The term "blood-brain crosstalk" refers to the approach by which systemic immune cells infiltrate into the brain parenchyma and interact with brain cells to affect central nervous system disease. Neutrophils are the first immune cells to cross the blood-brain barrier from the circulating blood to the site of disease and are one of the immune cells that play a major function in neuroinflammation. There are complex interactions between neutrophils and brain cells (such as neurons, microglia, astrocytes, and cerebral microvascular endothelial cells), which together are involved in the progression of neuroinflammatory diseases. Neutrophil-brain cell interactions can influence brain injury due to neuroinflammation. Therefore, this article summarizes the mechanism of neutrophil-brain cell interactions in neuroinflammatory diseases and the research progress of pharmacological intervention, with a view to providing a basis and reference for the future mediation through "blood-brain crosstalk" in the treatment of neuroinflammatory diseases.

Stem cells (SCs) are a class of self-renewing cells capable of differentiating into various human cell types. The field of stem cell regenerative medicine is flourishing, with China's first approved stem cell drug—Amimestrocel—bringing new hope to patients. Current research on stem cell mechanisms primarily focuses on biological and chemical factors; however, physical regulatory factors (e.g., mechanical forces, electromagnetic fields) offer distinct advantages, including non-invasiveness and temporally precise control. This review explores mechanisms by which physical factors regulate stem cell behavior. Dynamic mechanical forces influence stem cell differentiation and function by modulating key signaling pathways such as TGFβ/Smad, RAS/MAPK, and JAK/STAT, the activation of which plays crucial roles in organismal development and disease pathogenesis. Electromagnetic fields regulate stem cell proliferation, migration, and extracellular matrix remodeling through the modulation of membrane potential, ion channel activity, and gene expression, opening new research directions. Photobiomodulation enables stem cell regulation through adjustments to light parameters (wavelength, intensity, duration) or photosensitive materials, demonstrating significant effects in studies involving neural stem cells and bone marrow mesenchymal stem cells. Synergistic therapeutic strategies combining multiple physical factors with emerging technologies such as 3D printing and nanomaterials will further advance stem cell research and clinical applications.

Atherosclerosis is a multifaceted chronic inflammatory disorder intricately linked to various cardiovascular diseases, and its pathogenesis remains incompletely understood. Recent studies have highlighted the pivotal role of autophagy, an essential intracellular degradation and regeneration mechanism, in the onset and progression of atherosclerosis. Autophagy modulates the development of atherosclerosis by influencing the functionality of endothelial cells, smooth muscle cells, and macrophages. Current research indicates that autophagy not only contributes to the initiation and advancement of atherosclerosis, but may also unveil potential therapeutic targets through its impact on associated signaling pathways such as mTOR, AMPK, and NF-κB. This paper aims to review the mechanisms underlying autophagy in atherosclerosis, assess its effects on atherosis-related cell types, and explore future research directions to provide novel insights and strategies for preventing and treating atherosclerosis.

To address the growing global challenge of antibiotic resistance, the development of novel drug formulations with high specificity, excellent antibacterial activity, and the ability to reduce the generation of bacterial resistance has become an urgent and critical task. Recently, new molecular materials with aggregation-induced emission (AIE) properties have shown great potential in disease treatment due to their unique luminescent properties and high biocompatibility. By combining phage therapy with AIE-based photodynamic therapy (PDT), a new antibacterial strategy, known as AIE-armed phages, has been developed. This innovative approach not only enables bacterial imaging in vivo but also enhances targeted bacterial eradication through the synergistic effects of PDT and phage therapy. This review summarizes the research progress of AIE-armed phages, highlighting the unique targeting and visualization capabilities of this biological conjugate, demonstrating its potential to precisely locate and effectively eliminate bacteria, and discusses future directions for optimizing AIE-armed phages.

Alzheimer's disease (AD) is a neurodegenerative disorder with a variety of etiological hypotheses, among which the cholinergic hypothesis is one of the primary theories. Natural inhibitors of acetylcholinesterase (AChE), such as galantamine and rivastigmine, are clinically approved for the treatment of AD. Biologically active molecules like sesquiterpenoids exhibit significant structural diversity and are an important class of terpenoids with a wide range of biological activities. Previous studies have demonstrated the potential of various sesquiterpenoids to improve cholinergic transmission through inhibiting AChE. This review summarizes the research progress on the activity of various sesquiterpenoids against AChE over the past five years. Through analysis, it was found that several sesquiterpenoids possess AChE inhibitory activity to varying degrees, demonstrating their potential as candidate medicines for the treatment of AD.

UDP-2, 3-diacylglucosamine pyrophosphate hydrolase (LpxH) is a key enzyme in the lipid A biosynthesis pathway of Gram-negative bacteria. Lipid A is an indispensable component of lipopolysaccharide (LPS), and LPS plays an important biological function in Gram-negative bacteria. Therefore, LpxH is considered one of the important targets in the research of antibacterial drugs. Based on the function and structure of LpxH enzyme, this review systematically summarizes and analyzes the currently reported LpxH inhibitors, and offers perspectives on future research directions. This review will provide ideas for the design of LpxH inhibitors and contribute to the development of new antibacterial drugs targeting LpxH.

Octapeptin is a class of cyclic lipopeptides with broad-spectrum antibacterial activity, demonstrating significant potential against multidrug-resistant Gram-negative bacteria. This review provides a comprehensive overview of the sources, structure, synthesis methods, and antibacterial mechanisms of octapeptin, with a focus on the impact of structural modifications on their antibacterial activity. The unique structural features of octapeptin contribute to their strong antibacterial potential, and structural modifications can significantly enhance their activity, offering insights into the development of novel antibiotics.

The endogenous small molecule metabolites (such as neurotransmitters, fatty acids, carbohydrates, nucleotides, and amino acids) in the body played an important role in the development and progression of central nervous system diseases. With the emergence of precision medicine, an increasing demand for the precise localization and precise analysis of those metabolites, as well as a clearer understanding of their biological functions was presented. Mass spectrometry imaging (MSI) technology, a non-invasive in vivo imaging technique, could provide an accurate spatial distribution, and the concentrations of endogenous metabolites or pharmaceutical components in different organs. There were many advantages including no specific labeling, short duration, and wide detection range. Matrix-assisted laser desorption/ionization mass spectrometry imaging played a leading role in the study of the precise distribution of endogenous metabolites in the brain microregions of disease animal model such as Parkinson's disease, Alzheimer's disease, stroke, and insomnia. In addition, it was widely applied to study the distribution of central nervous system drugs in the brain microregions. This review systematically examines the fundamental principles underlying MSI and its groundbreaking technological innovations over the past decade.

