With the increased global demand for Chinese herbal medicines (CHM), the standardization and internationalization of CHM have been of interest to the scientific and research communities. However, there are some challenges in ensuring the quality of research and the acceptability globally since the complexity of the ingredients in CHM and their extracts. In this review, the international best practices in research reporting guidelines of CHM were summarized and discussed, including approaches to sourcing, cultivation and harvesting, extraction, preparation, bioactive constituents and quality control markers, as well as pesticide residues and safety, in vitro and in vivo studies, clinical trials, and the application of artificial intelligence. This review aims to enhance the reproducibility, and ensure precise interpretation of results, and hence, to improve research quality and transparent reporting, to facilitate the internationalization and standardization of CHM.

Objective To investigate the chemical constituents of Tibetan medicine Ligularia virgaurea. Methods The compounds were isolated and purified by silica gel column chromatography, Sephadex LH-20, ODS reversed-phase column chromatography and semi-preparative HPLC, and the structures were identified by spectral data (NMR and MS). Results A total of 19 compounds were isolated and identified as follows 1-[(1S,3aR,4R,7S,7aS)-4-hydroxy-4-hydroxymethyl-7-propan-2-yl-1,2,3,3a,5,6,7,7a-octahydroinden-1-yl]ethanone (1), 10α-hydroxy-1-oxoeremophila-7(11),8(9)-dien-12,8-olide (2), 6α,10α-dihydroxy-1-oxoeremophila-7(11),8(9)-dien-12,8-olide (3), 3-hydroxycacalolide (4), 6β-acetoxy-9-oxo-10αH-furanoeremophilane (5), pinoresinol (6), medioresinol (7), syringaresinol (8), p-hydroxybenzoic acid (9), methyl 3,4-dihydroxybenzoate (10), syringic acid (11), 3,4,5-trimethoxybenzoic acid (12), dimethy phthalate (13), syringin (14)、lupeol (15), obtusifoliol (16), ethyl linoleate (17), 15-vaccenic acid (18), 10-eicosenoic acid (19). Conclusion Compound 1 is a new compound, designated 14-hydroxy-oplopanone. Compounds 5—14, 16—19 are isolated from L. virgaurea for the first time.

Objective To investigate the chemical constituents of limonoid-type triterpenes from the 95% ethanol extract of Melia toosendan and their antitumor activity. Methods The compounds were isolated and purified using various chromatographic techniques, including normal-pressure column chromatography, high-pressure column chromatography, and gel column chromatography. The isolated compounds were structurally characterized by modern spectroscopic techniques. Their antitumor activity was evaluated using the CCK-8 assay. Results Eight limonoid-type triterpenoids were isolated from the ethyl acetate extract of M. toosendan. Structural elucidation by spectroscopic techniques identified them as 15-oxotrichilinin (1), 11,15-dioxotrichilinin (2), 12-O-ethyl nimbolinin B (3), mufolinoid U (4), trichilinin (5), trichilinin D (6), trichilinin E (7), and trichilinin G (8). Activity evaluation results showed that all compounds exhibited varying degrees of inhibitory effects on four tumor cell lines. Conclusion Compound 1 is a new compound, named 15-oxotrichilinin. Antitumor activity results indicated that compounds 1—8 all possess antitumor activity to varying extents, with some compounds showing favorable activity against HepG2 cells.

Objective To compare different varieties of Dioscoreae Rhizoma and reveal their chemical differences, providing a basis for targeted breeding of Dioscoreae Rhizoma varieties. Methods Eight batches of Dioscoreae Rhizoma materials with different germplasms were collected, their agronomic traits were measured and compared. Subsequently, ultra-high performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS)-based untargeted metabolomic approach was employed to systematically analyze their chemical compositions. Multivariate statistical methods, including principal component analysis (PCA), cluster heatmap analysis and correlation plots, were used to dissect the metabolic product differences among varieties and their associations with agronomic traits. Results The eight batches of Dioscoreae Rhizoma showed significant differences in indicators such as yield and tuber morphology. Based on high-resolution mass spectrometry, 164 chemical components were identified across the eight batches of Dioscoreae Rhizoma, including peptides, amino acids, flavonoids, and saponins. PCA indicated significant differences among the eight batches of Dioscoreae Rhizoma germplasm resources. Saponin levels were relatively high in Jin Dioscoreae Rhizoma, Taigu Dioscoreae Rhizoma, Shuangbang Dioscoreae Rhizoma, and Ma Dioscoreae Rhizoma, while Taigu Dioscoreae Rhizoma, Jin Dioscoreae Rhizoma, and Ma Dioscoreae Rhizoma exhibited similar chemical characteristics in terms of flavonoids. Cluster heatmap analysis revealed potential chemical markers, providing a basis for rapid discrimination among different varieties. The chemical differences between medicinal-edible Dioscoreae Rhizoma varieties such as Tiegun Dioscoreae Rhizoma and Shandong Ximao and other varieties may be related with amino acids and peptides. The correlation network indicated that stem diameter was significantly positively correlated with saponins, while bulbils weight showed positive correlations with saponins and flavonoids but negative correlations with amino acids and peptides. Conclusion The eight different batches of Dioscoreae Rhizoma possess distinct chemical characteristics, which provide a scientific basis for the targeted breeding of medicinal and edible-specific Dioscoreae Rhizoma varieties.

