Objective To investigate the mechanism of Wumei Pill (乌梅丸) in delaying the transformation of colitis cancer using microbiome and metabolomics. Methods A colitis cancer transformation animal model was constructed using azoxymethane (AOM)/dextran sulfate sodium (DSS). A total of 32 C57BL/6J mice were randomly divided into control group, model group, and Wumei Pill low-, high-dose (11.18, 22.36 g/kg) groups. Colonic tumor progression and histopathological changes were evaluated by intestinal endoscopic observation, macroscopic specimen assessment and hematoxylin-eosin (HE) staining. Levels of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) in serum, and levels of total bile acids (TBA), cholic acid (CA), deoxycholic acid (DCA), lithocholic acid (LCA) in serum and in feces were measured by ELISA. 16S rRNA sequencing technology was used to analyze gut microbial diversity and species abundance. The abundance of metabolites was assessed by non-targeted metabolomics analysis. qRT-PCR was used to verify the expression changes of Clostridium scindens (C. scindens), Clostridium hylemonae (C. hylemonae), Clostridium hiranonis (C. hiranonis) and the coenzyme A transferase encoding gene BaiF. Results Compared with model group, the intervention of Wumei Pill could delay the transformation of colitis cancer, and the therapeutic effect was better at low dose of Wumei Pill. Low dose Wumei Pill could significantly reduce the number of colon tumors (P < 0.001), lower colitis and tumor pathology scores (P < 0.05, 0.01), and lower levels of IL-6 and TNF-α in serum (P < 0.01, 0.001). The microbiological results showed that Wumei Pill could improve the structure of intestinal microbiota, reduce the abundance of opportunistic pathogenic bacteria such as Clostridium, and increase the abundance of probiotics such as Lactobacillus and Roseburia. The non-targeted metabolomics results showed that there were 82 differential metabolites between Wumei Pill low-dose group and model group, significantly enriched in secondary bile acid metabolism pathway. Wumei Pill could significantly reduce the levels of TBA, CA,and DCA in serum and feces (P < 0.05, 0.01, 0.001). qRT PCR results further confirmed that Wumei Pill could inhibit bacteria involved in DCA synthesis such as C. scindens and C. hylmonae (P < 0.05), and lead to a decrease in BaiF gene expression level (P < 0.001). Conclusion Wumei Pill could alleviate colitis and delay the progression of inflammatory cancer by modulating the bile acid homeostasis mediated by gut microbiota, reducing the abundance and dehydroxylation function of bacteria such as C. scindens, and decreasing the production of DCA.

Objective To identify the blood-absorbed constituents of ethanol extract of Tetrastigma hemsleyanum (SYQ) and investigate its anti-breast cancer activity and underlying mechanisms. Methods Blood-absorbed components of SYQ were identified using ultra-high performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry. The effects of SYQ drug containing serum on proliferation and migration of 4T1 cells were evaluated by MTT, colony formation and wound-healing assays. A 4T1 tumor-bearing mouse model was established, control group, model group, cyclophosphamide (20 mg/kg) group, SYQ low-, medium-, and high-dose (5, 10, 20 g/kg) groups were set up. After drug intervention, ELISA was used to detect the levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in serum. Hematoxylin-eosin (HE) staining was used to observe pathological changes in tumor tissue. Immunohistochemistry was used to detect CD68 and Ki67 expressions in tumor tissue. Western blotting was used to detect the expressions of TNF-α/protein kinase B (Akt)/signal transducer and activator of transcription 3 (STAT3) signaling axis and apoptosis related proteins in tumor tissues. Results A total of 19 blood components were identified by UHPLC-Q-TOF-MS/MS, mainly including flavonoids (rutin, tangeretin, narcissoside), phenylpropanoids, and other glycoside compounds (forsythoside E, darendoside B). The in vitro experimental results showed that SYQ drug containing serum significantly inhibited the proliferation, migration and colony formation of 4T1 cells (P < 0.01, 0.001). The in vivo experimental results showed that SYQ significantly inhibited tumor volume growth in tumor bearing mice (P < 0.001), with a tumor inhibition rate of 25.95% in high-dose group. SYQ reduced the levels of IL-6 and TNF-α in serum (P < 0.05, 0.001), induced tumor tissue necrosis and significantly reduced the expressions of CD68 and Ki67 (P < 0.001), while downregulated the expression levels of TNF-α, p-Akt/Akt, p-STAT3/STAT3 and B-cell lymphoma-2 (Bcl-2) in tumor tissue (P < 0.05, 0.01, 0.001). Conclusion SYQ could exerts anti-breast cancer effects by reducing the levels of inflammatory factors, inhibiting tumor cell proliferation and inducing tumor tissue necrosis. The mechanism may be related to the inhibition of TNF-α/Akt/STAT3 signaling axis.