Ulcerative colitis (UC) is a chronic, non-specific inflammatory disease of the colon that is characterized by complex etiologies, challenging treatment, and high recurrence rate. At present, the main clinical therapeutic agents include suppositories, foams, and liquid enema formulations. But they have problems such as low adhesion and short drug residence time. As a new dosage form, gel has the characteristics of high bioavailability, long drug retention time, high safety and appropriate biological adhesion. This article reviews the relevant literature at home and abroad in the past five years, and analyzes and summarizes the classification of gels, prescription design of various gels, matrix selection, preparation process and pharmacological effects. The results show that the gel can effectively prolong the action time of the drug and alleviate the inflammatory response, which can offer a promising therapeutic strategy for UC management.

Oral liver-targeting nano-drug delivery system is the most potential drug delivery method which has the advantages of good drug compliance, low side effects, and high safety. However, the biological and physiological barriers in the gastrointestinal tract and liver severely hinder the oral absorption and accumulation of drug carriers in the liver lesions. This review first describes the main biological and physiological barriers encountered by the oral liver-targeted nano-drug delivery system during the drug delivery period. Meanwhile, this article summarizes oral liver-targeting drug carriers in recent years. In addition, the research progress of oral liver-targeted nano-drug delivery system in the treatment of liver-related diseases is introduced, which provides ideas for the development of oral liver-targeting nano-drug delivery system in the liver-related disease treatment.

Traditional Chinese medicine contains numerous monomeric active ingredients, which have rich pharmacological effects. However, many of these active ingredients have poor physicochemical properties, making them challenging to use in clinical settings. Cocrystal technology has demonstrated significant potential in enhancing the solubility and bioavailability of these poorly soluble active ingredients. This article provides a comprehensive overview of the current research on cocrystals of active ingredients in traditional Chinese medicine and briefly discusses the relationship between the structures of these active ingredients and the cocrystal formers. The aim is to provide reference for further research on cocrystals of active ingredients of traditional Chinese medicine and their broader applications in the pharmaceutical industry.

The essential requirements of traditional Chinese medicine (TCM) and its identity of drug make it necessary to treat TCM preparations with considerations of clinical practice routines and drug attributes. Quality stability is an elementary drug quality attribute and lays the foundation to guarantee a consistent traditional Chinese medicines (TCMs) efficacy. In light of the complex ingredients of natural sources, the unique medical theory and the safety & efficacy proof from clinical practices, it's hardly possible to objectively evaluate and effectively control TCMs quality using qualitative and quantitative methods with chemical components as references. To ensure efficacy consistency, stable TCMs quality is a prerequisite and thus is regarded as an industrial focus in earnest need of regulatory attention and also the key points and challenge of TCM quality study and assessment. Since the initiation of drug evaluation system reform, Center for Drug Evaluation has devoted to TCM regulatory science study and issued a series of guidelines. These guidelines are established based on characteristics of TCM preparations, scientific principles and actual production situations. Meanwhile, traditional thinking and experience in TCM quality control and modern theories such as "Quality by Design" and "Whole-process Quality Control" were also adopted. As the characteristics of TCM preparations and quality control requirements, this paper analyses the above guidelines, and concludes that these documents have combined TCM clinical experience and drug attributes of safety, efficacy and quality control. They provide guidance for ensuring basic stability of TCM ingredients and reducing batch-to-batch variations. A quality control system that meets the above requirements is established to ensure consistent efficacy of TCM preparations.

Influenza A viruses (IAVs) exhibit rapid mutation and high infectivity, leading to a considerable number of cases and deaths globally. Thus, the development of broad-spectrum inhibitors is of paramount significance. In this study, targeting the hemagglutinin (HA) of IAVs, a set of high-affinity and specific aptamers, namely Apt(HA)-1, Apt(HA)-4, Apt(HA)-9, Apt(HA)-25, and Apt(HA)-93520, were obtained through CE-SELEX. The dissociation constants (K_D) of this group of aptamers [Apts(HA)] with the HA protein were determined by the SPR method to be 25.9, 44.8, 50.7, 54.0, and 45.2 nmol·L^-1, respectively. Apts(HA) can recognize and detect the internationally common influenza virus strains A/PR/8/34 (H1N1) and A/FM/1/47 (H1N1), as well as the clinically isolated Tamiflu-resistant strain A/Liaoningzhenxing/1109/2010 (H1N1), amantadine-resistant strains A/Hunanzhuhui/1222/2010 (H3N2) and A/Fujiantongan/196/2009 (H3N2), and possess broad-spectrum inhibitory effects on influenza viruses. These aptamer inhibitors feature strong in vitro accessibility, good stability, simple structure, and small size. This study not only developed novel broad-spectrum aptamers against IAVs, exerting extensive inhibitory effects on various IAVs, but also paved the way for an effective strategy in the development of universal virus inhibitors.

Seasonal influenza caused by the influenza virus poses a serious threat to human health. The rapid emergence of drug-resistant viral strains and the increasing incidence of novel infectious diseases underscore the urgent need for researchers to develop innovative anti-influenza strategies and drugs. The identification of effective drug combinations and the repurposing of existing drugs have gained traction as a potential avenue for antiviral therapy. This study employed a high-throughput screening system for influenza A virus (IAV) inhibitors to identify drug combinations that exhibit synergistic effects with baloxavir marboxil from FDA-approved and pharmacopeia-listed compound libraries. The results showed that triprolidine, hydroquinine, and mexenone, which individually display limited anti-influenza activity, exhibited synergistic effects when combined with baloxavir marboxil markedly enhancing its anti-IAV potency in cells. Mechanistic studies indicated the synergistic effects of these drug combinations with baloxavir marboxil were not attributed to the inhibition of IAV RNA dependent RNA polymerase (RdRp) activity, suggesting that alternative mechanisms may be responsible for their enhanced effects. This study provides preliminary evidence supporting the feasibility of this drug combination screening strategy and offers a theoretical basis for improving the therapeutic potential of baloxavir marboxil through combination therapy.

