The quality and safety of Chinese medicinal materials are the lifeline for the sustainable development of the traditional Chinese medicine (TCM) industry. Establishing a traceability system for Chinese medicinal materials is a core means to solve the problem of whole-chain quality control and achieve quality supervision, which is of great significance for maintaining the brand image of authentic medicinal materials and regulating the order of the medicinal materials market. At present, China’s traceability system for Chinese medicinal materials has integrated key technologies such as the Internet of Things, blockchain, big data, and AI, and has built a full-chain information management system covering “cultivation-processing-circulation-use”. Pilot projects in multiple regions have verified its application value, and the industry has entered a stable development stage of multi-technology collaboration. However, existing research and practice still face key controversies and deficiencies: technical coordination problems such as large differences in information dimensions among research and development institutions and disconnected traceability information across multiple links, subject awareness problems such as weak self-supervision awareness among business operators, cost and data problems such as high information collection costs and high barriers to participation for small and medium-sized entities, and industry standardization problems such as the lack of unified national data standards and imperfect regulatory mechanisms. These issues lead to limited coverage of the traceability system, the continued existence of counterfeit and substandard medicinal materials, and an urgent need to enhance the ability to trace back to the source and recall problematic products. The purpose of this study is to systematically review the development history and policy evolution of the traceability system for Chinese medicinal materials, summarize its core functions, key technology integration application logic, and typical pilot achievements, and propose targeted optimization suggestions for current deficiencies. The research structure analyzes the problem from the “root-amplification-ultimate” logical chain of Chinese medicinal material quality safety, then elaborates on the development and construction status of the traceability system, subsequently analyzes the challenges faced and proposes feasible countermeasures, and finally looks forward to future development directions. The study finds that the construction of the traceability system for Chinese medicinal materials needs to focus on standardization, intelligence, inclusiveness, and globalization as core directions. By formulating unified metadata standards, deepening multi-technology integration, constructing a “government subsidies + insurance linkage + brand premium” cost model, and improving diversified co-governance regulatory mechanisms, industry pain points can be resolved. In the future, further focus should be placed on differentiated traceability schemes for different categories of Chinese medicinal materials, low-cost implementation paths for multi-technology integration, correlation analysis between traceability data and clinical efficacy, and other directions, to promote deep integration of academic achievements and industrial applications, making the traceability system for Chinese medicinal materials a solid support for ensuring the quality and safety of Chinese medicinal materials and promoting the inheritance, innovation, and development of TCM.

Objective To investigate the chemical constituents of the n-butanol fraction of Patrinia villosa. Methods The compounds were isolated and purified by various chromatographic techniques including silica gel, ODS, Sephadex LH-20 gel, and semi-preparative HPLC, and their structures were elucidated by analysis of spectroscopic data. The anti-inflammatory activities were evaluated by against NO production from lipopolysaccharide (LPS)-stimulated RAW264.7 cells. Results A total of 14 compounds were isolated from the n-butanol fraction of P. villosa ethanol extract and identified as (2E,5R,6R)-5,8-dihydroxy-2,6-dimethyl-2-octenoic acid (1), (2E,5R,6R)-8-acetoxy-5-hydroxy-2,6-dimethyl-2-octenoic acid (2), (2E,6R)-8-hydroxy-2,6-dimethyl-2-octenoic acid (3), (6R,7E,9R)-9-hydroxy-4,7-megastigmadien-3-one (4), ethyl chlorogenate (5), ethyl caffeate (6), dihydrosyringenin (7), 7S,8S-threo-4,7,9,9'-tetrahydroxy-3,3'-dimethoxy-8-O-4'-neolignan (8), (+)-isolariciresinol (9), (7S,8R)-dihydrodehydrodiconiferyl alcohol (10), ceplignan (11), (+)-syringaresinol (12), salicifoliol (13), and 5-hydroxy-4-(4-hydroxyphenyl)-2(5H)-furanone hydroxy-butenolide (14). Compounds 1 and 2 are new compounds, and their NO inhibition rates at a concentration of 50 μmol/L were (6.16 ±1.41)% and (11.11 ±2.10)%, respectively. Conclusion Compounds 1 and 2 are new acyclic monoterpenoids, and namely patrivic acid B and 8-acetylpatrivic acid B. Compounds 3, 4, 10, 11 and 14 were isolated from P. villosa for the first time. Compounds 1 and 2 showed no significant anti-inflammatory activity.

Objective To investigate the chemical constituents from the inflorescences of Inula japonica and their in vitro anti-hepatocellular carcinoma activity. Methods The compounds were isolated and purified by multiple chromatographic techniques.. Their structures were identified by HRESIMS, NMR, IR, UV spectroscopic methods, and the absolute configurations of new compounds were determined by calculated NMR and ECD. The inhibitory effects of the compounds on the proliferation of HepG2 cells were evaluated by MTT assay and colony formation assay. Results Six sesquiterpenoid dimers were isolated from the ethyl acetate fraction of the 90% ethanol extract of I. japonica, among which compounds 1 (inujaponolide T) and 3 (inujaponolide U) were identified as new compounds. The bioactivity screening results showed that compounds 1—6exhibited potent cytotoxicity against HepG2 cells, with IC₅₀ values ranging from 2.61 to 13.94 μmol/L. Furthermore, compounds 1 and 3 inhibited the cell viability and reduced the colony formation of HepG2 cells in a dose-dependent manner. Conclusion Compounds 1 and 3 are new sesquiterpenoid dimers with anti-hepatocellular carcinoma activity, and may serve as potential antitumor lead compounds for further in-depth investigation.

Objective Utilize metabolomics techniques to analyze the changes in chemical components of raw and steamed Rhei Radix et Rhizoma and the differences in their blood-entering components, and to reveal the impact of processing on the material basis of Rhei Radix et Rhizoma. Methods UPLC-MS/MS technology was used to collect the chemical composition information of raw and steamed Rhei Radix et Rhizoma in vitro and in vivo. Identification was carried out by combining the characteristic fragment ions and fragmentation pathways of the compounds. Multivariate statistical analysis was combined to screen for differential markers. Results A total of 201 components were identified in both raw and steamed Rhei Radix et Rhizoma, including 45 anthraquinones, 43 tannins, 52 flavonoids, and 21 stilbenes, 12 phenolic acids, seven phenylpropanoids, three chromones, two aromatic aldehydes, and 16 other compounds. After steaming, 23 common components showed significant changes. Among them, seven components such as aloeemodin, gallic acid, and catechin were significantly upregulated (P < 0.05), and 16 components such as aloeemodin-1-O-β-D-glucopyranoside, sennoside A, and proanthocyanidins were significantly downregulated (P < 0.05). A total of 36 prototype components and 58 metabolites were identified in the drug-containing serum. 11 common components showed significant changes in raw and steamed Rhei Radix et Rhizoma compared with raw Rhei Radix et Rhizoma, the blood drug concentrations of some prototype and metabolite components in steamed Rhei Radix et Rhizoma increased. In particular, the blood concentration of the tannin monomer gallic acid, which is a prototype component entering the blood, and the blood concentration of prototype aglycone-type components increased. The relative content of some phase-Ⅱ metabolites of anthraquinones significantly increased. Conclusion The in-vitro and in-vivo metabolic profiles of raw and steamed Rhei Radix et Rhizoma have changed significantly. The differential components are mainly in anthraquinones, tannins, etc. Compared with raw Rhei Radix et Rhizoma, in steamed Rhei Radix et Rhizoma, most conjugated anthraquinones were converted into free anthraquinones, hydrolysable tannin polymers were decomposed into tannin monomers, and there was a dynamic transformation between condensed tannin components and flavonoid aglycone components. These conversions of components can provide a scientific basis for clarifying the material basis of the “different treatments for raw and steamed Rhei Radix et Rhizoma ”.

Objective To address the complex molecular structures and diverse ring systems characteristic of traditional Chinese medicine (TCM) compounds, this study aims to optimize the graph isomorphism network (GIN) to enhance its predictive capability for key molecular properties of TCM molecules. Methods Based on the GIN architecture, edge descriptors representing bond length strain and angle strain were introduced into the molecular graph, and multi-graph data were constructed by incorporating multiple favorable conformations, thereby developing a flexibility-ring enhanced graph neural network (FRGNN). The predictive performance of the proposed model on seven key molecular properties was evaluated using two TCM databases containing 37 822 molecules, and compared against three state-of-the-art (SOTA) graph neural network models and two basic graph neural network models. Results Compared with the second-best performing model, FRGNN achieved an average reduction of 8.63% in root mean square error across the seven property prediction tasks, for molecules containing polycyclic and macrocyclic structures, the predictive root mean square error was further reduced by 10.04%. Conclusion The proposed FRGNN model demonstrates superior performance over existing state-of-the-art models for small-molecule property prediction in the task of predicting key properties of TCM compounds, providing a novel and effective approach for the property prediction of complex natural products.

