Objective To explore the characteristics, medication patterns, and the mechanisms of action of traditional Chinese medicines for treating angle-closure glaucoma. Methods Literature and prescriptions related to the treatment of angle-closure glaucoma with traditional Chinese medicine were retrieved from databases including CNKI, Wanfang, VIP, China Biology Medicine (CBM), and the Ancient and Modern Medical Case Cloud Platform. Through data mining and analysis, core prescriptions were identified. Subsequently, a network pharmacology approach was employed. Active ingredients with oral bioavailability (OB) ≥ 30% and drug-likeness (DL) ≥ 0.18 were screened using the TCMSP and BATMAN-TCM 2.0 databases. Targets were uniformly annotated via the UniProt database and intersected with disease targets obtained by integrating and deduplicating data from four major databases (GeneCards, TTD, OMIM, and CTD) to identify pharmacodynamic targets. A “core prescription-active ingredient-pharmacodynamic target” interaction network was constructed using Cytoscape 3.10.2. The pharmacodynamic targets were then imported into the Metascape platform (restricted to human species, with a threshold of P < 0.05) for gene ontology (GO) (biological process, molecular function, cellular component) and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis, with the top 20 significant pathways and functions visualized. Additionally, a protein-protein interaction (PPI) network was constructed using the String database, and core targets were screened using the Cytoscape MCODE plugin (cut-off = 0.05) to elucidate the potential molecular mechanisms underlying the core prescriptions in treating angle-closure glaucoma. The binding energy between the active ingredient and the core target was investigated through molecular docking. Sodium dithionite was used to induce hypoxic injury in mouse retinal ganglion cells (RGCs) to establish an in vitro model. Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was employed to detect the effects of these active components on mRNA expression of core target genes. ELISA was used to measure the impact of these active components on tumor necrosis factor-α (TNF-α) levels. Immunofluorescence staining was applied to evaluate the effects of these active components on phosphorylated protein kinase B (p-AKT) and nuclear factor-κB (NF-κB) protein expression. Results The top three most frequently used traditional Chinese medicines for angle-closure glaucoma were Fuling (Poria), Danggui (Angelicae Sinensis Radix), and Chaihu (Bupleuri Radix). The core syndromes were mainly liver qi stagnation and liver-kidney deficiency. A core prescription consisted of Mudanpi (Moutan Cortex)-Angelicae Sinensis Radix-Zhizi (Gardeniae Fructus)-Bupleuri Radix-Juhua (Chrysanthemi Flos)-Baizhu (Atractylodis Macrocephalae Rhizoma)-Gancao (Glycyrrhizae Radix et Rhizoma)-Baishao (Paeoniae Radix Alba)-Poria. Cluster analysis yielded three clusters: modified Chaihu Shugan San (柴胡疏肝散), modified Yijing Buyang Huanwu Decoction (益精补阳还五汤), modified Qiju Dihuang Pills (杞菊地黄丸). Network pharmacology analysis showed that the core prescription regulated key targets such as AKT1, TNF-α, NF-κB by modulating signalling pathways such as TNF, phosphatidylinositol-3-hydroxykinase (PI3K)-Akt, and MAPK (mitogen-activated protein kinase), thereby participating in biological processes such as inflammatory response, and oxidative stress, and thus intervening in the pathological process of glaucoma. Quercetin and luteolin were identified as the core components of the core prescription for the intervention of angle-closure glaucoma through molecular docking, and in vitro experiments were conducted for verification. The results showed that all luteolin dose groups (5, 10, 20 μmol/L) upregulated AKT1 mRNA expression (P < 0.01), while the medium and high dose groups downregulated NF-κB and TNF-α mRNA expression (P < 0.05), significantly increased p-AKT protein expression, and decreased NF-κB protein and TNF-α levels (P < 0.05). The medium and high dose groups of quercetin (40, 80 μmol/L) also significantly upregulated AKT1 mRNA expression and downregulated NF-κB and TNF-α mRNA expression (P < 0.05). All low (20 μmol/L), medium, and high dose groups of quercetin increased p-AKT protein expression and decreased NF-κB protein expression (P < 0.05), while the medium and high dose groups significantly reduced TNF-α expression (P < 0.05). Conclusion Data mining techniques were systematically employed to analyze ancient and modern literature and medical records concerning the treatment of angle-closure glaucoma. From multiple dimensions including medication frequency, nature-flavor-meridian tropism, efficacy categories, association rules, and core combinations, the “high-frequency-core-differential” three-dimensional characteristics were progressively analyzed to extract core prescriptions and reveal their compatibility patterns. Network pharmacology analysis further confirmed that the above high-frequency prescriptions synergistically regulate key biological processes such as oxidative stress, inflammatory response, and apoptosis through a “multi-component-multi-target-multi-pathway” mechanism, reflecting the holistic therapeutic advantages of traditional Chinese medicine compound formulas, thereby providing scientific evidence for clinical syndrome differentiation and medication, with certain guiding significance. In vitro experiments simultaneously validated that, in the hypoxic injury model, both luteolin and quercetin significantly upregulated AKT1 mRNA and p-AKT protein expression, inhibited NF-κB and TNF-α mRNA and protein expression, effectively reduced levels of inflammation and apoptosis-related factors, and exerted protective effects on RGCs through a bidirectional mechanism of “anti-inflammatory and pro-survival” effects.