Objective A targeted nanoformulation (PEG-PTP/PPVI@IRMOF-1) was constructed by encapsulating the traditional Chinese medicine antitumor active component polyphyllin VI (PPVI) within isoreticular metal-organic framework-1 (IRMOF-1) and coating it with a cell-penetrating peptide. The study aims to characterize the nanoformulation and conduct a preliminary evaluation of its in vitro antitumor efficacy. Methods PEG-PTP/PPVI@IRMOF-1 was fabricated via the “triethylamine precipitation method” and “reaction coating method”; The physicochemical properties of the nanoformulation, including particle size, morphology, thermal stability, and specific surface area, were characterized by particle size and ζ-potential measurement, scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA), Brunauer-Emmett-Teller (BET) gas adsorption, and X-ray photoelectron spectroscopy (XPS). Hemolysis assay was used to assess the biosafety of nanomaterials. CCK-8 assay was adopted to evaluate the material safety and detect the effect of nanoparticles on the proliferation of human pancreatic cancer PANC-1 cells. The effect of materials and nanoparticles on the nuclear morphology of PANC-1 cells was observed by fluorescence microscopy; The targeting capability of the nanoparticles ex vivo was assessed by laser scanning confocal microscopy; The effects of nanoparticles on cell proliferation and migration were examined using colony formation and scratch assays, respectively; Flow cytometry was employed to determine the impact of the nanoparticles on the cellular apoptosis rate and intracellular reactive oxygen species (ROS) levels; The effect of nanoparticles on the mitochondrial membrane potential (MMP) of cells was examined by laser confocal microscopy. Results The targeted nanoformulation PEG-PTP/PPVI@IRMOF-1 was successfully fabricated as a dry white powder. The nanoparticles exhibited a quasi-spherical morphology with an average particle size of about (177.47 ± 3.97) nm and a ζ potential of (−7.94 ± 0.14) mV; The drug loading capacity (DLC) of PPVI@IRMOF-1 was (43.02 ± 0.08)%; The nanomaterial showed excellent biocompatibility; Notably, targeted peptide modification can significantly enhance the drug uptake of tumor cells, significantly inhibit cell proliferation, promote the production of ROS and the decrease of MMP in cells, and induce apoptosis of tumor cells. Conclusion The prepared PEG-PTP/PPVI@IRMOF-1 nanoparticles possess uniform morphology, high drug loading capacity, and good biocompatibility. Moreover, the nanoformulation significantly enhances the in vitro antitumor efficacy of PPVI, providing a reference for the development of PPVI-based antitumor formulations.

Objective To construct a Blumea balsamifera oil (BBO) Pickering emulsion (AMWE-BBO@PE) stabilized by soybean protein isolate (SPI) and loaded with Astragalus membranaceus water extract (AMWE), and evaluate its physicochemical properties, antibacterial activity, and effect on wound healing in mice. Methods AMWE-BBO@PE was prepared by ultrasonic emulsification method using SPI as the stabilizer and BBO as the oil phase. Key process partners including the concentration of AMWE, oil-water ratio, and ultrasonic power were optimized via single-factor experiments. The particle size, ζ potential, polydispersity index (PDI), and stability of the emulsion were characterized; its micromorphology was observed by transmission electron microscopy (TEM) and optical microscopy. Additionally, its transdermal penetration ability, antibacterial activity, and wound healing-promoting effect were evaluated. Results The optimal process partners were determined as follows: oil-water ratio of 4%, AMWE mass concentration of 0.020 g/mL, and ultrasonic power of 30 W. The prepared emulsion exhibited a regular elliptical shape with an average particle size of (422.2 ± 2.2) nm, PDI of 0.076 ± 0.019, and ζ potential of (−33.95 ± 0.54) mV, showing good stability. The cumulative transdermal amount within 48 h reached 4.857 mg, and the transdermal rate was approximately 6.3-fold higher than that of the BBO group. Furthermore, the emulsion displayed significant antibacterial activity against Escherichia coli and Staphylococcus aureus, accelerated the deposition and remodeling of collagen fibers, and remarkably promoted the healing of skin wounds in mice. Conclusion The AMWE-BBO@PE Pickering emulsion possesses excellent antibacterial activity and wound healing-promoting capacity, providing a novel strategy for the development of efficient and stable transdermal delivery systems for aqueous extracts of traditional Chinese medicines.

