Candida albicans (C. albicans) is the most prevalent conditionally pathogenic fungus implicated in clinical fungal infections. The formation of biofilms is a critical contributor to the pathogenicity of C. albicans, facilitating its invasion and the subsequent spread of infection. Biofilm-associated C. albicans exhibits reduced susceptibility to commonly used antifungal agents, leading to prolonged treatment durations, high rates of recurrence, and the emergence of drug resistance. Consequently, the development of antifungals that are both low in toxicity and effective in inhibiting biofilm formation is of paramount importance. Plant-derived natural products have emerged as a vital resource for drug discovery, owing to their diverse chemical structures and myriad biological activities. In this review, we provide a comprehensive summary of plant-derived natural products with demonstrated anti-biofilm activity against C. albicans, as reported over the past five years. This includes an examination of various classes of compounds such as alkaloids, polyphenols, terpenes, quinones, organic acids, and essential oils, thereby offering insights that may advance the development of novel anti-fungal therapeutics targeting C. albicans.

Anxiety disorder, as a psychiatric condition characterized by high prevalence, chronic progression, and frequent comorbidities, faces significant limitations with current clinical treatments such as selective serotonin reuptake inhibitors, serotonin-norepinephrine reuptake inhibitors, and benzodiazepines. These therapies are often constrained by limited efficacy, notable side effects, and issues of drug resistance. In recent years, the sigma-1 receptor has emerged as a promising target for novel anti-anxiety medications, with its ligands demonstrating significant anxiolytic effects in various animal models, offering new hope for addressing current therapeutic challenges. However, no anti-anxiety drugs targeting the sigma-1 receptor have advanced to clinical research stages. Therefore, this article reviews recent progress in understanding the molecular mechanisms of the sigma-1 receptor in anxiety disorders and summarizes the pharmacological efficacy of its ligands in animal models, aiming to provide a theoretical foundation and research directions for the development of innovative anti-anxiety drugs targeting the sigma-1 receptor.

Heart failure (HF) has become a major challenge in global public health, and its incidence and lethality are on the rise. While the effectiveness of clinical interventions in modern medicine, they are accompanied by rising adverse drug reactions, recurring diseases, and no reduction in long-term mortality. In this context, herb pairing methods informed by traditional Chinese medicine theories, through mechanisms of multi-component synergy and multi-target network regulation, have been vital in HF prevention and treatment. In this study, we have gathered and analyzed recent investigations on the pharmacological effects of traditional Chinese medicine for HF, resulting in a list of 16 notable and representative herb pairs. Their main functions include reducing inflammation, countering oxidative stress, preventing apoptosis, inhibiting cardiac fibrosis, regulating neuroendocrine disorders and myocardial microcirculation, and ultimately preventing or treating HF, thus broadening the scope of personalized clinical drug regimens and the development of new therapeutic agents.

Traditional Chinese medicine (TCM) databases provide a crucial platform for deepening TCM theory, exploring the relationship between disease and syndrome, revealing compatibility mechanism, and developing component-based Chinese medicine. The construction of TCM databases is centered around the multi-level "syndrome-formula-herb-component-target-pathway-function-disease" network. With the advancement of computer science and high-throughput experimental technologies, there is a growing trend of integration with artificial intelligence and multi-omics strategies. This paper reviews the current state of TCM databases, and summarizes the research progress from three aspects of data collection, data analysis, and network visualization. It systematically discusses the strategies and methods for building TCM databases. Additionally, it also reveals the challenges faced by the current TCM databases, and puts forward the future exploration direction and potential development trend, aiming at providing technical support for the construction of subsequent related databases and offering insights for research in related fields.