Intrahepatic cholangiocarcinoma (ICC), the second most common primary liver malignancy after hepatocellular carcinoma (HCC), has seen a dramatic rise in incidence in recent years without effective therapeutic drugs. Hu Qi San (HQS), an empirical formula developed by professor Qian Ying, a renowned traditional Chinese medicine expert, through years of clinical practice, has demonstrated clinical efficacy in chronic viral hepatitis and primary hepatocellular carcinoma. However, fundamental research on HQS for ICC remains uncleared. Thus, this study utilized a sgp19-KRas mouse model with typical ICC-like lesions and a QBC-939 cell model to evaluate the therapeutic effects of HQS on ICC in vitro and in vivo and elucidate its underlying mechanisms. First, crystal violet staining and EdU assays revealed that HQS significantly inhibited the proliferation and migration of QBC-939 cells (P < 0.05). Subsequently, sgp19-KRas mice were treated with daily oral administration of 20 g·kg^-1 HQS for three weeks. Results demonstrated that HQS effectively suppressed ICC growth and tumor cell proliferation in sgp19-KRas mice, while improving survival rates. Mechanistically, network pharmacology analysis predicted 303 overlapping targets between HQS's active components and ICC-related disease targets, with significant enrichment in the Hippo signaling pathway. The molecular docking results predicted that ononin, calycosin, gallic acid, and taxifolin in HQS can directly bind to Yes-associated protein (YAP). Experimental validation further demonstrated that these four components exhibit in vitro inhibitory effects on QBC-939 cells. Subsequent q-PCR and Western blot analyses of liver tissues from treated sgp19-KRas mice confirmed that HQS inhibited the transcription of YAP, downregulated YAP protein expression, and suppressed the transcription of YAP downstream genes, particularly those in the YAP-Notch pathway. In conclusion, this study demonstrates that HQS exhibits potential therapeutic effects on ICC, and its mechanism of action is closely associated with the YAP-Notch signaling pathway.

In colorectal cancer, tumor necrosis factor receptor-associated factor 2 (TRAF2) is a positive regulator of the Wnt/β-catenin signaling pathway and is closely related to the occurrence and development of colorectal cancer. This study aims to discover novel anti-colorectal cancer small molecules targeting TRAF2 by establishing a high-throughput TRAF2 affinity screening system. The study established and optimized a high-throughput TRAF2 screening system based on Dianthus, and conducted a large-scale screening of 3 716 traditional Chinese medicine (TCM) small molecules from a TCM small molecule library. Microscale thermophoresis (MST) and cellular thermal shift assay (CETSA) were further applied to verify the binding in both purified proteins and living cells. Among the candidates, the strongest binding compound, eupatilin (EUP), had an affinity of 7.87 μmol·L^-1 with TRAF2. TOPFlash assays showed that EUP can inhibit the activity of the Wnt/β-catenin signaling pathway and the growth of colorectal cancer cells and mouse small intestinal adenoma organoids. After TRAF2 was knocked out using CRISPR/Cas9 technology, the inhibitory effect of EUP on the Wnt signaling pathway and colon cancer cells was significantly weakened, indicating that TRAF2 is the functional target protein of EUP. In summary, the novel high-throughput screening method based on Dianthus constructed in this study can be used for the screening of small molecule targeted drugs, and it has been confirmed that EUP is a TRAF2-targeted small molecule that inhibits the development of colorectal cancer, providing a theoretical basis for the development of targeted TRAF2 anti-tumor drugs.

The intestinal barrier, as the primary interface between the body and the external environment, plays a crucial role in sepsis. Methylene blue, with its anti-inflammatory, antioxidant, and microcirculatory-improving properties, has been widely used in the treatment of sepsis. However, whether it can improve intestinal barrier function in sepsis and its underlying mechanisms remain unclear. In the present study, a sepsis mouse model was established via intraperitoneal injection of lipopolysaccharide (LPS). The effects of methylene blue (5-20 mg·kg^-1) on inflammation in septic mice were observed using HE staining, ELISA, and RT-PCR. Changes in intestinal barrier function in the jejunum and colon tissues were analyzed via Evans blue staining, immunofluorescence, and RT-PCR. Additionally, the effects of methylene blue (40 μg·mL^-1) on NF-κB and RhoA/ROCK signaling pathways in LPS-stimulated macrophages (RAW264.7) and intestinal epithelial cells (Caco-2) were evaluated. The animal study was approved by the Animal Ethics Committee of Southwest University (Approval No.: IACUC-20241125-04). Results showed that methylene blue (5-20 mg·kg^-1) significantly reduced the levels of inflammatory cytokines in plasma and intestinal tissues of septic mice and upregulated the expression of tight junction proteins in the jejunum and colon. In vitro experiments confirmed that methylene blue (40 μg·mL^-1) inhibited the increased secretion of inflammatory cytokines in LPS-stimulated RAW264.7 cells, a mechanism associated with NF-κB pathway inhibition. Methylene blue (40 μg·mL^-1) also reversed the LPS-induced decrease in transmembrane electrical resistance and cell migration in Caco-2 cells, a mechanism related to RhoA/ROCK pathway inhibition. In summary, the present study demonstrated that methylene blue can improve intestinal inflammation by inhibiting the NF-κB pathway and enhance intestinal barrier function via the RhoA/ROCK pathway, ultimately alleviating the progression of sepsis.

Enzalutamide resistance (ENZR) is the most common issue in castration-resistant prostate cancer (CRPC). After a period of treatment with the second-generation androgen receptor (AR) inhibitor enzalutamide (ENZ), patients develop resistance to ENZ, posing a serious threat to male health. However, there are currently no effective drugs for treating prostate cancer patients with ENZ resistance. Based on the structural optimization and modification of enzalutamide, and through molecular biological methods such as cell viability assays, colony formation, cell apoptosis detection, Western blot and other molecular biology techniques, it was discovered that the half-maximal inhibitory concentration values of LS-13 for prostate cancer LNCaP-ENZR and 22RV1 cells are 17.09 and 19.90 μmol·L^-1, respectively. Subsequently, after treating the resistant cell lines with 0.1, 1, 5, and 10 μmol·L^-1 of LS-13 for 12 days, the observation results indicated that LS-13 could inhibit the in vitro clonal formation of LNCaP-ENZR and 22RV1 cells in a dose-dependent manner. Furthermore, after treating the resistant cell lines with 1.25, 2.5, 5, and 10 μmol·L^-1 of LS-13 for 72 h, the results of flow cytometry also showed that LS-13 could promote apoptosis of the resistant cell lines in a dose-dependent manner. Furthermore, under the same concentration conditions described above, after LS-13 treatment of 22RV1 cells for 24 h, it was found that their migration speed was significantly slowed down (P < 0.05). These results collectively indicate that the small molecule compound LS-13 has a pronounced effect in inhibiting ENZR. Further mechanism studies found that LS-13 can target ARfl/ARv7 and promote its protein degradation, inhibit the transcription levels of AR-downstream target genes, thereby suppressing ENZR cell proliferation and promoting apoptosis. This study provides a candidate compound for targeting ARfl/ARv7 to overcome ENZ resistance.

