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 AsMAPK1 and AsMAPK2 are two mitogen-activated protein kinase (MAPK) genes with unknown functions in Aquilaria sinensis. To investigate their fundamental characteristics and potential functions in wound-induced agarwood formation, in this study, we performed cloning, expression analysis, and preliminary in vitro functional assays on the two genes. Methods Using cDNA from subcultured A. sinensis callus as the template, the coding sequences of AsMAPK1 and AsMAPK2 were obtained by PCR amplification. Bioinformatics analysis was performed using relevant online tools and softwares. The prokaryotic expression vectors pGEX4T-1-AsMAPK1/2 were constructed, transformed into E. coli BL21 for expression, and induction conditions were optimized. The AsMAPK1-GFP and AsMAPK2-GFP fusion expression vectors were constructed and transformed into Arabidopsis thaliana protoplasts to observe their subcellular localization using confocal microscopy. The kinase activity of the purified proteins was detected using an in vitro phosphorylation assay. Results The AsMAPK1 (1 188 bp) and AsMAPK2 (1 128 bp) genes were successfully cloned. Bioinformatics analysis revealed that both encoded proteins contain typical kinase domains and belong to the MAPK family. Phylogenetic analysis indicated that AsMAPK1 is most closely related to the AtMAPK6 homolog in A. thaliana and belongs to Group A, while AsMAPK2 clusters with Group B AtMAPK13. SDS-PAGE detection showed successful induction of soluble recombinant proteins of approximately 71 000 and 69 000. Subcellular localization results demonstrated that both AsMAPK1-GFP and AsMAPK2-GFP are localized in the cytoplasm and nucleus. Further kinase activity analysis confirmed that both AsMAPK1 and AsMAPK2 undergo autophosphorylation and can phosphorylate the universal substrate MBP. Conclusion This study confirms that AsMAPK1 and AsMAPK2 are active protein kinases localized in both the cytoplasm and nucleus. The cloning, expression analysis, and kinase activity detection of the two AsMAPKs provide a foundation for further elucidating their roles in the signal transduction mechanism underlying wound-induced agarwood formation in A. sinensis.

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 prepare naringenin micelles-loaded thermosensitive gel (NMTG), evaluate its biological safety and anti-inflammatory activity, and investigate its therapeutic effects on dry eye disease (DED) induced by benzalkonium chloride (BAC) in rats. Methods Naringenin micelles (NM) were prepared by the thin-film dispersion method. Quality evaluations of the prepared NM included appearance, particle size, ζ potential, transmission electron microscopy, encapsulation efficiency, drug loading and stability. Concentrations of Poloxamer 188 (P188) and Poloxamer 407 (P407) were optimized based on the phase transition temperatures (T_gel and T_{atd gel}) of NMTG before and after dilution with tear fluid. Optimal concentrations of P188 and P407 were dissolved in NM to form NMTG. Biological safety, anti-inflammatory activity, and therapeutic effects on BAC-induced DED rats were investigated. Results Prepared NM was clear and transparent. Transmission electron microscopy showed near-spherical particles with an average particle size of (21.56 ± 0.16) nm, a polydispersity index (PDI) value of 0.053 ± 0.033, and a ζ potential of (-18.63 ± 1.94) mV. Encapsulation efficiency and drug loading were (73.96 ± 0.24)% and (7.34 ± 0.38)%, respectively, and the stability was good within 14 d. The optimal NMTG was achieved with concentrations of 5.29% P188 and 16.24% P407. NMTG demonstrated good biological safety and anti-inflammatory activity. Pharmacological experiments indicated that NMTG showed favorable therapeutic effects on DED rats. Conclusion The successful preparation of NMTG improve the solubility of naringenin, which endow it with good DED therapeutic effects. This will provide support for new indications of naringenin in ocular applications and offer candidate solutions for the development of new naringenin dosage forms.

