAS staining were used to detect lung tissue lesions. The levels of apoptosis, oxidative stress and mitochondrial membrane potential in lung tissue, and peripheral blood immune cells (neutrophils, Treg cells, Th17 cells) were detected by flow cytometry. Transcriptomics analysis was used to screen core targets and signaling pathways. ELISA was applied to measure the levels of interleukin 17A (IL-17A), C-C chemokine ligand 7 (CCL7) and C-C motif chemokine ligand 20 (CCL20) in serum and lung tissue. qRT-PCR was used to detect the mRNA levels of IL-17A, CCL7 and CCL20 in lung tissue. IL-17A protein level was detected by Western blot. The results showed that Zhisou Huatan Wan significantly improved lung function, increased the level of blood oxygen partial pressure, mitochondrial membrane potential in lung tissue, and Treg cells in peripheral blood, promoted tracheal phenol red secretion, alleviated pathological lung injury, and reduced cough frequency, carbon dioxide partial pressure, the degree of pulmonary tissue edema, apoptosis and oxidative stress levels in lung tissue, as well as the levels of neutrophils and Th17 cells in peripheral blood. Zhisou Huatan Wan reduced the content of IL-17A, CCL7 and CCL20 in serum and lung tissue, and the corresponding mRNA levels in lung tissue, as well as the expression level of IL-17A in lung tissue. Transcriptomic analysis identified 78 differentially expressed genes, and primarily enriched in IL-17 signaling pathway, the chemokine signaling pathway and cytokine-cytokine receptor interaction. The results showed that Zhisou Huatan Wan had a significant improvement effect on COPD rats of phlegm-heat obstructing the lungs, and its mechanism may be related to the IL-17 signaling pathway.

This study investigated the mechanism by which notoginsenoside R1 (NGR1) alleviates neuroinflammation by regulating metabolic disorders in microglia through network pharmacology, in vitro experiments, and untargeted metabolomics. Neuroinflammation-related disease targets and NGR1 targets were obtained through multi-database mining, overlapping targets were screened using Venny, and GO and KEGG enrichment analyses as well as protein-protein interaction network construction were performed to screen core targets. Molecular docking software was used to evaluate the binding ability between NGR1 and the core targets. An inflammatory model was established by lipopolysaccharide-induced BV2 microglial cells, and Western blot was used to detect the effects of different concentrations of NGR1 on the expression of inflammation-related proteins in microglia, and to determine the effective concentrations of NGR1 affecting the NF-κB signaling pathway. Untargeted metabolomic analysis was conducted within the effective concentration range of NGR1, and differential metabolites were screened by combining multivariate statistical analysis with metabolic pathway enrichment analysis. Cytoscape and MetaboAnalyst platforms were further used to screen key metabolites closely associated with the NF-κB signaling pathway. Exogenous metabolite supplementation experiments were conducted to verify whether NGR1 could inhibit the activation of the NF-κB signaling pathway by regulating the levels of related metabolites. The results showed that there were 43 overlapping genes between neuroinflammation-related disease targets and NGR1 targets, among which 16 were core targets. KEGG enrichment results indicated that the overlapping targets were significantly enriched in inflammatory signaling pathways such as NF-κB. Molecular docking results showed that NGR1 had high binding affinity with multiple NF-κB signaling pathway proteins. Western blot results demonstrated that NGR1 could significantly regulate the expression of NF-κB signaling pathway-related proteins and inflammatory factors, and 25 and 50 μmol·L^-1 were the effective concentrations affecting the NF-κB signaling pathway. Metabolomic results showed that NGR1 could significantly affect multiple pathways such as glutathione metabolism. Metabolite-target association analysis results showed that NGR1 could reverse abnormal changes in metabolites closely related to NF-κB signaling pathway targets, including homocysteine, L-leucine, and prostaglandin E2 ethanolamide. Exogenous metabolite supplementation experiments showed that homocysteine could weaken the inhibitory effect of NGR1 on the NF-κB signaling pathway, suggesting that homocysteine may be involved in the NGR1-mediated NF-κB inhibitory process. In summary, NGR1 may inhibit the activation of the NF-κB signaling pathway through both direct inhibition and indirect regulation of homocysteine metabolism, thereby alleviating neuroinflammatory responses.