Objective This study applied the quality by design (QbD) principle to identify key attributes and elucidate their transfer relationships along the “spray-dried powder-ribbon-granule” manufacturing chain of Shanzha (Crataegi Fructus) formula granules. Methods A fractional factorial design was implemented to investigate seven major factors affecting the properties of the spray-dried powder, ribbons, and granules. The relevant properties of these intermediates were measured. Modeling and attribute identification were performed using analysis of variance, multiple linear regression, and orthogonal partial least squares-discriminant analysis (OPLS-DA). Results Excipient amount, inlet air temperature, and roller pressure were identified as critical process parameters. A key attribute transfer relationship was established, centered on glass transition temperature (T_g) and median particle size (D₅₀), and associated with supporting attributes including moisture content (H₁), bulk density (ρ_b1), tapped density (ρ_t1) of the spray-dried powder, ribbon yield, and granule yield. Among these, T_g served as a representative key indicator with both discriminative and transfer functions. Conclusion This research clarifies the identification and transfer relationships of key attributes in the dry granulation process of Crataegi Fructus formula granules, providing a basis for developing effective quality control strategies and supporting the high-quality development of the formula granules industry.

Objective To compare the levels of multiple components (phenolic acids and luteolin glycoside) in Jinyinhua (Lonicerae Japonicae Flos, LJF), along with changes in “colour, aroma and flavor”, under different drying methods to determine the optimal drying technique. Methods Quality control of phenolic acids and luteoloside in LJF was conducted using HPLC technology. Intelligent sensory technology was employed to study the color, aroma, and taste characteristics of LJF under different drying methods. The quality of LJF processed with various drying techniques was evaluated through correlation analysis of color-aroma-taste and multiple indicator components. Results Steam-blanched drying, freeze-vacuum drying, and microwave-blanched drying resulted in higher total phenolic acid content, while sun-drying, microwave-blanched drying, and steam-blanched drying yielded higher luteoloside levels. The differences in color-aroma-taste profiles of LJF under different drying methods were reflected in brightness (L^*), red-green chromaticity (a^*), and responses from W5S, W1W, W1S, W2S, W2W sensors, along with bitterness and astringency. Correlation analysis revealed that higher total phenolic acids correlated with darker green coloration, elevated W1W sensor responses, intensified astringency, and reduced bitterness. Conversely, increased luteoloside content was associated with higher brightness and diminished responses from W5S and W1W sensors. Freeze-vacuum drying produced optimal color, shade-drying achieved the most pronounced aroma, while steam-blanched and microwave-blanched drying generated distinctive bitter and astringent aftertastes. Conclusion From the perspectives of multi-component retention, chromatic characteristics, and taste profile, steam-blanched drying and microwave-blanched drying have been identified as the optimal drying processing methods for LJF. This study provides a novel methodology for optimizing drying techniques and enabling rapid quality assessment of LJF.

Objective This study aimed to investigate the optimal preparation process for asiaticoside microspheres using polyethylene glycol-polylactic acid-glycolic acid copolymer (mPEG-PLGA) as the carrier. Methods Key process parameters affecting microsphere preparation were first screened from factors including curing temperature, emulsifier PVA concentration, stirring speed, water-to-oil ratio, and mPEG-PLGA concentration. Subsequently, a Box-Behnken design-response surface methodology (BBD-RSM) experiment was employed to further optimize the preparation conditions for the drug-loaded microspheres. Their morphology, particle size distribution, drug loading, encapsulation efficiency, in vitro release behavior, and biocompatibility were comprehensively evaluated. Results Curing temperature, stirring speed, emulsifier PVA and mPEG-PLGA concentrations, and water-to-oil ratio all influenced the encapsulation efficiency and morphology of the microspheres. Among them, curing temperature and mPEG-PLGA concentration had a greater impact: both excessively high and low curing temperatures reduced encapsulation efficiency; increasing mPEG-PLGA concentration improved encapsulation efficiency, but excessively high concentrations led to adhesion and increased particle size. The optimized conditions were 40 ℃, 1 800 r/min, with appropriate emulsifier PVA and mPEG-PLGA concentrations. Using mPEG-PLGA concentration, water-to-oil ratio, and emulsifier PVA concentration as investigation factors in a 3-factor, 3-level BBD experimental design, the optimal preparation process for asiaticoside mPEG-PLGA microspheres was determined as: mPEG-PLGA concentration 123.8 g/L, PVA mass concentration 9.8 g/L, and water-to-oil ratio 12.7∶1. The resulting microspheres had uniform particle size (58.97 ± 4.44) μm, a drug loading of (8.43 ± 0.16)%, and an encapsulation efficiency of (62.66 ± 0.84)%. They released approximately 80% of asiaticoside at a stable rate within 96 h, and maintained over 90% cell viability after 72 h of co-culture with L929 cells. Conclusion Asiaticoside-loaded mPEG-PLGA microspheres were successfully prepared. The obtained microspheres exhibit uniform particle size, relatively high encapsulation efficiency and drug loading, enable sustained drug release, and possess good biocompatibility. This system utilizes the hydrophilicity of mPEG-PLGA to improve drug stability and release behavior, providing a new strategy for the efficient delivery of asiaticoside.

