Objective The HPLC fingerprint and the content determination method of the benchmark sample of Guizhi Gancao Decoction (GGD, 桂枝甘草汤) were established to study the rule of quantity-quality transmitting of GGD benchmark samples. Methods According to the method recorded in “Treatise on Febrile Diseases”, 15 batches of GGD benchmark samples were prepared and the HPLC fingerprint of 15 batches of benchmark samples was established. The peak attribution and similarity range in the fingerprint were clarified, and the quantity-quality transfer indexes such as the range of extract yield, the content range of index components and the range of transfer rate were analyzed. The quality control system for GGD benchmark samples was preliminarily established. Results A total of 40 common peaks were identified in the 15 batches of GGD benchmark samples, which with good similarity (> 0.99). The contents of protocatechuic acid, liquiritin, coumarin, cinnamyl alcohol, liquiritigenin, cinnamic acid, cinnamaldehyde, and glycyrrhizic acid in the benchmark sample were 0.080—0.087, 0.863—1.055, 0.431—0.560, 0.167—0.227, 0.087—0.127, 0.423—0.492, 0.160—0.199, and 2.437—2.904 mg/g, respectively. The transfer rates of the eight index components from decoction pieces to water decoction were 44.35%—54.84%, 40.56%—44.44%, 48.82%—56.00%, 22.84%—52.61%, 25.70%—30.02%, 33.75%—37.83%, 23.42%—28.98%, and 62.09%—65.13%, respectively. The transfer rates from the water decoction to the reference sample were 67.99%—84.39%, 62.15%—67.93%, 59.42%—74.22%, 76.20%—85.64%, 51.31%—60.73%, 78.80%—85.17%, 16.27%—22.19%, and 71.82%—76.58%, respectively. Overall, the transfer rates of components from decoction pieces to water decoction were relatively low, while the transfer rates of most components increased during the subsequent concentration and freeze-drying processes. Only cinnamaldehyde showed a significant loss during freeze-drying. The dry paste yield of the reference sample was stable, with an average transfer rate of 77.71 %. Conclusion Through the HPLC fingerprint of GGD benchmark samples and the quantity-quality transfer patterns of its active components, a preliminary quality control system based on index components was established. This provides a scientific basis for the standardization and quality consistency evaluation of GGD classical prescription preparations.

Objective To investigate the inhibitory effect and mechanism of pennogenin 3-O-β-chacotrioside (P3C), an active ingredient from Paris polyphylla, on the proliferation of Burkitt’s lymphoma (BL) cells, as well as the combined effect of P3C and the chemotherapeutic drug etoposide (VP-16) on BL cell proliferation. Methods The inhibitory effects of different concentrations of P3C alone or in combination with VP-16 on Raji and CA46 cells proliferation were assessed using CCK-8 assay. Flow cytometry was employed to analyze cell cycle distribution and apoptosis. Western blotting was used to detect the expressions of proteins related to cell cycle, apoptosis and autophagy. qRT-PCR was used to detect the expressions of autophagy-related genes. Using an immunodeficient mouse xenograft model, after P3C intervention, the effect of P3C on tumor growth in vivo was monitored via live imaging. Western blotting was performed to detect the expressions of Ki67 and apoptosis-related proteins cystein-asparate protease-3 (Caspase-3) and poly ADP-ribose polymerase (PARP) in tumor tissues. Results P3C inhibited the viability of Raji and CA46 cells in a dose-dependent manner, with half inhibitory concentration (IC₅₀) values of 1.12, 0.97 μmol/L, respectively, and induced G₀/G₁ phase cell cycle arrest. P3C significantly increased the proportion of apoptotic cells and elevated the expressions of apoptosis-related proteins such as Caspase-3, Caspase-9 and PARP, increased the expressions of autophagy-related genes and proteins such as microtubule-associated protein light chain 3 (LC3), autophagy-related gene 5 (ATG5) and ATG7, decreased the expression of p62, increased the phosphorylation level of p38, while decreased the phosphorylation level of extracellular regulated protein kinases (ERK), all of which were statistically significant compared with control group (P < 0.05, 0.01, 0.001). P3C and etoposide had a good synergistic effect, with a collaboration index (CI) < 1. Compared with the monotherapy group, the drug combination group had a stronger killing effect on Raji and CA46 cells, and further promoted p38 and reduced ERK phosphorylation levels (P < 0.05, 0.01, 0.001). In the in vivo experiments, compared with model group, P3C significantly inhibited tumor growth (P < 0.05, 0.01, 0.001), significantly reduced Ki67 protein expression in tumor tissue (P < 0.001), and significantly increases Caspase-3 and PARP protein expressions (P < 0.05), which was consistent with the results of cell experiments. Conclusion P3C could effectively inhibit the proliferation of Burkitt’s lymphoma cells, induce apoptosis, autophagy, and cell cycle G₀/G₁ arrest, as well as their growth and proliferation in vivo. At the same time, it could enhance the killing effect of chemotherapy drug etoposide on Burkitt’s lymphoma cells, and may achieve anti-tumor effects by regulating p38/ERK signaling pathway.