Objective In response to the traditional belief that the medicinal properties of Shengjiang (Zingiberis Rhizoma Recens, ZRR) change after processing, this study integrated electronic bionic sensing, gas chromatography-mass spectrometry (GC-MS), and molecular dynamics simulation techniques to systematically elucidate the dynamic changes in the volatile components of Sichuan-origin ZRR and its processed products [Ganjiang (Zingiberis Rhizoma, ZR), Paojiang (Zingiberis Rhizoma Praeparatum, ZRP), Jiangtan (Zingiberis Rhizoma Carbonisatum, ZRC)]. It also revealed the core molecular basis behind the pungency of ZRR and its interaction mechanism with the vanillin subtype 1 of the transient receptor potential vanilloid 1 (TRPV1). Methods Electronic tongue was utilized to establish a numerical evaluation model for the tastes of ZRR and its processed products, thereby clarifying the differences in odor between different samples. GC-MS was used to analyze the changes in volatile components of ZRR and its various processed products. Autodock-1.5.7 and Pymol were applied to perform molecular docking and visual analysis of the common components in these samples in relation to the pungent taste receptor TRPV1. Additionally, molecular dynamics simulations were conducted using the Gromacs 2022 software. Results Electronic tongue testing revealed significant differences in taste scores between ginger and its various processed products. GC-MS analysis demonstrated variations in the content and quantity of volatile components between ginger and its different processed products. A total of 15 chemical compounds were common to all products of ZRR, while 14 volatile compounds were unique to ZRR, eight to ZR, one to ZRP, and five to ZRC. Most of these compounds were terpenoids. The content of monoterpenoids decreased and the content of most sesquiterpenoids increased after ZRR was processed. the components of elemol and zingiberenol were converted into (−)-cis-β-elemene and α-curcumene. Molecular docking and dynamic simulation studies showed that compounds such as elemol and zingiberenol could form stable hydrogen bond networks with amino acid residues such as LEU-1383 and ILE-1441 on the 8GFA protein, with the binding mechanism being a combination of van der Waals forces, electrostatic interactions, and hydrogen bonds. Conclusion The pungency of ZRR significantly enhanced after processing, with sesquiterpenes likely serving as the primary material basis for this effect. These compounds bind to the TRPV1 receptor via stable hydrogen-bonding networks and van der Waals forces, thereby elucidating the scientific mechanism underlying the increased pungency post-processing.

Objective Based on the supramolecular “imprinting template” theory, this study integrates the matching frequency method, total statistical moment, and factor rotation method, combined with in vitro anti-tumor activity, to elucidate the pharmacodynamic material basis of Yuxingcao (Houttuynia cordata) volatile oil, providing a new approach for its quality evaluation and screening of potential pharmacodynamic-related component groups. Methods Volatile oil samples from 58 batches of H. cordata collected from different origins (S1—S58) were analyzed by GC-MS to establish fingerprints. The matching frequency method was employed to categorize “imprinting template” into structural “material units”. Total-statistical-moment parameters and information entropy were applied to verify the consistency of fingerprint characteristics before and after integration. Functional “material units” were extracted via factor rotation to construct a quality evaluation model to score each batch. The half maximal inhibitory concentration (IC₅₀) values of the 58 volatile oil batches against human lung adenocarcinoma A549 cells were determined using the CCK-8 assay, and Pearson correlation analysis was conducted to examine the relationship between the comprehensive scores and IC₅₀ values. Results The 58 batches were effectively reduced to an “imprinting template” consisting of 34 structural “material units” (A1—A34) by the matching frequency method. After integration, the average number of chromatographic peaks and information entropy decreased significantly (P < 0.01), while no significant differences were observed in the total zero-order moment, first-order moment, second-order moment, or information content. Factor rotation extracted eight characteristic common factors. Comprehensive scoring results showed higher scores for batches such as S16 and S54, and lower scores for batches such as S24 and S27. Pharmacodynamic experiments revealed significant differences in IC₅₀ values among batches (73.6—269.9 nL/mL). Correlation analysis confirmed a negative correlation between comprehensive scores and IC₅₀ values (r = −0.739, P < 0.01). Accordingly, structural “material units” with high contributions, including A34, A31, and A7, were preliminarily screened as potential pharmacodynamic component groups. Conclusion Based on the supramolecular “imprinting template” theory, this study categorized structural “material units” using the matching frequency method and verified the scientific validity of this division with the total statistical moment. Subsequently, functional “material units” were extracted via factor rotation to construct a comprehensive evaluation model, and the sample scores were correlated with anti-lung cancer activity. This approach preliminarily screened key structural “material units” in H. cordata volatile oil, indicating potential core pharmacodynamic component groups, and providing a basis for subsequent precise screening and validation.

Objective In order to improve the targeting release of gambogic acid, a folic acid-modified gambogic acid nanocrystals-phospholipid composite drug delivery system (GA-NCs@PL-FA) was designed and prepared, and its prescription process was optimized. The physicochemical properties, in vitro anti-hepatocarcinoma activity and in vivo tissue distribution characteristics of GA-NCs@PL-FA were investigated. Methods Gambogic acid nanocrystals (GA-NCs) were prepared by CO₂-assisted precipitation method, and then GA-NCs@PL-FA was constructed by thin film hydration method. The prescription process was optimized by single factor investigation and central composite design-response surface methodology (CCD-RSM). The microscopic morphology was observed by transmission electron microscopy (TEM), and the particle size, ζ potential, encapsulation efficiency, drug loading and stability were determined. The in vitro release behavior in pH 7.4, 6.5 phosphate buffer (PBS, containing 0.5% Tween 80) was investigated by dialysis bag method. The effects of different administration groups on the viability and migration ability of HepG2 cells were evaluated by cell counting kit-8 (CCK-8) method and cell scratch test. The content of gambogic acid in each tissue of tumor-bearing nude mice was determined by ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), and its tissue distribution and tumor tissue enrichment ability were analyzed. Results The GA-NCs@PL-FA was spherical, the particle size was (183.07 ±0.55) nm, the ζ potential was (−17.70 ±0.17) mV, the encapsulation efficiency and drug loading were (84.64 ±0.57)% and (4.33 ±0.07)%, respectively. The properties were stable after 7 d at 4, 25 ℃. In both release media, it showed obvious sustained release effect. In vitro experiments showed that compared with gambogic acid, GA-NCs and folic acid-free gambogic acid nanocrystals-phospholipid composite drug delivery system (GA-NCs@PL), GA-NCs@PL-FA had a stronger inhibitory effect on the proliferation and migration of HepG2 cells, and the half inhibitory concentration (IC₅₀) was reduced to 0.50 μg/mL. The tissue distribution test confirmed that GA-NCs@PL-FA could prolong the retention time of gambogic acid in vivo and promote its enrichment in tumor tissues, with good targeting. Conclusion The formulation and preparation process of GA-NCs@PL-FA were optimized. The system had uniform particle size, good drug loading performance, good sustained release effect and in vitro anti-hepatoma activity, and showed strong tumor tissue enrichment ability. It can provide reference for the development and clinical application of gambogic acid targeted preparations.