Metabolic dysfunction-associated steatohepatitis (MASH) has been identified as one of the leading factors contributing to the rising incidence of advanced liver disease, with its prevalence increasing annually. Currently, the primary treatment strategies focus on lifestyle modifications, including dietary adjustments and physical exercise, with weight loss being particularly crucial. Meanwhile, significant progress has been made in the development of pharmacological treatments, especially thyroid hormone receptor β (THR-β) agonists, which have garnered considerable attention due to their potential efficacy in MASH treatment. This review aims to provide an in-depth exploration of the latest research advancements in THR-β agonists for MASH treatment, offering new therapeutic strategies and perspectives for clinical practice.

This article reviews the secondary metabolites produced by marine microorganisms with anti-Pseudomonas aeruginosa activity in the past five years. The compounds are classified according to their structures, including macrolides, anthraquinones, peptides, alkaloids, terpenoids, and other kinds of structures. The chemical structure and antibacterial activity of these compounds are mainly introduced. This article provides a reference for further research and development of natural products from marine microorganisms with antibacterial activity.

Fingerprint refers to the chromatography, spectrum or other visual data that can identify the essential characteristics of certain substances or components with complex ingredients after proper processing and analysis. The analysis of effect components in biological samples is characterized by complex biological medium components, large differences in physical and chemical properties, complex and diverse metabolic reactions of effect components, and mainly trace amounts of prototypes and metabolites. Therefore, accurate and rapid determination of the effect components is of great significance. The advantages and disadvantages of each method in recent years are analyzed. The development trend of fingerprint technology in the analysis of effect components in biological samples is prospected in the future, so as to provide reference for the development and application of fingerprint technology in the field of biological sample detection.

This article focuses on the methods for determining vaccine potency, especially the feasibility and challenges of replacing traditional in vivo methods with in vitro methods, as well as related considerations. The in vivo method for determining vaccine potency has been used for a long time for vaccine quality control, but it has problems such as high variability, low experimental efficiency, and animal ethics issues. With the 3R principle and technological progress, in vitro methods have gradually become a trend to replace in vivo methods. In vitro testing methods have advantages in evaluating the consistency of vaccine production and monitoring changes during the production process, and can more effectively evaluate the stability and immune effects of vaccines. Although the mechanisms of in vitro and in vivo methods are different, and the quality control attributes of the two methods are not completely consistent, in vitro methods can still effectively replace in vivo potency assays, especially for quality control during the early stages of vaccine development. Common in vitro alternative methods include immunoassay, physicochemical analysis, and molecular biological assay. The selection of appropriate methods depends on the type of vaccine and its mechanism of action. In addition, the article emphasizes that the establishment of in vitro methods should be included in the vaccine development process as early as possible, and ensure its stability, reproducibility, and correlation with clinical efficacy. Although the promotion of in vitro alternative methods faces challenges, with technological progress, it is expected to further replace animal experiments in the future, improving the efficiency and ethicality of vaccine quality control.

Compared with normal cells, tumor cells exhibit unique metabolic characteristics. The therapeutic strategies formulated based on the energy metabolic features of tumors aim to block the energy and material supply of tumor cells at the source, remodel the immunosuppressive tumor microenvironment (TME), and create favorable conditions for immune cell infiltration and function. In recent years, Chinese herbal medicine-derived extracellular vesicles have attracted attention due to their unique metabolic regulatory capabilities and targeting properties. These extracellular vesicles can not only precisely deliver multi-component active molecules but also reprogram the metabolic processes in the tumor microenvironment, reverse immunosuppressive conditions, and thereby enhance antitumor immune responses. This review systematically summarizes the metabolic regulatory mechanisms of TCM-derived vesicles and their synergistic effects in immunotherapy, and looks forward to their clinical transformation potential and future research directions.