Tumor-associated macrophages (TAMs) have a dual role in the tumor microenvironment, mediating both immune activation and suppression. M1-TAMs kill tumor cells, while M2-TAMs are immunosuppressive and promote tumor growth. The activation of TAMs and enhancement of their anti-tumor immune function is a major goal in cancer immunotherapy. Here, we report a novel pyrazole derivative, IHMT-140, which reprograms tumor-promoting M2 macrophages into tumor-suppressing M1 macrophages, thereby inhibiting tumor growth. Mechanistically, IHMT-140 raises lysosomal pH in M2 macrophages, blocking autophagy and activating MAPK and NF-κB signaling, which induces M2-TAMs to M1-TAMs and mediates the anti-tumor response. As a potential immunomodulator, IHMT-140 provides not only a new tool for studying TAM reprogramming, but also a foundation for developing related immunotherapies. All animal experimental protocols were approved by the Hefei Institutes of Physical Science, Chinese Academy of Sciences, with approval number DWLL-2023-116.

Based on the cellular proliferation tracing technique (ProTracer), this study explored the role of rifampicin (RIF) in promoting liver regeneration in a mouse model with zone-specific liver injury. The ProTracer system enables in vivo recording of cell proliferation by labeling proliferating cells with GFP fluorescence. Carbon tetrachloride (CCl₄)-induced injury and bile duct ligation (BDL) models were established, followed by RIF or vehicle treatment. Liver function was assessed by measuring serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), total bile acids (TBA), total bilirubin (TBIL), and direct bilirubin (DBIL). Liver injury area was evaluated by H&E staining, while whole-tissue fluorescence imaging was used to assess liver regeneration. Immunofluorescence co-staining was performed to quantify proliferating hepatocytes, identify cell types, and analyze zonal distribution. RNA sequencing was conducted to identify key differentially expressed genes and signaling pathways, with Western blot used for protein-level validation. In vitro validation was performed using the HepG2 hepatocyte cell line. Results showed that RIF significantly reduced serum ALT and AST levels in CCl₄-induced liver injury and decreased TBA, TBIL, and DBIL levels in the BDL-induced liver injury, effectively reducing the area of liver damage. Whole-tissue fluorescence imaging revealed enhanced GFP signals and increased proliferating cells in the RIF-treated group. Immunofluorescence analysis indicated that RIF primarily promoted hepatocyte proliferation, with a marked increase in proliferating cells in zone 2 of the liver lobule (approximately 2.5-fold), and also significant increased in zones 1 and 3. RNA-Seq and Western blot analyses demonstrated upregulation of cell cycle and proliferation-related genes such as the Mcm family, Ccnd1, Ccna2, and Mki67, with enrichment in the cell cycle pathway. In vitro experiments showed that RIF treatment enhanced HepG2 cell viability and increased the proportion of Ki67-positive cells. In conclusion, RIF promotes hepatocyte proliferation and enhances liver regeneration following zone-specific liver injury by activating cell cycle signaling pathways. Animal experiments were conducted with approval from the Animal Ethics Committee of Binzhou Medical University (approval number: 2025-2018).

This study aimed to construct a stable cell line of human embryonic kidney epithelial cells 293 (HEK293) expressing recombinant human coagulation factor Ⅸ (rhFⅨ) and to identify the procoagulant biological activity of the expressed product. The recombinant plasmid pcDNA3.1-FⅨ was transfected into HEK293 cells, and a monoclonal stable transfection cell line was obtained through geneticin (G418) pressure screening. Reverse transcription-polymerase chain reaction (RT-PCR) was used to determine whether the FⅨ gene was transcribed and expressed in the monoclonal stable transfection cell line. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot were used to identify the expression of rhFⅨ; the concentration of rhFⅨ was determined by bicinchoninic acid assay (BCA), and the procoagulant activity of rhFⅨ was determined by activated partial thromboplastin time (APTT). The results showed that after G418 (200 µg·mL^-1) pressure screening, 5 monoclonal stable cell lines could be normally transcribed, and the protein expression could be achieved through the transcription-translation process; the protein in the cell supernatant was purified and showed a single band of rhFⅨ; the expression level of rhFⅨ reached (1.58 ± 0.10) mg·L^-1, and the maximum procoagulant activity was (66.8 ± 0.5)%. The established HEK293 monoclonal stable transfection cell line can efficiently and stably express rhFⅨ and has good procoagulant activity, providing a cell source for promoting the pharmacological evaluation and preclinical research of rhFⅨ.