Objective To develop a puerarin-loaded supramolecular gel based on chitosan-modified β-cyclodextrin (Pur@CS-β-CD Gel), optimize its formulation, and evaluate its potential for nasal delivery and anti-migraine efficacy. Methods The Pur@CS-β-CD Gel formulation was optimized using single-factor experiments and Box-Behnken design-response surface methodology (BBD-RSM). The optimized Pur@CS-β-CD Gel was characterized for its microstructure, pH value, stability, and in vitro release profile. A chronic migraine rat model was established. The therapeutic efficacy was assessed through behavioral indicators and by measuring the levels of calcitonin gene-related peptide (CGRP), 5-hydroxytryptamine (5-HT), and interleukin-1β (IL-1β) in brain tissue. Results The optimal formulation consisted of CS-β-CD and sodium carboxymethylcellulose at a mass ratio of 1:9, a total polymer concentration of 2.6%, and 40 mg of puerarin. The resulting Pur@CS-β-CD Gel exhibited a three-dimensional porous structure, a pH of 6.5, good stability, and a biphasic in vitro release profile characterized by an initial rapid release (82.75% within 4 h) followed by sustained release, with no obvious nasal-mucosal irritation. Pharmacodynamic studies demonstrated that Pur@CS-β-CD Gel significantly improved migraine-related behavioral performance in rats, reduced CGRP and IL-1β levels, and increased 5-HT content in the brain. Conclusion The Pur@CS-β-CD Gel demonstrates a feasible preparation process, favorable properties for nasal administration, and effective anti-migraine activity, providing an experimental basis for the development of novel nasal delivery systems and migraine therapies.

Objective To prepare diosgenin-rebaudioside A self-assembled nanomicelles (Dio-Reb A-SNM), and evaluate its oral pharmacokinetics and hypoglycemic effects in vivo. Methods Single factor experiment was employed to investigate the prescriptions of Dio-Reb A-SNM. The amounts ratio of rebaudioside A to diosgenin, diosgenin concentration and ultrasonic time were selected as main influencing factor, the overall desirability of envelopment efficiency and drug loading was employed as optimization index, and Box-Behnken response surface design method (BBD-RSM) was employed to optimize prescriptions of Dio-Reb A-SNM. Transmission electron microscope (TEM) was employed to observe the microscopic appearance of Dio-Reb A-SNM, and X-ray powder diffraction (XRPD) was employed to analyze the crystal form of diosgenin in Dio-Reb A-SNM powder. The saturated solubility and release behavior of Dio-Reb A-SNM were determined in different pH media. SD rats were administered intragastrically with diosgenin and Dio-Reb A-SNM, respectively. Blood drug concentration was determined, and the relative oral bioavailability of Dio-Reb A-SNM was calculated. The model of type 2 diabetes mellitus (T2DM) in rats was established, and the effects of diosgenin and Dio-Reb A-SNM on hypoglycemic effects, serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), urea nitrogen and creatinine level were compared. The histopathological state of liver and kidney was observed by HE staining method. Results The optimal formulations of Dio-Reb A-SNM: amounts ratio of rebaudioside A to diosgenin was 12.32:1, diosgenin concentration was 1.97 mg/mL and ultrasonic time was 20.20 min. Envelopment efficiency, drug loading, particle size and ζ potential were (93.59 ±0.63)%, (5.65 ±0.07)%, (25.66 ±1.76) nm and (−24.59 ±1.18) mV, respectively. The appearance of Dio-Reb A-SNM were spherical. The crystal form of diosgenin changed into an amorphous form in Dio-Reb A-SNM powder. Dio-Reb A-SNM greatly improved the saturated solubility of diosgenin in different pH media. Drug release behavior of Dio-Reb A-SNM in vitro had obvious sustained-release characteristics, the cumulative release rate was increased to 90.94% in 18 h, and the drug release process conformed to Weibull model. Pharmacokinetics showed that t_max of Dio-Reb A-SNM was shortened to (2.06 ±0.29) h, t_1/2 was increased to (8.39 ±1.94) h, C_max and oral relative bioavailability were increased to 3.59-fold and 6.82-fold, respectively. Compared with diosgenin group (30 mg/kg), Dio-Reb A-SNM (30 mg/kg) could significantly reduce the blood glucose of T2DM rats at the fourth week (P < 0.01), and effectively reduced the AST, ALT, urea nitrogen and creatinine level in serum (P < 0.01). The pathological damage of liver and kidney was further alleviated by Dio-Reb A-SNM. Conclusion Dio-Reb A-SNM significantly promoted the oral absorption of diosgenin and improved hypoglycemic efficacy in vivo.

Objective To analyze and predict the quality markers (Q-Markers) of Mujingye (Viticis Negundo Folium, VNF) standard decoction based on characteristic chromatogram, network pharmacology, and quantitative analysis, and to construct a multi-dimensional quality evaluation system for VNF standard decoction. Methods UPLC was used to establish the characteristic chromatogram of VNF standard decoction. The chromatographic conditions were as follows: Cortecs C₁₈ column (100 mm×2.1 mm, 2.7 μm); the mobile phase consisted of acetonitrile (A) and 0.1% phosphoric acid (B), using gradient elution (0—9 min, 3%—16% A; 9—21 min, 16%—20% A; 21—22 min, 20%—65% A; 22—23 min, 65% A; 23—29 min, 65%—95% A); the detection wavelength was 270 nm; the column temperature was 35 ℃; the flow rate was 0.3 mL/min; and the injection volume was 2 μL. Network pharmacology was employed to obtain the key targets and key pathways of candidate Q-Markers, and the “candidate Q-Marker-key target-key pathway” network was constructed. Quantitative analysis was performed on the candidate Q-Markers of VNF to clarify the rules of quantity transfer of candidate Q-Markers from decoction pieces to standard decoction. Results The characteristic chromatogram of VNF standard decoction was established, and nine characteristic peaks were marked. Among them, four chromatographic peaks (peaks 1, 3, 5, and 8) were identified by reference substances as p-hydroxybenzoic acid, isoorientin, isovitexin, and apigenin-7-O-β-D-glucuronide (AG). Network pharmacology analysis revealed that p-hydroxybenzoic acid, isoorientin, isovitexin, and AG in VNF standard decoction could act on key targets such as matrix metalloproteinase 2 (MMP2), v-Rel avian reticuloendotheliosis virus oncogene homolog A(RELA), and histone deacetylase 1 (HDAC1), and regulate key pathways including cellular senescence, advanced glycation end products-receptor for advanced glycation end products (AGE-RAGE) signaling pathway, and phosphatidylinositol 3-kinase-protein kinase B (PI3K-Akt) signaling pathway to exert therapeutic effects against bronchitis. Quantitative analysis of these four components showed that their average contents in 15 batches of VNF standard decoction were 18.98, 9.13, 4.72, 1.64 mg/g, respectively, while their average contents in the decoction pieces were 4.98, 11.44, 2.38, 2.60 mg/g, respectively. The average transfer rates from decoction pieces to standard decoction were 118.83%, 24.61%, 54.75%, 19.81%, respectively. p-Hydroxybenzoic acid, isoorientin, isovitexin, and AG were preliminarily identified as the Q-Markers of VNF for treating bronchitis. Conclusion The established quality evaluation method is accurate and reliable, with uniform and stable quantity transfer. Combined with the Q-Markers screened by network pharmacology, it can provide a basis for the quality control of VNF standard decoction and support for the subsequent elucidation of the mechanism of action of its pharmacodynamic material basis.

Objective To investigate the effect and potential mechanism of cuttlebone (CB)/bovine serum albumin (BSA) photocrosslinked hydrogel on rats with critical sized bone defects (CSBD) model in skull tissues. Methods SD rats were randomly divided into control group, positive control group, BSA group, 0.5% CB/BSA group, 1.5% CB/BSA group and 3.0% CB/BSA group, with six rats in each group. A rat CSBD model was established using a 5 mm outer diameter ring drill, while the control group only underwent modeling without implants, positive control group implanted with Bio-Oss^® collagen bone filling material, CB/BSA photocrosslinked hydrogels with CB mass fraction of 0, 0.5%, 1.5% and 3% were implanted in BSA group, 0.5% CB/BSA group, 1.5% CB/BSA group and 3.0% CB/BSA group, respectively. After eight weeks after surgery, the microstructural changes of bone tissue were analyzed using micro computed tomography (Micro CT). Hematoxylin-eosin (HE) and Masson staining were used to observe pathological changes in skull tissue. Immunohistochemistry and immunofluorescence were used to detect the positive expressions and fluorescence intensity of collagen type I alpha 1 (COL1A1), platelet endothelial cell adhesion molecule-1 (PECAM-1) and osteocalcin (OCN) in skull tissue, respectively. qRT-PCR and Western blotting were used to detect the gene and protein expressions of Wnt family members 3A (WNT3A), β-catenin, vascular endothelial growth factor A (VEGFA), COL1A1, PECAM-1 and OCN in skull tissue, respectively. In order to evaluate the safety of hydrogel in vivo, the abdominal aorta blood of rats was collected to analyze the changes of blood routine and serum liver and kidney function indexes. Results Micro-CT results showed that no new bone formation was observed in the bone defect area of rats in control group, indicating that bone healing was not achieved during the observation period. The pathological tissue section results showed that the defect area in control group was mainly composed of loose fibrous tissue. In CB/BSA photocrosslinked hydrogel group, new bone formation was observed. Compared with control group, bone volume/tissue volume (BV/TV), bone mineral density (BMD and trabecular thickness (Tb.Th) of rats in CB/BSA photocrosslinked hydrogel group were significantly increased (P < 0.05), bone surface/bone volume (BS/BV) was significantly decreased (P < 0.05), the positive expressions and fluorescence intensity of COL1A1, PECAM-1, OCN in skull tissue were significantly increased (P < 0.05), and the expression levels of COL1A1, PECAM-1, β-catenin, WNT3A, VEGFA, OCN were significantly increased (P < 0.05), among which, the 3.0% CB/BSA photocrosslinked hydrogel group had the best comprehensive effect. No significant abnormalities were found in the blood routine and serum liver and kidney function indicators of rats in each group, and no pathological changes such as inflammation, necrosis or fibrosis were observed in the main organs. Conclusion CB/BSA photocrosslinked hydrogel shows good bone repair effect and safety in vivo, and its osteogenic effect may be mediated by activation of Wnt/β-catenin signaling pathway.