Tripterygium wilfordii Hook F (TWHF), specifically triptolide (TP), is an active compound derived from the plant with significant anti-rheumatoid arthritis (RA) effects. Studies show that TP exerts potent anti-inflammatory and antioxidant properties by inhibiting the expression of several inflammatory cytokines, such as TNF-α, IL-1β, and IL-6. In terms of immune regulation, TP modulates the activity of immune cells like T cells and neutrophils, reducing inflammation and tissue damage. Additionally, TP can regulate multiple signaling pathways, including NF-κB, JAK/STAT, and MAPK, to suppress the proliferation and invasion of synovial cells in RA, thereby mitigating joint damage. With the advancement of nanotechnology, the development of TP-based nanomedicine has gained attention in recent years. Innovative nanocarriers, such as polymer nanoparticles, exosomes, and inorganic nanoparticle drug delivery systems, have significantly improved TP's bioavailability and targeting capabilities while reducing its toxicity. By modifying the surface of these nanomaterials, their ability to target and release TP at inflammation sites is enhanced, leading to more precise drug delivery. Moreover, nanomedicines extend TP's circulation time in the body and reduce early-stage toxicity, thus improving the therapeutic outcomes for RA. In conclusion, TP and its nanomedicine formulations show great promise in RA treatment. Future research should continue to explore their mechanisms and clinical applications, offering new perspectives for the treatment of rheumatoid arthritis.

During the decoction process of traditional Chinese medicine (TCM), not only do chemical bonds interact, but non-covalent interactions also occur. In recent years, studies have discovered the presence of aggregates connected by non-covalent bonds in TCM decoctions, collectively referred to as supramolecules of traditional Chinese medicine (STCM). This review summarizes the literature on STCM, discussing research methods, formation mechanisms, and research significance from four perspectives: individual components, single herbs, medicinal pairs and compound prescriptions. The aim is to provide a reference for future research on STCM. Due to the complexity and diversity of TCM components, the research on the formation mechanism of STCM in compound formulas, drug pairs and single herb decoction liquids is not perfect, which is also a key and difficult point in future STCM research. The perspective of self-assembled supramolecular chemistry provides a new idea for the study of the pharmacodynamic material basis of TCM, and pushes the research in the field of TCM to a new height.

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) has shown significant potential in cancer therapy due to its ability to selectively induce apoptosis in tumor cells while minimally affecting normal cells. Currently, developed TRAIL-based cancer therapies have demonstrated better therapeutic effects in preclinical trials. However, their clinical application has been limited by issues such as poor stability, insufficient targeting, short half-life in vivo, and low bioavailability. In recent years, nanocarriers, including liposomes, nanoparticles, and micelles, have been widely utilized in the field of drug delivery. Combining TRAIL with these nanocarriers can help overcome these limitations and enhance its antitumor efficacy. This review summarizes the latest research progress on the combination of TRAIL with liposomes, nanoparticles, and micelles in cancer treatment. It explores the mechanisms by which these nanocarriers improve the stability, targeting, and bioavailability of TRAIL, as well as the therapeutic advancements of this combined approach at both the animal and clinical levels. The aim is to provide valuable insights for the development and clinical application of TRAIL-based cancer therapies.

With the introduction of the concept of precision medicine, pharmaceutical treatment is shifting from 'one-size-fits-all' to personalization, and 3D printing, as an emerging formulation technology, can significantly enhance the complexity and flexibility of formulations, solve the many problems of the traditional process, and achieve personalized drug therapy. In recent years, with the rapid development of 3D printing technology, various 3D printing technologies have been used to develop oral solid dosage forms. Given the differences in the types and characteristics of 3D printing technologies, it is necessary to select appropriate printing materials according to the application characteristics of printing technology, and the choice of materials can significantly impact the personalized shaping and precise release of oral solid dosage forms. This paper lists the principles and characteristics of fused deposition molding technology, semi-solid extrusion technology, melt extrusion deposition technology, and binder jetting technology, which is suitable for the 3D printing of oral solid preparations. It explores the selection and application of printing materials for commonly used 3D printing technologies in detail to guide the choice of materials to prepare different principles of 3D printing of oral solid preparations.