The traditional cigarette smoke-induced chronic obstructive pulmonary disease (COPD) mouse model has a long period and its stability is not easily guaranteed. Based on the intratracheal instillation of cigarette smoke extract (CSE), this study aims to establish a CSE-induced COPD mouse model. C57BL/6J mice were used as model animals, except for the normal control group, animals in each group received intratracheal instillation of CSE. The airway reactivity and pulmonary function of the mice were measured using a small animal respiratory pulmonary function testing system, the leukocyte classification count in the bronchoalveolar lavage fluid (BALF) of the mice was determined using a blood and body fluid analyzer, the levels of inflammatory factors and neutrophil extracellular traps (NETs) in BALF were measured using enzyme-linked immunosorbent assay (ELISA), and the histopathological changes in the mouse lung tissue were detected using H&E staining method. The experiments were approved by the animal care and welfare committee of Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College (approval No. 00006301). The results of the study showed that after intratracheal instillation of CSE, the model group mice exhibited significant COPD-like pathological and functional changes, characterized by tachypnea, slow weight gain. H&E staining showed pathologic changes such as massive inflammatory infiltration of lung tissue, alveolar wall collapse and alveolar lumen dilatation. The leukocyte classification count (total leukocytes, neutrophils, eosinophils), the levels of inflammatory factors such as tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and NETs levels in the BALF were significantly increased compared to the normal control group (P < 0.01, P < 0.001). The index of airway reactivity, specific resistance, airway (sRaw) was significantly increased (P < 0.05), pulmonary function indicators such as peak expiratory flow (PEF), forced expiratory flow at 75% of forced vital capacity (FEF₇₅), dynamic lung compliance (Cdyn), forced expiratory volume in the first 100 ms (FEV₁₀₀), and the ratio of forced expiratory volume in the first 100 ms to forced vital capacity (FEV₁₀₀/FVC) were significantly reduced (P < 0.01), while the respiratory index (RI) was significantly increased (P < 0.05). Compared to the model group, roflumilast can significantly improve the inflammatory pathological changes and pulmonary function damage in model animals. In summary, the intratracheal instillation of CSE can rapidly establish a COPD mouse model, replicating the airway inflammation and decreased pulmonary function of COPD to a certain extent, and provides a new candidate animal model for the study of the pathological mechanism and drug screening evaluation of COPD.

The objective of this study was to explore the therapeutic effects and underlying mechanisms of Guizhi Fuling Wan (GZFLW) in the treatment of hepatocellular carcinoma (HCC). Firstly, the therapeutic effect of GZFLW on HCC was evaluated by constructing a subcutaneous xenograft mouse model of H22 hepatoma cell line and combining with cell experiments. Liquid chromatography tandem mass spectrometry (LC-MS/MS) was used to analyze the drug components of GZFLW, and network pharmacology and transcriptomics methods were combined to screen out the potential core targets and signaling pathways of GZFLW in the treatment of HCC, and the core targets were verified by molecular experiments. Bioinformatics analysis results showed that, GZFLW may regulate p53, vascular endothelial growth factor A (VEGF-A) and hypoxia inducible factor-1α (HIF-1α), which may play an important role in the regulation of GZFLW. HIF-1α and other key targets, which have a certain impact on angiogenesis and ferroptosis in HCC. The results of in vitro and in vivo experiments showed that GZFLW could significantly inhibit the growth of subcutaneous transplanted tumor, and the serum containing GZFLW could inhibit the proliferation of H22 cells in vitro. The results of immunofluorescence and immunohistochemistry showed that GZFLW could down-regulate the expression of vascular endothelial markers cluster designation 31 (CD31) and cluster designation 34 (CD34). It also reduced the levels of VEGF-A and HIF-1α. Western blot analysis further showed that after GZFLW treatment, the expression of ferroptosis-related proteins solute carrier family 7 member 11 (SCL7A11) and glutathione peroxidase 4 (GPX4) was significantly decreased. However, the expression of p53 was significantly increased. The comprehensive analysis results suggest that GZFLW may induce ferroptosis in HCC by regulating the p53/SLC7A11/GPX4 signaling pathway, and inhibit tumor angiogenesis by inhibiting the expression of HIF-1α and VEGF-A, thereby effectively inhibiting the occurrence and development of HCC. This experiment has been approved by the Animal Experiment Ethics Committee of Shaanxi University of Chinese Medicine (ethics approval number: SUCMDL20230915001).

In this study, a series of 4-phenoxypyridine derivatives containing imidazole-4-formamido and 1,2,4-triazole were designed and synthesized as novel inhibitors of Fms-like tyrosine kinase 3 (FLT3). The results showed that the series of compounds had good antiproliferative activity. Among them, the preferred compound 10c was in two cell lines containing FLT3-ITD (internal tandem duplication, ITD) mutations, namely human myeloid core solo leukemia cells (MV4-11) and human acute myeloid leukemia cells (Molm-13), with IC₅₀ values of 1.90 and 5.78 μmol·L^-1, respectively. Apoptosis and cycle results showed that compound 10c could promote apoptosis and prevent cells in S phase. Molecular docking results showed that compound 10c formed two π-π stacking effects with PHE691 and HIS809 of FLT3, and the pyridine parent nucleus and Linker part formed important hydrogen bonds with LYS644 and CYS694 of FLT3, respectively. In summary, compound 10c exhibited good anti-proliferation activity in vitro, which is of value for further in-depth research, and also lays the foundation for the development of more novel FLT3 inhibitors with novel structures.

Microtubule-targeting drugs play a significant role in the treatment of metastatic and drug-resistant cancers. However, the emergence of resistance to microtubule-targeting drugs in clinical settings necessitates the development of molecules with novel modes of action and mechanisms to overcome drug resistance. The multi-target drug molecule strategy is an approach that has been developed in recent years to address complex diseases and drug resistance issues. A series of compounds based on tirbanibulin with increased Fsp3 have been prepared, which have been identified with ¹H NMR, ¹³C NMR and MS. The antiproliferative activities have been evaluated against HeLa and HepG2 with MTT method. The effect on microtubule and cell cycle distribution of compound 22 has been evaluated with immunofluorescence and flow cytometry. IC₅₀ of compound 22 against HeLa and HepG2 were 45 and 51 nmol·L^-1. And it destroys microfilament and arrests cell cycle at G2/M. The SAR show that pyridine moiety in tirbanibulin cannot be replaced with 2-imidazolidinone. Compound 22 was identified as a lead in the further research.

