Objective To explore the potential active components, targets and molecular mechanism of Tianma Gouteng Yin (天麻钩藤饮) in improving vascular endothelial function in hypertension based on network pharmacology, molecular docking and experimental verification, and to clarify whether it exerts effects via phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/endothelial nitric oxide synthase (eNOS) signaling pathway. Methods Active components and targets of Tianma Gouteng Yin were screened through HERB and SwissTargetPrediction databases. Hypertension-related targets were obtained from OMIM and GeneCards databases. A protein-protein interaction (PPI) network was constructed, and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis was performed. Molecular docking was used to verify the binding affinity between core components and key targets in PI3K/Akt/eNOS pathway. Human umbilical vein endothelial cells (HUVECs) injured by angiotensin Ⅱ (Ang-Ⅱ) were used as the cell model. CCK-8, qRT-PCR, Western blotting, reactive oxygen species (ROS) and nitric oxide (NO) assays were applied to evaluate the effects of Tianma Gouteng Yin on cell viability, oxidative stress, inflammatory response and PI3K/Akt/eNOS pathway. Results Network pharmacology identified 228 active components and 969 overlapping hypertension-related targets of Tianma Gouteng Yin. Core targets included serine/threonine kinase 1 (AKT1) and PI3K family members [phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA), phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit beta (PIK3CB), etc.]. KEGG enrichment indicated that PI3K/Akt pathway was the key pathway. Molecular docking confirmed that core components such as kaempferol and apigenin showed strong binding affinity to PI3K/Akt pathway targets. The results of cell experiment showed that 25-100 mg/mL Tianma Gouteng Yin had no cytotoxicity in normal HUVECs, and dose-dependently attenuated Ang-Ⅱ-induced reduction in cell viability (P < 0.001), significantly inhibited the expressions of angiotensin-Ⅱ type 1 receptor (AT1R) and NADPH oxidase 4 (NOX4), as well as ROS production (P < 0.001), upregulated the levels of antioxidant enzymes including superoxide dismutase 1 (SOD1), catalase (CAT) and glutathione peroxidase 1 (GPX1) (P < 0.05, 0.01, 0.001), reduced the expressions of inflammatory factors including interleukin-6 (IL-6), IL-1β, tumor necrosis factor-α (TNF-α), intercellular adhesion molecule 1 (ICAM1) and vascular cell adhesion molecule 1 (VCAM1) (P < 0.01, 0.001), increased NO production and suppressed endothelin-1 (ET-1) expression (P < 0.01, 0.001), upregulated the protein expressions of p-PI3K, p-Akt and p-eNOS (P < 0.001). After administering the PI3K specific inhibitor Wortmannin, the activation effect of Tianma Gouteng Yin on PI3K/Akt pathway was significantly weakened, indicating that its endothelial protective effect depends on PI3K/Akt/eNOS signaling pathway. Conclusion Tianma Gouteng Yin may activate PI3K/Akt pathway through multi-target synergistic effects of core components such as kaempferol and apigenin, improve oxidative stress and inflammatory response, regulate the balance of NO and ET-1, and protect endothelial function in hypertension.