This study was to investigate the protective effect of Saposhnikovia divaricata (Turcz.) Schischk. (SD) on ulcerative colitis (UC) and its relationship with aryl hydrocarbon receptor (AHR)/NOD-like receptor family, pyrin domain containing 3 (NLRP3) pathway. The acute colitis model of C57BL/6J mice was established by continuous drinking water with 2.5% dextran sulfate sodium (DSS) for 7 days. At the same time, the ethanol extract of Saposhnikovia divaricata (2 and 5 g·kg^-1·d^-1) or 5-aminosalicylic acid (5-ASA, 100 mg·kg^-1·d^-1) was given by gavage. Body weight and disease activity index (DAI) were recorded. Colon length, hematoxylin-eosin (HE) and Alcian blue-periodic acid-Schiff staining (AB-PAS) staining and immunofluorescence were used to evaluate colon tissue damage and changes of goblet cells and tight junction proteins. ELISA, RT-qPCR and Western blot were used to analyze the expression of inflammatory factors, tight junction proteins and AHR/NLRP3-related molecules in serum and colon tissues. The use of animals in this study was approved by the Ethics Committee of Shanghai University of Traditional Chinese Medicine (approval number: PZSHUTCM2510310022). The results showed that compared with the DSS model group, Saposhnikovia divaricata could dose-dependently reduce DSS-induced weight loss and DAI increase, prolong colon length, and reduce mucosal pathological damage. At the same time, it down-regulated the expression of inflammatory factors such as TNF-α, IL-1β and IL-6 in serum and colon tissues, up-regulated the mRNA and protein levels of zonula occludens-1 (ZO-1) and occludin tight junction proteins, and increased the expression of AHR protein, and inhibited NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC), Caspase-1 and IL-1β. In AHR knockout mice, the alleviation effect of the ethanol extract of Saposhnikovia divaricata on DSS-induced acute ulcerative colitis in mice was not observed. In summary, Saposhnikovia divaricata may inhibit the activation of downstream NLRP3 inflammasome by activating AHR, thereby maintaining intestinal mucosal barrier function and reducing intestinal inflammation, providing experimental basis for its application in the prevention and treatment of UC.

Alzheimer's disease (AD) is a chronic neurodegenerative disorder. Current medications can only alleviate symptoms but have not yet achieved a cure. Given its multifactorial pathogenesis, the development of small-molecule compounds that act simultaneously on multiple targets is particularly important. Traditional Chinese medicine (TCM) small molecules, which can regulate multiple biological targets and signaling pathways, offer unique advantages in AD treatment. This study utilized genipin, a TCM small molecule with potential neuroprotective effects, as the lead compound to design and synthesize a series of multi-target anti-AD small molecule compounds. Among these, compounds JD-5 and JD-7 exhibited significantly enhanced GSK-3β inhibitory activity (JD-5, IC₅₀ = 3.54 μmol·L^-1; JD-7, IC₅₀ = 2.47 μmol·L^-1), substantially outperforming the lead compound genipin (IC₅₀ = 28.16 μmol·L^-1). Furthermore, both JD-5 and JD-7 demonstrated significant neuroprotective effects in SH-SY5Y cell damage experiments induced by tBHP and Aβ_25-35. Further investigation revealed that compounds JD-5 and JD-7 could significantly reduce Tau phosphorylation (S396) levels in primary neuronal cells, with dose-dependent effects. All animal experiments were approved by the Experimental Animal Ethics Committee of Nanjing Ramda Pharmaceutical Co., Ltd (approval number: IACUC-20220202). Based on these results, it is hypothesized that compound JD-5 likely exerts neuroprotective and antioxidant stress effects by inhibiting GSK-3β activity and reducing Tau protein phosphorylation levels, providing a reference for the development of multifunctional anti-AD drugs.