Mechanistic modeling, which describes cellular metabolic behavior through mathematical equations, is an important theoretical tool for optimizing and predicting biological processes. In this study, a cell culture mechanistic model for biosimilar trastuzumab was constructed based on the Monod growth metabolism model. To address the high complexity of differential equation parameter fitting and the tendency to fall into local optima, particle swarm optimization (PSO), grey wolf optimization (GWO), and differential evolution (DE) algorithms were introduced for model parameter fitting. The results show that GWO outperforms the others in global search capability and fitting accuracy, demonstrating higher stability and adaptability in handling multi-parameter fitting problems. Furthermore, improvements were made to the position and velocity update methods of the GWO algorithm. The improved grey wolf optimization (IGWO) significantly enhanced the model's predictive ability, enabling accurate predictions of cell density, metabolite concentration, and antibody titer. This study provides important theoretical support for improving the production efficiency of biopharmaceuticals, reducing development costs, and optimizing cell culture processes.

With the widespread dissemination of the plasmid-mediated colistin resistance gene mcr-1, colistin resistance has spread globally, severely threatening the last line of defense in global anti-infective therapy. This study aimed to evaluate the resistance-reversing effects of linalool combined with colistin against mcr-1-positive Escherichia coli (E. coli) and to elucidate the underlying mechanisms, aiming to propose a therapeutic strategy to combat colistin resistance. The synergistic antibacterial effects in vitro were determined using the broth microdilution checkerboard method and time-kill curves, and biofilm assays. Further investigations elucidated the underlying mechanisms by assessing inner/outer membrane permeability, membrane potential (ΔΨ), dehydrogenase activity (DHA), proton gradient (ΔpH), ATP levels, and mcr-1 gene expression. Finally, we assessed the in vivo efficacy of linalool in combination with colistin against Galleria mellonella infection and mouse acute peritonitis models. Results demonstrated that linalool-colistin combination exhibited synergistic antibacterial effects against mcr-1-positive E. coli both in vitro and in vivo. Mechanistically, linalool directly damaged the bacterial outer membrane, reduced ΔΨ, and increased inner membrane permeability. Moreover, linalool inhibited DHA activity, disrupted ΔpH establishment, ultimately impairing ATP production. Additionally, linalool significantly downregulated the mcr-1 expression, suppressing lipid A modification in resistant strains. These effects collectively enhanced colistin penetration, triggering membrane disruption and energy metabolism collapse, ultimately achieving synergistic antibacterial efficacy. In vivo, combination therapy significantly improved survival rates in both infection models. In conclusion, linalool restores colistin susceptibility in mcr-1-positive E. coli through synergistic mechanisms, providing a promising therapeutic strategy against colistin resistance. All animal experimental procedures were reviewed and approved by the Animal Welfare and Ethics Committee of Xiangnan University (approval No.: 2023DWLL029).

Neurological diseases, cardiovascular diseases, and pulmonary diseases have always been major threats to human health. How to intuitively, efficiently, and rapidly screen out drugs with therapeutic effects is a key scientific issue in the field of biomedicine that urgently needs to be addressed. In this study, aiming at the pathological characteristics of these three types of diseases, key gene promoters such as those of nerve growth factor, vascular regeneration and repair factors and pneumonia factors were cloned and linked to reporter genes for quantification. Finally, a molecularly targeted drug screening model based on the stem cell differentiation system was constructed, and its function was verified by applying known pathway activators. This model not only has the universal characteristic of predicting the efficacy of compounds, but also can achieve dynamic analysis of the relationship between time and efficacy as well as the relationship between dosage and efficacy in view of the complex characteristics of traditional Chinese medicines and their compound ingredients, effectively revealing the material basis of their efficacy and providing important technical support for the quality control of subsequent clinical medications. While maintaining the advantages of traditional screening methods, this model breaks through the limitations of existing technologies. Finally, we discuss the technical characteristics of existing drug screening models and conduct a prospective discussion on the application prospects of this model in the field of drug research and development. The animal welfare and experimental procedures of the work were approved by the Medical Ethics Committee of Southwest Jiaotong University [approval No. SWJTU-2403-NSFC(026)].