Objective To explore the pharmacological effect of Ejiao (Colla Corii Asini, CCA) on delaying D-galactose-induced skin aging and maintaining microvascular homeostasis, and to clarify the molecular mechanism of its regulation of vascular maturity through tyrosine kinase with immunoglobulin-like and EGF-like domains 2 (Tie2)/angiopoietin (Ang)/vascular endothelial cadherin (VE-cadherin) signal axis. Methods The model of premature senility in mice was established by sc D-galactose, and CCA (270.3 mg/kg) was administered ig for eight weeks. The back blood flow was monitored by laser speckle contrast imaging (LSCI), the microvessel density was observed by optical coherence tomography angiography (OCTA), the skin tissue morphology and collagen content were evaluated by hematoxylin-eosin staining (HE) and Masson staining, the expressions of vascular endothelial marker CD31 and pericyte marker platelet-derived growth factor receptor β (PDGFRβ) were detected by immunofluorescence and immunohistochemistry, the Ang1/Ang2 value in serum was detected by ELISA, the expressions of interleukin-1β (IL-1β), C-X-C motif chemokine ligand 1 (CXCL1) and tumor necrosis factor-α (TNF-α) mRNA in skin tissue was detected by qRT-PCR. Human umbilical vein endothelial cells (HUVEC) were induced by hydrogen peroxide (H₂O₂) to construct a stress premature aging model, cells were pretreated with 62.5—250.0 μg/mL CCA for 24 h. Cell viability was detected by CCK-8. Senescence-associated β-galactosidase (SA-β-gal) staining and reactive oxygen species (ROS) level were used to assess the degree of aging. qRT-PCR was used to detect the senescence-associated secretory phenotype (SASP) and inflammation related gene expressions. Western blotting was used to detect tumor protein p53 (p53), cyclin dependent kinase inhibitor 1 A (p21), p-Tie2, Tie2, VE-cadherin and vascular endothelial growth factor A (VEGFA) protein expressions. Ang1/Ang2 value was detected by ELISA. Vascular stability was evaluated through pericyte recruitment and endothelial permeability experiments. Results In the animal experiment, CCA significantly improved the skin fold of aging mice, increased the thickness of epidermis and dermis and collagen volume fraction (P < 0.01, 0.001), restored skin blood flow perfusion and reduced disordered angiogenesis, increased the expression of PDGFRβ in skin tissue and the value of Ang1/Ang2 in serum (P < 0.001), decreased the expressions of inflammatory factors (P < 0.001), and enhanced vascular maturity. In the cell experiment, CCA significantly reduced the positive rate of SA-β-gal and ROS level in HUVEC (P < 0.001), down-regulated the expressions of SASP and inflammation related genes (P < 0.01, 0.001), up-regulated the protein expressions of p-Tie2 and VE-cadherin (P < 0.001), down-regulated the protein expressions of p53, p21, Tie2 and VEGFA (P < 0.05, 0.001), and increased the value of Ang1/Ang2 (P < 0.05, 0.01), significantly enhancing the ability of endothelial cells to recruit pericytes and the integrity of vascular endothelial barrier. Conclusion CCA can improve the aging phenotype of aging skin model induced by D-galactose, improve microvascular maturity and maintain vascular homeostasis, which may be related to the regulation of Tie2/Ang/VE-cadherin signal axis.

Objective To investigate the therapeutic effect and mechanism of iridoid fraction from Morinda officinalis on aging-related sarcopenia. Methods Using monotropein and deacetylasperulosidic acid as analytical markers, effects of different ethanol concentrations on extraction efficiency of active iridoids fraction from M. officinalis via heated reflux extraction was investigated. The extract was concentrated under reduced pressure and subsequently enriched and purified using macroporous adsorption resin to obtain the total active iridoid fraction. By comparing the elution efficiencies of solvents with varying polarities, purification parameters were optimized. Following the establishment and validation of a high performance liquid chromatography (HPLC) analytical method, the content of major iridoid components in active fraction was quantified. The purified fraction was then used for subsequent pharmacodynamic evaluations. The C2C12 cell differentiation model and a D-galactose (D-gal)-induced C2C12 myotube atrophy model were employed to assess the effects of iridoid fraction on promoting myotube differentiation and ameliorating myotube atrophy, with MYHC protein expression serving as the indicator. A D-gal-induced accelerated aging mouse model was utilized to evaluate the therapeutic effects of iridoid fraction through grip strength tests, hindlimb muscle mass measurements, and hematoxylin-eosin (HE) staining of tibialis anterior and gastrocnemius muscles. Differentially expressed genes in gastrocnemius muscle of mice in each group was analyzed by transcriptome sequencing, and the results were further verified by Western blotting and qRT-PCR for the expression changes of related differentially expressed genes indicated by transcriptome sequencing, in combination with network pharmacology analysis. Results The extraction of iridoid components from M. officinalis was optimized using a 60% ethanol solution under heated reflux conditions, resulting in the highest iridoid content. In the purification process, H103 macroporous adsorption resin demonstrated superior adsorption performance, with water as the eluent resulting in the highest elution rate of target constituents. The iridoid fraction from M. officinalis significantly ameliorated D-gal-induced atrophy in C2C12 myotubes (P < 0.001), and promoted myotube differentiation (P < 0.05, 0.01, 0.001). The iridoid fraction could significantly enhance the limb pulling force and ratio of hind limb muscle mass to body mass of accelerated aging mice (P < 0.05, 0.01, 0.001), significantly increase the cross-sectional area of the tibialis anterior muscle and gastrocnemius muscle in aging mice (P < 0.01, 0.001), thereby improving the phenotype of age-related sarcopenia. Further mechanism studies revealed that iridoid fraction could up-regulate the expression levels of key proteins in phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathway (P < 0.05, 0.01, 0.001), and significantly up-regulate the expression levels of phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA), serine/threonine kinase 1 (AKT1) and mTOR genes (P < 0.05, 0.01, 0.001). And PI3K inhibitor LY294002 significantly inhibited the upregulation of PI3K/Akt/mTOR signaling pathway related protein expressions by iridoid fraction (P < 0.05). Conclusion The iridoid fraction of M. officinalis can significantly improve sarcopenia caused by aging, and its mechanism is related to the regulation of PI3K/Akt/mTOR signaling pathway.