Objective To investigate the potential targets and mechanism by which curcumin regulates cholesterol metabolism in intestinal epithelial cells based on both in vivo and in vitro experiments. Methods A high-cholesterol model was established in ApoE^-/- mice fed a high-fat diet, followed by intervention with curcumin, lipid levels in serum were measured. Hematoxylin-eosin (HE) staining was used to observe pathological changes in small intestine tissue. Western blotting was used to detect the protein expressions of low-density lipoprotein receptor (LDLR), NPC1 like intracellular cholesterol transporter 1 (NPC1L1), adenosine triphosphate binding cassette transporters G8 (ABCG8), ABCG5 and liver X receptor α (LXRα) in small intestine tissue. An in vitro high-cholesterol model was constructed in rat intestinal epithelial cells-6 (IEC-6) using cholesterol micelle stimulation. After intervention with curcumin or LXRα inhibitor GSK2033, lipid levels were detected by oil red O staining and immunofluorescence. Transwell experiment was used to detect cholesterol transport. Western blotting and immunofluorescence were used to detect cholesterol metabolism-related protein expressions. Results In vivo experiments showed that 100, 200 mg/kg curcumin caused no damage to small intestine tissue structure of ApoE^-/- mice fed with high-fat diet, and could reduce the body weight and lipid levels in serum of mice (P < 0.05), regulate the expressions of proteins related to cholesterol uptake and intestinal cholesterol excretion (TICE) pathway (P < 0.05, 0.01). The regulation results of protein profile in vitro experiments were consistent with the in vivo model, and curcumin could reduce the lipid levels of intestinal epithelial cells in a high cholesterol environment and inhibit cholesterol transport (P < 0.05, 0.01). After further intervention with LXRα inhibitor GSK2033, there was a significant reversal in intracellular lipid levels and expressions of TICE pathway related proteins such as ABCG5, ABCG8 and NPC1L1 (P < 0.05). Conclusion Intestinal epithelial LXRα is an important target of curcumin for promoting cholesterol efflux and inhibiting cholesterol uptake, thereby contributing to the maintenance of systemic cholesterol homeostasis, and its effects may also involve other regulatory pathways.

Objective To investigate the therapeutic effect and potential mechanism of Penthorum chinense (PCP) on primary sclerosing cholangitis (PSC). Methods The chemical constituents of PCP extract were analyzed using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). A total of 50 mice were randomly divided into control group, model group, obeticholic acid (6.5 mg/kg) group and PCP low-, high-dose (2, 4 g/kg) groups, with 10 mice in each group. Mice were fed with 0.1% 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC) feed to induce PSC model. After 14 d of intervention, the efficacy of PCP was evaluated by measuring serum biochemical indicators, liver histopathological changes and liver fibrosis markers. To further explore the mechanisms, transcriptomic sequencing, qRT-PCR experiment and quantitative analysis of bile acid profile were performed. Results A total of 331 chemical constituents were identified in PCP extract. Pharmacodynamic results showed that compared with model group, high-dose PCP significantly reduced liver index and levels of liver injury markers in serum (P < 0.05, 0.01, 0.001), and effectively alleviated cholestasis and fibrosis in liver tissue, with effects superior to those of positive drug obeticholic acid. Transcriptomic analysis identified 51 differentially expressed genes. Enrichment analysis suggested that the effect of PCP was associated with pathways such as bile secretion. Mechanistic validation revealed that compared with model group, high-dose PCP significantly down-regulated mRNA expressions of key enzymes for primary bile acid synthesis in liver tissue (P < 0.05), while up-regulated expression of bile acid‑conjugating enzyme (P < 0.001). Serum and liver bile acid profiling revealed that high-dose PCP intervention significantly increased the ratio of conjugated to unconjugated bile acids (P < 0.001). Conclusion PCP ameliorates DDC‑induced cholestatic liver injury and exerts a therapeutic effect on PSC by inhibiting primary bile acid synthesis, promoting bile acid conjugation and excretion.