To investigate whether pretreatment with cake-separated moxibustion can alleviate the injury of aspirin to the gastric mucosa and reveal its potential mechanism, Sprague-Dawley rats (male and female) were randomly divided into the blank control group, model group, pretreatment acupoint group, and pretreatment control point group, 20 rats in each group. This experiment was approved by the Institutional Animal Care and Use Committee (IACUC) of Hunan Slack Jingda Laboratory Animal Co., Ltd. (approval No.: IACUC-SJA18038). From day 1 to day 8, the rats in the blank control and model groups were tied for 30 min each day, and the rats in the pretreatment acupoint group, and pretreatment control point group were tied and treated with moxibustion for 30 min each day. The rats in each group were fasted without water for 24 h after tying on days 2, 4, 6 and 8, respectively, and then gavaged at 150 mg·kg^-1 aspirin, and the gavage volume of 1 mL per 100 g body weight was injected into the stomach of the rats in the model group, pretreatment acupoint group, and pretreatment control point group, and the normal saline was injected into the stomach of the rats in the blank control group with a gavage needle at a dose of 1 mL per 100 g body weight. Gastric mucosa injury index was observed, and histology of gastric mucosa was observed by hematoxylin-eosin staining. Superoxide dismutase (SOD) and malondialdehyde (MDA) content in serum and gastric mucosa were detected by enzyme-linked immunosorbent assay. The levels of Kelch-like ECH-associated protein 1 (Keap1), nuclear factor-erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), catalase (CAT), glutathione S-transferase (GST), and NAD(P)H: quinone oxidoreductase 1 (NQO1) in gastric mucosa were detected by quantitative reverse transcription polymerase chain reaction (RT-qPCR) and Western blot. The results showed that compared with the blank control group, the rats in the model group showed increased behavior status, emotional response, skin hair, diet status and stool status scores (P < 0.05), significantly higher gastric mucosal injury index (P < 0.05), significantly lower SOD (P < 0.05), and significantly higher MDA (P < 0.05), and significantly upregulated Keap1 expression in gastric mucosal tissues (P < 0.05), while the expression of Nrf2, HO-1, CAT, GST, and NQO1 was significantly downregulated (P < 0.05). Compared with the model group, the emotional response and sleep status scores of the rats in the pretreatment acupoint group were reduced (P < 0.05), and the gastric mucosal injury index was significantly reduced (P < 0.05), and the gastric mucosal injury in pretreatment control point group tended to be reduced, but there was no statistical difference (P > 0.05), and SOD was significantly increased (P < 0.05), while MDA was significantly decreased (P < 0.05) and the level of Keap1 was significantly decreased (P < 0.05), while the expression of Nrf2, HO-1, CAT, GST, and NQO1 was significantly upregulated (P < 0.05) in the pretreatment acupoint group and pretreatment control point group. Compared with the pretreatment control point group, the gastric mucosal injury index was significantly lower (P < 0.05), SOD was significantly higher (P < 0.05), while MDA was significantly lower (P < 0.05), Keap1 was significantly lower (P < 0.05) and Nrf2, HO-1, CAT, GST, and NQO1 were significantly higher (P < 0.05) in the pretreatment acupoint group. The above results suggested that the pretreatment of cake-separated moxibustion could activate Nrf2/antioxidant response element (ARE)/HO-1 signaling pathway, enhance the antioxidant stress ability of gastric mucosa cells, and reduce the damage of gastric mucosa caused by aspirin.