Objective To investigate the therapeutic effect of schisandrin B (Sch B) on ulcerative colitis (UC) in rats induced by 2,4,6-trinitrobenzenesulfonic acid (TNBS), and explore its molecular mechanism in alleviating pyroptosis of colonic epithelial cells by regulating autophagy-related proteins as well as NOD-like receptor thermal protein domain associated protein 3 (NLRP3)/cysteine-aspartic protease-1 (Caspase-1)/gasdermin D (GSDMD) signaling pathway. Methods SD rats were randomly divided into control group, model group, mesalazine (100 mg/kg) group, Sch B low-, medium- and high-dose (10, 20, 50 mg/kg) groups, with 10 rats in each group, half male and half female. A TNBS-induced UC model was constructed, after 14 consecutive days of administration, the general conditions of rats were observed. The disease activity index (DAI) score, colon length and colon mucosa damage index (CMDI) score were detected. Histopathological injury of colonic tissue was evaluated by hematoxylin-eosin (HE) staining. Levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and IL-1β in serum were measured by ELISA. The expressions of autophagy related protein 16 like protein 1 (ATG16L1) and GSDMD proteins in colonic tissue were detected by immunohistochemistry. Western blotting was performed to detect the expressions of autophagy-related proteins (Beclin-1, microtubule-associated protein light chain 3B (LC3B), ATG16L1, p62) and pyroptosis pathway-related proteins (NLRP3, Caspase-1, GSDMD) in colonic tissue. Results Compared with model group, Sch B significantly ameliorated body weight loss, hematochezia and other symptoms in UC rats, reduced DAI and CMDI scores (P < 0.05, 0.01), increased colon length (P < 0.05, 0.01), alleviated colonic mucosal congestion, edema, ulcer formation and inflammatory cell infiltration (P < 0.05, 0.01), decreased levels of IL-6, TNF-α and IL-1β in serum (P < 0.05, 0.01), upregulated Beclin-1 protein expression and LC3-Ⅱ/Ⅰ ratio in colonic tissue (P < 0.01), and downregulated ATG16L1, p62, NLRP3, Caspase-1, GSDMD protein expressions (P < 0.05, 0.01). Conclusion Sch B exerts a remarkable therapeutic effect on TNBS-induced UC in rats. Its mechanism may be associated with maintaining autophagy flux homeostasis, inhibiting colonic epithelial cell pyroptosis mediated by NLRP3/Caspase-1/GSDMD signaling pathway, reducing the release of pro-inflammatory factors and thereby alleviating intestinal inflammatory response. Normalization of autophagy flux may be a key regulatory link for Sch B to inhibit colonic epithelial cell pyroptosis.

Objective To investigate the therapeutic effect and molecular mechanism of aconitine combined with paeoniflorin on knee osteoarthritis (KOA) based on Indian hedgehog (Ihh)-glioma-associated oncogene homolog (Gli) signaling pathway, as well as to evaluate the safety of this combined administration. Methods Anterior cruciate ligament transection was performed to establish a rat KOA model. Sham group, model group, celecoxib (24 mg/kg) group, and aconitine-paeoniflorin low- (5 + 50 μg/kg), medium- (10 + 100 μg/kg), and high-(20 + 200 μg/kg) dose groups were set up, with 10 rats in each group. Behavioral tests were conducted to assess analgesic efficacy. Hematoxylin-eosin (HE) staining and micro-computed tomography (Micro-CT) were used to observe morphological changes of articular cartilage and bone microarchitecture. ELISA was used to detect levels of matrix metalloproteinase 13 (MMP13) and type Ⅱ collagen (Col Ⅱ) in serum. Immunofluorescence staining and qRT-PCR were applied to determine the protein and gene expressions of Ihh, Gli, patched 1 (Ptch1) and MMP13 in knee joint tissues. Molecular docking was performed to verify the binding affinity between aconitine, paeoniflorin and core targets of Ihh-Gli signaling pathway. Activity of aspartate aminotransferase and levels creatinine, blood urea nitrogen, urinary protein in serum were detected, and morphological analysis on heart, liver and kidney tissues of rats in each group were observed by HE staining to evaluate the safety of aconitine combined with paeoniflorin. Results During the administration period, no death or toxic symptoms were observed of rats in each group. There were no significant abnormalities in serum liver and kidney function indicators, as well as in the morphology of heart, liver and kidney tissues, indicating that the combination of aconitine and paeoniflorin had good safety. Compared with sham group, the mechanical pain threshold and thermal pain threshold of rats in model group were significantly reduced (P < 0.01, 0.001), the articular cartilage surface was damaged, the tidemark was incomplete, the subchondral bone was lost, and the distribution of bone trabeculae was disordered. The level of MMP13 in serum was significantly increased (P < 0.001), while the level of Col II was significantly decreased (P < 0.001). The protein and gene expressions of Ihh, Gli, Ptch1 and MMP13 in knee joint tissue were significantly upregulated (P < 0.001), indicating abnormal activation of Ihh-Gli pathway in KOA state. Compared with model group, the combination of aconitine and paeoniflorin could increase the mechanical pain threshold and thermal pain threshold of rats (P < 0.05, 0.01, 0.001), repair cartilage matrix, improve bone microstructure, increase MMP13 level and decrease Col II level in serum (P < 0.05, 0.01, 0.001), and significantly downregulate the protein and gene expressions of Ihh, Gli, Ptch1 and MMP13 in knee joint tissue (P < 0.05, 0.01, 0.001). The molecular docking results showed that the binding energies of aconitine and paeoniflorin to Ihh, Gli, a disintegrin and metalloprotease with thrombospondin motifs 5 (ADAMTS5) and MMP13 were all ≤ −5 kcal/mol, indicating that aconitine and paeoniflorin have strong binding abilities to key targets of Ihh-Gli pathway. Conclusion The combination of aconitine and paeoniflorin can inhibit abnormal activation of Ihh-Gli signaling pathway, downregulate MMP13 expression and upregulate Col II expression, inhibit cartilage matrix degradation, improve joint function and cartilage pathological damage in KOA rats, and has a good safety, providing experimental evidence for the treatment of cold dampness obstruction type KOA with the herb pair of Wutou Decoction.

Objective To investigate the effects of ig administration and ip injection on the pharmacokinetic behavior and tissue distribution of tetrahydropalmatine (THP) in neuropathic pain model rats, and to evaluate the analgesic effects of the two administration routes against acute thermal pain in mice using the hot-plate test. Methods A rapid and sensitive high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was established for determining THP concentrations in the plasma and tissues (including heart, liver, brain, lung, kidney) of neuropathic pain model rats. The pharmacokinetic parameters were fitted using the non-compartmental model with WinNonlin 7.0 software, and the distribution of THP in various rat tissues at different time points was investigated. Furthermore, the analgesic effects of THP via ig and ip routes in mice were explored, using the licking latency induced by warm thermal stimulation as the evaluation index. Results A quantitative method for determining THP was successfully established. The methodological validation results showed a good linear relationship (r = 0.999 3) between 0.5—1 250.0 ng/mL, the precision, accuracy and matrix effects all met the requirements of biological analysis. Pharmacokinetic results showed that compared with ig administration, ip administration led to faster absorption and higher bioavailability. The time to peak concentration (t_max) for the low-dose ip administration group (0.21 ±0.08) h was comparable to that of ig administration group (0.25 ±0.00) h, but its area under the plasma concentration-time curve (AUC_0—t) was significantly higher than that of ig administration group at 4 mg/kg dose. Regarding tissue distribution, THP was widely distributed in major rat tissues. At multiple time points, concentrations in the liver, kidney and other tissues, as well as plasma, were higher in ip administration group than in ig administration group. The liver and kidney were identified as the primary drug accumulation organs, and the brain tissue also showed relatively high exposure. Pharmacodynamics results indicated that both ig and ip administration significantly prolonged the licking latency in mice induced by warm thermal stimulation, with the ip administration group showing the best analgesic effect. Conclusion Compared to ig administration, ip administration offers advantages including faster absorption, higher bioavailability and more rapid tissue distribution. Both ig and ip administration routes exhibit significant analgesic effects, with ip administration showing the best efficacy. This verifies the consistency between pharmacodynamics and pharmacokinetics.