Objective To explore the effect of usnic acid on inflammatory injury in mice with intracerebral hemorrhage (ICH) based on high mobility group box 1 (HMGB1)/receptor for advanced glycation end products (RAGE) pathway. Methods C57BL/6 mice were randomly divided into sham group, model group, HMGB1 inhibitor ethyl pyruvate (40 mg/kg) group, usnic acid low-, high-dose (20, 30 mg/kg) groups and usnic acid (30 mg/kg) + rHMGB1 (8 μg/kg) group, with 18 mice in each group. The ICH model was constructed by intracranial injection of 0.5 μL collagenase. After continuous administration for 3 d, the modified neurological severity score (mNSS) and left turn rate of mice in each group were evaluated. The water rate of brain tissue in mice was determined. ELISA was used to measure the levels of interleukin-6 (IL-6), IL-1β and IL-18 in brain tissue. Hematoxylin-eosin (HE) staining was used to observe pathological changes in brain tissue. Immunofluorescence was used to observe microglia activation in brain tissue. TUNEL staining was used to detect cell apoptosis of brain tissue. Western blotting was used to detect the expressions of HMGB1/RAGE pathway related proteins in brain tissue. Results Compared with sham group, the striatum tissue of mice in model group was severely damaged, mNSS, left cerebral hemisphere water rate, ionized calcium-binding adapter molecule 1 (Iba1) positive cells number, inflammatory factors levels, cell apoptosis rate, HMGB1, RAGE, phosphorylated nuclear factor-κB p65 (p-NF-κB p65)/NF-κB p65 and TNF-α protein expression levels were significantly increased (P < 0.05), and the left turn rate was significantly decreased (P < 0.05). Compared with model group, the damage degree of striatum tissue in usnic acid low-, high-dose group and ethyl pyruvate group was alleviated (P < 0.05), mNSS, left cerebral hemisphere water rate, Iba1 positive cells number, inflammatory factors levels, cell apoptosis rate and HMGB1/RAGE pathway related proteins expression levels were significantly decreased (P < 0.05), and the left turn rate was significantly increased (P < 0.05). rHMGB1 significantly reversed the inhibitory effect of usnic acid on inflammatory injury in ICH mice (P < 0.05). Conclusion Usnic acid may alleviate inflammatory injury in ICH mice by inhibiting HMGB1/RAGE pathway.

With the wide application of traditional Chinese medicine (TCM) at home and abroad, the problem of toxicity reduction of toxic Chinese materia medica needs to be solved urgently. Rational compatibility of TCM is the basis for safe use of Chinese medicinal formulas, and herb pair is the smallest unit of TCM compatibility. By starting from toxicity reduction through herb pair compatibility, this study summarizes the mechanism of action of “toxic” TCM, the differences between ancient and modern dosages, and the typical theories of herb pair compatibility, to provide ideas and references for research on toxicity reduction in herb pairs. In addition, this study summarizes studies on chemical composition changes of individual herbs after pairing, in vivo pharmacokinetics, and pharmacological mechanisms of herb pair compatibility. It is concluded that under the guidance of basic theories of TCM, herb pairs can be studied at multiple levels and from multiple perspectives by combining multiple methods to investigate the mechanisms of their combinations in reducing toxicity. This approach helps to clarify the compatibility rules of Chinese medicinal formulas and provides a scientific reference for in-depth studies on the mechanisms of compatibility and detoxification of herb pairs, as well as for the clinical safe use of TCM.