Objective To explore the differences in the in vivo drug metabolism of raw and processed Dahuang (Rhei Radix et Rhizoma) in a rat model of middle cerebral artery occlusion and reperfusion (MCAO/R). Methods The MCAO/R rat model was established by the suture method. The successfully modeled rats were randomly divided into model group, raw Rhei Radix et Rhizoma (2.50 g/kg) group and processed Rhei Radix et Rhizoma group, with six rats in each group. Rats were administered continuously for 7 d. 12 h before the last administration, the rats were fasted, and 1 h after administration, blood and brain tissues were collected. Ultra-high performance liquid chromatography-quadrupole electrostatic field orbitrap linear ion trap mass spectrometry (UPLC-Orbitrap Fusion Lumos Tribrid-MS) was used to characterize the prototypes and metabolites of raw and processed Rhei Radix et Rhizoma in plasma and brain tissues of MCAO/R rats. Multivariate statistical analysis was used to screen the differential components, and desorption electrospray ionization mass spectrometry imaging (DESI-MSI) technique was combined to visualize the in situ spatial distribution of the differential components of raw and processed Rhei Radix et Rhizoma in brain tissue. Results A total of 109 prototype components and metabolites were identified in MCAO/R rats. According to variable important in projection (VIP) > 1 and P < 0.05, 23 and eight differential components of raw and processed Rhei Radix et Rhizoma were screened from the plasma and brain tissues, respectively. In the plasma of processed Rhei Radix et Rhizoma group, anthraquinone glycosides were decreased, while in the brain tissue, aglycones and metabolites (such as gallic acid, M31) were increased, and were mainly distributed in hippocampus, midbrain and cortex of brain tissue. The common differential components gallic acid and metabolite M31 were significantly increased in the processed Rhei Radix et Rhizoma group and were specifically enriched in the striatum. Conclusion This study characterized and spatially located the differential components of raw and processed Rhei Radix et Rhizoma in MCAO/R rats and found that processed Rhei Radix et Rhizoma increased the exposure levels of active aglycones and metabolites in the brain, enhancing its protective effect on brain tissue, providing a scientific basis for the “different treatment for raw and processed” processing theory.

Objective This study aims to systematically analyze the intellectual property landscape of traditional Chinese medicine (TCM) globally, with a focus on comparing the patent layout characteristics and competitive dynamics in five key countries: China, the United States, Japan, South Korea, and Germany. The goal is to provide strategic references for promoting the high-quality development and internationalization of China's TCM industry. Methods Leveraging the Patyee database, this research retrieved and obtained TCM-related patent data globally and from the aforementioned five countries. Employing a combination of quantitative and qualitative analysis methods, an in-depth examination was conducted from multiple dimensions, including patent application trends, geographical distribution, applicant types, core technological topics, and international layout strategies. Results The research findings indicate that despite a vast number of domestic TCM patent applications (approximately 360 000), the proportion of international applications filed via the Patent Cooperation Treaty (PCT) is relatively low (only 1.37%), revealing a significant deficiency in overseas layout. Regarding the distribution of technological topics, patents related to TCM quality control and standardization technologies are relatively scarce globally (including in China), leaving a gap in meeting the high standards of quality controllability required for modern industrialization and internationalization. In terms of applicant types, 44% of TCM patents in China are held by individuals, while enterprises account for only 34%, reflecting a pattern of individual-led inheritance with enterprise-driven R&D yet to be enhanced. In contrast, TCM-related patent activities in the United States, Japan, and Germany are predominantly driven by multinational pharmaceutical companies or consumer goods giants with complete industrial chains. Conclusion To achieve high-quality internationalization of TCM intellectual property, China urgently needs to establish a systematic overseas patent layout strategy, forming a patent protection system covering the entire industrial chain of R&D, quality control, production, and marketing. The core task is to transform domestic technological advantages into internationally recognized technical standards and to promote the transformation of TCM from a traditional empirical model to a modern, evidence-based innovation paradigm by strengthening collaborative innovation among industry, academia, research, and users.