Objective To study the anti-anxiety effect of baicalein based on zebrafish model. Methods The zebrafish were divided into control group, model group, baicalein group and diazepam group. Except for the control group, all other groups underwent the induction of a zebrafish anxiety model through unpredictable chronic stress (UCS). After drug intervention, the behavior changes of zebrafish were evaluated through novel tank test. ELISA was used to measure the levels of stress response biomarkers cortisol and brain neurotransmitter serotonin 5-hydroxytryptamine (5-HT). qRT-PCR was employed to detect the gene expressions of tryptophan hydroxylase 2 (TPH2), a rate-limiting enzyme for 5-HT synthesis in brain tissue, 5-hydroxytryptamine 1ab receptors (5-HT_1abR), interleukin-1β (IL-1β), nuclear factor-κB (NF-κB), as well as intestinal IL-1β, IL-6, tumor necrosis factor-α (TNF-α) and NF-κB. Western blotting was used to detect the protein expressions of 5-hydroxytryptamine 1A receptors (5-HT_1AR) in brain tissue. The composition of intestinal microbiota was analyzed using 16S rRNA microbial diversity sequencing technology. Results After UCS treatment, zebrafish exhibited anxiety-like behavior, with a decrease in the total number and diversity of intestinal microbiota species, and an increase in the proportion of Aeromonas. Compared with model group, after intervention with baicalein, the dwell time of zebrafish at the bottom of the tank was significantly reduced (P < 0.001), cortisol level was significantly decreased (P < 0.001), 5-HT level in brain tissue was significantly increased (P < 0.001), gene expression of TPH2 in brain tissue was significantly increased (P < 0.05), gene expressions of IL-1β, NF-κB in brain tissue and IL-1β, IL-6, TNF-α, NF-κB in intestine were significantly decreased (P < 0.05, 0.01, 0.001), the protein expression of 5-HT_1AR in brain tissue was decreased (P < 0.001), species richness and diversity of intestinal microbiota species were increased, the gene expression of 5-HT_1abR in brain tissue was significantly reduced in 2 mg/L baicalein group (P < 0.01), while the gene expression of 5-HT_1abR in brain tissue was significantly increased in 1, 4 mg/L baicalein groups (P < 0.05, 0.001). Conclusion Baicalein has a certain protective effect on UCS-induced anxiety-like behavior in zebrafish, which may be achieved by anti-inflammatory and regulating gut microbiota, inhibiting excessive production of cortisol in zebrafish, maintaining normal secretion of 5-HT, and exerting anti-anxiety effects.

