This study aimed to elucidate the role of the G protein-coupled receptor 35 (GPR35)/AMP-activated protein kinase (AMPK)/mechanistic target of rapamycin (mTOR) signaling axis in mediating mitochondrial energy metabolism in radiation-induced myocardial fibrosis (RIMF) and to evaluate the interventional effect of astragaloside Ⅳ (AS-Ⅳ). Forty Wistar rats were randomly divided into four groups (n=10 per group). Except for the blank group, all rats were anesthetized and subjected to a single 40-Gy local thoracic X-ray irradiation to establish the RIMF model. After irradiation, AS-Ⅳ was administered intragastrically once daily at doses of 80 and 160 mg·kg^-1 for 30 consecutive days. After 30 days of continuous administration, the serum levels of N-terminal pro-B-type natriuretic peptide (NT-proBNP), cardiac troponinI (cTnI), and creatine kinase-MB (CK-MB) were quantified by ELISA. Hematoxylin-eosin (HE) and Masson staining were used to observe myocardial histopathological changes, and TUNEL staining was performed to detect cardiomyocyte apoptosis. The levels of adenosine triphosphate (ATP), adenosine diphosphate (ADP), and adenosine monophosphate (AMP), as well as the activities of mitochondrial respiratory chain complexes Ⅰ-Ⅳ, were measured colorimetrically. The fluorescence intensity of Ca²⁺ in myocardial tissue was detected using a fluorescent probe, and ultrastructural changes were observed by transmission electron microscopy. The fluorescence intensity and protein expression levels of GPR35, phosphorylated AMPK (p-AMPK), phosphorylated mTOR (p-mTOR), Caspase-3, Caspase-9, B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax), type Ⅰ collagen (COL-Ⅰ), and α-smooth muscle actin (α-SMA) in myocardial tissue were analyzed by immunofluorescence and Western blot. Results showed that compared with the blank group, serum NT-proBNP, cTnI, and CK-MB levels in the model group were significantly increased. Myocardial tissue exhibited focal myocyte necrosis and dissolution, connective tissue hyperplasia, and an increased area of collagen fiber positivity. The cardiomyocyte apoptosis rate was elevated. Myocardial ATP levels were significantly decreased, while ADP and AMP levels were significantly increased. The activities of mitochondrial respiratory chain complexes Ⅰ-Ⅳ were markedly reduced. The Ca²⁺ fluorescence intensity in myocardial cells was significantly enhanced. Severe mitochondrial damage was observed, showing flocculent changes, increased numbers of damaged mitochondria, swelling, rupture, disordered arrangement, disrupted cristae, and partial vacuolation. The fluorescence intensity and protein expression levels of GPR35, p-AMPK, Bax, Caspase-3, Caspase-9, COL-Ⅰ, and α-SMA in myocardial tissue were significantly increased, whereas those of p-mTOR and Bcl-2 were decreased. Compared with the model group, AS-Ⅳ significantly reduced serum NT-proBNP, cTnI, and CK-MB levels, markedly decreased the cardiomyocyte apoptosis rate, significantly increased ATP levels while reducing ADP and AMP content, enhanced the activities of mitochondrial respiratory chain complexes Ⅰ-Ⅳ, diminished Ca²⁺ fluorescence intensity, and alleviated mitochondrial damage, with mitochondria arranged more regularly. The fluorescence intensity and protein expression levels of GPR35, p-AMPK, Bax, Caspase-3, Caspase-9, COL-Ⅰ, and α-SMA were significantly decreased, while those of p-mTOR and Bcl-2 were increased. In conclusion, AS-Ⅳ exerts an inhibitory effect on RIMF, and its mechanism may be associated with the regulation of the GPR35/AMPK/mTOR signaling axis to improve mitochondrial energy metabolism.