Objective To investigate the targets and mechanisms of Huachansu in alleviating oxaliplatin-induced peripheral neurotoxicity using bioinformatics and network pharmacology. Methods The targets of Huachansu were predicted via SwissTarget Prediction, PharmMapper, and SuperPred databases. Targets associated with oxaliplatin-induced peripheral neurotoxicity were retrieved from GeneCards, OMIM, and Harmonizome 3.0 databases. Combined with differential genes after oxaliplatin treatment from the GEO database, common targets were obtained through intersection analysis. The DAVID database was used for gene ontology (GO) function and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment. Core targets were screened via the PPI network and machine learning. Single-cell analysis was performed to explore the specific expression landscape of the core targets. AutoDock and Gromacs were employed for molecular docking and molecular dynamics simulation, with visualization assisted by PyMOL, QtGrace 2.6 and Python 3.7. Results A total of 37 active components of Huachansu were identified (e.g., telocinobufagin, hellebrigenol). Forty-five common targets were obtained. CCND1, PDGFRB, and other core targets were jointly determined by PPI network and machine learning. Molecular docking showed that telocinobufagin and other components exhibited binding energies of < -5 kcal/mol with Cyclin D1 (CCND1) and platelet-derived growth factor receptor beta (PDGFRB). Molecular dynamics simulation confirmed the stable binding between PDGFRB and telocinobufagin. Single-cell analysis suggested that Huachansu targets PDGFRB in stromal cells to reshape the neuro-supportive microenvironment. Conclusion Huachansu exerts neuroprotective effects at the acute OIPN stage by targeting PDGFRB⁺ neural stromal cells and activating the phosphatidylinositol-3-hydroxykinase (PI3K)-protein kinase B (Akt) signaling pathway, providing a theoretical basis for the early prevention of chemotherapy-induced neurotoxicity.

Objective A comprehensive analysis of the research hotspots and prospective trends of Croton tiglium from the perspectives of knowledge graphs and global patents is conducted to provide theoretical basis and reference direction for clinical practice and subsequent in-depth exploration of C. tiglium. Methods C. tiglium was used as the keyword to search four databases: CNKI, Wanfang, VIP, and Web of Science (WOS), and the literature was imported into the NoteExpress literature management software for duplication checking and screening. Excel, CiteSpace, VOSviewer and other softwares were used to conduct a visual analysis of the current status and development trends of C. tiglium research at home and abroad from the dimensions of publication trends, publishing countries, institutions, authors, and keywords. The Incopat patent database was used to search and analyze C. tiglium-related global patent applications from aspects such as patent application trends, global geographical distribution, patent applicants and patent technology fields. Results A total of 1 320 Chinese literature and 1 528 English literature that met the criteria were retrieved. After a preliminary search, a total of 11 004 patents were obtained in the Incopat patent database. After the merger process, 5 196 patents were finally retained. Among them, there are 2 346 domestic patents, accounting for 45.15% of the total, occupying a leading position. In general, the research heat in the field of C. tiglium continues to rise, the number of Chinese and English publications is comparable, and the academic influence at home and abroad is balanced. Among them, the United States has the highest number of publication volume in the world, and China is in the forefront, which are important research force in this field. In terms of scientific research cooperation, Chinese medicine colleges and affiliated medical institutions are the main body at home, with a single research structure and loose institutional cooperation, and there are obvious research barriers. International research relies on transnational scientific research subjects, with close cooperation networks and mature collaboration systems. In terms of research content, the domestic focus is on the traditional pharmacology and clinical application of C. tiglium, with an emphasis on the inheritance of traditional medicinal properties. The international focus is on fundamental and cutting-edge research such as active ingredients and mechanisms of action On the whole, C. tiglium research continues to break through the traditional category, relying on multidisciplinary cross to achieve innovative development, forming a development trend of inheritance and innovation. Conclusion Research in the field of C. tiglium is constantly expanding and deepening. Its research paradigm is undergoing a significant transformation, gradually shifting from the application of traditional Chinese medicine to multi-disciplinary interdisciplinary research and in-depth analysis of molecular mechanisms. China dominates the patent layout among various countries in the C. tiglium field. However, its patent system has significant weaknesses, mainly manifested in the premature expiration of a large number of patents, low technology industrialization, and insufficient commercial expansion capabilities. The research hotspots in Chinese literature focus on the anti-inflammatory pharmacological effects and processing techniques for reducing toxicity of C. tiglium, gradually extending to agricultural applications. Meanwhile, English research hotspots are concentrated on antioxidant activity, chemical taxonomy, and natural product chemistry, with antioxidant research showing significant growth in recent years.

