Objective To investigate the protective effect and mechanism of small RNAs (sRNA) derived from Xuanfei Baidu Formula (宣肺败毒方, XFBD) targeting angiotensin-converting enzyme (ACE) in lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice. Methods The modified CTAB method was employed to extract sRNA from XFBD and a library was constructed, followed by prediction and screening of sRNAs targeting ACE. The targeting specificity was verified using a dual-luciferase reporter system, and sRNAs capable of suppressing ACE expression were screened in human pulmonary microvascular endothelial cells (HPMEC). The effects of sRNA on angiotensin Ⅱ (Ang II) generation, inhibitor of inhibitor of nuclear factor-κB α (IκBα) and inflammatory cytokine expressions were examined by ELISA, Western blotting and qRT-PCR. An LPS-induced ALI mice model was established, control group, model group, captopril (10 mg/kg) group, XFBD (9.2 g/kg) group, NC-sRNA (10 nmol/animal) group, ACE-sRNA-1 (10 nmol/animal) group and ACE-sRNA-26 (10 nmol/animal) group were set up, with six mice in each group. Hematoxylin-eosin (HE) staining and Micro CT were used to evaluate the pathological and imaging changes of lung tissue; The number of white blood cells and neutrophils in peripheral blood, as well as the total protein concentration, white blood cells, neutrophils and lymphocytes numbers in bronchoalveolar lavage fluid (BALF) were detected; Immunohistochemistry was used to detect the expressions of vascular endothelial-cadherin (VE-cadherin) and intercellular adhesion molecule-1 (ICAM-1) in lung tissue; Western blotting and qRT-PCR were used to detect the expressions of ACE-Ang Ⅱ-Ang Ⅱ type 1 receptor (AT1R) pathway, IκBα and inflammatory factors in lung tissue; ELISA was used to detect the levels of Ang II and inflammatory factors in serum. Results A total of 50 potential ACE-targeting sRNAs were screened from XFBD, with 26 sRNAs validated to exhibit targeting effects, among which 12 sRNAs significantly inhibited ACE expression in HPMEC cells (P < 0.05, 0.01, 0.001). Seven sRNAs significantly suppressed Ang II generation, with ACE-sRNA-1/26 demonstrating the most potent effect (P < 0.001), inhibited IκBα protein and inflammatory factor expressions (P < 0.01, 0.001). In animal experiments, mice in model group had severe lung injury (P < 0.001); Compared with model group, ACE-sRNA-1 and ACE-sRNA-26 significantly improved lung injury in mice (P < 0.05, 0.01, 0.001), up-regulated VE-cadherin expression in lung tissue (P < 0.001), down-regulated ICAM-1, ACE, AT1R, IκBα and inflammatory factor expressions in lung tissue (P < 0.05, 0.01, 0.001), and reduced Ang II and inflammatory factor levels in serum (P < 0.05, 0.01, 0.001). Conclusion ACE-sRNA-1 and ACE-sRNA-26 derived from XFBD could target the inhibition of ACE expression and activity, alleviate pulmonary endothelial inflammation and barrier damage, and improve LPS-induced ALI in mice. The mechanism may be related to the regulation of ACE-Ang Ⅱ-AT1R pathway.