This study employs silica gel column chromatography and thin-layer chromatography to study the chemical constituents of the effective substance groups from Artemisia ordosica, and analyze the spectral data of the separated compounds to determine their structures. Nine alkynes, including ordosin (1), capillene (2), capillin (3), capillinol (4), trans-dehydromatricaria ester (5), capillarin (6), o-hydroxycapillene (7), capillaridin (8), o-hydroxycapillin (9), and one dihydroflavone, 5-hydroxy-7,4′-dimethoxydihydroflavone (10), were isolated and identified from the effective substance groups of Artemisia ordosica. Among these, compound 1 is a new alkyne, while 2-4, 6, 8, and 9 were isolated for the first time from Artemisia ordosica. By examining the effects of compounds 1-4, 6, 8, and 9 on the release of nitric oxide (NO) in the culture medium of RAW264.7 macrophages induced by lipopolysaccharide (LPS), the inhibitory effect on inflammatory response was evaluated. The results revealed that compounds 1-4, 6, 8, and 9 exhibited NO inhibitory activity at a concentration of 10 μmol·L^-1, with inhibition rates of 96.7%, 88.4%, 91.6%, 77.8%, 98.7%, 86.7% and 75.1%, respectively.

The ethyl acetate fraction of the 95% ethanol extract from black mulberry branches was isolated and purified by comprehensive application of chromatographic techniques, including normal-phase silica gel column chromatography, Sephadex LH-20 gel column chromatography, and semi-preparative high-performance liquid chromatography (semi-prep HPLC). Fourteen compounds were obtained, and their structures were identified using modern spectroscopic techniques (UV, IR, MS, NMR). These compounds were identified as nigrasin M (1), nigrasin N (2), sanggenol F (3), sanggenon A (4), nigrasin K (5), nigrasin H (6), petalopurpurenol (7), solophenol D (8), nigragenon A (9), nigragenon I (10), nigragenon J (11), notabilisin A (12), notabilisin C (13), and licoflavone C (14). Among them, compounds 1 and 2 are new flavonoids, while compounds 13 and 14 were isolated from Morus nigra branches for the first time. To evaluate the hypoglycemic potential of compounds 1-13, a glucose uptake assay was performed using an L6 cell model. The results showed that compounds 6, 9, 11, and 12 significantly enhanced glucose uptake in L6 cells, suggesting their potential hypoglycemic activity.

Nine sesquiterpenoids were isolated from the dichloromethane fraction of the 95% ethanol extract from the roots of Lindera glauca by using a combination of separation techniques, including silica gel, ODS and Sephadex LH-20 column chromatographies, as well as semi-preparative HPLC. Based on a comprehensive analysis of modern spectroscopic data, such as IR, UV, NMR, HRESIMS and ECD, the structures of isolated compounds were identified as glaucatotone M (1), glaucatotone K (2), glaucatotone G (3), pseudoguaianelactone C (4), pseudoguaianelactone B (5), elema-1,3,11(13)-trien-12-oic acid (6), α-costic acid (7), 3β-hydroxycostic acid (8) and rupestonic acid I (9), respectively. Compound 1 was a new guaiane-type sesquiterpenoid, and compounds 6 and 7 were first reported from the genus Lindera. The inhibitory activity of compounds 1-7 and 9 against both the proliferation of TNF-α-induced rheumatoid arthritis fibroblast-like synoviocytes cell line MH7A and the release of pro-inflammatory cytokines (IL-6 and IL-1β) were evaluated. The results showed that compound 1 had a significant inhibitory effect against TNF-α-induced MH7A cell proliferation and reduced the production of IL-6 and IL-1β effectively.

The 95% ethanol extract of Curcuma longa was subjected to isolation and purification using silica gel column chromatography, C₁₈ column chromatography, and semi-preparative HPLC. The planar structures of the isolated compounds were elucidated by HR-MS, IR, and NMR spectroscopy. The absolute configurations of the new compounds were determined by calculated electronic circular dichroism (ECD) and DP4+ NMR analysis. Finally, three bisabolane-type sesquiterpenoids were isolated and identified from C. longa as (1R,7S)-15-norbisabolane-2-ene-11-ol-4,9-dione (1), (7S)-15-norbisabolane-1,3,5,10-tetraene-4-ol (2), and 8-hydroxyl-ar-turmerone (3). Compounds 1 and 2 were two novel 15-norbisabolane-type sesquiterpenoids.