Objective To evaluate the clinical efficacy, pharmacoeconomic value, and potential for reducing antibiotic use of Liushen Capsules in the treatment of acute tonsillitis. Methods A multicenter, prospective cohort study was conducted, enrolling 875 patients with acute tonsillitis (660 in the exposed group, 215 in the non-exposed group). Comparisons were made between the two groups regarding sore throat Visual Analog Scale (VAS) scores, disease recovery rate, efficacy of traditional Chinese medicine (TCM) syndrome differentiation, as well as antibiotic and antipyretic-analgesic usage rates. Pharmacoeconomic evaluation and multivariate logistic regression analysis were also performed. Results After seven days of treatment, the exposed group showed significantly better outcomes than the non-exposed group in terms of reduction in sore throat VAS score, improvement in TCM syndrome recovery rate, and reduction in antibiotic use rate (P<0.05).. Pharmacoeconomic cost-minimizationt analysis indicated that the exposed group was more cost-effective than the non-exposed group. Conclusion Liushen Capsules can effectively alleviate symptoms, improve clinical efficacy, significantly reduce antibiotic use, and demonstrate favorable pharmacoeconomic value in the treatment of acute tonsillitis.

Objective To explore the syndrome differentiation and medication, and formulation rules of renowned traditional Chinese medicine (TCM) practitioners in treating type 2 diabetes mellitus (T2DM) using data mining technology, and validate the efficacy of the identified core formula using an animal model. Methods A total of 200 medical cases of T2DM treated by famous TCM practitioners were collected from databases such as China National Knowledge Infrastructure (CNKI), Wangfang, VIP. A database was established using the TCM Inheritance Computing Platform (V3.5). The silhouette coefficient was introduced to evaluate K-means clustering performance, and hierarchical clustering was used for cross-validation to screen for core formulas. For the experimental validation, 40 male Sprague-Dawley (SD) rats were randomly divided into four groups (n = 10 each): control, model, core formula, and metformin. Except for the control group, T2DM models with “qi-yin deficiency” syndrome were induced by a 4-week high-fat and high-sugar diet combined with daily oral administration of Qingpi Fuzi decoction (15.75 g/kg), followed by an intraperitoneal injection of streptozotocin (STZ, 30 mg/kg). After successful modeling, the core formula group received the core formula (10.5 g/kg), and the Metformin group received metformin (200 mg/kg) via gavage daily; Control and Model groups received equal volumes of saline, each group was given the drug once a day for four weeks of intervention. Body weight (BW) and fasting blood glucose (FBG) were monitored during the administration period. Serum levels of insulin (INS), glycated hemoglobin (IFCC-HbA1c), interleukin-1β (IL-1β), IL-6, superoxide dismutase (SOD), and malondialdehyde (MDA) were measured by ELISA. The proportion of insulin-positive areas was assessed via immunohistochemical staining. Oil red O staining was used to observe lipid deposition in the liver. PAS staining was used to evaluate the pathological changes of renal tissue. Protein expressions of the phosphatidylinositol-3-hydroxykinase (PI3K)/protein kinase B (Akt), nuclear factor-κB (NF-κB), nuclear factor erythroid-2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) pathways in liver tissues were detected by Western blotting. Results Data mining revealed that qi-yin deficiency was the primary syndrome of T2DM. High-frequency herbs included Huangqi (Astragali Radix), Huanglian (Coptidis Rhizoma), and Fuling (Poria). The medicinal properties were predominantly cold, warm, and neutral, while flavors were mainly sweet, bitter, and pungent, targeting the spleen, lung, and liver meridians. These types of drugs such as tonifying deficiency, clearing heat, promoting diuresis, and activating blood circulation were most prevalent. Association rule analysis identified Astragali Radix-Gengen (Puerariae Lobatae Radix) and Astragali Radix-Coptidis Rhizoma as core herb pairs. Clustering identified a core formula consisting of Astragali Radix, Shanyao (Dioscoreae Rhizoma), Puerariae Lobatae Radix, Shanzhuyu (Corni Fructus), and Poria. In vivo experiments showed that the core formula group had significantly lower FBG, INS, and IFCC-HbA1c levels compared to the Model group (P < 0.05, 0.01, 0.001), with a gradual recovery in BW. Furthermore, IL-1β and IL-6 were significantly decreased (P < 0.01, 0.001), SOD was increased (P < 0.01), and MDA was decreased (P < 0.05). Pathologically, the core formula group showed a significantly higher ratio of insulin-positive area (P < 0.05), reduced hepatic lipid deposition (P < 0.01), and lower renal pathological injury scores (P < 0.05). Western blotting analysis indicated that core formula significantly up-regulated the expression of p-PI3K/PI3K, p-Akt/Akt, Nrf2, and HO-1 (P < 0.05, 0.01), while down-regulating p-NF-κB p65/NF-κB p65 (P < 0.05). Conclusion The core formula exhibits a definite hypoglycemic effect. Its mechanism of action is associated with activating the PI3K/Akt signaling pathway to enhance insulin sensitivity, inhibiting NF-κB-mediated pro-inflammatory responses, and up-regulating the Nrf2/HO-1 pathway to ameliorate oxidative stress. This formula effectively maintains the number and structure of pancreatic islet cells and protects liver and kidney tissues from diabetic pathological damage, providing a scientific basis for the prevention and treatment of T2DM with TCM.