This study aimed to elucidate the mechanisms of diosgenin, a natural compound derived from traditional Chinese medicine, on the triple-negative breast cancer cell line MDA-MB-231, with a focus on its induction of cell cycle arrest and inhibition of cell migration. Multiple experimental approaches were employed, including RNA sequencing analysis, flow cytometry for cell cycle detection, wound healing assays to evaluate migration capacity, F-actin staining to observe cytoskeletal changes, transmission electron microscopy (TEM) to examine cellular morphology and filopodia alterations, as well as Western blot and immunofluorescence staining to detect the expression of related proteins. RNA sequencing revealed significant gene expression differences between the diosgenin-treated group and the control group, with KEGG and GO analyses showing enrichment of differentially expressed genes mainly in pathways related to cell cycle and migration. Flow cytometry demonstrated that diosgenin induced G₀/G₁ phase arrest in MDA-MB-231 cells in a dose-dependent manner. Further Western blot and immunofluorescence assays indicated that diosgenin regulated the cell cycle via the CDC25A-CDK2/4/6 axis. Wound healing assays showed that diosgenin significantly inhibited cell migration. F-actin staining indicated that diosgenin suppressed F-actin formation and disrupted its structure. TEM observations revealed a reduction in filopodia formation on the cell surface after diosgenin treatment. Moreover, Western blot and immunofluorescence results demonstrated that diosgenin inhibited migration by suppressing epithelial-mesenchymal transition (EMT) and the RhoA-ROCK1 signaling pathway. In conclusion, diosgenin induces G₀/G₁ phase arrest in MDA-MB-231 cells by regulating cell cycle-related proteins and significantly inhibits cell migration by suppressing F-actin formation, reducing filopodia on the cell surface, and inhibiting EMT and the RhoA-ROCK1 pathway.

Based on the holistic regulation and multi-target characteristics of TCM, this study investigated the mechanism of Panlongqi Tablets (PLQT) in ameliorating rheumatoid arthritis (RA) by regulating key proteins, including glycogen synthase kinase-3β (GSK-3β), phosphatidylinositol 3-kinase catalytic subunit alpha (PIK3CA), and protein kinase B3 (AKT3), within the PI3K-AKT signaling pathway. An adjuvant-induced arthritis (AIA) rat model was established by intradermal injection of type Ⅱ collagen-Freund's complete adjuvant into the right hind paw, and rats received different doses of PLQT for intervention. Pharmacodynamic effects were evaluated by monitoring changes in body weight, measuring paw swelling, observing ankle synovial tissue pathology via hematoxylin-eosin (HE) staining, and detecting serum levels of inflammatory cytokines, including tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β), using ELISA. Targets identified through prior network pharmacology screening were integrated with therapy-related biomarkers from untargeted metabolomics for key pathway enrichment and identification of critical targets. Western blot and RT-qPCR were subsequently used to validate the expression of these key targets. Animal experimental results showed that, compared with the model group, all PLQT dose groups significantly improved weight loss, reduced paw swelling, inhibited synovial tissue fibrosis hyperplasia and inflammatory cell infiltration, and decreased serum levels of pro-inflammatory cytokines such as TNF-α, IL-6, and IL-1β in RA rats, exhibiting a partial dose-dependent manner. Metabolomics analysis revealed that PLQT exerted regulatory effects on 179 of 276 RA-related biomarkers. Enrichment analysis revealed that GSK-3β, PIK3CA, and AKT3 proteins in the PI3K-AKT signaling pathway were the key targets for PLQT intervention in RA. Validation by Western blot and RT-qPCR demonstrated that PLQT significantly modulated the expression of these three proteins. In conclusion, the therapeutic effects of PLQT on RA are likely associated with regulation of GSK-3β, PIK3CA, and AKT3 within the PI3K-AKT signaling pathway.