Objective To investigate the preparation process of the benchmark sample of Yinchenhao Decoction (YD, 茵陈蒿汤). Methods An HPLC method was established to simultaneous determine the content of 11 components in YD, including chlorogenic acid, 1-caffeoylquinic acid (1-CQA), geniposide, shanzhiside, genipin gentiobioside (GG), crocin I, crocin II, jasminoside B, aloe emodin-8-O-glucopyranoside (AE8G), rhein-8-O-glucoside (R8G) and gallic acid. Using the extraction amount or transfer rate of these 11 components as indicators, single factor investigations were conducted on the preparation of benchmark sample of YD, including the decocting methods, quantity of water addition, decoction time, decoction times, and concentration temperature. Taking the quantity of water addition, decoction time, and concentration temperature as the evaluation factors, the two levels set for the single factor investigations mentioned above were optimized and combined by the orthogonal experimental method, and the optimized process was verified through amplification experiments. Results The single-factor investigation indicated that the extraction amounts of six, nine, and 10 components were higher under the conditions of co-decoction, decocting with 18-fold and 15-fold water for 30 min and 20 min, respectively, compared to the conditions of pre-decocting Yinchen (Artemisiae Scopariae Herba) followed by decocting with 12-fold and 10-fold water for 60 min and 40 min (P < 0.05, 0.01). Compared with the total amount extracted three times, the extraction amount of each component in the first two decoctions were all close to or exceeded 70%. The transfer rates of two components were significantly higher under reduced-pressure concentration at 50  ℃ than at 80  ℃ (P < 0.01). The optimal preparation process using orthogonal experiment for the benchmark sample of YD was as follows: decocting the three herbs together twice, with 18-fold and 15-fold volume water, for 30 and 20 min, respectively, then concentrating at 50 ℃ under reduced pressure. The results of the validation experiment magnified by 10 was consistent with lab-scale results. Conclusion The optimized process is stable, simple, and repeatable, which can be used for the preparation of substance benchmark of classical prescription YD.

Objective To investigate the protective effects and underlying molecular mechanisms of p-coumaric acid (p-CA) against acute myocardial ischemia (AMI) through inhibition of inflammatory signaling. Methods Common targets of p-CA and AMI were predicted and screened using databases such as PubChem. These common targets were imported into the STRING database to construct a protein-protein interaction (PPI) network, followed by topological analysis. Gene ontology (GO) function and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis were performed using DAVID database and bioinformatics platform. The binding affinity and stability between p-CA and myeloid differentiation 2 (MD2), a co-receptor of Toll-like receptor 4 (TLR4), were analyzed using molecular docking and molecular dynamics simulations. An AMI mouse model was established by ligating the left anterior descending coronary artery. The mice were randomly assigned into sham group, model group, p-CA (100 mg/kg) group, carvedilol (7.6 mg/kg) group and TAK-242 (3 mg/kg) group. After seven consecutive days of treatment, echocardiography was performed to evaluate cardiac function. Myocardial histopathological damage was observed using hematoxylin-eosin (HE) and Masson staining. Activity of creatine kinase-MB, level of cardiac troponin I (cTnI) in serum and levels of inflammatory factors in myocardial tissue were detected. The expressions of proteins related to TLR4/nuclear factor-κB (NF-κB) inflammatory pathway in myocardial tissue were measured by Western blotting. A hypoxia-inflammatory complex injury model in H9c2 cardiomyocytes was constructed to further evaluate the regulatory mechanism of p-CA on TLR4/NF-κB inflammatory signaling pathway. Results Network pharmacology analysis indicated that p-CA could act on core targets such as TLR4 and regulate TLR and NF-κB pathways to treat AMI. Molecular docking results demonstrated that p-CA exhibited favorable binding affinity with MD2, a co-receptor of TLR4. Molecular dynamics simulations further verified that the p-CA-MD2 complex possessed good structural stability. The results of animal experiments showed that p-CA significantly improved cardiac function in AMI mice (P < 0.01, 0.001), alleviated inflammatory cell infiltration in myocardial tissue, inhibited collagen deposition and myocardial fibrosis, and reduced activity of CK-MB and level of cTnI in serum (P < 0.01, 0.001). Furthermore, p-CA downregulated TLR4, p-NF-κB/NF-κB, NOD-like receptor thermal protein domain associated protein 3 (NLRP3), gasdermin D (GSDMD), interleukin-1β (IL-1β) protein expressions and IL-6 level in myocardial tissue (P < 0.05, 0.01, 0.001). The results of cell experiments confirmed that p-CA significantly suppressed the protein expression levels of TLR4, p-NF-κB/NF-κB and NLRP3 (P < 0.05, 0.01). Conclusion p-CA could improve cardiac function in AMI, inhibit myocardial inflammatory response and fibrosis, and its mechanism is associated with the regulation of TLR4/NF-κB inflammatory pathway.