Objective To investigate the effects of Niuxi (Achyranthis Bidentatae Radix, ABR) on tissue distribution, plasma protein binding rate and receptor expression of marker components (amygdalin, hydroxysafflor yellow A, ferulic acid, paeoniflorin, naringin) from Xuefu Zhuyu Decoction (血府逐瘀汤, XFZYD) in rats with metabolic-associated fatty liver disease (MAFLD) of qi stagnation and blood stasis syndrome (QS-BSS), and to further clarify the mechanism underlying its meridian-guiding effect. Methods A rat model of MAFLD with QS-BSS was established by high-fat diet combined with adrenaline injection. After successful modeling, rats were stratified and grouped by body weight, followed by intragastric administration of XFZYD (full formula), XFZYD without Jiegeng (Platycodonis Radix, PR), XFZYD without ABR, and XFZYD without both PR and ABR. For the plasma protein binding rate and receptor expression experiments, abdominal aortic blood was collected at 10, 30 min after a single administration, after which rats were euthanized for the collection of brain, heart, lung, liver, and kidney tissues. For the tissue distribution experiment, rats were euthanized at 10, 30, 60 min after a single administration, with the same tissues collected as above. The concentrations of marker components in tissues were determined by high-performance liquid chromatography-mass spectrometry (HPLC-MS). Qualitative and quantitative analyses of these components in plasma were performed to determine their plasma protein binding rates. The expression levels of platelet-activating factor (PAF) and receptors including adenosine A1 receptor (A1R), P2Y12 receptor, and endothelin A receptor (ETAR) in tissues were measured by ELISA. Results ABR decreased the plasma protein binding rates of amygdalin, hydroxysafflor yellow A, ferulic acid, and naringin in model rats. Meanwhile, ABR increased the distribution of amygdalin and paeoniflorin in the brain and liver, hydroxysafflor yellow A in the liver and kidney, ferulic acid in the brain, heart, liver, and kidney, and naringin in the liver and kidney. Furthermore, ABR upregulated the expression levels of A1R in brain, heart, and liver as well as P2Y12 receptor in the brain, heart, and kidney, while downregulating the expression level of PAF in brain, heart, liver, and kidney and the expression level of ETAR in brain, heart and lung. Conclusion ABR could reduce the plasma protein binding rates of amygdalin, hydroxysafflor yellow A, ferulic acid and naringin in MAFLD-QS-BSS rats, thereby increasing the distribution of these components in the corresponding tissues, enhancing the effect of each component on the related receptors, and ultimately regulating the upregulation or downregulation of receptor expression in various tissues, which synergistically exerts the therapeutic effect against QS-BSS. This may be one of the mechanisms by which ABR exerts its meridian-guiding effect in XFZYD.

Objective To investigate the mechanism of Qingke Pingchuan Granules (清咳平喘颗粒) in the treatment of rats with acute exacerbation of chronic obstructive pulmonary disease (AECOPD) based on the effect of autophagy on cyclic GMP-AMP synthase (cGAS)/stimulator of interferon genes (STING) pathway. Methods Male SD rats were randomly divided into control group, model group, dexamethasone (4 mg/kg) group, Qingke Pingchuan Granules low-, high-dose (2.5, 5.0 g/kg) groups and autophagy activator rapamycin (1 mg/kg) group, with 10 rats in each group. An AECOPD rat model was established using cigarette smoke exposure combined with lipopolysaccharide (LPS) airway instillation. After drug intervention, the general status of rats in each group was observed. The pulmonary function and pathological changes in lung tissue were detected. The numbers of inflammatory cells in peripheral blood and bronchoalveolar lavage fluid (BALF) were analyzed. The levels of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and IL-1β in BALF were measured by ELISA. The mRNA expressions of interferon-β (IFN-β) and chemokine C-X-C motif ligand 10 (CXCL10) in lung tissue were detected by qRT-PCR. The protein expressions of cGAS, STING, p-STING, TANK binding kinase 1 (TBK1), p-TBK1, nuclear factor-κB p65 (NF-κB p65), p-NF-κB p65, microtubule-associated protein light chain 3 (LC3) and autophagy-related gene 5 (ATG5) in lung tissue were determined by Western blotting. Results Compared with model group, the pathological damage and lung function of rats in each treatment group were improved (P < 0.01, 0.001), the numbers of inflammatory cells in peripheral blood and BALF were significantly reduced (P < 0.05, 0.01, 0.001), and the levels of inflammatory factors in BALF were significantly reduced (P < 0.05, 0.01, 0.001). The mRNA expression levels of IFN-β and CXCL10 in lung tissue of rats in Qingke Pingchuan Granules high-dose group and rapamycin group were significantly reduced (P < 0.01, 0.001), the protein expression levels of cGAS, p-STING, p-TBK1 and p-NF-κB p65 were significantly reduced (P < 0.05, 0.01, 0.001), the protein expression levels of LC3-II/I and ATG5 were significantly increased (P < 0.01, 0.001). Conclusion Qingke Pingchuan Granules inhibit cGAS/STING pathway by activating autophagy, thereby alleviating the persistent inflammatory response of AECOPD.