Ischemic stroke (IS) is a disease with high mortality and disability rate worldwide, and there are few clinical treatment drugs. The natural active ingredients of traditional Chinese medicine, such as cinnamaldehyde and anisaldehyde, have antioxidant and anti-inflammatory effects, which can reduce the oxidative stress injury caused by ischemic stroke. In addition, studies have found that L-arginine is a substrate of nitric oxide synthase (NOS), which can be converted into NO in the body to dilate blood vessels and increase blood flow in the cerebral ischemic area. In this paper, 10 new L-arginine/active aldimine derivatives were designed and synthesized, and the neuroprotective effects of ischemic stroke were evaluated biologically. The results of oxygen glucose deprivation/reperfusion (OGD/R) cell model showed that the cell survival rate of the arginine imine derivative AI-2 group formed by L-arginine and 2-methoxycinnamaldehyde was higher than that of the positive drug butylphthalide (NBP) group, and it could effectively release L-arginine, significantly reduce the levels of reactive oxygen species and malondialdehyde, increase the level of superoxide dismutase, and had a high antioxidant effect. The results of in vivo experiments showed that AI-2 (5 mg·kg^-1) could significantly improve the neurological function score of transient middle cerebral artery occlusion model in rats and effectively reduce cerebral infarction, which was superior to butylphthalide. In this study, a class of L-arginine/active aldimine derivatives with neuroprotective activity were found, which provided lead compounds for the treatment of ischemic stroke. All operations in the experiment were approved by the Animal Ethics Committee of China Pharmaceutical University, approval No. YSL-202503102.

A total synthesis of dehydrocurvularin (DCV) demethylated derivatives and structure modification have been completed using a Friedel-Crafts reaction and a highly olefin metathesis reaction. All compounds were identified by NMR, HRMS, etc. Their antitumor activity with high expression of ATP-citrate lyase was evaluated by CCK-8 assay in MDA-MB-231 and HepG2 cells. The structure activity relationship studies were also investigated. Among them, compound 7e showed better antitumor activity than DCV, and could significantly induce apoptosis in MDA-MB-231 cells. These results represent a step toward the development of novel and improved DCV derivatives.

Colorectal cancer (CRC) poses a serious threat to human life and health. The intestinal pathogenic bacteria Fusobacterium nucleatum (Fn) and enterotoxigenic Bacteroides fragilis (ETBF) are closely associated with the occurrence and progression of CRC. The development of novel compounds targeting Fn and ETBF holds significant clinical importance for the treatment of CRC. In this study, broth microdilution method was employed to screen 3 400 compounds in a compound library for in vitro antibacterial activity against the two bacteria. Using a criterion of MIC₅₀ ≤ 64 μg·mL^-1, 52 compounds with anti-Fn activity and 44 compounds with anti-ETBF activity were identified. Among them, compound G7 exhibited the best inhibitory activity against both two bacteria (Fn, MIC₅₀ = 1 μg·mL^-1; ETBF, MIC₅₀ = 4 μg·mL^-1) and could effectively inhibit the migration of colorectal cancer cells by inhibiting bacteria. Preliminary mechanistic studies revealed that G7 exerts its antibacterial effects by inhibiting biofilm formation and the expression of multiple genes related to adhesion and hyphal formation.

From an aqueous extract of the stems with hooks of Uncaria rhynchophylla, eight pairs of scalemic enantiomers of minor lignanoid (+)-/(-)-1 - (+)-/(-)-6、(+)-/(-)-9, and (+)-/(-)-10 as well as two pairs of racemic enantiomers (+)-/(-)-7 and (+)-/(-)-8 were isolated and chirally separated by column chromatographic techniques over macroporous adsorbent resin, MCI resin, Toyopearl HW-40, silica gel, and Sephadex LH-20, together with flash and chiral HPLC techniques. Their structures including the absolute configurations were determined by spectroscopic data analysis and theoretic calculations of electronic circular dichroism (ECD) spectra. Among them, chiral separation and absolute configurational determination of the optical active compounds (+)-/(-)-1 - (+)-/(-)-7 are reported for the first time, while the occurrence of scalemic enantiomers (+)-/(-)-9 and (+)-/(-)-10 in nature are discovered for the first time. Preliminary bioassay showed that (+)-8 and (-)-8 inhibited acetaminophen (APAP) induced HepG2 cell damage at a concentration of 10 μmol·L^-1.

The chemical constituents of the aerial parts of Euphorbia antiquorum (family Euphorbiaceae) were investigated. From the EtOAc partition of the 90% EtOH extract of the title species, thirteen compounds were separated by silica gel, octadecylsilyl silica gel and Sephadex LH-20 column chromatographies, as well as semi-preparative HPLC. The structures of these isolates were characterized as five pimarane-type (1-5), two abietane-type (6, 7) and six atisane-type (8-13) diterpenoids on the basis of spectroscopic analyses (MS, NMR, ECD, etc.). Among them, compounds 1 and 6 were identified as two previously unreported ones. The α-glucosidase inhibitory activity of selective diterpenoids was further evaluated in the current work. Compound 1 exhibited moderate inhibition against the enzyme, with an IC₅₀ of 150.03 ± 3.41 μmol·L^-1, being lower than that of the reference drug acarbose.

This study employed plant morphology, histology, and DNA barcoding techniques to identify the species of wild Cistanche plant in Kenya, and HPLC to determine the contents of its main active components, phenylethanoid glycosides, sugar alcohols, and betaine, alongside its fingerprint. The results showed that the wild Cistanche plant collected from Kenya was taxonomically identified as Cistanche senegalensis (Reut.) Beck. Compared to cultivated C. tubulosa from China, Kenyan C. senegalensis exhibited lower levels of echinacoside, cistanoside A, acteoside, isoacteoside, and 2′-acetylacteoside, but higher concentrations than those in C. deserticola. Betaine content was comparable to C. deserticola, while glucose was undetectable. The fingerprint of Kenyan C. senegalensis showed closer similarity to C. tubulosa. This work establishes a foundation for conserving Kenyan Cistanche resources and supports further research into their potential applications.