Thirteen compounds were isolated from the twigs and leaves of Meiogyne hainanensis (Merr.) N. T. Ban by silica gel column chromatography, sephadex LH-20, and preparative HPLC. Their structures were respectively determined as chienhainanin N (1), (S)-15, 16-didehydrocoriolic acid methyl ester (2), 13-hydroxy-9Z, 11E, 15E-octadecatrienoic acid (3), (S)-coriolic acid (4), 13-oxo-(9Z, 11E)-octadecadienic acid (5), 13-oxo-(9E, 11E)-octadecadienic acid (6), 9-oxo-(10E, 12Z)-octadecadienic acid (7), 9-oxo-(10E, 12E)-octadecadienic acid (8), α-tocopherol (9), vitamin E quinone (10), α-tocospiro A (11), 17c-ethoxypheaophorbide A (12), mustakone (13) by spectral data analysis. Among them, compound 1 is a new compound, compounds 2, 3, 5-8, 10-12 are isolated from Annonaceae plants for the first time, and compounds 4, 9 and 13 are isolated from the genus Meiogyne for the first time. Their antimicrobial activities were evaluated by the broth microdilution method. Compounds 2, 3, 5-7 showed weak antimicrobial effect against MRSA with MIC values from 64 to128 μg·mL^-1, respectively. Compound 13 showed weak antimicrobial effects against Cryptococcus neoformans with the MIC value of 128 μg·mL^-1.

This study investigated cultivated Astragalus from three major production regions-Shanxi, Neimenggu, and Gansu-using a pseudo-targeted metabolomics approach based on UHPLC-Q-TRAP-MS. After systematic optimization of collision energy (CE) and declustering potential (DP) parameters, relative quantification of 109 compounds in 16 batches was performed. Combined with multivariate statistical analysis, chemical differences among samples from different origins were assessed. The results revealed certain differences in the overall chemical composition among the three origins. Approximately 60% of the detected components showed no significant content differences across regions, while the differential compounds-primarily secondary metabolites-were mainly observed between Shanxi Astragalus and samples from Neimenggu and Gansu. Compared to Neimenggu and Gansu samples, Shanxi Astragalus was characterized by higher contents of acylated flavonoid glycosides and flavonoid glycosides, along with lower levels of flavonoid aglycones. Thus, the flavonoid glycoside/aglycone ratio can serve as a reliable indicator to distinguish Shanxi Astragalus. These characteristic differences may serve as potential markers for geographical traceability and authenticity evaluation of cultivated Astragalus, providing a scientific basis for constructing a quality assessment system.

To investigate the oral bioaccessible contents of lead (Pb) and arsenic (As) in the animal-derived traditional Chinese medicine earthworm and to assess the associated health risks, the contents of Pb and As in earthworm were determined by inductively coupled plasma mass spectrometry (ICP-MS). An in vitro digestion/intestinal organ-on-a-chip model suitable for the matrix of traditional Chinese medicine was established for the first time to simulate the gastrointestinal digestion and absorption processes of Pb and As from earthworm, and their bioaccessible contents was determined. The health risks were assessed using the hazard index (HI) method based on both the total contents and the bioavailable contents of the heavy metals. The results indicated that the total contents of Pb and As in 9 batches of earthworm ranged from 0.16-12.10 and 0.15-1.20 mg·kg^-1, respectively. After transported by the intestinal organ-on-a-chip, the bioavailable concentrations ranged from 0.02-0.92 mg·kg^-1 for Pb and 0.01-0.09 mg·kg^-1 for As. The risk assessment based on total contents revealed that the HI for Pb and As in multiple batches exceeded 1, indicating an unacceptable risk. In contrast, the HI values based on bioavailable concentrations were all below 1, suggesting an acceptable risk. The in vitro digestion/intestinal organ-on-a-chip model established in this study for the first time can more accurately reflect the actual exposure risk of heavy metals in traditional Chinese medicine following oral intake, providing a methodological foundation for the safety evaluation and standard setting of traditional Chinese medicines.