Objective To investigate the components of Isodon lophanthoides var. gerardianus aqueous extract (ILAE) and its protective effects against ethanol-induced alcoholic liver disease (ALD) in zebrafish, and to analyze its potential active components and targets using network pharmacology. Methods Ultra-performance liquid chromatography-quadrupole/time-of-flight mass spectrometry (UPLC-Q-TOF/MS) was employed to identify the components of ILAE. Wild-type AB strain and transgenic neutrophil zebrafish Tg (lyz: DsRed) at 4 d post-fertilization (4 dpf) were treated with 36, 73, 146 μg/mL ILAE or 70 μg/mL silymarin for 16 h, followed by induction of ALD with 2% ethanol for 32 h. The protective effects of ILAE on liver were assessed by observing developmental status, measuring biochemical indicators and conducting behavioral tests. Network pharmacology was used to analyze the potential components and targets of ILAE in treating ALD. Validation was performed through molecular docking and Western blotting. Results A total of 56 components were identified in ILAE, primarily flavonoids and phenolic acids. ILAE significantly alleviated ethanol-induced developmental impairment in zebrafish, the delayed absorption area of yolk sac and the area of liver enlargement were significantly reduced (P < 0.05, 0.001), lipid accumulation was significantly reduced (P < 0.001), liver pathological damage was relieved, liver cells were arranged in an orderly manner, and fat vacuoles were reduced. The activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were significantly decreased (P < 0.05, 0.01, 0.001), while the activity of alcohol dehydrogenase (ADH) and reduced glutathione (GSH) level were significantly increased (P < 0.05, 0.01, 0.001). The levels of reactive oxygen species (ROS), malondialdehyde (MDA) and triglycerides (TG) were significantly decreased (P < 0.05, 0.01, 0.001), and the number of neutrophils was significantly reduced (P < 0.01, 0.001). The excitement state of zebrafish was decreased, and the rapid movement trajectory and total movement distance were decreased (P < 0.05, 0.001). The core targets associated with ILAE and ALD were protein kinase Bα (AKT1), nuclear factor-κB subunit 1 (NF-κB1), signal transducer and activator of transcription 3 (STAT3), key components were chrysin, eupatilin and rosmarinic acid, which exhibited strong binding affinity to the targets. Kyoto encyclopedia of genes and genomes (KEGG) analysis and gene ontology (GO) analysis showed that the process involved lipid metabolism, insulin resistance, bile acid secretion, and exogenous substance metabolism pathways. Western blotting results showed that ILAE significantly reduced the expression levels of NF-κB1, STAT3 proteins and their phosphorylated forms in ethanol-induced zebrafish (P < 0.05, 0.01, 0.001), and increased the expression levels of AKT1 protein and its phosphorylated forms (P < 0.05, 0.01). Conclusion ILAE may improve ALD by targeting AKT1, NF-κB1, STAT3 through rosmarinic acid, chrysin, caffeic acid, 7-hydroxycoumarin and other components. Its protective effect may be related to reducing inflammatory response, decreasing lipid accumulation and enhancing antioxidant activity.

Objective To investigate the mechanism by which sodium new houttuyfonate (SNH) inhibits hepatocellular carcinoma (HCC) progression through autophagy regulation via ubiquitin-specific peptidase 22 (USP22)-mediated silent mating type information regulation 2 homolog 1 (SIRT1) deubiquitination. Methods Human liver cancer HepG2 cells were treated with SNH or sorafenib for 24 h, cell viability was detected by CCK-8 assay, cell apoptosis was detected by flow cytometry, and cell invasion ability was detected by Transwell assay. After plasmid transfection induced overexpression of USP22, the USP22-SIRT1 interaction was verified by co-immunoprecipitation (Co-IP). USP22 and SIRT1 protein expressions, as well as SIRT1 ubiquitination were detected by Western blotting. After treatment with SIRT1 activator, autophagy activity was evaluated through mRFP-GFP-LC3 dual fluorescence labeling, adenosine triphosphate (ATP) level measurement, lactate production assay, and analysis of USP22, SIRT1, microtubule-associated protein light chain 3 (LC3)-II/I and p62 protein expressions. For in vivo experiments, 40 BALB/c-nu nude mice were eutectopically transplanted with HepG2 cells and intervened with SNH or sorafenib for 12 d. Tumor