Objective To systematically identify the candidate core herb combination and potential therapeutic targets for ischemic stroke by integrating patent mining, spatiotemporal multi-omics analysis, and computational simulation, and elucidate the scientific connotation of the traditional Chinese medicine therapeutic principle of “Tongfu Xingshen” based on the patent database of traditional Chinese medicine compound prescriptions. Methods The China National Intellectual Property Administration patent database was searched. Cluster analysis was used to screen the candidate core herb combination. Network pharmacology, molecular docking, and molecular dynamics simulations were then performed to identify and evaluate candidate therapeutic targets of traditional Chinese medicine compound prescriptions for ischemic stroke. Single-cell RNA sequencing, spatial transcriptomics, and CellOracle-based in silico knockdown analysis were further integrated to perform multidimensional association analysis. Results A candidate core herb combination consisting of Yuanzhi (Polygalae Radix), Shichangpu (Acori Tatarinowii Rhizoma), Dahuang (Rhei Radix et Rhizoma), and Yujin (Curcumae Radix) was identified. Nine representative active components, including 1-hydroxyacoronene, aristolone, calamendiol, β-asarone, α-asarone, kaempferol, thymol, eugenol, and caffeic acid, and six candidate therapeutic targets, including tumor necrosis factor (TNF), interleukin-6 (IL-6), protein kinase B1 (AKT1), epidermal growth factor receptor (EGFR), mitogen-activated protein kinase 1 (MAPK1), and prostaglandin-endoperoxide synthase 2 (PTGS2), were obtained. Molecular docking and molecular dynamics simulations suggested that some representative active components exhibited certain binding tendencies with the candidate target proteins. Among them, the kaempferol-PTGS2 and kaempferol-EGFR complexes showed a degree of stability under simulation conditions. Single-cell and spatial transcriptomic analyses further showed that the regions with high expression of candidate targets were spatially consistent with the distribution of macrophages, astrocytes, and other cell populations. CellOracle analysis suggested that Jun/Fos, signal transducer and activator of transcription 3 (STAT3), and other related transcription factors may be involved in post-ischemic inflammatory responses, cell-state transitions, and repair-related regulatory processes. Conclusion This study identified a candidate core herb combination consisting of Polygalae Radix, Acori Tatarinowii Rhizoma, Rhei Radix et Rhizoma, and Curcumae Radix from national patent traditional Chinese medicine compound prescriptions, and proposed that PTGS2, IL-6, EGFR, and related targets may serve as important candidate targets in the regulation of the inflammatory microenvironment in ischemic stroke. These findings preliminarily construct a mechanistic evidence chain integrating the core herb combination, core components, candidate targets, and spatiotemporal validation, and provide a reference for the modern mechanism explanation and subsequent experimental verification of the “Tongfu Xingshen” therapeutic principle in ischemic stroke from the perspectives of inflammatory microenvironment regulation, cell-cell communication, and neurovascular remodeling.

Objective Based on the traditional Chinese medicine (TCM) theory of meridian tropism, this study aimed to systematically elucidate the organ-specific therapeutic preference of Salvia miltiorrhiza in heart failure (HF) and its key molecular mechanisms by integrating multi-level evidence from genetic regulation, causal inference, and transcriptomics. Methods Major active components of S. miltiorrhiza were screened through TCMSP, ETCM, HERB, and document literature, and potential target genes were predicted. Genotype-tissue expression (GTEx) multi-tissue expression quantitative trait loci (eQTL) data and heart failure genome-wide association study (GWAS) data were then integrated, and a two-sample Mendelian randomization (MR) framework was applied to evaluate the tissue-specific causal effects of target genes in the heart, liver, lung, and kidney. The volume of interest (VOI) was further constructed to quantify organ-oriented characteristics, and HF transcriptomic data were used to analyze the expression patterns of key genes. Results Multi-tissue MR showed that S. miltiorrhiza target genes exhibited certain genetic effect patterns in the heart, liver, and lung. VOI analysis indicated that genes such as aldehyde dehydrogenase 2 (ALDH2), pyruvate dehydrogenase E1 beta subunit (PDHB), and cholesteryl ester transfer protein (CETP) contributed more prominently in the heart and liver, whereas genes such as protein kinase C alpha (PRKCA) also showed relatively strong tissue preference in the lung. The cross-organ consistently protective key gene Caspase-7 (CASP7) showed a directional effect associated with reduced HF risk in MR analysis and was significantly downregulated in HF transcriptomic data. Conclusion This study established an integrated framework linking meridian tropism, organ regulation, and disease-related causal effects. The results suggest that S. miltiorrhiza-related target genes exhibit a multi-organ action pattern dominated by the heart and liver while also involving lung tissue, providing new quantitative clues for the traditional meridian tropism theory of S. miltiorrhiza and suggesting that CASP7 may be an important candidate target mediating its cross-organ anti-HF effects.

Objective To apply bibliometric methods to review the research progress, hotspots and frontiers of traditional Chinese medicine (TCM) in regulating mitochondrial reactive oxygen species (mtROS), summarize the regulatory mechanisms, and provide data support and directional guidance for subsequent research and research planning. Methods Relevant Chinese and English literature were retrieved from CNKI, Wanfang, VIP and Web of Science Core Collection using TCM and mtROS as search terms or keywords, with the search period covering from database inception to March 31, 2026. CiteSpace, VOSviewer and Bibliometrix were applied to complete the visualization presentation and construction of the knowledge graph. Statistical analyses were performed on annual publication volumes, countries, institutions, authors, journals and keywords. Scientific research collaboration networks, co-occurrence clustering and keyword burst detection analyses were also carried out. Results A total of 1 026 Chinese articles and 649 English articles were included. Knowledge maps showed a sustained growth in annual publication output. China dominated this research field, while domestic and international cooperation networks were not well-developed. Core research institutions included Heilongjiang University of Chinese Medicine, Henan University of Chinese Medicine and Guangxi University of Chinese Medicine. The most productive journals were Chinese Journal of Experimental Traditional Medical Formulae and Journal of Ethnopharmacology. The top five keywords in Chinese literature were oxidative stress, mitochondria, TCM, Chinese herbal medicine and mechanisms of action; the top five keywords in English literature were oxidative stress, apoptosis, activation, TCM and reactive oxygen species. Keyword burst analysis indicated that the research frontier is shifting from basic antioxidant mechanisms toward precise mechanisms such as network pharmacology, molecular docking and ferroptosis. Conclusion Research on TCM regulating mtROS has become a growing research hotspot in recent years and will draw more academic attention, while interdisciplinary cooperation needs to be further strengthened. The combined application of network pharmacology and molecular docking is gradually becoming an emerging trend in this field.

Objective To comprehensively analyse the research status and hotspots of sinomenine by using bibliometric methods, and provide theoretical guidance for the future in-depth research and application of sinomenine. Methods A total of two databases of China Knowledge Network (CNKI) and Web of Science (WOS) were searched using sinomenine as the keyword, and the literature was imported into NoteExpress literature management software for screening. The analysis employed tools such as Excel, CiteSpace, and VOSviewer to visualize the current status of domestic and international research on sinomenine across various dimensions, including publication trends and national distribution, publishing institutions, publishing authors and keywords. Results A total of 710 Chinese and 471 English literature were retrieved that met the requirements. The research of sinomenine is mainly in China, and the international attention has also increased year by year in recent years, and the overall research has shown a continuous development trend. The core team of sinomenine research has been initially formed, and the cooperation is mostly carried out within the institution or in the same region, with less cross-regional cooperation. Both Chinese and English literature in the field of sinomenine focus on the pharmacological effects and mechanisms of sinomenine, such as anti-arthritis, while Chinese literature also focuses on the extraction and separation of sinomenine, quality standards, and formulation development. Conclusion The research of sinomenine is still in the limelight, and multi-omics-driven mechanism analysis, precision medicine-orientated dosage form innovation, and the expansion of new indications may be the hot spots of sinomenine research in the future. In addition, the research of sinomenine should strengthen multidisciplinary co-operation and apply new technologies in order to fully explore the potential of sinomenine.