Objective To establish a method for differentiating gelatinous Chinese medicines (GCMs) based on supramolecular sensing technology. Methods An indicator displacement assay (IDA) strategy was employed where test sample solutions of GCMs served as analytes and macrocyclic molecule-dye host-guest pairs with specific fluorescence signals were used as sensing pairs to construct a sensor array. The fluorescence intensity changes resulting from the interaction between different GCMs and the sensing units were detected and combined with pattern recognition methods for differentiation. Results Through screening host-guest sensing units with selective fluorescence responses to GCMs from a supramolecular library consisting of cyclodextrins, cucurbiturils, calixarenes, and biphen[n]arenes hosts combined with dyes, a quaternary supramolecular sensor array was constructed including quaternary ammonium salt calix[5]arene (QAC5A)·eosin Y (EY) (0.4 μmol/L vs 0.5 μmol/L), quaternary ammonium modified azocalix[4]arene (QAAC4A)·EY (1 μmol/L vs 1 μmol/L), dodecyl quaternary ammonium salt azo calix[4]arene (QAAC4A12C)·EY (0.6 μmol/L vs 1 μmol/L), and terphen[3]arene (WTP3)·fluorescein (Fl) (0.3 μmol/L vs 0.6 μmol/L). Principal component analysis (PCA) demonstrated that Ejiao (Colla Corii Asini), Xin’ejiao (Colla Corii Suilli), Biejiajiao (Colla Carapacis Trionychis), Lugujiao (Colla Ossis Cervi), Guijiajiao (Colla Carapacis et Plastri Testudinis) or Lujiajiao (Colla Cornus Cervi) could be significantly distinguished within a 95% confidence interval. Conclusion This study developed a quaternary supramolecular sensor array integrated with pattern recognition analysis which enables rapid discrimination of various GCMs.

Objective To screen for the most effective antimicrobial carbon dots from the traditional Chinese medicines Baiji (Bletilla striata), Cili (Rosa roxburghii), Lingzhi (Ganoderma lucidum) and Tianma (Gastrodia elata), and to systematically investigate the physicochemical properties, photothermal performance, antibacterial activity and mechanism of manganese-doped G. lucidum carbon dots (Mn-GLCDs). Methods Four kinds of herbal carbon dots derived from B. striata, R. roxburghii, G. lucidum and G. elata were prepared by a conventional hydrothermal method. The carbon dots with optimal antibacterial activity were screened out, and the morphology, structure and in vitro photothermal performance of the manganese-doped carbon dots were characterized. The antibacterial performance against Staphylococcus aureus and Escherichia coli under near-infrared (NIR) irradiation was evaluated through zone of inhibition assays, bacterial co-culture experiments, and biofilm inhibition and disruption tests. The antibacterial mechanism was preliminarily investigated by intracellular active oxygen fluorescence detection using a DCFH-DA probe. Biosafety was assessed by hemolysis assay and cytotoxicity test. Intracellular reactive oxygen species (ROS) fluorescence measurement using the 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA) probe was adopted to preliminarily elucidate its antibacterial mechanism. Hemolysis assays and cytotoxicity evaluations were conducted to verify their biological safety. Results GLCDs possessed remarkably stronger antibacterial activity than carbon dots prepared from B. striata, R. roxburghii and G. elata carbon dots. Mn-GLCDs had a particle size of approximately 5 nm, and their surfaces were rich in hydrophilic functional groups including -OH, -NH₂, C=O, C-N-C and C-O-C. X-ray photoelectron spectroscopy (XPS) and inductively coupled plasma mass spectrometry (ICP-MS) confirmed that the mass fraction of Mn²⁺ reached 0.83%, and Mn²⁺ was stably present in the carbon dot framework in a coordinated form. The red shift of the UV-Vis absorption peak and the Raman I_D/I_G > 1 indicated that Mn²⁺ doping narrowed the bandgap and increased the density of structural defects. Photothermal performance evaluation showed that Mn-GLCDs could rapidly heat up under 808 nm laser irradiation (1.5 W/cm²), with a photothermal conversion efficiency of 35.03%. The maximum temperature fluctuation in six cycles of heating-cooling stability test was less than 6%, indicating good photothermal stability. Fluorescence performance test showed a fluorescence quantum yield of 20.51% and excitation-dependent fluorescence emission. In vitro antibacterial experiments demonstrated that, compared with undoped G. lucidum carbon dots, Mn-GLCDs exhibited the strongest antibacterial effect against S. aureus and E. coli under