Using separation methods such as silica gel, MCI, ODS, Sephadex LH-20 gel column chromatography, and high-performance liquid chromatography, two C21 steroid components were isolated and identified from the traditional medicinal plant Munronia unifoliolata Oliv. (commonly known as "Aituotuo"), which is used for treating rheumatic joint pain and traumatic injuries. The structures of these compounds were identified as 2α, 3α, 15β-trihydroxy-20(S)-benzoyl-pregnane (1) and 2α, 3α, 15β-trihydroxy-20(S)-tigloyl-pregnane (2) using comprehensive structural identification methods, including UV, IR, high-resolution mass spectrometry, and one-dimensional and two-dimensional nuclear magnetic resonance. Compounds 1 and 2 were reported for the first time as pregnane-type components from M. unifoliolata, with compound 1 being a new compound. The results of the bioactivity screening indicated that both compound 1 and compound 2 did not inhibit nitric oxide (NO) production and showed no activity in reversing multidrug resistance.

The rice fermentation products of the fungus Scedosporium aurantiacum GXIMD 02519 were isolated and purified by silica gel column chromatography and semi-preparative liquid chromatography, etc. Their structures were characterized by various spectroscopic methods, including HR-ESIMS and NMR, etc. The inhibitory activity of nuclear factor kappa-B (NF-κB) and osteoclast differentiation of these compounds were determined by approaches of NF-κB luciferase reporter gene and tartrate-resistant acidic phosphatase (TRAP) staining. A new azaphilone, isochromophilone M (1), along with nine known compounds, were isolated and identified from the fungus S. aurantiacum GXIMD 02519, which were identified as 6-((1E, 3E)-3, 5-dimethylhepta-1, 3-dien-1-yl)-2, 4-dihydroxy-3-methylbenzaldehyde (2), barbatinic acid (3), botryorhodine B (4), 2, 6-dihydroxy-3, 4-dimethylbenzoic acid methyl ester (5), 2, 4-dihydroxy-3, 6-dimethylbenzoic acid (6), atranol (7), 4-methoxyphenylacetic acid (8), 4-hydroxybenzaldehyde (9), and scopararane C (10). Compounds 3 and 10 showed inhibitions of lipopolysaccharide (LPS)-induced NF-κB activation in RAW264.7 cells. Moreover, compound 10 could further inhibit receptor activator of nuclear factor κB ligand (RANKL)-induced osteoclast differentiation in bone marrow macrophages without observed cytotoxicity. This is the first report of the diterpene compound (10) as a potential inhibitor of osteoclast differentiation.

Seventeen compounds were isolated from the ethyl acetate extract of the tubers of dry Alisma plantago-aquatica by silica gel, Sephadex LH-20 and high performance liquid chromatography. All of them were identified as the prototerpene triterpenoids by the analysis of mass spectrometry and nuclear magnetic resonance spectroscopy. Compounds 1 and 2 are the new compounds. Among of them, 16 compounds were screened for α-glucosidase inhibitory activity, however, none of them was active.

Gliomas represent highly malignant brain tumors with high recurrence rates, susceptible to drug resistance and difficult to treat, therefore finding and screening new therapeutic candidates is extremely significant. In this study, we firstly used UHPLC-Q/TOF-MS method to identify the chemical constituents of Scutellaria barbata extract, and a total of 38 constituents were identified. A microfluidic chip model containing six independent U251 cell units was constructed, each U251 cell unit consisted of three parallel channels, the middle channel was used for culturing matrigel-encapsulated U251 cells, and the channel of medium or drug-containing medium was on both sides, the U251 cells formed a multicellular sphere in the matrigel, and were able to mimic the microenvironment of tumor hypoxia to a certain extent. Based on the constructed glioma microfluidic chip model and the perfusion flow mode for the screening of active components of Scutellaria barbata which had antitumor activity, the perfusion samples of Scutellaria barbata were analyzed by using UHPLC-Q/TOF-MS, and the relative concentrations of potential active components were calculated by the internal standard method, and it was found that the constituents of the samples containing scutellarin, apigenin-7-O-β-D-glucuronide, luteolin, scutellarein, naringenin and apigenin were significantly decreased (P < 0.05), suggesting that these six compounds have potential anti-glioma activity. Using CCK-8 assay, scratch assay and apoptosis assay to validate the medicinal efficacy of the six potential active components obtained from the screening, it was observed that all the six potential active components could reduce the proliferation of U251 cells, decrease the migration rate of the cells, and kill the cells by apoptosis. The sensitization of temozolomide performed by the six potential active compounds was further investigated, which indicated that naringenin, apigenin and luteolin in combination with temozolomide individually increased the sensitivity of U251 cells to temozolomide as well as decreased the cell viability. These results will provide technical and scientific evidence for treating glioma and further solving the anti-drug resistance phenomenon of glioma treatment by Scutellaria barbata and its active components.

This study investigated the effects of acteoside (ACT) on serum metabolites in purinomycin aminonucleoside (PAN)-induced chronic glomerulonephritis (CGN) in young rats using non-targeted metabolomics and lipidomics, and initially explored its potential mechanism of efficacy. The experiment was approved by the Animal Welfare Ethics Committee of Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences (No. 2022B152). Firstly, ELISA kits were used to detect serum cytokines including IP-10, IL-6, IL-1β, and TNF-α in rats of each group to evaluate the efficacy of ACT. Non-targeted metabolomics and lipidomics techniques were used to determine metabolites and lipids in serum, and integrated analysis was conducted to reveal the mechanism. The results showed that compared with the model group, the levels of IP-10, IL-6, IL-1β, and TNF-α in the serum of the ACT groups were significantly reduced, indicating a significant pharmacological effect of ACT. Non-targeted metabolomics results revealed that 23 different metabolites were identified between the control and model groups, involving pathways such as biosynthesis of unsaturated fatty acids, biosynthesis and degradation of valine, leucine, and isoleucine, and linoleic acid metabolism. Between the ACT and model groups, 25 different metabolites were identified, involving pathways such as biosynthesis of unsaturated fatty acids, the TCA cycle, and linoleic acid metabolism. Lipidomics results showed that 68 significantly different lipids were identified between the control and model groups, involving pathways such as omega-3/omega-6 fatty acid synthesis, T-cell receptor and costimulatory signaling, and G-protein-coupled receptor activation of PKC. After ACT treatment, significant changes were observed in 67 differential lipids, with notable reversal in the aforementioned pathways. It also regulated PPARα modulation of lipid metabolism and related pathways such as α-linolenic acid and linoleic acid metabolism. Correlation analysis of differential metabolites and lipids indicated that metabolites such as homovanillic acid, palmitic acid, indoxyl sulfate, as well as triglycerides and phosphatidylcholines were critical. The commonly regulated pathways by non-targeted metabolomics and lipidomics were biosynthesis of unsaturated fatty acids and linoleic acid metabolism. In conclusion, ACT primarily treats CGN in young rats by regulating the biosynthesis of unsaturated fatty acids and linoleic acid metabolism pathways through differential lipids such as triglycerides and some phosphatidylcholines, and differential metabolites such as homovanillic acid, palmitic acid, and indoxyl sulfate. This study initially elucidated the intervention mechanism of ACT in the treatment of pediatric chronic glomerulonephritis from the perspective of serum metabolism, laying a solid foundation for the rational clinical application of ACT in children with CGN.