Objective The study employed bioinformatics to identify prognostic genes associated with pancreatic adenocarcinoma (PAAD) and predict Chinese medicines and active ingredients that regulate these genes. Methods The integration of data from the TCGA, GTEx, and GEO databases was conducted, then screening the prognostic genes through differential gene analysis, COX regression analysis, and survival analysis. The subsequent step involves prognostic efficacy analysis and a verification of the expression trends of the identified key genes. Concurrently perform gene ontology (GO) functional and Kyoto encyclopedia of genes and genomes (KEGG) pathway analyses, as well as gene set enrichment analysis (GSEA), to identify potential pathways and biological processes. The impact of feature genes on the infiltration status of immune cells was analyzed, and their correlation with immune checkpoints was further investigated, and tumor immune escape capacity and tumor mutation burden were predicted. Validation of gene expression patterns at the single-cell level. The DSigDB, CTD, and HERB databases are used to predict the potential active ingredients of Chinese medicine and perform molecular docking. Additionally, pharmacokinetic analysis is conducted using Swiss ADME and prediction and analysis of Chinese medicine by Coremine Medical database. Cellular experiments were conducted to validate the regulatory effect of the key active ingredient on the characteristic gene. Results The investigation ultimately led to the identification of the key gene interferon alpha inducible protein 27 (IFI27), which exhibited significantly elevated expression levels in PAAD tissues. Patients exhibiting low IFI27 gene expression demonstrated superior overall survival. Enrichment analysis indicated its potential involvement in various biological processes, including apoptosis, ECM-receptor interaction, regulation of T cell activation, macrophage migration, and signaling pathways such as PI3K-Akt and NF-κB. Immune infiltration analysis indicates that IFI27 may regulate the infiltration status of various immune cells, such as B cells naive, T cells CD8⁺, T cells regulatory (Tregs), and macrophages M0, suggesting that it has an important function in regulating the tumor immune microenvironment. Meanwhile, the expression of IFI27 showed significant positive correlations with multiple immune checkpoint molecules. Further studies have revealed that low expression of IFI27 is significantly associated with the response to immunotherapy, supporting its value as a therapeutic target. Single-cell RNA sequencing verified IFI27 upregulation in PAAD, pinpointing its expression to macrophages and other immune cells. Experiments confirmed that fisetin significantly downregulated both mRNA and protein expression levels of IFI27 in pancreatic cancer cells. Furthermore, a total of 22 Chinese medicines, including Jianghuang (Curcumae Longae Rhizoma), Zaojiaoci (Gleditsiae Spina), and Yujin (Curcumae Radix), were predicted. The four qi encompass cold and warm, and the five flavors are primarily bitter, sweet, and pungent, with a focus on the liver, spleen, and stomach meridians. The main effects are activating blood and resolving stasis, moving qi to relieve pain, and clearing heat and detoxifying, combined with tonifying and inducing diuresis. Conclusion The results of this study imply that IFI27 may serve as a prognostic gene related to PAAD immunity, while fisetin and various Chinese medicines may serve as potential therapeutic drugs.