Objective To investigate the preventive and therapeutic effects of Banxia Xiexin Decoction (半夏泻心汤, BXD) on colorectal cancer (CRC) and its underlying mechanism based on “farnesoid X receptor (FXR)-bile acid-immune microenvironment” axis. Methods A mouse CRC model was established by intraperitoneal injection of azoxymethane combined with oral administration of dextran sulfate sodium (DSS). Control group, model group, BXD low-, medium-, high-dose (3.915, 7.830, 15.660 g/kg) groups and mesalazine (0.6 g/kg) group were set up, with 10 mice in each group. Drugs were continuously given for eight weeks, general condition of mice was recorded. Histopathological changes in colon tissue were observed. Immunofluorescence staining was used to examine the distribution and expressions of CD11c, CD80 and CD86 in colon tissue. Flow cytometry was employed to detect the expressions of CD11b, CD11c, CD80, CD86 and CD206 in spleen tissue. Targeted metabolomics was used to analyze the levels of bile acids in feces of mice. Levels of interferon-γ (IFN-γ), interleukin-12 (IL-12), IL-18, total bile acids in serum and total bile acids in bile were detected. qRT-PCR was performed to measure the mRNA expressions of FXR, fibroblast growth factor 15 (FGF15), G protein-coupled receptor 41 (GPR41), GPR43, GPR109A, histone deacetylase 3 (HDAC3), HDAC7, inducible nitric oxide synthase (iNOS), IL-12, epidermal growth factor (EGF) and transforming growth factor-β1 (TGF-β1) in colon tissue. Western blotting was used to detect the protein expressions of FXR, iNOS, IL-12, EGF, TGF-β1, FGF15, GPR41, GPR43 and GPR109A in colon tissue. Results Compared with control group, the model group showed significant weight loss, increased number and size of colon tumors, severe disruption of glandular structure, irregular hyperplasia and distortion, formation of gland-like or nest-like structures, accompanied by marked inflammatory cell infiltration and tissue damage. PAS-positive substances were focally aggregated, distributed disorderly with irregular morphology. CD11c⁺ cells were extensively aggregated in colon tumor tissue, while CD80⁺ cells showed almost no fluorescence. The expressions of CD11b, CD11c and CD80 in spleen tissue was significantly increased (P < 0.01). Levels of deoxycholic acid (DCA), lithocholic acid (LCA) and taurocholic acid (TCA) in fecal were significantly elevated (P < 0.01). Protein expressions of iNOS, IL-12, EGF, TGF-β1, HDAC7 and HDAC3 in colon tissue were significantly increased (P < 0.01), while protein expressions of FXR, FGF15, GPR41, GPR43 and GPR109A were significantly decreased (P < 0.01). Compared with model group, the changes in the above indicators were reversed in both BXD and mesalazine groups, and the changes were more pronounced in BXD high-dose group (P < 0.05, 0.01). Conclusion BXD may exert its preventive and therapeutic effects on colorectal cancer by activating FXR signaling pathway, regulating bile acid metabolic homeostasis, and subsequently reshaping the tumor immune microenvironment.