Objective Dried flower buds or newly opened flowers of Lonicera macranthoides are used as Shanyinhua (Lonicerae Flos) for medicinal purposes. The Xianglei cultivar, developed from a natural mutant, exhibits a distinctive phenotype characterized by non-opening corolla and prolonged bud stage. Aiming to elucidate the cellular and molecular mechanisms underlying this unique corolla phenotype. Methods The flower of conventional cultivar (WT) and the Xianglei cultivar (XL) of L. macranthoides were used as materials. Corolla adaxial epidermal cells and longitudinal sections were observed using light microscopy. The expression patterns of xyloglucan endotransglucosylase/hydrolase genes (LmXTH6, LmXTH7, LmXTH33), expansin gene (LmEXP7), and pectin methylesterase genes (LmPME28, LmPME45) were analyzed by qRT-PCR. Protein-protein interaction networks were predicted using the STRING database. Ruthenium red staining coupled with Image J quantitative analysis was performed to assess the degree of pectin de-methylesterification in corolla cell walls. Results In the WT, the adaxial cells of corolla displayed a spatially sequential expansion, with the basal cells expanding first, followed by the middle cells. Moreover, the expansion rate of the adaxial cells is higher than that of the abaxial cells. In contrast, the expansion rate of the adaxial cells is significantly decreased in Xianglei, and the extend of cell expansion is not evident. Molecular analyses revealed that LmXTH6, LmXTH7, and LmXTH33 were highly expressed during the pre-anthesis and anthesis stages in WT, but were significantly suppressed in Xianglei. Expression of LmEXP7 peaked just before corolla opening in WT but remained low in Xianglei at the same stage. Furthermore, ruthenium staining showed a higher overall degree of pectin de-methylesterification in Xianglei compared with WT, which is consistent with the expressin patterns of LmPME28 and LmPME45. Conclusion The non-opening of the corolla in the L. macranthoides cultivar Xianglei is closely associated with inhibited expansion of corolla cells. The low expression of XTHs and EXP collectively contributes to the reduced expansion capacity of corolla cells in Xianglei, while the up-regulation of PMEs increases pectin de-methylesterification in the cell wall, potentially enhancing cell wall stiffness. The synergistic effect of XTH/EXP and PME may be an important factor leading to corolla non-opening in Xianglei. These findings provide experimental evidence for understanding the regulatory mechanisms underlying the superior phenotype of the Xianglei cultivar and offer new insights into the cell wall regulatory mechanisms governing floral organ morphogenesis in plants.

Objective To analyze the diversity and functional potential of endophytic bacterial communities in different tissues of Duchesnea indica and explore their potential relationship with the medicinal activity of the host plant. Methods Endophytic bacteria from the root, stem, leaf and fruit tissues of D. indica were studied. DNA fragments of the microbial communities were subjected to paired-end sequencing using Illumina high-throughput technology. Sequences were processed using DADA2 and Vsearch, followed by bioinformatics analysis and functional potential prediction. Results High-throughput sequencing of the endophytic bacterial genomes yielded a total of 1 914 360 raw sequences. At the phylum level, Proteobacteria was the dominant group. Among different tissues, the stem exhibited the highest richness of endophytic bacteria, while the root showed the highest diversity. Sphingomonas was identified as a key biomarker distinguishing intergroup differences and showed a significant positive correlation with Pseudomonas. Both bacterial genuses were major contributors to the N₁₀-formyltetrahydrofolate biosynthesis pathway. Conclusion The endophytic bacterial communities in the leaf, fruit and stem tissues of D. indica exhibited high similarity but were significantly distinct from those in the root. Pseudomonas and Sphingomonas may play synergistic roles in the synthesis of secondary metabolites or in enhancing host stress resistance.

Objective To screen and identify members of the bHLH transcription factor family based on genomic data from Forsythia suspensa and characterize their features and functions. Methods Using the genomic data of F. suspensa, the bHLH transcription factor family was screened and identified, analysing their physicochemical properties and phylogenetic relationships, and characterizing the bHLH genes by analysing their chromosomal locations, gene structures, functional annotations, and cis-acting elements. In addition, F. suspensa leaves were treated with exogenous methyl jasmonate (MeJA) and expression pattern of FsbHLHs within 48 h after treatment was detected by qRT-PCR. Correlation analysis was conducted in combination with the changes in phillygenin content. Results A total of 170 bHLH transcription factors with complete HLH conserved domains were identified, and these factors were distributed across 14 chromosomes. Their amino acid lengths ranged from 67 to 885 aa, relative molecular masses from 7 910.58 to 98 854.78, and theoretical isoelectric points (pI) from 4.71 to 10.44. Based on phylogenetic analysis, FsbHLHs were grouped into 13 subfamilies, with subfamily III being the largest. Through cis-acting element analysis, it was found that FsbHLHs contained multiple elements associated with light regulation, hormone response, and stress response. Subsequently, the correlation between MeJA-induced expression of eight FsbHLH genes and phillygenin content was analysed, and these results suggested that FsbHLH26 and FsbHLH139 were potentially critical regulators of phillygenin biosynthesis and accumulation. Conclusion This study identified 170 members of the FsbHLH gene family. Through systematic bioinformatics analysis and gene expression pattern analysis following MeJA treatment, FsbHLH26 and FsbHLH139 may be the key transcription factors involved in the biosynthesis of phillygenin, laying the foundation for future research into the molecular mechanisms underlying phillygenin biosynthesis.

Objective To investigate the differences in sterol contents and gene transcription levels in different tissues of Stellaria dichotoma var. lanceolata, and explore key enzyme genes related to the biosynthetic pathway of sterols in this plant. Methods The contents of sterols in the roots, stems, leaves and flowers of S. dichotoma var. lanceolata was determined by high-performance liquid chromatography (HPLC). Combined with full-length transcriptome sequencing and comparative transcriptomic analysis, weighted gene coexpression network analysis (WGCNA) was used to mine key enzyme genes associated with sterol biosynthesis. The expression levels of these genes were further validated using RT-qPCR. Results The sterols contents in the roots, stems, leaves, and flowers of S. dichotoma var. lanceolata exhibited significant differences. A total of 372 483 high-quality full-length transcripts were obtained by transcriptome sequencing, of which 128 646 full-length transcripts were successfully annotated. Differential gene analysis revealed that 43 354 genes were expressed across the four tissues (roots, stems, leaves and flowers), accounting for 50.02% of all total genes. There were 9 647 differentially expressed genes (DEGs) between roots and flowers, 12 143 DEGs between roots and leaves, and only 388 DEGs between leaves and stems. A total of 119 key enzyme genes involved in sterol biosynthesis were identified, and 43 gene co-expression modules were identified by WGCNA. In the MEblue module, six key enzyme genes involved in sterol biosynthesis were identified, including NP_NY_transcript_168676 (FPPS), NP_NY_transcript_335811(SQS), NP_NY_transcript_44001(SQS), NP_NY_ transcript_246835 (CAS), NP_NY_transcript_328932(CAS) and NP_NY_transcript_ 183565(GPPS). The expression levels of the above-mentioned key enzyme genes were validated by RT-qPCR, and the results showed expression trends consistent with those obtained from the transcriptome data. Conclusion This study revealed the key enzyme genes involved in sterol biosynthesis in S. dichotoma L.var. lanceolate, providing a research basis for analyzing the biosynthetic pathways and molecular mechanisms that regulate its active ingredients.

Objective To explore the botanical origin of traditional Chinese medicine Tong-guan-teng and establish a semi-quantitative analysis method, and provide a scientific basis for the quality control and criterion improvement of the medicinal material. Methods Reference sample of Tong-guan-teng, market-available materials, and vine samples of two possible origin plants (Marsdenia tenacissima and M. cavaleriei) were collected. Ultra-high-performance liquid chromatography coupled with charged aerosol detector (UHPLC-CAD) was employed, and an ACQUITY BEH C₁₈column (100 mm×2.1 mm, 1.7 μm) was used. The mobile phase consisted of 0.1% formic acid aqueous solution and acetonitrile, at a flow rate of 0.5 mL/min, with an injection volume of 2 μL. The similarity of sample chromatograms was analyzed by the “Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine (2012 edition)”. Based on the chromatographic peak areas of eight main components in the UHPLC-CAD fingerprint, semi-quantitative analysis of relative contents and total relative contents among batches was carried out. Results The established fingerprint was validated by methodology, with demonstrated satisfactory precision, stability and repeatability, which met the requirements of traditional Chinese medicine fingerprint analysis. The similarity of fingerprints among the reference sample and 12 batches of commercial Tong-guan-teng samples ranged from 0.927 to 0.988. Significant differences were observed between the reference fingerprint of Tong-guan-teng and those of M. tenacissima stems, while a high degree of consistency was achieved with M. cavaleriei stems (similarity > 0.85) and the total contents of major components were relatively consistent. The relative contents of the eight major components followed the order: tenacissoside A > marsdenoside H > marsdenoside K > tenacissoside B > tenacissoside H > tenacissoside D > tenacissoside I > tenacissoside E. Conclusion In addition to tenacissoside H (the index component specified in the 2025 edition of the Chinese Pharmacopoeia), tenacissoside A, marsdenoside H, marsdenoside K and tenacissoside B have relatively high contents, which can serve as potential quality markers and provide a theoretical basis for the botanical origin identification of the traditional Chinese medicine Tong-guan-teng from the perspective of chemical composition.