Gouteng Xuanshen Granules, as an institutional preparation of Shandong University of Traditional Chinese Medicine Affiliated Hospital, have shown significant efficacy in reducing blood pressure, but their mechanisms of action are not yet fully understood. This study used HPLC-MS to perform non-targeted metabolomics analysis on serum samples from subjects, including those with prehypertension (PHT) and healthy individuals. TCMSP was used to identify the active components of Gouteng Xuanshen Granules. Databases like Gene Cards and OMIM were used to predict potential targets related to normal high blood pressure and to explore the antihypertensive effects of the granules on PHT. The study recruited 20 patients with prehypertension meeting the diagnostic criteria into the PHT group (Group P) and 20 healthy volunteers into the control group (Group C). The prehypertension group was administered Gouteng Xuanshen Granules (Group GT). The study was approved by the Ethics Committee of Shandong University of Traditional Chinese Medicine Affiliated Hospital, and informed consent was obtained from all participants [ethical review approval number: (2022) Lun Review No. (036)-KY]. All subjects underwent four lipid tests and blood pressure measurements. Blood samples were collected once before and after administration of the granules in the prehypertension group, and once at enrollment in the control group. HPLC/MS was used to analyze serum samples from all subjects. The results showed that after administration of the granules, patients with prehypertension had a significant decrease in blood pressure and changes in the four lipid parameters. Metabolomics analysis revealed significant changes in purine metabolism, arginine biosynthesis, and sphingolipid metabolism after granule administration. Network pharmacology analysis suggested that the granules primarily exert their antihypertensive effects by acting on targets such as SIRT1, STAT3, and AKT1. This study provides preliminary insights into the metabolic differences among healthy individuals, those with prehypertension, and the treatment group, offering valuable references for the prevention and treatment of prehypertension. The study preliminarily analyzed the metabolic differences among healthy individuals, patients with elevated blood pressure within the normal range, and those treated with Gouteng Xuanshen Granules, providing valuable reference for the prevention and treatment of elevated blood pressure within the normal range.

In this study, 17 new unreported pyrrolidine derivatives 5a-5p, 6awere synthesized in a fully synthetic manner using the FDA-approved drug captopril as the basic structure, and the antimicrobial activity was tested in vitro against clinically isolated Klebsiella pneumoniae carrying the New Delhi metal beta lactamase (NDM-1) gene and Gram-negative E. coli carrying the Klebsiella pneumoniae carbacyticase-2 (KPC-2) gene. The results showed that 5jcould reduce the minimum inhibitory concentration (MIC) of meropenem (MEM) against clinically isolated Klebsiella pneumoniae containing NDM-1 to 32 μg·mL^-1, which was better than that of the positive drug captopril; 5l, 5m, and 5oreduce the MIC of MEM against KPC-2-containing Gram-negative E. coli to 2 μg·mL^-1, which was better than that of the positive drug captopril; the virtual molecular docking results of some pyrrolidine derivatives suggested the possible binding mode of pyrrolidine derivatives to NDM-1 and KPC-2.

In this study, a series of novel biaryl amide derivatives featuring an acrylamide covalent warhead were designed and synthesized. The antiviral efficacy of these synthesized compounds against the human β-coronavirus HCoV-OC43 were evaluated using the CCK-8 assay. Among them, compound 6b, containing a 2-fluoroacrylamide covalent warhead, exhibited significant antiviral activity with an EC₅₀ value of 2.52 μmol·L^-1 and a selectivity index (SI) of 29, suggesting a reasonable therapeutic window. Molecular docking studies revealed that compound 6bcould inhibit the activities of host transmembrane serine protease 2 (TMPRSS2) and viral 3-chymotrypsin like protease (3CL^pro) through covalent bond formation, thereby exerting its antiviral effects. These findings not only expand the structural diversity of this class of compounds but also highlight their potential applications in antiviral therapy. Compound 6brepresents an ideal lead compound for the development of a new class of covalent antiviral drugs against coronaviruses.