Rhubarb is a Chinese medicinal herb of multiple origins, there are differences in the purgative efficacy of three origins of rhubarb, and the active components of the purgative efficacy of rhubarb is not yet clear. The study reveals the differences in the purgative efficacy of three origins of rhubarb by method of pseudo-targeted metabolomics combined with pharmacological experiments (approval number: BUCM-4-2022080301-3026), and screens out the active components of the purgative efficacy of rhubarb. Analyzing the differences in the purgative efficacy of three origins of rhubarb through animal pharmacological experiments. Subsequently, potential active components of purgative efficacy were identified through literature review, and corresponding ion pairs were obtained by fragmentation pathways and software optimization, and the peak areas of each component were obtained by using dyMRM mode of QQQ-MS. Multivariate statistical analysis was used to analyze the potential active components of purgative efficacy in rhubarb with different origins, thereby achieving pseudo-targeted metabolomics analysis, and the active components can be screened with pharmacological experimental results. The pharmacological experimental results indicate that the purgative efficacy of Rheum tanguticum Maxim. ex Regel is significantly higher than that of Rheum palmatum L. and Rheum officinale Baill. The results of PCA showed that there were significant differences in the potential active components of purgative efficacy among three origins of rhubarb. 97 differential potential active components of purgative efficacy were screened through OPLS-DA combined with Kruskal Wallis test; combined with the results of metabolomics, 23 active components of purgative efficacy were screened, among which the compounds with clear chemical structures were sennoside A, sennoside B, rheinosides A, rheinosides C, rheinosides D, physcion-8-O-glucoside, rhein-8-O-glucoside, chrysophanol-1-O-glucoside. The above results suggest that the method of pseudo-targeted metabolomics combined with pharmacological experiments has been used to clarify the relationship of purgative efficacy between the three origins of rhubarb, reveal the distribution of potential active components of purgative efficacy in three origins of rhubarb, and screen out the active components of purgative efficacy. This not only provides a theoretical basis for the quality evaluation and precise clinical use of rhubarb, but also provides a new idea for traditional Chinese medicine to search for efficacy markers.

A new vardenafil analogue has been discovered in a sexual enhancement health beverage through the application of ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC/Q-TOF MS) and high-performance liquid chromatography-diode array detection (HPLC-DAD). This analogue was then purified via preparative HPLC (Prep-HPLC), and its chemical structure was determined using NMR spectroscopy. Spectral data indicated that the vardenafil analogue possesses an n-propyloxy group in place of the ethoxy group on the aromatic ring, and a hydroxyethyl group in substitution for the ethyl group on the piperazine ring. The compound, termed "propoxyphenyl hydroxyvardenafil", has not been reported in China.

This study aimed to investigate the effects of colloidal silica nanoparticles Aerosil 200 (A200) on intestinal barrier and mucosal immunity in childhood mice. Three-week-old BALB/c mice were administered A200 (20, 50, 200 mg·kg^-1) via gavage for 7 days. Intestinal morphology, barrier function, immune cell phenotypes, and oral tolerance were systematically evaluated. Histopathological analysis of intestinal morphology revealed that no obvious pathological damage in the intestine. However, 200 mg·kg^-1 A200 increased ileal villus length, reduced crypt depth. Functional assessments of the intestinal barrier demonstrated that 200 mg·kg^-1 A200 upregulated ZO-1 in the ileum but downregulated it in the colon; The intestinal stem cell marker Lgr5 remained unchanged, while Mucin-2 increased and lysozyme decreased in the ileum. Analysis of immune cell phenotypes revealed that the 200 mg·kg^-1 group exhibited elevated proportions of Treg and Th2 cells in the mesenteric lymph nodes, while serum and intestinal levels of TNF-α and IL-6 remained unaffected. Oral tolerance assays revealed no alteration in OVA-specific IgG, IgG1, or IgG2a antibody levels. Animal welfare and the animal experimental protocols were strictly consistent with related ethics regulations of Peking University Health Science Center (Approval No.: BCJB0076). In conclusion, A200 is generally safe for childhood mice but may modulate epithelial barrier proteins and immune cell phenotypes at high doses, which may provide insights into its pediatric application.

To investigate the transformation of wettability of phosphoric acid-treated pollen after treatment with different reagents, to optimize the preparation of the wettability-transformed pollen sporopollenin carrier system to find the optimal alkali treatment conditions, and to characterize its structure and surface morphology, we first analyzed the wettability of the original pollen and the acid-dissolved pollen, measured the water contact angle of the acid-dissolved pollen treated with each reagent with the help of a contact angle tester, conducted a one-way test on the alkali treatment conditions that could transform the wettability of the pollen, explored the influence of the optimization of the conditions for the preparation of the hydrophilic pollen sporopollenin carriers by using a response surface methodology and the glycerol absorption time as the response value, and carried out a validation and verification test on the process parameters. The process parameters were verified and validation tests were conducted, and finally the sporopollenin was characterized by FTIR and SEM. The contact angle test showed that the water contact angle of pollen after acid digestion was 135.1°, which was changed to 16° after NaOH treatment, and the contact angle of glycerol was changed from 140.8° to 9.6°. The best alkali treatment process was obtained through the optimization of one-factor test and response surface method, and the final optimization conditions were obtained according to the actual experimental process: the volume of NaOH was 105 mL, i.e., 15 times the volume of pollen, and the concentration of NaOH was 1.5 mol·L^-1. The stirring time was 18 h, and the stirring temperature was 85 ℃. Under these conditions, three parallel experiments were carried out to verify the wettability of Codonopsis pollen and Lotus pollen respectively, in which the average value of Codonopsis pollen results was 58 s, and the average value of Lotus pollen results was 49 s, and the RSD was less than 5%. The wettability study revealed that pollen sporopollenin changed from hydrophobicity to hydrophilicity after NaOH treatment, and according to the results of response surface optimization, the prepared alkali-treated pollen sporopollenin showed a significantly shorter time for complete glycerol absorption and a better hydrophilicity with a water contact angle of close to 0°, which is a reproducible process and a stable and reliable preparative process, and theoretically, will be more conducive to the loading of hydrophilic drugs.