Objective To improve the YOLOv8 model and provide a high-accuracy and high-efficiency automated detection solution for Chinese herbal slices in practical scenarios such as production, dispensing, and teaching. Methods Taking YOLOv8 as the baseline model, a lightweight residual depthwise-attention Bottleneck (RDA-Bottleneck) is proposed to replace the Bottleneck blocks in C2f, reduce redundant computations and enhance the expression of channel features. To strengthen the discriminative capability of the detector during multi-scale feature learning, a frequency-aware spatial attention (FASA) module is introduced to replace the Conv blocks in the Backbone and Neck of YOLOv8. To evaluate the performance of the improved model, a dedicated dataset containing 10 categories of Chinese herbal slices [e.g., Banlangen (Isatidis Radix) and Gancao (Glycyrrhizae Radix et Rhizoma)] is constructed, comprising 8 281 images. Results Compared with YOLOv8, the proposed model reduces parameters by 39.5% and floating point operations (FLOPs) by 34.1%, while improving single-threshold average accuracy mAP50 by 0.2% and multi-threshold average accuracy mAP50-95 by 0.2%. Conclusion The improved model achieves higher detection accuracy and inference efficiency for Chinese herbal slice detection under complex backgrounds and diverse appearances, providing an effective method for automatic detection of Chinese herbal slices.

Objective To reveal the dynamic evolution of the global innovation cooperation network in the field of pentacyclic triterpenoids, and analyze the collaborative evolution mechanism of network structure, national innovation strategies and individual behaviors at different stages of the technology lifecycle. Methods Based on global patent data from 1945 to 2025, social network analysis method is used to construct and analyze the innovation cooperation network in this field. According to the theory of technology lifecycle, four development stages are divided for dynamic comparison. Results The cooperative network structure presents a phased evolution, from a closed “core small world” in its infancy to a long-term expansion, ultimately converging into a “strategic focused” tight structure; The national innovation strategy is divided into three types of paths: “sustained openness”“first rising and then falling”“low-level stability”. The intensity of international cooperation decreases in an “inverted U-shape” as technology matures; There is a significant “structural fragmentation” among micro entities, with the industry forming a high-density collaborative closed loop, while key academic institutions are often in a “bilateral isolation” state at the edge of the network, leading to an ecological imbalance of “industry academia decoupling”. Conclusion This study systematically reveals the dynamic evolution mechanism of innovation cooperation networks in the field of pentacyclic triterpenoids from three levels: network structure, national strategy, and subject behavior. It deepens the theoretical understanding of cooperation and competition relationships in the technology lifecycle and provides empirical evidence and policy implications for building a more resilient innovation ecosystem.