volume and weight were monitored every 3 d. Hematoxylin-eosin (HE) staining and TUNEL staining were performed on tumor tissues, and immunohistochemistry was used to detect the protein expressions of nuclear proliferation antigens Ki67, USP22 and SIRT1. Results SNH dose-dependently suppressed HepG2 cell viability and invasion (P < 0.01, 0.001), induced cell apoptosis (P < 0.001). The effect of high-dose SNH was comparable to sorafenib. Co-IP confirmed USP22-SIRT1 protein interaction, which was significantly reduced by high-dose SNH (P < 0.01). High-dose SNH significantly down-regulated USP22 and SIRT1 protein expressions (P < 0.01, 0.001), increased SIRT1 ubiquitination (P < 0.001). These effects were reversed by USP22 overexpression (P < 0.05, 0.001). SNH inhibited autophagy in HepG2 cells, manifested by enhanced GFP/mRFP fluorescence signal (P < 0.01), decreased ATP level (P < 0.001), increased lactate level (P < 0.001), increased p62 expression and decreased expression levels of USP22, SIRT1 and LC3-II/I proteins (P < 0.05, 0.001). SIRT1 activation partially counteracted the autophagic inhibition of SNH (P < 0.05, 0.01, 0.001). In vivo, high-dose SNH significantly reduced tumor volume, weight and malignancy degree (P < 0.001), induced tumor cell apoptosis and decreased Ki67, USP22, and SIRT1 expressions (P < 0.001). Conclusion SNH inhibits HCC progression by suppressing autophagy through USP22-mediated regulation of SIRT1 deubiquitination, providing a potential therapeutic strategy for HCC.

Objective To screen the core active components of Shenfu Injection (参附注射液, SFI) using UHPLC-MS and molecular docking techniques, and to verify the effects of SFI on expressions of post-translational modifications of tubulin in chronic heart failure of heart-yang deficiency syndrome through in vivo and in vitro experiments. Methods UHPLC-MS was applied to identify the main components of SFI, and molecular docking was performed to analyze their interactions with tubulin-related targets. SD rats were randomly divided into control group, model group, colchicine (0.1 mg/kg) group, SFI low- and high-dose (3, 6 mL/kg) groups, with eight rats in each group. The CHF heart-yang deficiency model was replicated using sc isoproterenol, and drug intervention was given for two weeks. Echocardiography was used to detect cardiac function. ELISA was used to detect N-terminal pro-B-type natriuretic peptide (NT-proBNP) level in serum. Hematoxylin-eosin (HE) staining was used to detect pathological changes in myocardial tissue. Immunofluorescence was used to detect the expressions of acetyl α-tubulin, polyglut α-tubulin and detyr α-tubulin in myocardial tissue. Immunohistochemistry was used to detect the expressions of tubulin tyrosine ligase (TTL) and histone deacetylase 6 (HDAC6) in myocardial tissue. H9c2 cardiomyocytes were treated with isoproterenol to establish a cell injury model, and after intervention with SFI, the level of microtubule protein modification was detected. Results A total of 12 main active ingredients in SFI were obtained through UHPLC-MS screening, and molecular docking results showed strong binding affinity with tubulin and modification related proteins. The animal experiment results showed that compared with model group, SFI could significantly improve cardiac function of rats (P < 0.01), reduce the level of NT-proBNP in serum (P < 0.01), alleviate the degree of myocardial interstitial fibrosis, reduce the expressions of microtubule protein α-tubulin and microtubule density in myocardial tissue (P < 0.05, 0.01), reduce the expressions of microtubule protein tyrosine and glutamate, increase acetylation expression, down-regulate HDAC6 positive expression (P < 0.01), and up-regulate TTL positive expression (P < 0.01). The results of cell experiments were consistent with animal experiments, further verifying that SFI could improve post-translational modifications of microtubule proteins. Conclusion SFI could effectively reduce the levels of tyrosinization and glutamatzation of microtubules, increase acetylation levels, correct the imbalance of post-translational modifications of microtubules, thereby improving the abnormal microtubule network in cardiac myocytes of rats with CHF heart-yang deficiency, enhancing microtubule stability and improving heart function. This provides experimental evidence for the molecular mechanism of SFI treatment for CHF heart-yang deficiency.