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 Through multidimensional dynamic correlation analysis of the “appearance, nature, odor, and texture” during the processing of Mifuzhiqiao (honey bran-fried Aurantii Fructus, hbAF), the multidimensional characterization variation patterns during the processing of hbAF were explored. Methods Headspace-gas chromatography-mass spectrometry (HS-GC-MS) and HPLC were employed to identify and analyze the volatile components, drying ingredients and effective ingredients of Chinese herbal decoction pieces. Intelligent sensory technologies such as colorimeters, high-resolution cameras, texture analyzers, and electronic noses were combined with methods including principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) to evaluate indicators of the herbal decoction pieces, such as color, texture, and odor, and convert these indicators into objective and quantitative characterization information. Pearson correlation analysis combined with the Mantel test was used to analyze the relationships between key quality parameters of the “external characteristics-internal components” of the herbal decoction pieces. Results The study found that the color of the herbal decoction pieces at the optimal processing time (100—125 s) was consistent with the results of experience-based identification by traditional senior herbal processing technicians, with the color parameters showing a^* 5.05—5.50, b^* 38.01—39.25, and a moderate texture (65.15—69.06 N). During herbal processing, volatile components undergo phased evolution, with their characteristic odors gradually changing from fruity and baking aromas to a scorched aroma. Quantitative component analysis indicates that drying ingredients are significantly reduced after processing, while the components responsible for the “regulating vital energy and eliminating flatulence” effect have relatively higher contents at the properly processing stage, which corroborate the processing theory of “reducing dryness and enhancing efficacy”. Pearson correlation analysis showed that the external quality indicators of the samples, such as color and texture, were significantly correlated with the contents of components including 1,3,8-menthatriene, (+)-α-magnolene, γ-magnolene, and 3,5,6,7,8,3′,4′-heptemthoxyflavone. Conclusion Through the comprehensive analysis of objective characteristics such as the “appearance, nature, odor, and texture” of the samples, the law of quality changes of hbAF throughout the entire processing process can be preliminarily evaluated, which provides a reference for the rapid identification of the processing degree and quality control of hbAF.

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 prepare hyaluronic acid-modified polyphyllin VII-loaded metal-organic framework material IRMOF-10 targeted nanoparticles (HA@IRMOF-10@PPVII), systematically evaluate their physicochemical properties, safety and in vitro anti-triple negative breast cancer (TNBC) activity. Methods Metal-organic framework material IRMOF-10 was synthesized by triethylamine method. PPVII was encapsulated by the heterotopic encapsulation method, and the surface of the drug-loaded nanoparticles was modified with hyaluronic acid to construct the targeted nanoparticles HA@IRMOF-10@PPVII. The drug loading was determined by HPLC. The morphology, structure and thermal stability of nanoparticles were characterized by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), thermogravimetric analysis (TG) and BET specific surface area method. Its blood safety was evaluated by hemolysis test, and its cell targeting was verified by laser confocal microscopy. CCK-8, DAPI staining, Annexin V/PI double staining, migration assay, clone formation assay, cell uptake, reactive oxygen species (ROS) detection, mitochondrial membrane potential (MMP) detection and other methods were used to verify its inhibitory effect on TNBC 4T1 cells. Results The drug loading capacity of HA@IRMOF-10@PPVII was (39.56% ± 1.78)% (n = 3). HA@IRMOF-10@PPVII had good biosafety, which could significantly reduce the hemolysis risk of PPVII. This nanoparticle can effectively improve the uptake of PPVII in 4T1 cells, and significantly inhibits cell migration and clone formation. Its anti-tumor mechanism is closely related to inducing an increase in intracellular ROS levels, a decrease in MMP triggering apoptosis, and inhibiting the migration and cloning of 4T1 cells. Conclusion The HA@IRMOF-10@PPVII nanoparticles were successfully prepared, which exhibit excellent targeting ability and anti-TNBC activity. They can significantly enhance the efficacy of PPVII, providing experimental evidence for the development of PPVII-based anti-tumor formulations.

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 To investigate the biphenyl compounds in the ethyl acetate-soluble fraction of the twigs of Garcinia yunnanensis. Methods Based on the UV spectral characteristics absorption of biphenyl derivatives, targeted separation and purification were performed using various chromatography techniques, including silica gel column chromatography, Sephadex LH-20 gel column and semi-preparative HPLC. The structures of the isolated compounds were elucidated by means of physicochemical properties and spectral data of the monomer compounds, in combination with relevant literature. The anti-inflammatory activity of the new compound was evaluated by measuring its inhibition effect of nitric oxide (NO) production. Results Eight bipheny derivatives were obtained from the ethyl acetate fraction of G. yunnanensis, elucidated as (R)-5-(4-hydroxyphenyl)-2-(2-hydroxypropan-2-yl)-2,3-dihydrobenzofuran-7-ol (1 ), garcibiphenyl C (2 ), cylindrobiphenyl B (3 ), multibiphenyl C (4 ), doitungbiphenyls B (5 ), multiflorabiphenyl B (6 ), honokiol (7 ), and magnolol (8 ). Conclusion Compound 1 is a new compound named yunnabiphenyl A, while compounds 2 —8 were reported from this plant for the first time. Compound 1 showed moderate anti-inflammatory activity, with a median inhibition concentration (IC₅₀) value of 21.5 μmol/L.