This study aims to investigate the potential active components and mechanism of action of "Ginseng-Schisandra" herb pair in the treatment of hepatocellular carcinoma (HCC). The blood-entering components of the "Ginseng-Schisandra" herb pair were identified using high pressure liquid chromatography-quadrupole-time of flight mass spectrometry (HPLC-QTOF/MS). The alterations of bile acid (BA) levels in the plasma, liver, and feces of HCC rats following intervention with the "Ginseng-Schisandra" herb pair were comprehensively analyzed by LC-MS. The biomarkers of HCC were screened by integrating orthogonal partial least squares-discriminant analysis (OPLS-DA) and Student's t-test. Finally, the key pharmacodynamic substances of "Ginseng-Schisandra" herb pair in the treatment of HCC were identified based on correlation analysis and further verified through molecular docking. The animal experiments were approved by the Ethics Committee of Shenyang Pharmaceutical University (approval No.: CSE202106002). As a result, 22 blood-entering prototype components of the "Ginseng-Schisandra" herb pair were identified. This study demonstrated that the "Ginseng-Schisandra" herb pair markedly restored dysregulated BA metabolism in the plasma, liver, and feces of HCC rats, and 9 bile acids were successfully characterized as HCC biomarkers. Correlation analysis and molecular docking demonstrated that ginsenoside Rb1 and schisandrin A, which exhibited strong binding affinities to key BA regulatory proteins [farnesoid X receptor (FXR), cytochrome P450 family 7 subfamily A member 1 (CYP7A1), bile salt export pump (BSEP), and organic anion transporting polypeptide 1B1 (OATP1B1)], were crucial pharmacodynamic constituents of the "Ginseng-Schisandra" herb pair in the treatment of HCC. Collectively, this study established a scientific foundation for the clinical application of the "Ginseng-Schisandra" herb pair in the treatment of HCC.

Ultra-high performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry (UPLC-QTOF-MS/MS) was used to analyze and identify the chemical components of Dabufei Decoction (DBFT). Combined with network pharmacology, the mechanism of action of DBFT in the stable phase of chronic obstructive pulmonary disease (COPD) was explored. Under positive and negative ion modes, the chemical components of DBFT were identified based on information such as the accurate molecular weight of compounds, MS/MS fragmentation rules, and literature-reported data. Then, combined with the TCMSP database and literature mining, the active components and action targets of DBFT were obtained. Targets related to the stable phase of COPD were retrieved from the OMIM and Gene Cards databases. The overlapping targets between drug targets and disease targets were screened out, imported into the STRING database and Cytoscape 3.10.3 to construct a protein-protein interaction (PPI) network, and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses of core targets were performed using the Metascape database. The results showed that a total of 132 chemical components were identified in DBFT, among which 87 components were derived from the "Yong" group of herbs, 49 from the "Ti" group of herbs, 4 from the "Hua" group of herbs, and 8 components were shared by the "Ti" and "Yong" groups of herbs. A total of 84 potential active components of DBFT were screened out based on the Swiss ADME gastrointestinal absorption and five principles of druggability. There were 294 common targets with the stable phase of COPD, and 87 core targets were obtained through calculation using Cytoscape. The KEGG-enriched pathways mainly included the PI3K-AKT signaling pathway, HIF-1α signaling pathway, AGE-RAGE signaling pathway, EGFR tyrosine kinase inhibitor resistance, and MAPK signaling pathway, which played a role in the treatment of the stable phase of COPD. Among them, the "Ti" group of herbs mainly acted on the C-type lectin receptor signaling pathway, the "Yong" group of herbs acted on the PI3K-AKT signaling pathway, and the "Hua" group of herbs acted on the differentiation process of Th17 cells. The research results provide a scientific basis and support for the clinical application and in-depth study of this prescription.