near-infrared irradiation, and could significantly inhibit and disrupt bacterial biofilms. Intracellular active oxygen detection showed that upon near-infrared irradiation, Mn-GLCDs elevated the active oxygen levels in S. aureus and E. coli to 6.75-fold and 5.44-fold of the negative control group, respectively. Biosafety evaluation showed that the hemolysis rate of Mn-GLCDs was below 5% within the antibacterial concentration range, and Mn-GLCDs exhibited no obvious toxicity to L929 and RAW264.7 cells. Conclusion This study constructs for the first time Mn-GLCDs, which significantly enhance the photothermal conversion capability and antibacterial activity. The mechanism is related to the induction of intracellular active oxygen. This work provides an experimental basis for the development of novel traditional Chinese medicine-based nanoscale antibacterial agents.

Objective To analyze the structural characteristics of water-eluted fraction of Sanghuangporus vaninii polysaccharides (SHP-W) and explore the mechanism of SHP-W on improving carbon tetrachloride (CCl₄)-induced liver fibrosis in mice by regulating gut microbiota. Methods SHP-W was extracted from S. vaninii using water extraction and ethanol precipitation, followed by purification through weak anion-exchange chromatography. The total sugar, uronic acid and protein contents of SHP-W were determined by phenol sulfuric acid method, m-hydroxybiphenyl method and BCA method. The structural characterization of SHP-W was carried out by high performance size exclusion chromatography (HPSEC), Fourier transform infrared spectrophotometer (FT-IR), ion chromatography (IC) and methylation combined gas chromatography-mass spectrometry (GC-MS). C57BL/6J mice were prepared with a 15% CCl₄ olive oil solution via ip for six weeks to establish a liver fibrosis model. SHP-W (25, 50, 100 mg/kg) or sorafenib (10 mg/kg) were administered for three weeks, and the pathological morphological changes in liver tissues were observed using hematoxylin-eosin (HE), sirius red and Masson staining. A fully automated biochemical analyzer was used to analyze the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bile acids (TBA), total bilirubin (TBIL) and direct bilirubin (DBIL) in serum. Immunohistochemistry was used to detect the expressions of collagen-I (Col-I) and α-smooth muscle actin (α-SMA) in liver tissue. Western blotting was used to detect the expressions of Col-I, α-SMA and transforming growth factor-β1 (TGF-β1) proteins in liver tissue. 16S rDNA sequencing was used to analyze the gut microbiota, and antibiotic interference was used to verify the role of the microbiota. Results SHP-W was a refined polysaccharide composed of two polysaccharides with different relative molecular weights (1.818 × 10⁴, 2.040 × 10³), with a total sugar content of (88.27 ± 3.76)%, and without uronic acid and protein. The monosaccharide composition results showed that SHP-W was composed of galactose, mannose, fucose and glucose, with a molar ratio of 34.6∶23.0∶22.7∶19.7, and also contained 3-O-methylgalactose. Methylation analysis showed that SHP-W was composed of 14 glycosidic bonds, including 1,3,6-Glcp.The animal experiment results showed that compared with control group, the levels of AST, ALT, TBIL, DBIL and TBA in serum of mice in model group were significantly increased (P < 0.001), the arrangement of liver cell cords was disordered, inflammatory cell infiltration was obvious, and a large amount of fibrous tissue proliferation was appeared, the expressions of Col-I, α-SMA and TGF-β1 protein in liver tissue was significantly up-regulated (P < 0.05, 0.001). Compared with model group, the collagen fiber deposition in pseudo lobules and portal area of liver tissue of mice in each treatment group was improved to varying degrees, and the serum liver function level was significantly reduced (P < 0.05, 0.01, 0.001), the expressions of Col-I, α-SMA and TGF-β1 protein in liver tissue was down-regulated (P < 0.05, 0.01, 0.001). After antibiotic interference, the effect of SHP-W on improving liver fibrosis was significantly inhibited. Conclusion SHP-W were prepared from S. vaninii fruiting bodies, the determination of uronic acid content, infrared spectroscopy and monosaccharide composition showed that SHP-W were neutral polysaccharide. SHP-W could significantly improve serum liver function, inhibit HSCs activation, reduce collagen fiber deposition, and thus improve CCl₄-induced liver fibrosis in mice. Its anti-fibrotic effect depends on the regulation of gut microbiota.