Objective To investigate the mechanism by which Tongguanteng Injection (通关藤注射液, TGT) modulates macrophage polarization to synergize with doxorubicin (DOX) in inhibiting triple-negative breast cancer (TNBC) growth. Methods A 4T1 TNBC- bearing mouse model was constructed, randomly divided into model group, TGT group, DOX group and TGT combined with DOX treatment group. Tumor volume and weight were observed to evaluate in vivo antitumor efficacy. Hematoxylin-eosin (HE), TUNEL and immunohistochemical staining were employed to examine histopathological morphological changes in tumor tissues, tumor cell apoptosis, and protein expressions of cleaved cystein-asparate protease-3 (Caspase-3), B-cell lymphoma-2 (Bcl-2), and Bcl-2 associated X protein (Bax). Immune organ indices and routine blood analysis were conducted to assess immune function in mice. ELISA, qRT-PCR and immunofluorescence staining were performed to measure inflammatory cytokine levels and expressions of macrophage polarization markers CD86, inducible nitric oxide synthase (iNOS), CD206, arginase-1 (Arg-1) in tumor tissues. Flow cytometry and immunohistochemical analysis were used to determine the infiltration ratios of CD4⁺ T and CD8⁺ T cells, while qRT-PCR was employed to detect mRNA expression levels of T cell exhaustion-related factors and cytotoxic factors. Transcriptome sequencing and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis were conducted to identify core signaling pathways and perform validation. Results Compared with model group, the tumor volume and weight were significantly reduced in TGT group, DOX group and TGT combined with DOX treatment group (P < 0.01, 0.001), varying degrees of necrosis were observed in tumor tissues, the apoptosis rate of tumor cells was significantly increased (P < 0.001), these changes were more pronounced in TGT + DOX combination therapy group (P < 0.05, 0.01, 0.001). Compared with model group, the expression levels of pro-inflammatory cytokines tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were significantly increased in TGT group (P < 0.01, 0.001), while the expression levels of anti-inflammatory cytokines IL-4 and IL-10 were significantly decreased (P < 0.05, 0.01). The proportion of M1-type macrophages (CD86⁺/F4/80⁺) was increased (P < 0.001), with significantly elevated mRNA expression levels of markers CD86 and iNOS (P < 0.05, 0.01). The proportion of M2-type macrophages (CD206⁺/F4/80⁺) was decreased (P < 0.001), with significantly reduced mRNA expression levels of markers CD206 and Arg-1 (P < 0.01, 0.001). The proportion of CD8⁺ T cells was significantly increased (P < 0.05, 0.001), while the proportion of CD4⁺ T cells was significantly decreased (P < 0.01). The mRNA expression levels of T cell exhaustion markers programmed cell death protein-1 (PD-1), T cell immunoglobulin and mucin domain-containing protein-3 (Tim-3) and lymphocyte activation gene-3 (Lag-3) were significantly downregulated (P < 0.05, 0.01), whereas the mRNA expression levels of T cell cytotoxic factors perforin, interferon-γ (IFN-γ) and granzyme B were significantly upregulated (P < 0.05, 0.001). These changes were more pronounced in TGT combined with DOX treatment group (P < 0.05, 0.01, 0.001). Transcriptome and KEGG enrichment analyses revealed that, compared with model group, differentially expressed genes in the combination therapy group were significantly enriched in Janus kinase (JAK)-signal transducer and activator of transcription (STAT) signaling pathway. Immunohistochemical staining results indicated that TGT combined with DOX significantly downregulated the expressions of p-JAK2 and p-STAT3 proteins in tumor cells (P < 0.001). Conclusion TGT combined with DOX significantly inhibits the growth of TNBC, promotes the polarization of tumor associated macrophages from M2 to M1, promotes CD8⁺ T cell infiltration and reduces their exhaustion. Its mechanism is related to the inhibition of phosphorylation of JAK-STAT signaling pathway.

Objective To systematically evaluate the therapeutic effects of Jinzhen Oral Liquid (金振口服液, JZOL) on a pathological model of chronic obstructive pulmonary disease (COPD) complicated with phlegm-heat syndrome, and to elucidate its pharmacological mechanisms from multiple dimensions, including improvement of pulmonary function, suppression of systemic inflammation, regulation of airway mucus secretion, restoration of Th1/Th2 and Th17/Treg immune balance, and modulation of transcriptomic expression profiles. Methods A rat COPD model was established through eight weeks of cigarette smoke exposure combined with intranasal lipopolysaccharide (LPS) instillation. Subsequently, a compound model of COPD with phlegm-heat syndrome was induced by 9 d of wind-heat exposure combined with LPS instillation. Model rats were randomly assigned to model group, dexamethasone (0.2 mg/kg) group and JZOL low-, medium-, high-dose (0.8, 1.6, 3.2 g/kg) groups, and received continuous oral administration for 7 d. Therapeutic efficacy was comprehensively evaluated by assessing pulmonary function parameters, systemic symptoms and signs, inflammatory factors in serum, lung histopathological changes and proportions of lung T cell subsets in lung tissue. Furthermore, transcriptomic sequencing and qRT-PCR were employed to validate the regulatory effects of JZOL on key differentially expressed genes, including lipocalin-2 (Lcn2), indoleamine 2,3-dioxygenase 1 (Ido1), cyclin E1 (Ccne1), homeobox protein B8 (Hoxb8) and G protein-coupled receptor 17 (Gpr17). Results The compound modeling successfully induced severe airflow limitation, fever, constipation, and histopathological and cytopathological alterations in rats. Compared with model group, JZOL significantly improved lung function (P < 0.05, 0.01, 0.001), significantly reduced levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), C-reactive protein (CRP) and neutrophil elastase (NE) in serum (P < 0.001), significantly reduced bronchial epithelial goblet cell proliferation and acid mucin secretion (P < 0.05, 0.001), and corrected imbalanced Th1/Th2 and Th17/Treg cells ratios (P < 0.05, 0.001). The transcriptome and qRT-PCR validation results showed that JZOL improved COPD through multiple mechanisms, including downregulation of immune microenvironment and protease imbalance related genes, inhibition of abnormal cell cycle and airway remodeling genes, upregulation of key lung regeneration factors and regulation of developmental factors, as well as activation of tissue repair and inflammation resolution pathways. Conclusion JZOL effectively improves lung function and systemic symptoms in COPD rats with phlegm heat syndrome by inhibiting systemic and local inflammatory responses through multiple targets, regulating T cell immune balance.