Objective To systematically investigate the intestinal absorption characteristics of baicalin, glycyrrhizic acid, aloe-emodin, wogonin, emodin and chrysophanol in Jinzhen Oral Liquid (金振口服液) using the rat in situ single-pass intestinal perfusion model and Caco-2 cell model. Methods In vitro experiments were conducted using Caco-2 monolayer cell model to preliminarily evaluate the absorption capacity of the six components through apparent permeability coefficient (P_app). In situ experiments were conducted using rats in situ intestinal perfusion to further verify the absorption of the six components in rat intestinal segments (duodenum, jejunum, ileum, and colon) through the absorption rate constant (K_a) and effective permeability coefficient (P_eff). The in vitro and in situ absorption rules were compared and analyzed to clarify their absorption mechanisms. Results The in vitro and in situ experimental results showed good consistency. Under low-, medium-, and high-concentration perfusion conditions, all six components were absorbable in various intestinal segments of rats. Among them, the P_eff values in the duodenum, jejunum, and colon were all greater than 1.2 × 10⁻³ cm/min, indicating that the six components were well absorbed in these three intestinal segments. In the ileum segment, the P_eff values of baicalin, glycyrrhizic acid and wogonin were 1.8 × 10⁻⁴—1.2 × 10⁻³ cm/min, indicating moderate absorption of these three components in the ileum. The P_eff values of aloe-emodin, emodin and chrysophanol were greater than 1.2 × 10⁻³ cm/min, indicating good absorption of these three components in the ileum. In addition, there were differences in the absorption mechanisms of these six components, and most of them exhibited absorption characteristics that required carrier involvement, such as active transport and diffusion promotion (some of which were concentration dependent or saturated). At the same time, there were also components that mainly participate in passive diffusion or both active and passive transport. Conclusion In situ and in vitro intestinal absorption experiments reveal the segment-specific absorption patterns of six major components in Jinzhen Oral Liquid, with good overall absorption that meets the requirements for oral preparations.

Objective To analyze the formulation patterns of health foods containing Sangshen (Mori Fructus) and investigate their potential mechanisms using data mining, network pharmacology, and molecular docking techniques. Methods Information on Mori Fructus-related health foods up to September 13, 2025, was collected from the Special Food Information Query Platform of the State Administration for Market Regulation and Yaozhi.com. Formulation patterns were analyzed using frequency statistics, cluster analysis, and association rule analysis. Active ingredients of Mori Fructus were retrieved from databases such as the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP). Potential targets were predicted using SwissTargetPrediction, while disease targets related to primary health functions were retrieved from databases like GeneCards database. Target gene names were standardized using the UniProt database. Potential targets were screened using Venny and STRING databases. Protein-protein interaction (PPI) network analysis was then performed, and core targets were screened based on degree values. Gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analyses were conducted using the DAVID database. Finally, molecular docking was employed to validate the binding affinity between core targets and key components. Results A total of 243 Mori Fructus-containing health foods meeting the inclusion criteria were included, involving 177 traditional Chinese medicine ingredients, primarily heat-clearing drugs and tonifying deficiency drugs. The medicinal properties were mainly warm and neutral; the medicinal flavor was predominantly sweet; the meridians entered the lung, heart, and liver channels. The most common dosage form was capsules. The top two health functions were helping to enhance immunity and helping to alleviate physical fatigue. For immunity enhancement, 453 potential targets and 10 core targets were identified. For alleviating physical fatigue, 166 potential targets and 10 core targets were identified. Mori Fructus exerts its effects on enhancing immunity and alleviating physical fatigue through signaling pathways such as the TNF signaling pathway and the lipid and atherosclerosis pathway. Molecular docking results indicated that the key components of Mori Fructus could spontaneously and stably bind to the core targets. Conclusion This study, utilizing data mining, network pharmacology, and molecular docking, analyzed information related to Mori Fructus-containing health foods and preliminarily revealed the potential targets and mechanistic pathways associated with the main health functions of Mori Fructus, providing a theoretical basis for the subsequent development of Mori Fructus-based products.