Objective To clone the full-length cDNA sequence of the CoMYB62 gene from Cornus officinalis, and analyze the structural characteristics, phylogenetic relationships, subcellular localization, and tissue expression patterns of its encoded protein, thereby providing a molecular basis for elucidating the potential roles of MYB transcription factors in the growth, development, and environmental adaptation of C. officinalis. Methods Based on transcriptome data of C. officinalis, the CoMYB62 gene sequence was screened and its full-length cDNA sequence was cloned using RT-PCR. Bioinformatics analyses were conducted to predict the physicochemical properties, conserved domains, secondary and tertiary structures, and phylogenetic relationships of the CoMYB62-encoded protein. A pCAMBIA1300-CoMYB62-GFP fusion expression vector was constructed and transiently expressed in tobacco leaves, and the subcellular localization of the CoMYB62 protein was observed using a laser confocal inverted microscope. In addition, quantitative real-time PCR (qRT-PCR) was performed to analyze the expression patterns of CoMYB62 in four tissues, including root, stem, leaf, and fruit. Results The full-length cDNA of the CoMYB62 gene was successfully cloned. Sequence analysis indicated that the encoded protein belonged to a typical R2R3-MYB transcription factor and contained a highly conserved DNA-binding domain at the N-terminus, which was predicted to be mainly localized in the nucleus. Phylogenetic analysis revealed that CoMYB62 shared high evolutionary conservation with MYB homologous proteins from various plant species. Laser confocal microscopy further confirmed that the CoMYB62 protein was localized in the nucleus. The qRT-PCR results showed that CoMYB62 was expressed in all examined tissues of C. officinalis, although the expression levels varied significantly among different tissues. Conclusion CoMYB62 is a conserved MYB transcription factor in C. officinalis with tissue-specific expression characteristics, which may participate in the regulation of plant growth, development, and stress responses to provide a foundation for further biological functional analysis of CoMYB62 in C. officinalis.

Type 2 bitter taste receptors (TAS2Rs), a subfamily of G protein-coupled receptors, comprise 25 receptor-members. Besides being expressed on taste buds to perceive bitterness, TAS2Rs are also distributed in multiple systems, including the respiratory, digestive, immune systems, where they regulate various physiological processes such as airway defense, gastrointestinal motility, and inflammation regulation. In traditional Chinese medicines (TCMs) theory, the bitter TCMs are able to purge, dry and firm. Their functional substances often serve as both taste substances that evoke bitter perception and the key component responsible for the medicinal properties and efficacy of bitter herbs. These components may exert various pharmacological effects, such as regulating glucolipid metabolism and inflammatory responses, by activating TAS2Rs expressed in different tissues and organs. This article systematically expounds the association between the efficacy of bitter TCMs and the function of TAS2Rs, as well as the tissue distribution, physiological functions, structural characteristics and signal transduction mechanisms of TAS2Rs, and analyses the problems and challenges in research between bitter TCMs and TAS2Rs pharmacology. It’s expected to reveal the scientific essence of the “property-effect-substance” correlation of bitter TCMs.