Objective To investigate the ameliorative effect and underlying mechanism of Compound Danshen Soft Capsules (复方丹参软胶囊, CDSC) on isoproterenol (ISO)-induced myocardial ischemia in mice through the integration of serum metabolomics and network pharmacology of absorbed bioactive components. Methods A mouse model of myocardial ischemia was established by ip ISO. After CDSC intervention, the pathological changes in myocardial tissue were evaluated using hematoxylin-eosin (HE) and Masson staining. The activity of energy metabolism enzymes in myocardial tissue were measured. ELISA was used to measure the levels of myocardial injury markers in myocardial tissue. Identification and analysis of CDSC blood components were performed using ultra-high performance liquid chromatography-time-of-flight mass spectrometry (UHPLC-TOF-MS). CDSC differential blood components for network pharmacology analysis was used to explore potential mechanisms. Serum untargeted metabolomics was used to analyze the effect of CDSC on serum metabolic levels in mice with myocardial ischemia. Western blotting was used to validate the protein expressions of related pathways. Results CDSC significantly alleviated the pathological damage of myocardial tissue in ISO-induced myocardial ischemia mice, increased the activities of energy metabolism enzymes in myocardial tissue (P < 0.001), and reduced the levels of myocardial injury markers (P < 0.001). A total of 17 differential blood components were identified, and network pharmacology analysis screened 187 intersecting targets related to myocardial injury. Kyoto encyclopedia of genes and genomes enrichment analysis suggested that the effect of CDSCs in improving myocardial ischemia may be related to signaling pathways such as resistance to epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors, AMP-activated protein kinase (AMPK) and advanced glycation end products-receptor for advanced glycation end products (AGE-RAGE). Serum metabolomics analysis suggested that the improvement effect of CDSC on myocardial ischemia mice may be related to multiple metabolic pathways such as glucagon, carbon metabolism, pyruvate metabolism and purine metabolism. Western blotting results further confirmed that CDSC could dose-dependently regulate the phosphorylation levels of AMPK and mammalian target of rapamycin (mTOR) protein (P < 0.01, 0.001). Conclusion CDSC improves myocardial ischemia by regulating energy metabolism and other mechanisms, and its mechanism may be related to AMPK/mTOR signaling pathway.

Objective To investigate the protective effect and mechanism of Buyang Huanwu Decoction (补阳还五汤, BYHWT) on intestinal mucosal barrier injury following cerebral ischemia-reperfusion injury (CIRI) in rats. Methods Network pharmacology was employed to screen potential targets of BYHWT for treating intestinal mucosal injury post-CIRI, followed by enrichment analysis. The improved Zea Longa suture method was used to prepare a rat CIRI model, which was randomly divided into model group and BYHWD low-, medium-, and high-dose (1.29, 2.57, 5.14 g/kg) groups, with six rats in each group, and six rats in sham group. After continuous administration for 5 d, the neurological deficit score and cerebral infarction area were measured. Hematoxylin-eosin (HE), TUNEL staining and electron microscopy were used to observe the pathological, apoptotic and ultrastructural changes of colon tissue. Immunohistochemistry and Western blotting were used to detect the protein expressions of Occludin and zonula occludin-1 (ZO-1) in colon tissue. Non-targeted metabolomics was performed on intestinal contents, and the results were integrated with network pharmacology to construct a “metabolite-reaction-enzyme-gene” network. Key pathways were validated using molecular docking, ELISA and Western blotting. Results Network pharmacology identified 22 core targets of BYHWT for treating post-CIRI intestinal mucosal injury, involving pathways such as lipid metabolism and mitogen activated protein kinase (MAPK). Compared with sham group, model group showed aggravated neurological damage (P < 0.001), increased cerebral infarction area (P < 0.05), disrupted colon mucosal structure, decreased expressions of Occludin and ZO-1 (P < 0.01), and increased cell apoptosis (P < 0.05). Compared with model group, BYHWT significantly improved the above indicators (P < 0.05, 0.01). Metabolomics discovered nine differential metabolite downregulation pathways, mainly enriched in steroid hormones, bile acids and arachidonic acid metabolism pathways. The integrated analysis of metabolomics and network pharmacology suggested that BYHWT exerted its effect by regulating arachidonic acid/dihydro-γ-linolenic acid prostaglandin metabolic pathway. Molecular docking showed that prostaglandin E₁ (PGE₁) bind well to prostaglandin endoperoxide synthase 2 (PTGS2). After BYHWT intervention, the level of PGE₁ in colon tissue was significantly increased (P < 0.05), while the expression of PTGS2 protein was significantly decreased (P < 0.05). Conclusion BYHWT may alleviate intestinal mucosal barrier injury following CIRI by modulating arachidonic acid/dihomo-γ-linolenic acid-PTGS2-prostaglandin metabolic network.