In this study, we employed astragalus polysaccharides (APS), derived from the traditional Chinese medicine Astragalus, as a natural carrier, and curcumol (CUL), a hydrophobic component with antitumor activity in the traditional Chinese medicine Curcuma longa, as a model drug, to construct a drug-loaded integrated nanoimmune drug delivery system, CUL@AHNP, and systematically evaluate its in vitro properties, oral absorption properties, and antitumor immune activity. The CUL@AHNP was prepared by high-pressure homogenization, resulting in a particle size of approximately 116.4 nm. It exhibited a spherical structure, a negatively charged surface, and good dispersion and stability. In vitro experiments demonstrated that CUL@AHNP could significantly enhance the inhibitory effect of CUL on breast cancer 4T1 cells and effectively induce immunogenic cell death (ICD) and activate anti-tumor immune responses. Assays measuring cellular uptake and tumor sphere penetration verified the sub stance's excellent transmembrane transport and tissue penetration ability. In vivo studies demonstrated that the oral administration of CUL@AHNP led to a substantial inhibition of tumor growth and an enhancement in the infiltration density of CD8⁺ and CD4⁺ T cells. The combination of CUL@AHNP and the chemotherapeutic drug doxorubicin (DOX) exhibited a certain degree of synergistic antitumor effect, without causing significant liver or kidney toxicity. All animal experiments had been approved by the Experimental Animal Ethics Committee of Tianjin University of Traditional Chinese Medicine (No. 2023114). In conclusion, this study constructed an oral nanoimmune drug delivery system based on astragalus polysaccharides, which provides a safer and more efficient therapeutic option for the treatment of breast cancer with traditional Chinese medicine.

To achieve precise drug delivery responsive to the tumor microenvironment (TME), this study designed and constructed a multifunctional nanotherapeutic system. The system employs spherical manganese dioxide (MnO₂) as the core, which is loaded with gold nanoparticles (Au nanoparticles, GNPs) and the chemotherapeutic drug 5-fluorouracil (5-FU). It successfully integrates the near-infrared photothermal effect of GNPs and the responsiveness of MnO₂ to high glutathione concentration and acidic pH, enabling on-demand drug release under multiple stimuli. Experimental results demonstrate that the as-constructed MnO₂/GNPs@5-FU nano-system exhibits high drug loading capacity, excellent photothermal conversion efficiency, and remarkable TME-dependent drug release behavior. In vitro cell experiments demonstrated that the system, by combining MnO₂-mediated chemodynamic therapy and 5-FU chemotherapy, achieved a highly efficient synergistic killing effect against 4T1 tumor cells. This study provides preliminary in vitro experimental evidence and novel insights for the development of intelligent nanodrug delivery platforms for tumor-targeted therapy.

Citrus plants are economically and medicinally important crops worldwide. However, due to the highly complex genetic background of the genus, the accurate identification of medicinal species within this genus remains a major challenge. In this study, we employed the analysis of whole-GEnome (AGE), also known as the Shizhen Method, to conduct bioinformatics analysis on 49 genomes from 15 Citrus species, successfully constructing a candidate species-specific target sequence library that covers these species. The number of specific target sequences per species predominantly ranges from 10 000 to 100 000. From this candidate library, species-specific target sequences were selected for the corresponding species. Their high specificity was subsequently verified using Sanger sequencing and CRISPR-Cas12a technology, which confirmed their efficacy in identifying medicinal Citrus species. Based on this foundation, we established an AGE-based identification system for closely related Citrus species, which enabled the precise authentication of 30 commercial samples of Citri Reticulatae Semen and its decoction pieces. This study demonstrates that the AGE can be used for the accurate identification of medicinal Citrus species, which breaks through the bottlenecks of traditional methods and effectively resolves the identification challenges. It thereby provides technical support for quality control of Chinese herbal medicine and clinical medication safety.

To identify the key genes involved in the synthesis of polyphyllin within downstream pathways, this study employed high-performance liquid chromatography (HPLC) to quantify the content of eight types of polyphyllins in two groups of Paris polyphylla var. yunnanensis rhizomes, followed by transcriptome sequencing analysis. The results revealed significant differences in the content of the eight types of polyphyllins between the two groups. Specifically, the total saponin content in the high saponin content group (HS) was 24.81 mg·g^-1, whereas that in the low saponin content group (LS) was 5.99 mg·g^-1. Notably, polyphyllin Ⅱ, Ⅲ, and Ⅴ were undetectable in the LS group. The substantial difference in total saponin content between the two groups can be primarily attributed to the considerable accumulation of diosgenin-type saponins in the HS group. A total of 11 399 differentially expressed genes were identified through transcriptome sequencing, the comparison between HS and LS, with 5 708 genes exhibiting upregulation and 5 691 genes exhibiting downregulation. This study successfully screened seven candidate genes involved in the synthesis of polyphyllin and conducted protein structure prediction for these genes. Through interaction network analysis, 80 transcription factors with a correlation coefficient of 0.95 or higher with the candidate genes were identified, among which a total of 8 transcription factors were correlated with two or more candidate genes. The findings provide a theoretical foundation for predicting genes related to polyphyllin synthesis, exploring the biosynthetic pathway of polyphyllin, and understanding their accumulation patterns.