Objective To clarify the comprehensive clinical value, advantages, characteristics and therapeutic positioning of Ningmitai Capsules (宁泌泰胶囊) in the treatment of chronic prostatitis (damp-heat accumulation syndrome), and to provide references for decision-making of national medical and health administrative departments and rational clinical drug use. Methods An evaluation system was constructed based on the “6 + 1” dimensions of safety, effectiveness, economy, innovation, suitability, accessibility, and traditional Chinese medicine (TCM) characteristics. Multi-source evidence including questionnaire surveys, official website data, secondary evaluation of literature, and real-world data was integrated. The multi-criteria decision analysis (MCDA) model was used to measure each dimension, and the total value score was calculated based on TCM clinical evidence using software CSCv2.0. Results Ningmitai Capsules were rated Grade A for both safety and effectiveness (no serious adverse reactions, significant effects in anti-infection, relieving pain and alleviating urinary abnormalities); Grade B for economy, innovation, suitability, accessibility, and TCM characteristics (supported by economic evidence, obtained 1 invention patent, high acceptance among target populations, good resource accessibility, and a unique Miao medicine formula); the comprehensive evaluation of clinical value was Grade A. Conclusion Ningmitai Capsules have significant clinical value in the treatment of chronic prostatitis. It is recommended to further conduct pharmacological mechanism research and high-quality evidence-based research, optimize the production process, improve the drug insert, and give full play to its clinical advantages.

Objective This study investigated the regulatory mechanism by which methyl jasmonate (MeJA) modulates triterpenoid saponin biosynthesis in Jixuecao (Centella asiatica). Methods Plants were treated with 200 μmol/L MeJA, and temporal changes in leaf asiaticoside and madecassoside contents were quantified by high-performance liquid chromatography (HPLC). Transcriptome data were analyzed using weighted gene co-expression network analysis (WGCNA) to identify modules significantly correlated with asiaticoside accumulation, followed by Mfuzz clustering to refine gene subclusters. Expression patterns of candidate key genes were validated by quantitative real-time PCR (qRT-PCR). Results MeJA treatment markedly promoted asiaticoside and madecassoside accumulation, increasing their contents by 18.28% and 14.58%, respectively. WGCNA revealed 33 co-expression modules, with the darkred module displaying a highly significant positive correlation with asiaticoside content (P < 0.01). Mfuzz clustering subdivided the 286 genes in this module into four subclusters; Notably, 239 genes in Clusters 1-3 were significantly enriched in the terpenoid backbone biosynthesis, sesquiterpenoid biosynthesis, and triterpenoid biosynthesis pathways. Twenty-eight structural genes directly involved in asiaticoside synthesis were identified, exhibiting expression profiles closely aligned with saponin accumulation. qRT-PCR confirmed significant MeJA-induced upregulation of 11 key structural genes associated with triterpenoid backbone synthesis and modification, consistent with the transcriptome findings. Conclusion MeJA enhances triterpenoid saponin biosynthesis in C. asiatica through coordinated upregulation of these 11 key structural genes within the darkred module. By integrating multi-omics profiling with experimental validation, this work systematically elucidates the molecular mechanisms underlying MeJA-induced triterpenoid saponin accumulation, offering valuable candidate genes for future functional characterization and genetic enhancement of C. asiatica.

