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.