Objective To comprehensively analyze the research status and hotspots of Guizhi (Cinnamomi Ramulus) from the perspective of bibliometrics, so as to provide theoretical guidance for the clinical application and in-depth research of Cinnamomi Ramulus in the future. Methods The four databases, namely CNKI, Wanfang, VIP, and Web of Science (WOS), were searched with Cinnamomi Ramulus as the keyword. The retrieval time was set from January 1, 1990 to March 31, 2025. The literatures were imported into NoteExpress literature management software for duplicate checking and screening. Software such as Excel, CiteSpace, and VOSviewer were used to conduct visual analysis on domestic and foreign researches on Cinnamomi Ramulus from the dimensions of publication trends and national distribution, publication institutions, publication authors, keywords, etc. Results A total of 2 476 Chinese literatures and 591 English literatures met the inclusion criteria. In the field of Cinnamomi Ramulus research, the China Academy of Chinese Medical Sciences ranks first in the number of published papers in both Chinese and English. Wang Zhenzhong and Chen Lidian respectively hold the highest number of publications in Chinese and English in this field. Research on Cinnamomi Ramulus is currently in a sustained development phase with high overall research heat, showing an upward trend. China is the primary contributing country, with gradually increasing international attention in recent years. Core research teams in this field have initially formed, butcooperative effect among institutions has not yet been fully highlight, the cooperation model is more often manifested as internal collaboration between medical colleges and their affiliated hospitals, as well as among research institutions within the same region. Deeper cooperation across regions and disciplines still needs to be further expanded. In the field of Cinnamomi Ramulus research, both Chinese and English literatures have focused on aspects such as its active components and mechanisms of action. From the perspective of keyword analysis, Chinese studies have paid more attention to classic famous prescriptions related to Cinnamomi Ramulus, the inheritance of medical cases, and clinical disease applications, while English studies have emphasized the exploration of its modern compound compatibility, pharmacological mechanisms, and network regulatory pathways. Conclusion Research in the field of Cinnamomi Ramulus is continuously expanding and deepening, evolving from early focus on traditional Chinese medical therapies and classical formulas to multi-faceted investigations covering component research, clinical diseases, action mechanisms, and modern technological applications.

Objective As key water-soluble components in traditional Chinese medicine (TCM) formulas, polysaccharides play a critical role in elucidating the mechanisms of these formulas. Clarifying their functions is a key breakthrough to decipher the mysteries of TCM formulas and advance its modernization. This study employs bibliometric methods to comprehensively analyze the current status, hotspots, and trends of research on polysaccharides from TCM formulas between 2005 and 2025, aiming to provide a reference for future development in this field. Methods Relevant Chinese and English literature were retrieved from the China national knowledge infrastructure (CNKI) and Web of Science (WOS) databases, respectively. Bibliometric tools including CiteSpace and VOSviewer were employed to analyze publication output, journals, authors/institutions, citation frequency, and keywords of both Chinese and English literature, respectively. Results Over the past two decades, a total of 227 literature that met the requirements were retrieved. The annual publication output showed a fluctuating upward trend, with Chinese literature (199 papers, 87.7%) dominating, underscoring the distinct characteristics and advantage of China in this field. The most prolific institutions were Shihezi University (40 articles) for Chinese publications and Shanghai Jiao Tong University (six articles) for English publications. Gu Xinli ranked first among Chinese authors with 40 publications, while Li Xiaobo and Peng Ying tied for the lead among English authors, each with six publications. Keyword analysis revealed that research evolution within the past 20 years can be divided into three main phases: establishment of basic technologies, expansion of activity studies, and the intervention of microecological functions. Hot topics included optimization of extraction processes, characterization of basic properties, investigations into classic formulas such as Sijunzi Tang and Yupingfeng San, along with studies on immunomodulation, antioxidant and antitumor effects, and gut microbiota-mediated mechanisms. Conclusion Research on polysaccharides in TCM formulas constitutes a relatively niche yet steadily evolving area, marked by distinctive Chinese characteristics and regional advantages. However, limitations remain, including insufficient research coverage, limited research depth, and inadequate exploration of synergistic mechanisms. Future breakthroughs will require interdisciplinary integration to tackle these core challenges, thereby potentially accelerating the modernization and internationalization of TCM formulas.