Non-alcoholic fatty liver disease (NAFLD) is the most prevalent chronic liver disease worldwide, characterized primarily by hepatic inflammation and dysregulated lipid metabolism. As a classic representative formula of the "bitter-purging method", Modified Wushi Xiexin Decoction has demonstrated definite clinical efficacy; however, its anti-inflammatory effects and underlying mechanisms in the treatment of NAFLD remain unclear. This study combines network pharmacology with animal experimental validation to evaluate the therapeutic effect of Modified Wushi Xiexin Decoction on NAFLD and reveal its potential mechanisms of action. Drug targets were collected from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP). NAFLD-related disease targets were retrieved from the Therapeutic Target Database (TTD), GeneCards, Online Mendelian Inheritance in Man (OMIM), Pharmacogenomics Knowledge Base (PharmGKB), and Comparative Toxicogenomics Database (CTD). R software was used to screen the intersection targets of the two sets. Core target analysis and network visualization were performed by using the Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) database and Cytoscape 3.8.2 software. R software was applied for Gene Ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Core targets were analyzed based on the Gene Expression Omnibus (GEO) database, and receiver operating characteristic (ROC) curves were plotted to evaluate the diagnostic efficacy of the core targets. For animal experimental validation, a NAFLD mouse model was established by feeding a 60% high-fat diet combined with intraperitoneal injection of streptozotocin. Pathological improvements were assessed by hematoxylin-eosin (HE) staining and oil red O staining. Enzyme-linked immunosorbent assay (ELISA) was used to detect the level of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β) in liver tissue. Immunohistochemistry (IHC) and Western blot (WB) were performed to measure the expression of Toll-like receptor 4 (TLR4), nuclear factor-κB (NF-κB), and NOD-like receptor protein 3 (NLRP3) proteins in mouse liver tissue. Network pharmacology identified 97 active components, 108 intersection targets, and 10 core targets of Modified Wushi Xiexin Decoction. GO enrichment analysis indicated that anti-NAFLD effects were exerted by regulating biological processes such as oxidative stress and nutritional levels. KEGG enrichment analysis mainly involved signaling pathways, including TLR, NF-κB, and TNF. The GEO database indicates that there is a significant difference in TLR4 expression between healthy individuals and NAFLD patients (P<0.05), with the area under the ROC curve (AUC) being greater than 0.810, suggesting a high diagnostic efficacy. Animal experiments showed that Modified Wushi Xiexin Decoction could alleviate pathological damage in the liver tissue of NAFLD mice, reduce blood lipids, improve liver function, and downregulate the expression of inflammatory factors in liver tissue. IHC and WB confirmed that Modified Wushi Xiexin Decoction could downregulate the TLR4 expression, inhibit NF-κB activity, and suppress NLRP3 inflammasome activation. In conclusion, Modified Wushi Xiexin Decoction may alleviate inflammatory responses and improve liver function and blood lipids in NAFLD by regulating the TLR4/NF-κB signaling pathway and inhibiting NLRP3 inflammasome activation, thereby exerting therapeutic effects on NAFLD.

This study is aimed to investigate the therapeutic effects of Junren Bufei Yixin Granules (JRBF) on hypoxic pulmonary hypertension (HPH) and explore the therapeutic mechanism through the polypyrimidine tract-binding protein 1 (PTBP1)/pyruvate kinase isozyme type M2 (PKM2) signaling pathway. Sixty SPF-grade C57BL/6 male mice were randomized into normal, model, sildenafil, and low/medium/high-dose JRBF groups. HPH was induced by hypoxic exposure (10% O₂, 8 h/day) for 4 weeks. The right ventricular systolic pressure (RVSP) was measured by right cardiac catheterization. Echocardiography was employed record the pulmonary artery acceleration time (PAAT), pulmonary ejection time (PET), tricuspid annular plane systolic excursion (TAPSE), and right ventricular anterior wall thickness at diastole (RVAWd). The histopathological changes were observed by hematoxylin-eosin (HE) and Masson staining. Western blot was employed to measure the expression levels of PTBP1, PKM2, and PKM1 in the lung tissue. The immunofluorescence assay was employed to detect the co-localization of PTBP1 with the marker alpha-smooth muscle actin (α-SMA) of pulmonary arterial smooth muscle cells (PASMCs). The lactate assay kit was used to measure the lactate levels in the serum and lung tissue. In the cell experiments, the hypoxia model was established by exposing human pulmonary artery smooth muscle cells (hPASMCs) to 1% O₂ for 48 h. The optimal (10%) JRBF concentration for intervention was determined by the cell-counting kit-8 (CCK-8). The cell proliferation (EdU), migration (Transwell), PTBP1/PKM2/PKM1, and lactate production were analyzed. The results showed that JRBF significantly reduced the RVSP, improved the PAAT/PET ratio and TAPSE, and attenuated pulmonary arteriolar remodeling and myocardial fibrosis. Furthermore, JRBF suppressed PTBP1 and PKM2 expression and reduced the lactate accumulation in the lung tissue and PASMCs. The cell experiment results showed that the 10% JRBF-containing serum inhibited the proliferation and migration of hPASMCs, while decreasing the PTBP1/PKM2 expression and lactate production. In conclusion, JRBF mitigates HPH progression by suppressing the PTBP1/PKM2 axis and restoring the glycolytic flux.