Twelve compounds were isolated and purified from Fissistigma polyanthum by using various modern separation techniques such as silica gel, MCI, ODS, and Sephadex LH-20 column chromatography, as well as preparative HPLC. The structures of these compounds were determined by spectroscopic methods and identified as fissistigmaoid A (1), (10S)-11-hydroxy-β-cyperone (2), 11-hydroxy-8-oxo-β-cyperone (3), (4S, 5E, 10R)-7-oxo-tri-nor-eudesm-5-en-4β-ol (4), oxyphyllanene B (5), teuhetone (6), teuhetenone A (7), trans-4, 5-dihydroxycorocalane (8), 4-hydroxy-4, 7-dimethyl-1-tetralone (9), epicatechin (10), dihydroquercetin (11), and piperolactam C (12). Among them, compound 1 is a new nor-eudesmane-type sesquiterpene, while compounds 1-9 were isolated from F. polyanthum for the first time. Compounds 8 and 12 exhibited NO inhibitory activity in an LPS-induced RAW 264.7 murine macrophage inflammation model, with IC₅₀ values of 30.5 ± 1.5 and 10.5 ± 1.0 μmol·L^-1 respectively.

This study aimed to screen and identify bioactive peptides from Tabanus, characterize their structural and functional features, and provide a theoretical basis for the development of novel natural peptide-based therapeutics. Initially, bioactive peptides were extracted using phosphate buffered saline (PBS), followed by protein precipitation with cold ethanol and purification via solid-phase extraction (SPE). Peptide identification and sequence analysis were performed based on LC-MS/MS data using PEAKS Studio software. Peptide activity potential was then predicted using Peptide Ranker, and a comprehensive bioinformatics analysis was conducted to evaluate key functional characteristics, including active sites, solubility, stability, toxicity, and intestinal absorption capacity. A total of 10 378 peptide sequences were identified, with lengths predominantly ranging from 6 to 26 amino acids. Most peptides had a molecular weight below 1 500 Da, and 97.49% were below 2 500 Da. Based on Peptide Ranker score (≥ 0.80), 95 candidate bioactive peptides were selected. Among them, 59 peptides were identified as having neuroregulatory potential via the BIOPEP-UWM database, due to the presence of key functional residues. Bioinformatics prediction further revealed that among these 59 peptides, 29 exhibited good water solubility, 22 were predicted to be stable, 43 had favorable intestinal absorption potential, and none were toxic, highlighting their suitability for further functional studies and therapeutic development. Molecular docking analysis demonstrated that FPPRDF, the highest-ranking peptide according to Peptide Ranker, formed stable interactions with dipeptidyl peptidase Ⅲ (DPP Ⅲ), suggesting its potential neuroregulatory and antitumor activities. Overall, this study established a systematic and efficient workflow for the screening and functional prediction of bioactive peptides from natural sources. The proposed strategy provides robust technical support for subsequent experimental validation, mechanistic studies, and the development of novel peptide-based drugs.

A method was developed for the determination of toosendanin in Toosendan Fructus by HPLC coupled with charged aerosol detector (CAD) in the present study. Solid phase extraction (SPE) was employed for sample pretreatment to remove the impurities and enrich the target compounds. The methanal extracts of Toosendan Fructus were dissolved in 5 mL of 30% methanol and loaded to a C18 SPE column, followed by eluting with 6 mL of 30% methanol and 6 mL of methanol, respectively; and the components eluted with methanol was collected and used for preparation of the sample solution. Samples were separated on an Agilent ZOBAX SB C₁₈ column (4.6 mm × 250 mm, 5 μm) using the mixture of acetonitrile and water (33∶67) as the mobile phase, the column temperature was 30 ℃. The nitrogen inlet pressure of the CAD detector was 55 psi and the nebulizer chamber temperature was 35 ℃. As results, the HPLC-CAD spectrum of Toosendan Fructus sample was clear with little interference, and the separation of the target compound toosendanin was good. The established method was well validated, and showed high sensitivity, precision, good repeatability, and satisfactory stability. The limit of detection of toosendanin was 24.0 ng, the RSD values for precision, repeatability, and stability tests were all less than 5.0%; the recoveries of toosendanin at high, middle, and low levels were ranged from 95.9% to 99.7%. The established HPLC-CAD method was applied for quantification of toosendanin in 10 batches of Toosendan Fructus samples and compared with those of the LC-MS method. Pearson correlation analysis showed that the contents of toosendanin determined by the two methods were very similar and was highly positively correlated (the correlation coefficient was 0.995 0, P < 0.001). In conclusion, a method for determination of toosendanin in Toosendan Fructus was established by HPLC-CAD coupled with SPE, providing references for the quality control and evaluation of Toosendan Fructus.