Objective To systematically identify the R2R3-MYB transcription factor family in Xuanmugua(Chaenomeles speciose) based on transcriptomic and metabolomic data, and to screen key candidate genes potentially involved in the regulation of flavonoid biosynthesis. Methods Transcriptome and metabolome databases were constructed using different tissues of C. speciosa. The R2R3-MYB transcription factor family was systematically identified based on full-length transcriptome data. Subsequently, the sequence characteristics, phylogenetic relationships, and tissue-specific expression patterns of these genes were analyzed. Results A total of 31 R2R3-type CsMYB genes were identified from the transcriptome data of C. speciosa. Based on phylogenetic analysis with Ninanjie(Arabidopsis thaliana) and Yueji (Rosa chinensis), we classified these genes into 11 subfamilies. Members within the same subfamily showed similar conserved motifs and gene structures. Flavonoids, including catechin and epicatechin, were significantly enriched in the pulp and peel. The 31 CsMYB genes exhibited differential expression patterns across various tissues of C. speciosa, among which 14 CsMYB genes, such as CsMYB1, CsMYB21, and CsMYB25, were highly expressed in the peel or pulp. Further correlation analysis revealed that six candidate genes, including CsMYB1, CsMYB2, and CsMYB14, were significantly and positively correlated with the accumulation of multiple flavonoids, such as catechin and epicatechin. These results were further validated by RT-qPCR. Conclusion This study provides the first systematic identification of the R2R3-MYB transcription factor family in C. speciosa and screens key CsMYB genes potentially involved in the regulation of flavonoid biosynthesis. These findings provide a reference for subsequent functional verification of candidate genes and lay a foundation for elucidating the molecular mechanisms underlying the quality formation of C. speciosa.

Objective This study aims to clone the LbbHLH94-like gene from Gouqi (Lycium barbarum), analyze its spatiotemporal expression, stress and hormone response patterns, and clarify its regulatory function in carotenoid biosynthesis, so as to provide a theoretical basis for revealing the molecular mechanism of goji berry fruit quality formation. Methods Using Lycium barbarum cv. ‘Ningqi 1’ as material, we employed techniques such as PCR and qRT-PCR to clone and analyze the expression patterns of LbbHLH94-like in different tissues, fruit developmental stages, abiotic stresses and exogenous hormone treatments. The overexpression vector was constructed and transformed into goji plants, and the expression of key genes, enzyme activities and total carotenoid content in carotenoid metabolism were determined. Results The LbbHLH94-like gene was successfully cloned, with an 888 bp open reading frame encoding 296 amino acids, and the sequence was identical to the genome reference sequence. This gene showed obvious spatiotemporal specificity, with the highest expression level at fruit ripening stage. It responded to high and low temperature, salt, drought stresses as well as 6-BA, IBA and GA₃ induction, showing a general pattern of promotion at low concentration and inhibition at high concentration. Overexpression of LbbHLH94-like significantly up-regulated the expression of PDS, enhanced the enzyme activities of PSY and PDS, inhibited the expression and enzyme activity of CCD4, and remarkably increased total carotenoid content in fruits. Conclusion LbbHLH94-like is a key positive regulator of carotenoid biosynthesis in goji berry, and also participates in abiotic stress response and hormone signaling. The results provide important gene resources and theoretical support for quality improvement and precise molecular breeding of Lycium barbarum.

Objective To establish an accurate and efficient molecular identification method for specific DNA Signature Sequence Tags (DSS) of Tianshanxuelian (Saussurea involucrate) and its adulterants Shuimuxuetuzi (S. medusa) and Miantouxuetuzi (S. laniceps). Methods The Chloroplast Genome Information Resource (CGIR) were used to obtain the complete chloroplast genome sequences of Saussurea DC, the IdenDSS software was used to screen for specific DNA signature sequence tags (DSS) of the S. involucrata. Five sets of DSS markers were selected, and the corresponding specific primer pairs were designed using IdenDSS. Through DNA extraction from the sample, PCR amplification, Sanger sequencing and sequence alignment verification, the presence of DSS or its reverse complementary sequence of the target species S. involucrata in the test sample was detected for original identification. Results By comparing the selected five pairs of DSS sequences with the authentic source and its adulterants sequences of S. involucrata, all five sets of DNA Signature Sequence Tags could be used for identification of S. involucrata were finally determined. The above sequences were uploaded to the standard database of the Traditional Chinese Medicine Molecular Identification Platform (www.herbsdna.com), and combined with the DSS identification function of the database, accurate and efficient identification of S. involucrata origin can be achieved. Conclusion The five selected DNA Signature Sequence Tags had strong specificity and high accuracy, which can effectively solve the problem of original identification of S. involucrata.