This study aims to investigate the effects of Naotaifang (NTF) on neuronal ferroptosis and pyroptosis in cerebral ischemia-reperfusion injury (CIRI), with a focus on nuclear factor erythroid 2-related factor 2 (Nrf2). A rat model of CIRI was established by middle cerebral artery occlusion/reperfusion (MCAO/R). Sprague-Dawley rats were randomly assigned to a sham group, a model group, low/medium/high-dose (4.5, 9, 18 g·kg^-1, respectively) NTF groups, and a butylphthalide (60 mg·kg^-1) group (n=15 per group). After modeling, the treatment groups received corresponding agents by gavage once daily for 7 days, and the sham and model groups received an equal volume of normal saline. After treatment, the neurological function was assessed based on the Zea-Longa score, the infarct volume by TTC staining, and cellular morphology by HE staining. The levels of Fe²⁺, interleukin-1β (IL-1β), and interleukin-18 (IL-18) were measured by ELISA, and those of malondialdehyde (MDA) and glutathione (GSH) by biochemical assays. Western blot was employed to examine the expression levels of Nrf2, glutathione peroxidase 4 (GPX4), NOD-like receptor family pyrin domain-containing 3 (NLRP3), and the N-terminal fragment of gasdermin D (GSDMD-N). In the cell experiment, oxygen-glucose deprivation/reperfusion (OGD/R) was used to model CIRI in SH-SY5Y cells. siRNA-mediated Nrf2 knockdown (si-Nrf2) was employed to probe into the role of Nrf2 in OGD/R-induced ferroptosis and pyroptosis and the intervention effect of NTF. Membrane damage was evaluated by Hoechst/propidium iodide staining and LDH release, and GPX4 and GSDMD immunofluorescence was employed to indicate ferroptosis and pyroptosis, respectively. Fe²⁺, MDA, and GSH were quantified biochemically, and the levels of IL-1β and IL-18 by ELISA. The expression of Nrf2, GPX4, NLRP3, and GSDMD-N was quantified by Western blot. The cellular ultrastructure was observed by transmission electron microscopy. The results of the animal experiment showed that NTF promoted the expression of Nrf2 to alleviate neuronal ferroptosis and pyroptosis, thus producing significant neuroprotection against CIRI in a dose-dependent manner. The results of the cell experiment showed that the neuroprotective effects of NTF after CIRI were mediated by Nrf2-dependent suppression of neuronal ferroptosis and pyroptosis. Taken together, NTF attenuates CIRI by suppressing neuronal ferroptosis and pyroptosis through Nrf2-mediated regulation.

Gardeniae Fructus (GF) is one of the most commonly used medicinal and edible resources in China, renowned for its anti-inflammatory and antipyretic effects. However, studies of the chemical characteristics, metabolic transformation and pharmacokinetic behavior of GF in vivo remain incomplete. This study utilized mass defect filtering (MDF) combined with UHPLC-ESI-Q-TOF-MS/MS to analyze the in vitro chemical constituents of GF aqueous extracts and its metabolites in vivo in rats, comprehensively characterizing 114 chemical constituents and 181 endogenous substances, including 18 prototype constituents and 163 metabolites (21 phase Ⅰ and 142 phase Ⅱ metabolites). Additionally, a quantitative method using UHPLC-QQQ-MS/MS was developed to analyze the in vivo dynamics of seven bioactive compounds, and the pharmacokinetic parameters of the main active components in the GF aqueous extracts, namely geniposidic acid, deacetyl asperulosidic acid methyl ester, gardenoside, genipin, genipin 1-β-D-gentiobioside, geniposide, and crocetin, were obtained. This study conducted a comprehensive and systematic investigation into the potential material basis and the in vivo biotransformation profile of GF aqueous extracts based on UHPLC-ESI-Q-TOF-MS/MS coupled with MDF technology and UHPLC-QQQ-MS/MS, providing crucial evidence for further elucidating its pharmacological mechanisms.

This study aims to systematically retrieve and sort out the clinical research evidence of oral Chinese patent medicines in the treatment of vascular dementia (VaD) and understand the distribution and quality of the existing evidence in this field. PubMed, Web of Science, Cochrane Library, EMbase, CNKI, VIP, Wanfang, and SinoMed databases were retrieved from their establishment to November 5, 2024. Evidence diagrams were used to comprehensively present the data distribution characteristics. A total of 211 pieces of literature were included, among which 176 were RCTs, 11 were non-RCTs, 3 were retrospective studies, 1 was cohort study, 17 were systematic reviews/Meta-analyses, and 3 were expert recommendations or consensuses. It involves 45 kinds of Chinese patent medicines. Among them, Ginkgo Leave Tablets/Capsules are the most widely used. Most of the research periods are 8-12 weeks, and the sample size is 51-100 cases. The main outcome indicators include clinical efficacy, cognitive function evaluation, quality of life evaluation, safety indicators, neurological function indicators, hemorheological indicators, etc. The literature quality of RCTs, non-RCTs, and systematic reviews/Meta-analyses are generally not high, and the quality level of evidence is relatively low. Most studies have shown that oral Chinese patent medicines have potential efficacy in the treatment of VaD, but there is a lack of high-quality clinical evidence. Chinese patent medicines have advantages in the treatment of VaD, but there is still a lack of high-quality studies. This suggests that in future clinical research, large-sample and multi-center RCTs need to be carried out. Meanwhile, clinical research design and reporting should be standardized in accordance with relevant guidelines or tools to explore the characteristic advantages of oral Chinese patent medicines in the treatment of VaD, select reasonable outcome indicators, and strive to improve the quality of clinical research in TCM. This will help standardize the research plan, produce advanced evidence-based medical evidence with rigor and scientificity, and provide strong support for the prevention and treatment of VaD with TCM.