Objective Based on transcriptome data from roots of Panax notoginseng under arsenic (As) stress, the tonoplast intrinsic protein gene PnTIP1;3 was cloned. Its functional characteristics in As transport were investigated through bioinformatic analysis and heterologous expression in yeast. Methods Bioinformatic tools were used to analyze the sequence features and promoter cis-acting elements of PnTIP1;3. A yeast heterologous expression system was constructed, and the role of PnTIP1;3 in cellular As transport was systematically evaluated through phenotypic assays, intracellular As content and subcellular distribution measurement by ICP-MS, and analysis of relevant antioxidant indicators. Results The tonoplast intrinsic protein gene PnTIP1;3 was cloned, with an open reading frame of 750 bp encoding 250 amino acids. Phylogenetic analysis indicated that it belongs to the TIP1 subfamily. Promoter analysis revealed the presence of multiple cis-acting elements related to hormones, growth and development, and stress responses. As tolerance assays showed that the half-maximal effective concentration (EC₅₀) of yeast transformants heterologously expressing PnTIP1;3 (TPnTIP1;3) was 5.36 mmol/L, higher than that of the empty vector control (5.03 mmol/L). As treatment inhibited the growth of both strains, but the empty vector control was more severely affected. ICP-MS analysis indicated that total As accumulation was significantly higher in TPnTIP1;3 than in the control, and the subcellular distribution of As followed the pattern: cell wall > vacuole > cytoplasm. Under As stress, reactive oxygen species (ROS) levels were significantly decreased (P < 0.001) in TPnTIP1;3, while the activities of superoxide dismutase (SOD) and catalase (CAT), as well as the contents of glutathione (GSH) and metallothionein (MT), were significantly increased (P < 0.05). Correlation analysis further demonstrated significant positive relationships between these antioxidant indicators and both total As content and cell wall As concentration. Conclusion The P. notoginseng tonoplast intrinsic protein gene PnTIP1;3, a member of the TIP1 subfamily, enhances yeast tolerance to As stress by promoting As accumulation and vacuolar sequestration, and through activation of the host antioxidant system.

Objective To identify members of the WRKY transcription factor family in Codonopsis pilosula at the genome-wide level and analyze their characteristics, and explore their potential roles in response to salt stress, thereby laying a foundation for functional studies of C. pilosula WRKY transcription factors. Methods Potential gene family members were screened across the entire genome based on conserved protein domains using the Hidden Markov model (HMM). Protein sequence alignment, conserved domain analysis, phylogenetic tree construction, gene synteny analysis, and cis-acting element analysis in the promoter region were performed using software and online tools including MAFFT, IQ-TREE, MEME, Plant CARE and TBtools. Expression patterns of WRKY genes were analyzed based on transcriptome data from different tissues (roots, stems, leaves, and flowers) and qRT-PCR was used to determine their relative expression levels. In addition, the transcriptome data of C. pilosula treated with 0, 60, and 120 mmol/L NaCl were analyzed to examine the expression changes of WRKY genes under salt stress. Results A total of 50 WRKY transcription factor members were identified in C. pilosula. Motif analysis showed that all members contained the typical WRKY domain with the conserved sequence WRKYGQK. Phylogenetic analysis with WRKY transcription factors from Arabidopsis thaliana and Oryza sativa classified the 50 C. pilosula WRKY transcription factors into three groups, with group II further divided into five subgroups. The amino acid lengths of the identified WRKY proteins ranged from 103 to 986 residues, with isoelectric points (pI) of 4.87−9.99 and molecular weights (MW) of 12 435.14−107 946.28. Gene expression analysis revealed significant differences in the expression levels of CpWRKY26, CpWRKY48, CpWRKY2, and CpWRKY49 among roots, stems, leaves, and flowers of C. pilosula. The results of salt stress treatment indicated that most CpWRKY genes showed differential expression trends under different concentrations of NaCl, and some members cluster phylogenetically with previously reported salt-tolerant WRKY genes, suggesting that they may be involved in the regulation of C. pilosula salt stress responses. Conclusion This study represents the first systematic identification and bioinformatics analysis of the WRKY gene family in C. pilosula at the genome-wide level. By combining tissue expression and salt stress response analyses, the study reveales that the WRKY genes in C. pilosula may be involved in physiological processes such as growth and development, secondary metabolite biosynthesis, and stress responses. This lays a theoretical foundation for further research on the functions of WRKY genes and their roles in regulating medicinal components, and provides gene resources and scientific evidence for molecular-assisted breeding of C. pilosula.