China has recently proposed the “artificial intelligence +” strategy. Machine vision technology (MVT), which serves as the visual perception interface for artificial intelligence, has demonstrated broad application potential across multiple domains. This review provides a systematic overview of MVT applications throughout the entire industrial chain of Chinese medicinal herbs. By acquiring conventional visual images and multi-source spectral images, and by integrating both traditional machine learning and deep learning algorithms, MVT has been widely implemented in critical processes including seed quality evaluation and sorting, suitability analysis and regional zoning of cultivation environments, plant growth monitoring and field management, harvesting decision-making, primary processing control at the place of origin, and product quality assessment. These applications have significantly enhanced the intelligence level of production processes and the capability for quality control. Nevertheless, prominent challenges remain, such as technological fragmentation, data silos, lab-bound models, and high implementation costs. Guided by the “artificial intelligence +” strategy, this paper further proposes strategic directions including establishing intelligent agent systems covering the entire industrial chain, creating collaborative data resource ecosystems, developing lightweight adaptive algorithm models, and innovating inclusive business models. These measures aim to promote deep integration and large-scale application of MVT in the Chinese medicinal herb industry, thereby providing technical support for intelligent, high-quality development of the entire value chain.

Natural products, characterized by structural diversity and broad biological activities, have become a major source for drug discovery and lead compound identification. However, unclear targets have seriously hindered the research and development of many natural products, making target identification a critical bottleneck and a major focus in this field. In recent years, extensive efforts have been devoted to the discovery of natural product targets, and a relatively systematic methodological framework has gradually been established. According to whether chemical modification is required, current target identification strategies for natural products can be broadly classified into two categories:labeled and label-free approaches. This review systematically summarizes the major target identification techniques within these two strategic frameworks, with particular emphasis on their underlying principles, technical advantages, limitations, applicable scenarios, and representative applications, in order to provide references and insights for mechanistic studies and target discovery of natural products.

The incidence of arthritis has steadily increased in recent years, accompanied by complex pathological changes. Arthritis is among the leading causes of disability worldwide. However, research on its pathogenesis and therapeutic targets remains insufficient, necessitating novel approaches to elucidate disease mechanisms. Single-cell RNA sequencing (scRNA-seq) enables not only the construction of cellular atlases of joint tissues but also the identification of key cell subpopulations and molecular targets driving disease progression, thereby resolving gene expression heterogeneity at the single-cell level. Increasingly applied in arthritis research, scRNA-seq offers new avenues for exploring pathogenesis and developing therapeutic targets. This review systematically summarizes the technical advantages of scRNA-seq, discusses its application prospects and challenges in arthritis pathogenesis and research on treatment with traditional Chinese medicine, and provides a reference for in-depth mechanistic studies and the development of novel Chinese herbal therapeutics.

High altitude disease (HAD) refers to disorders triggered by specific environmental factors such as low oxygen levels, low atmospheric pressure, cold and dry conditions, and strong ultraviolet radiation. Its pathogenesis is complex and often linked to qi deficiency and blood stasis. Hongjingtian (Rhodiolae Crenulatae Radix et Rhizoma), a renowned traditional Chinese medicinal herb, contains diverse active ingredients including phenylethanoid glycosides, flavonoids, phenylpropanoids, volatile oils, organic acids and other active components. It has the effects of nourishing qi, promoting blood circulation, clearing meridians and relieving asthma, exhibiting multiple biological effects in prevention and treatment of HAD. Modern pharmacological studies have confirmed that Rhodiolae Crenulatae Radix et Rhizoma and its active ingredients play a preventive and therapeutic role in HAD through anti-inflammatory, anti-oxidative stress, anti-fatigue, regulation of programmed cell death, improvement of glucose and lipid metabolism, and intestinal flora regulation. This review summarizes the mechanisms and applications of Rhodiolae Crenulatae Radix et Rhizoma and its active ingredients in prevention and treatment of HAD in recent years, providing a reference for clinical research on Rhodiolae Crenulatae Radix et Rhizoma in treatment of HAD.

Qingdai (Indigo Naturalis) is a dried powder, lump, or granule prepared from the leaves or stems and leaves of Baphicacanthus cusia, Polygonum tinctorium or Isatis indigotica. Indigo Naturalis contains inorganic components such as calcium carbonate and silicon dioxide, as well as organic components such as alkaloids, organic acids, diterpenoids, triterpenoids, steroids, etc. Indigo Naturalis has anti-viral, anti-inflammatory, antibacterial, antioxidant, antitumor, and immunomodulatory effects. Indigo Naturalis is clinically used for ulcerative colitis, psoriasis, acute promyelocytic leukemia, atopic dermatitis, etc. Long-term and excessive consumption of Indigo Naturalis can cause toxic side effects such as liver damage, pulmonary arterial hypertension, acute colitis, intussusception, and kidney damage. This article reviews the research progress in the chemical composition, pharmacological effects, clinical applications, and toxic side effects of Indigo Naturalis, providing a reference for the in-depth research and development of Indigo Naturalis.

The treatment of brain diseases is largely hindered by the blood-brain barrier (BBB), which limits drug delivery to the brain and contributes to nonspecific distribution and immune clearance. Although synthetic nanocarriers can improve drug pharmacokinetics, their long-term stability and biosafety remain uncertain. Plant-derived nanovesicles (PDNVs) have emerged as promising therapeutic and delivery platforms owing to their natural lipid bilayers, good biocompatibility, low immunogenicity, scalability, and BBB-crossing potential. Carrying lipids, proteins, nucleic acids, and bioactive small molecules, PDNVs integrate delivery capability with intrinsic therapeutic activity. Component-structure theory interprets the functions of multicomponent systems through elemental structure, quantitative-ratio structure, and phase structure. From this perspective, PDNVs can be regarded as natural composite nanovesicles formed by the ordered integration of endogenous components within membrane-confined spaces. This review discusses the relationships among PDNVs composition, membrane architecture, BBB-crossing delivery, and therapeutic effects in brain diseases, and summarizes engineering strategies for functional optimization. It provides an analytical framework for the rational design, quality control, and optimization of PDNVs as potential central nervous system drug delivery platforms.

Ulcerative colitis (UC) is a chronic, non-specific inflammatory bowel disease. A vicious cycle driven by gut microbiota dysbiosis and disruption of the intestinal barrier is considered a key mechanism underlying its recurrent and refractory nature. Reduced microbial diversity, decreased beneficial bacteria, and increased pathogenic taxa lead to disturbances in metabolites such as short-chain fatty acids and tryptophan, which subsequently impair the mucus layer, tight junction proteins, and immune barrier function, further aggravating mucosal inflammation. According to traditional Chinese medicine (TCM), core pathogenic factors such as “turbidity‑toxin theory” play central roles in UC. Guided by these principles, TCM therapies intervene through multiple approaches, including reshaping gut microbial composition, enhancing the production of beneficial metabolites, and upregulating tight junction proteins. These actions contribute to breaking the aforementioned vicious cycle. This review systematically summarizes the interactions between gut microbiota and the intestinal barrier in UC, as well as the theoretical basis and modern biological mechanisms by which TCM exerts therapeutic effects. The findings provide a theoretical reference for the application of TCM in UC management and highlight its potential clinical value.

The toxicity of traditional Chinese medicines (TCMs), along with the four natures and five flavors, ascending, descending, floating, and sinking, and channel tropism, belongs to the category of pharmacological theory and together constitutes the fundamental principles guiding clinical TCM practices. However, the complex composition of highly toxic medicinal materials makes it difficult to fully elucidate their toxic components and mechanisms of action. Additionally, the diversity of clinical medication regimens and significant individual differences pose considerable challenges to related research. Based on the characteristics of highly toxic medicinal materials, this study systematically reviews the highly toxic medicinal materials included in the 2025 edition of the Chinese Pharmacopoeia. By integrating herbal textual research with modern pharmacological studies, this paper provides an in-depth analysis from multiple perspectives, including common toxic components and pathways, the bidirectional relationship between toxicity and efficacy, and strategies for reducing toxicity while preserving efficacy.Furthermore, it reviews the current application of modern analytical detection techniques in the quality control of these medicinal materials and discusses other potentially toxic substances in TCMs along with risk management strategies. This review aims to serve as a reference for promoting safe and rational clinical use of highly toxic medicinal materials and strengthening the prevention and control of toxicity risks.