Objective High-throughput sequencing of Rehmannia chrysantha L.was performed to elucidate the structure of its chloroplast genome and its phylogenetic relationship with related species. Methods Codon preference analysis of chloroplasts of R. chrysantha was performed using CodonW v1.3 software. The codon usage bias was analyzed by using CodonW v1.3 software. Then, the obtained data were imported into IRscope and mVISTA to clarify sequence variations between R. chrysantha and related species. This study constructed a phylogenetic tree of the genus Rehmannia and its related genera on MEGA11 software. Results The chloroplast genome of R. chrysantha was a typical circular double-stranded molecule with a quadripartite structure and a total length of 153 789 bp. It has 28 high-frequency codons. Neutral plot analysis, ENC-plot analysis, and PR2-plot analysis all indicated that natural selection was the main cause of codon usage bias in R. chrysantha. It was also found that there were varying degrees of variation in the inverted repeat (IR) boundaries of the Rehmannia genus, and these changes were mainly concentrated in JLA and JLB. The genes rpl2, psbI, petN, psbZ, ycf1, and ndhB were identified as potential DNA barcodes for distinguishing closely related genus Rehmannia. In addition, phylogenetic analysis revealed a close evolutionary relationship between R. chrysantha and R. glutinosa L. Conclusion Protein-coding genes in the chloroplast genomes of genus Rehmannia exhibited a preferential use of codons ending in A/U, with natural selection being the primary factor influencing this codon usage bias. These findings will provide a foundation for future studies on molecular evolution, phylogeny, and chloroplast genetic engineering within this genus.

Objective To identify members of the R2R3-MYB gene family (named CiaMYB) in Chrysanthemum indicum var. aromaticum, a folk medicinal plant from Shennongjia, analyze their genetic evolutionary characteristics and expression patterns in response to UV-B stress, and explore the molecular mechanisms by which they regulate flavonoid synthesis. Methods Based on the whole-genome sequence of C. indicum var. aromaticum, bioinformatics analyses were performed to comprehensively identify CiaMYB members, and systematic analyses were conducted on their protein physicochemical properties, chromosomal localization, gene structure, collinearity relationships, and composition of cis-acting elements. Combined with transcriptome sequencing and correlation analysis, CiaMYB genes involved in flavonoid biosynthesis were screened. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to verify the expression patterns of key genes, and the functional localization of the proteins was analyzed. Results A total of 140 CiaMYB genes were identified, encoding hydrophilic proteins consisting of 206—868 amino acids, which are unevenly distributed on nine chromosomes. The promoter regions are enriched with cis-acting elements related to light response, abscisic acid (ABA) and methyl jasmonate (MeJA) signaling pathways. UV-B irradiation significantly induces differential expression of CiaMYB genes and accumulation of total flavonoids in leaves. Among them, CiaMYB040 and CiaMYB066 are highly homologous to the S7 subfamily, which is critical for flavonol synthesis in Arabidopsis thaliana, and their expression levels show a strong positive correlation with total flavonoid content, suggesting that they play a key transcriptional regulatory role in the flavonoid synthesis pathway. Subcellular localization confirmed that CiaMYB040 and CiaMYB066 proteins are localized in the nucleus. Conclusion This study is the first to systematically reveal the evolutionary characteristics of R2R3-MYB family members in C. indicum var. aromaticum, and preliminarily screen CiaMYB genes that respond to UV-B stress and are involved in regulating flavonoid synthesis. It provides important candidate gene resources for elucidating the molecular mechanism of plant high-altitude stress adaptation, as well as for flavonoid metabolism regulation and stress-resistant genetic improvement in Asteraceae plants.