Acute myeloid leukemia (AML), characterized by rapid progression, therapeutic resistance, and high relapse rates, remains a critical threat to patient survival. The complex leukemic microenvironment often diminishes the efficacy of conventional monotherapies and single-target approaches. Drug delivery strategies based on microenvironment characteristics and multi-mechanism synergy are expected to have better therapeutic effects on AML. Therefore, we developed a biomimetic supramolecular nanoparticle (Fe-GA@CM) through homologous targeting strategy and metal-polyphenol coordination for co-delivery of gambogic acid (GA) and Fe³⁺. The particle size, zeta potential, and release performance of the system were characterized. Furthermore, the efficacy and mechanism of action of this system in treating AML was explored. The results showed that the particle size of the nano system was approximately 270 nm and positively charged. After being specifically taken up by AML cells, it is reduced by high concentrations of glutathione in the tumor microenvironment and released Fe²⁺ and GA in a responsive manner. The combination therapy for AML is achieved by inducing ferroptosis with Fe²⁺ and inhibiting glycolysis in AML cells with GA. In summary, the formulated Fe-GA@CM provides novel ideas for targeted therapy and combination therapy for AML.

Pig hoof nails represent an underutilized byproduct of pork processing. According to traditional Chinese medical theory, pig hoof nails are believed to contain compounds that may exhibit pharmacological properties such as promoting hemostasis, aiding coagulation, and reducing inflammation, which could potentially benefit the healing of skin wounds. In this study, different molecular masses of hoof nail peptides (HNPs) were extracted from porcine hoof nails. Wound healing assays were employed to identify HNPs that significantly promote fibroblast proliferation and collagen synthesis across three molecular weight ranges: less than 10 kDa (HNP Ⅰ), 3-10 kDa (HNP Ⅱ), and more than 10 kDa (HNP Ⅲ). We formulated HNP Ⅱ into a temperature-sensitive gel designed to conform to uneven wound surfaces. This supports the development and application of hoof nail peptide formulations. The animal operation was approved by the Laboratory Animal Ethics Committee of Southwest University (No: IACUC-20231218-01). All experiments were conducted in accordance with relevant guidelines and regulations.

Simvastatin is a class of hydroxymethylglutaryl coenzyme A reductase inhibitors used to lower blood lipids and prevent atherosclerosis, but its poor water solubility and low bioavailability seriously affect clinical efficacy. In this experiment, simvastatin solid dispersions were prepared using supercritical CO₂ antisolvent method, and the prescription process parameters were optimized by Box-Benhnken response surface methodology, to improve the dissolution characteristics. The Box-Benhnken response surface methodology optimization experimental scheme took in vitro cumulative dissolution degree of simvastatin (dependent variable) as the evaluation index, investigated the influence of four factors (independent variables) including pressure, temperature, carrier type, and CO₂ outflow rate on the preparation of simvastatin solid dispersions by supercritical CO₂ antisolvent technology. Next, the particle size (PZ), infrared (IR) absorption spectrum, differential scanning calorimetry (DSC), X-ray diffraction (XRD), and scanning electron microscopy (SEM) characterization were performed on the preferred process conditions for simvastatin dispersion. Based on these findings, further research was conducted on the in vitro dissolution properties of the product as well as the relative bioavailability after oral administration to rats. Animal experiments were approved by the Animal Ethics Committee of Jiangsu College of Food and Drug Engineering (No. JSSPYPCAC2024). The results showed that the optimal process parameters for preparing simvastatin solid dispersions by critical CO₂ antisolvent technology were pressure 12.00 Mpa, temperature 35.0 ℃, CO₂ flow rate 3.3 L·min^-1, F-127/PVP K23-27 (1∶10, w/w). The prepared solid dispersion was in a loose powder form, which appears as overlapping flaky aggregates under SEM. The laser particle size analysis showed a D50 value of 0.78 μm. DSC and XRD indicated that simvastatin existed in an amorphous form within the dispersion. The in vitro cumulative dissolution value after 60 minutes was 89.87%, and the relative bioavailability of oral administration to rats was 226.6%. In summary, this study successfully applied supercritical CO₂ antisolvent method to prepare simvastatin solid dispersions, simplified the experimental steps and optimized process routes using Box-Benhnken response surface methodology. The prepared simvastatin solid dispersions exhibited fast dissolution rates and high cumulative dissolution degrees, which can be used as a prescription strategy for developing novel oral simvastatin formulations with high bioavailability.

Chinese medicine supramolecular is an important material form and effective form of Chinese medicine compound. The form and activity of supramolecular are affected by many factors, and the mixing order is one of the key factors. In this study, dynamic light scattering (DLS), scanning electron microscopy (SEM), turbidimeter and Tyndall effect were used to observe the differences of micromorphology and physical properties of three different groups of Coptidis Rhizoma group + Gardeniae Fructus + Scutellariae Radix (CGS), Coptidis Rhizoma group + Scutellariae Radix + Gardeniae Fructus (CSG) and Scutellariae Radix + Gardeniae Fructus + Coptidis Rhizoma group (SGC). The main components of the three groups of decoction were analyzed by high performance liquid chromatography (HPLC). The antibacterial activity of the three groups of decoction was preliminarily compared by in vitro Staphylococcus model, and the dynamic formation process of supramolecules in the three groups of decoction was preliminarily discussed by molecular dynamics simulation (MDS) technology. The results showed that different mixing sequences led to changes in the morphology of supramolecular parts of Huanglian Jiedu Decoction, with significant differences in macroscopic phenomena, electron microscopy, particle size, potential and turbidity, and significantly affected the original antibacterial activity. In addition, the results of molecular dynamics simulation also show that the different mixing order of the main components of Huanglian Jiedu Decoction will significantly affect its self-assembly behavior. This study is to further explore the factors affecting the formation of supramolecular substance basis of Huanglian Jiedu Decoction, also provides a reference for the clinical rational administration of Huanglian Jiedu Decoction formula granules.