Published in 1856, Catalogus Medicamentorum Sinensium stands as the earliest known modern pharmacological work in existence. Compiled by the Russian scholar Alexander Tatarinov during the 19th century, this groundbreaking study systematically documented and identified the botanical origins of 495 Chinese medicinal materials procured from Beijing, supplemented with plant specimens collected in China. It represents the first application of modern scientific nomenclature, combining Latin binomial names with specified medicinal parts, for the systematic classification of Chinese medicinal materials. This research conducted a comprehensive analysis of the cataloged substances through meticulous examination of relevant literature, archival records, and reference materials. By using methods such as materia medica verification, data analysis, etc., the information of the listed drugs was verified, classified, organized, and examined, and compared with the origins recorded in modern standards or specialized works. The compendium documented 495 medicinal entries (including 33 duplicate medicinal materials), comprising 396 botanical, 35 mineral, 40 zoological, and 24 other substances. This book identified a total of 190 Chinese medicinal materials whose origin was consistent with that recorded in the 2020 edition of the Pharmacopoeia of the People's Republic of China or Chinese Materia Medica. There were 76 cases with completely different origins. Another 189 cases had their origins not fully identified, but only partial information was marked. Lastly, there were 40 cases that were either untested or could not be matched with current Chinese medicinal materials. The research results indicate that this book is the first in modern times to use Linnaean binomial nomenclature to name the origin of Chinese medicinal materials. It laid the foundation for subsequent research on the origin of Chinese medicinal materials and established the methods and annotation formats for such research. It has been subsequently adopted and referenced by subsequent TCM researchers both in China and abroad. The publication of this book can be regarded as a historical turning point in the evolution of the definition of the origin of Chinese medicinal materials from traditional to modern times. It has advanced the description of the name and reality of Chinese medicinal materials in the form of text and pictures to the definition method using modern natural classification systems and labeled with biological Latin names, which is a pioneering work. This research can provide a reference for sorting out the development process of modern research on the origin of Chinese medicinal materials.

Based on extensive research into the current status of understory medicinal herb cultivation and an in-depth analysis of existing problems, this study proposes policy and technical recommendations to promote sustainable and high-quality development. The research team conducted field investigations in advantageous cultivation regions across seven provinces (autonomous regions), including Guangxi, Yunnan, Fujian, and Jilin, supplemented by policy reviews and literature analysis. The findings reveal that although understory medicinal herb cultivation benefits from ecological advantages and policy support, it still faces a series of problems, such as an underdeveloped service system, insufficient regulatory capacity, low market alignment, and immature technical systems. It should be particularly noted that understory cultivation commonly encounters specific challenges, including the economic dilemma of high input and low output, mismatches between forest canopy density and the light requirements of medicinal plants, ambiguous boundaries between ecological protection and utilization in policy implementation, and weak research support, leading to high trial-and-error costs in technology adoption. The following development recommendations are proposed: ① establish a coordination mechanism between forestry and traditional Chinese medicine authorities to integrate planning and policy synergy; ② enhance the alignment of understory medicinal herb cultivation with market demand; ③ develop an independent technical system and standards for understory medicinal herb cultivation, such as establishing a variety suitability framework based on canopy density classification and ecological cultivation protocols; ④ strengthen research focused on industrial development, with priority given to large-scale plantations (e. g., pine and fir forests), dual-purpose food-medicine species (Polygonatum, Gastrodia elata), vine species, and intercropping effects; ⑤ establish a quality certification system and corresponding regulatory framework for understory-cultivated medicinal materials to promote premium pricing; ⑥ improve land-use policies and supporting infrastructure, clarifying utilization rights for different forest types and simplifying approval procedures for temporary facilities.