Fritillaria ussuriensis Maxim is a valuable medicinal plant in Northeast China. Based on previous chloroplast genome sequencing results, this study selected the chloroplast genome fragments atpF and petB as specific DNA barcodes to identify germplasm resources of commercially available F. ussuriensis. Additionally, the content of peiminine was analyzed using a UV spectrophotometer, aiming to clarify the main germplasm resources and quality of F. ussuriensis in the market. In this study, 74 samples of commercially available F. ussuriensis were collected from 52 cities across 23 provinces. Based on the combined analysis of atpF and petB sequences, a total of 10 haplotypes (h2, h3, h4, h5, h6, h11, h13, h16, h19, and h20) were identified, among which h19 and h20 were reported for the first time. Haplotype h4 was the predominant haplotype, accounting for 44.59% of the total samples, followed by haplotype h3. The content determination results showed that 60 batches (81.08%) of commercially available F. ussuriensis samples met the standards of the 2025 edition of the Pharmacopoeia of the People's Republic of China. The content of peiminine in the samples ranged from 0.044% to 0.075%, and the sample quality was uneven; among them, haplotype h16 had the highest average content of peiminine at 0.063%, while haplotype h6 had the lowest at 0.044%. Statistical analysis indicates that, overall, there is no statistically significant difference between haplotypes and the content of peiminine. Given its highest peiminine content, haplotype h16 shows potential value for selective breeding. This study is conducive to guiding the circulation of F. ussuriensis samples in the market and rational clinical use, and also provides a reference for protecting the genetic diversity of F. ussuriensis and for its sustainable development and utilisation.

Rhei Radix et Rhizoma is one of the genuine medicinal materials in Sichuan. Its original species are Rheum palmatum, Rh. tanguticum, and Rh. officinale, recorded by Chinese Pharmacopoeia (2025 Edition). However, there are many cases of multi-origin mixing and adulterant incorporation in the current commercially available Rhei Radix et Rhizoma, and the variety origin needs to be clarified urgently. In this study, the ndhF-rpl32 region of chloroplast genomes, as specific DNA barcode reported previously, was selected and confirmed. The VPCR method was established to identify the authenticity and origin of rhubarb medicinal materials, decoction pieces and Chinese patent medicines, in the process of optimizing DNA extraction procedure, amplification system and amplification procedure. Compared with the traditional PCR method, the amplification procedure has been refined through a VPCR heating model, curtailing the detection from 1.5 h to 30 min. The 85 batches of samples collected from the circulation area were tested by VPCR method, including 30 batches of medicinal materials, 40 batches of decoction pieces and 15 batches of Chinese patent medicines which contain Rhei Radix et Rhizoma. The experimental results showed that 31 batches are Rh. tanguticum (including 18 batches of medicinal materials, 10 batches of decoction pieces and 3 batches of Chinese patent medicines), 36 batches are Rh. officinale (including 1 batches of medicinal materials, 25 batches of decoction pieces and 10 batches of Chinese patent medicines) and 13 batches are Rh. palmatum (including 8 batches of medicinal materials and 5 batches of decoction pieces). The pass rate is 94.1%. Three batches of rhubarb medicinal materials (No. CD1, CD2, CD3) and two batches of Chinese patent medicines (No.Z4 tablet, No.Z12 granule) were determined to be Rh. franzenbachii Munt, as one of counterfeit Rhei Radix et Rhizoma varieties. The VPCR method established in this study can be used as supplement to rhapontin TLC inspection test of Rhei Radix et Rhizoma in Chinese Pharmacopoeia (2025 Edition) and standardize the market for this clinical medication as well as improve the safety of Rhei Radix et Rhizoma.