This study initially explored the influence of different brewing methods of Rhei Radix et Rhizoma-Coptidis Rhizoma formula granules on the "bitter and cold" bias (formula granule-cold mixing, FG-CM; formula granule-hot mix and reheating, FG-HMR). We characterized the differences in morphology, molecular thermodynamics, spectroscopy and diarrhea phenotype of mice of the two brewing methods decoctions, in order to analyze the potential impact of different brewing methods on the clinical efficacy of Rhei Radix et Rhizoma-Coptidis Rhizoma formula granules. The research found that there were a large number of flocs in the FG-CM system under macroscopic observation, and the FG-HMR system was in a uniform turbid state. Similarly, under the scanning electron microscope, the particle size of FG-HMR was more uniform and the scale was smaller. The ITC experiment found that the interaction between the formula particles of Rhei Radix et Rhizoma-Coptidis Rhizoma was obvious. However, the characterization results of UV-vis, IR and HPLC indicated that the chemical types and contents of FG-CM and FG-HMR were basically consistent, suggesting that the brewing method did not change the chemical composition but there was molecular interaction. Through the mouse diarrhea model, it was found that compared with the FG-CM group, the "bitter and cold" bias in the FG-HMR group was more moderate. It could better maintain the normal intestinal homeostasis and intestinal tissue structure of mice, reduce the rate of soft stools in mice, and alleviate the side effects of diarrhea in mice. This study started from the perspective of material basis and diarrhea phenotype, with the expectation of providing a reference for the clinical administration of formula granules containing Rhei Radix et Rhizoma-Coptidis Rhizoma. The animal experiment was approved by the Ethics Committee of Beijing University of Chinese Medicine (Approval No.: BUCM2024-093-M).

Pathogenesis related protein 1 (PR1) plays an important role in plant resistance to stress and growth and development. This study identified and systematically analyzed 8 AsPR1 genes in Aquilaria sinensis using bioinformatics methods. The predicted molecular weight of encoded AsPR1 proteins is between 12.95 kD and 43.41 kD. The two-dimensional structure and conserved motif analysis of the AsPR1 proteins demonstrated that the similarity of 8 AsPR1 proteins was 51.38%, and all 8 AsPR1 proteins had conserved CAP-PR1 domains. Cis-acting elements analysis showed that the AsPR1 gene family proteins have cis-acting elements that respond to plant hormones, abiotic stress, and biological processes. Chromosomal mapping showed that the AsPR1 gene was distributed on chromosomes 1, 3, 4 and 8. The full-length AsPR1-4 sequence was successfully cloned according to the transcriptome sequencing analysis from A. sinensis, which contains a 516 bp ORF that encodes 171 amino acids. Furthermore, we successfully expressed AsPR1-4-GST fusion protein. The subcellular localization experiments indicated that AsPR1-4 is located in the cytoplasm and chloroplast. The real-time fluorescence quantitative PCR results showed that AsPR1-4 had the highest expression level in stems, followed by roots, and the lowest expression level in leaves. Exogenous MeJA and SA significantly increased the expression of AsPR1-4 in Calli. Additionally, the transcripts level of AsPR1-4 remarkably decreased under four abiotic stresses, including salt, low temperature, heavy metals, and drought stress. This study lays the foundation for further elucidating the function of PR1 protein in the defense response of A. sinensis and its application in agarwood production.