Objective To clone the CtFD1 and CtFD2 genes from the genome of Carthamus tinctorius and perform bioinformatics, expression specificity, and subcellular localization analyses. Methods The CtFD1 and CtFD2 genes were screened and cloned based on the C. tinctorius genome. Bioinformatics online tools were used to analyze physicochemical properties, conserved domains, and other features. Quantitative real-time polymerase chain reaction (qRT-PCR) was employed to examine tissue-specific expression patterns. Subcellular localization was observed using a tobacco transient expression system. Soluble expression of the proteins was achieved by optimizing inducer concentration and temperature. Results The coding sequences (CDS) of CtFD1 and CtFD2 were 726 bp and 540 bp in length, encoding 241 and 179 amino acids, respectively. Both proteins contained a typical bZIP transcription factor binding domain, which were hydrophilic, and localized to the nucleus. Phylogenetic analysis indicated a close evolutionary relationship with Artemisia annua. Tissue expression analysis showed that CtFD1 and CtFD2 were most highly expressed in shoot apical meristem (SAM). Prokaryotic expression analysis revealed that both proteins were abundantly expressed in the supernatant at 16  ℃ with an IPTG concentration of 0.2 mmol/L. Conclusion The CtFD1 and CtFD2 genes belong to the bZIP family and exhibit the highest expression levels in the SAM of C. tinctorius, suggesting that they may play important roles in regulating flowering. This study provides a scientific basis for further functional characterization of these genes and elucidation of the flowering mechanism in C. tinctorius.

Objective To establish a detection method that combines the appearance characteristics (color, odor, taste) and internal chemical components of Baihe (Lilii Bulbus) to conduct a comprehensive evaluation of the overall quality of lily medicinal materials. Methods Using intelligent sensory technology to detect the color, odor, and taste of 15 batches Lilii Bulbus from the perspectives of vision, smell, and taste. At the same time, HPLC fingerprint spectra and multi-index content determinations were established to compare the content differences of specific components (regaloside C, regaloside A, 2-acetate regaloside A and regaloside E) among medicinal materials from the perspective of chemical composition. Multivariate statistical analysis methods such as hierarchical cluster analysis (HCA), principal component analysis (PCA), and partial least squares discriminant analysis (PLS-DA) were used to analyze the color, odor, taste, and fingerprint spectra of Lilii Bulbus from different origins, and screen for differential components. Pearson correlation analysis was used to reveal the correlation between appearance color, odor, taste, and exclusive ingredients. Results The results of the colorimeter showed that the colors of the 15 batches of Lilii Bulbus medicinal materials were all bright white and lighter, and Zhejiang and Hunan were whiter than those of Jiangsu. The electronic nose analysis results indicated that sensors W3S, W5C, W6S, and W1S, corresponding to 15 batches of Lilii Bulbus medicinal materials contained relatively high levels of alkanes and aromatic compounds. However, Zhejiang Lilii Bulbus also exhibited elevated hydrides. Electronic tongue results revealed that all taste indicators showed obvious responses except sourness. Intelligent sensory analysis demonstrated that Lilii Bulbus from different regions exhibited significant differences in color, odor and taste. A total of 15 common peaks were identified in the fingerprint spectra. Content determination results indicated obvious quality differences among Lilii Bulbus samples from different origins. Among them, Peak 13, peak 6 (regaloside A) and peak 11 (regaloside E) were the main marker components causing compositional differences of Lilii Bulbus from different producing areas. The correlation analysis results indicated that yellow blue chromaticity(b^*), total color value(E^*ab), W1S, W6S, sweetness, astringency, aftertaste-A in Lilii Bulbus were correlated with regaloside C, regaloside A, 2-acetate regaloside A and regaloside E. Conclusion The method of integrating intelligent sensory technology with chemical composition research for comprehensive quality evaluation of Lilii Bulbus medicinal materials is feasible. It can be used to evaluate the comprehensive quality of Lilii Bulbus from different origins, providing reference for the quality control and application of Lilii Bulbus.