Objective To investigate the medication rules, core formula, and multi-dimensional mechanisms of traditional Chinese medicine (TCM) in the treatment of cancer cachexia (CC). Methods Medication rules and the core formula were screened from 211 publications through data mining. Network pharmacology was subsequently applied to identify the active components and potential targets of the core formula, followed by gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analyses and molecular docking validation. Finally, the therapeutic efficacy and mechanisms were validated in vivo using an A549 tumor-bearing nude mouse CC model. Evaluation indicators included tumor-free body weight, food intake, muscle function, inflammatory cytokines [interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α)], and key proteins [muscle atrophy F-box protein (Atrogin-1), muscle RING-finger protein-1 (MuRF1), protein kinase B1 (Akt1), and hypoxia-inducible factor-1α (HIF-1α)]. Results A total of 67 prescriptions involving 171 drugs were identified, and the core formula was determined as Fuling (Poria), Baizhu (Atractylodis Macrocephalae Rhizoma), Huangqi (Astragali Radix), Shanyao (Dioscoreae Rhizoma), and Zhigancao (Glycyrrhizae Radix et Rhizoma Praeparata cum Melle). Network pharmacology predicted 149 potential targets involved in the PI3K-Akt, HIF-1α, p53, and apoptosis signaling pathways. Molecular docking showed that core components, such as quercetin, exhibited strong binding affinities with key targets. In vivo experiments demonstrated that the high-dose core formula exhibited the best therapeutic effect, significantly improving anorexia, maintaining tumor-free body weight, and restoring peak grip strength and quadriceps wet weight (all P < 0.05). Mechanistically, the core formula significantly reduced serum levels of IL-6 and TNF-α. Furthermore, it markedly inhibited the expression of muscle-atrophy-related proteins Atrogin-1 and MuRF1 in the gastrocnemius muscle. It also upregulated the expression of the anabolic protein Akt1 and anti-apoptotic protein B-cell lymphoma-2 (Bcl-2), while downregulating the expression of metabolic reprogramming and pro-apoptotic proteins, including HIF-1α, tumor protein p53 (TP53), and cellular-Myc (MYC) (all P < 0.05). These results were consistent with the regulatory patterns predicted for the HIF-1α and PI3K-Akt signaling pathways. Conclusion The core TCM formula for treating CC follows the therapeutic principles of invigorating the spleen, resolving dampness, supplementing qi, and strengthening healthy qi. Its mechanism likely involves the modulation of the PI3K-Akt/HIF-1α signaling axis to comprehensively regulate metabolic reprogramming, inflammatory response, and cell apoptosis while effectively inhibiting the ubiquitin-proteasome pathway, thereby exerting a multi-dimensional and holistic anti-cachexia effect.

Objective To systematically evaluate the current evidence from randomized controlled trials (RCTs) on traditional Chinese medicine (TCM) for pulmonary nodules (including early-stage lung cancer), and to provide evidence-based recommendations for future clinical practice and research. Methods Comprehensive searches were conducted across the following databases up to September 2025: Intelligent Clinical Evidence Database of Chinese Patent Medicine, China national knowledge infrastructur (CNKI), VIP Journal Database, Wanfang Data Knowledge Service Platform, Chinese biomedical literature service system (SinoMed), PubMed, Web of Science, Embase, and the Cochrane Library. Extracted data included key characteristics of participants and studies, such as disease subtype, TCM syndrome, year of publication, sample size, TCM therapeutic principles, details of treatment and control interventions, duration of treatment and follow-up, methodological details, and outcome measures. For studies involving Chinese patent medicines, extracted data were categorized using the Aireview Agent and subsequently verified manually. Data from other TCM studies were extracted and verified manually. Results A total of 70 RCTs were included, with the greatest number published in 2024. The studies showed that deficiency of both qi and yin syndrome and phlegm-stasis mutual binding syndrome were the most common TCM syndromes. The main therapeutic principles were resolving phlegm, tonifying qi, and dispersing nodules. In the treatment groups, Chinese herbal decoctions were the predominant intervention. Clinical efficacy was the most commonly reported outcome measure. In the risk-of-bias assessment, the measurement of the outcome domain was most frequently rated at high risk of bias. Conclusion RCTs of TCM interventions for pulmonary nodules (including early-stage lung cancer) exhibited several critical limitations, such as inadequate specification of disease subtypes in participant eligibility criteria, inconsistent TCM syndrome differentiation, failure to report or analyze primary endpoints, and a high risk of bias—particularly in outcome measurement. We recommend developing a core outcome set for TCM interventions in pulmonary nodules. Future clinical trials—whether decentralized or large-scale, multicenter randomized controlled trials should be stratified by disease subtype, use standardized protocols, and strictly adhere to established follow-up guidelines to generate high-quality evidence, particularly for nodules at elevated risk of malignancy.