Objective To investigate the effect and underlying mechanism of Prunella vulgaris-derived exosomes loaded with orlistat (PVENs-Orl) on lipid metabolism and malignant biological behaviors of hepatoma cells. Methods PVENs were extracted by ultracentrifugation. After characterization by transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA), a PVENs-Orl drug delivery system was constructed. The inhibitory effects of PVENs-Orl on the proliferation, migration and invasion of HepG2 cells were evaluated through CCK-8, cloning, scratch and Transwell assays. The free fatty acids (FFA) uptake, fatty acid synthase (FASN) activity, triglyceride (TG) content, lipid peroxidation indicators and mitochondrial membrane potential and morphology were detected. The expressions of genes related to lipid metabolism and ferroptosis [acyl-CoA synthetase long-chain family member 4 (ACSL4), glutathione peroxidase 4 (GPX4), solute carrier family 7 member 11 (SLC7A11), B-cell lymphoma-2 interacting protein 3 (BNIP3L), lysophosphatidylcholine acyltransferase 3 (LPCAT3), transferrin receptor (TFRC)] was detected by qRT-PCR. Results The PVENs were successfully extracted and characterized, and the constructed PVENs-Orl could be effectively internalized by HepG2 cells. PVENs and PVENs-Orl exhibited dose-dependent inhibition of the proliferation, migration and invasion of HepG2 cells (P < 0.001). In terms of lipid metabolism phenotype, PVENs-Orl could inhibit FASN activity (P < 0.01), and simultaneously cause intracellular accumulation of FFA and TG (P < 0.01). PVENs-Orl significantly increased the levels of intracellular reactive oxygen species (ROS) and malondialdehyde (MDA) (P < 0.01, 0.001), and increased the release of lactate dehydrogenase (LDH) (P < 0.001). PVENs-Orl significantly induced a decrease in mitochondrial membrane potential (P < 0.01) and triggered mitochondrial dysfunction (P < 0.001). At the molecular mechanism level, PVENs-Orl upregulated the expressions of genes related to lipid peroxidation and ferroptosis promotion (P < 0.001), and downregulated the expressions of genes related to antioxidant and ferroptosis inhibition (P < 0.001). Conclusion PVENs could effectively deliver orlistat, disrupt lipid homeostasis by inhibiting FASN activity, induce mitochondrial dysfunction and lipid peroxidation, and synergistically regulate the expressions of key genes involved in ferroptosis, thereby inhibiting the growth of liver cancer cells. This provides experimental evidence for targeted lipid metabolism therapy based on natural exosomes.

Objective To integrate the correlation between the administration routes of traditional Chinese medicine essential oils and diseases, as well as their core biomarkers in disease treatment, analyze their pathways and mechanisms of action on the body, and promote the rational application and innovative development of traditional Chinese medicine essential oils. Methods China National Knowledge Infrastructure(CNKI), VIP Information(VIP), Wanfang Data, Web of Science were used to analyze the distribution of administration routes for traditional Chinese herbal essential oils and the types of diseases treated. The correlation between different administration routes and disease types was examined, along with frequency statistics of biomarkers involved in the therapeutic process. After identifying the most closely associated biomarkers, their mechanisms of action in disease treatment by traditional Chinese herbal essential oils were analyzed. Results Among all administration routes, oral administration accounted for 66.23%, inhalation for 21.37%, topical application for 8.18%, intraperitoneal injection for 3.17%, and intravenous injection for 1.06%. The disease types most strongly associated with oral and inhalation routes were neurological and psychiatric disorders, while those most associated with topical application were skin diseases and immune-related conditions. Intraperitoneal injection showed the strongest association with tumor diseases. Among biomarkers appearing more than 10 times, those related to oxidative stress regulation included malondialdehyde (MDA), nitric oxide (NO), and superoxide dismutase (SOD); those involved in inflammatory response regulation included tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), IL-1β, and IL-8; those associated with apoptosis regulation included B-cell lymphoma-2 associated X protein (Bax), B-cell lymphoma-2 (Bcl-2), and cysteinasparate protease-3 (Caspase-3); and those involved in neural regulation included 5-hydroxytryptamine (5-HT), γ-aminobutyric acid (GABA), norepinephrine (NE), and acetylcholinesterase (AchE). Conclusion Traditional Chinese medicine essential oils enter the human body through three primary routes: inhalation, transdermal absorption and oral administratio to exert their biological effects. They can exert effects via by regulating multiple mechanisms such as oxidative stress, neurotransmitter, inflammatory responses, and apoptosis, demonstrating significant advantages in disease prevention and treatment.