Objective To identify potential therapeutic targets for idiopathic pulmonary fibrosis (IPF) and predict related herbal medicines by integrating bioinformatics, Mendelian randomization (MR), and machine learning approaches. Methods IPF microarray datasets were obtained from the GEO database to identify differentially expressed genes (DEGs). Using expression quantitative trait loci (eQTL) data and genome-wide association study (GWAS) data, MR analysis was conducted to screen for genes associated with IPF. The risk genes identified from MR analysis were intersected with DEGs to filter core IPF-related genes. Subsequent evaluations included functional enrichment analysis, gene set enrichment analysis (GSEA), immune cell infiltration analysis, and single-cell RNA sequencing. Machine learning algorithms were applied to select optimal diagnostic feature genes. An independent GEO cohort was used for differential expression validation and receiver operating characteristic (ROC) analysis. Results A total of 916 IPF-associated DEGs were identified. Intersection with 224 MR risk genes yielded seven key genes: IRF7, TTC32, IFI6, and ISG15 (risk genes), along with ZNF204P, ISOC1, and CTSK (protective genes). These genes were primarily enriched in pathways related to interferon-beta production, RIG-I-like receptor signaling, Toll-like receptor signaling, and type I interferon signaling. Immune infiltration analysis revealed a decrease in M1/M0 macrophages and resting mast cells, alongside an increase in activated mast cells in IPF tissues. Single-cell RNA sequencing demonstrated specific expression patterns of these genes within epithelial cell subpopulations. Machine learning algorithms identified ZNF204P and IRF7 as the optimal diagnostic genes. Validation in the dataset confirmed their significant differential expression in IPF and high diagnostic accuracy. A total of 385 traditional Chinese medicines (TCMs) related to the key genes were predicted. The primary properties of these TCMs were cold, warm, and neutral (four natures); their main flavors were bitter, sweet, and pungent (five flavors); the principal meridian tropisms were the liver, lung, stomach, spleen, and kidney meridians; and the major classifications were heat-clearing drugs, tonifying drugs, blood-activating and stasis-resolving drugs, exterior-releasing drugs, and dampness-draining diuretics. Molecular docking simulations revealed that these chemical components could form stable interactions with the core proteins. Conclusion This study identified seven key genes associated with IPF. Machine learning screened ZNF204P and IRF7 as robust diagnostic biomarkers with therapeutic target potential. Furthermore, TCMs such as Zhizi (Gardeniae Fructus), Chaihu (Bupleuri Radix), Roucongrong (Cistanches Herba), and Huangqi (Astragali Radix) might be potential TCMs that targets core genes associated with IPF.

Objective Discovering the ancient and modern Chinese compound prescriptions containing Suanzaoren (Ziziphi Spinosae Semen)-(Renshen) Ginseng Radix et Rhizoma herb pair (ZSS-GRR) and further analyzing the diseases commonly treated by the pairs and their therapeutic mechanisms. Methods Data mining method was applied to collect the compound formulas containing ZSS-GRR and count the commonly treated diseases and pharmacological effects, network pharmacology was applied to predict the mechanism of ZSS-GRR herb pair, and molecular docking technique was used to verify the binding ability of candidate compounds to potential targets, and finally enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry were used to validate the discovered mechanisms of ZSS-GRR for insomnia and nervous system function. Results A total of 596 compound formulations containing the ZSS-GRR were searched. Statistical analysis of the pharmacological effects of these formulations revealed that the ZSS-GRR primarily exerts sedative and hypnotic effects and is commonly used to treat conditions such as insomnia. Network pharmacology analysis indicates that the ZSS-GRR may exert its sedative and hypnotic effects by regulating neuroactive ligand-receptor interactions, gamma-aminobutyric acid (GABA) ergic synapses, and inflammation-related pathways. Molecular docking results suggest that multiple core active components exhibit strong binding affinity with key targets. Experimental results demonstrate that the ZSS-GRR combination can alleviate glutamate-induced damage in SH-SY5Y cells, increase levels of relevant neurotransmitters in rats with an insomnia model, reduce levels of inflammatory factors, and promote the expression of gamma-aminobutyric acid type A receptor subunit alpha 1 (GABRA1) and gamma-aminobutyric acid type A receptor subunit gamma 2 (GABRG2) in the hippocampus and hypothalamus. Conclusion Through the exploration of the commonly treated diseases and the sedative-hypnotic mechanism of ZSS-GRR, the main direction and mechanism of action of ZSS-GRR have been revealed, which provides certain valuable references for the subsequent in-depth research and clinical application of the pair.

Objective To perform a visualized bibliometric analysis of research on the Dai medicine Clerodendranthus spicatus in the field of traditional Chinese medicine based on the CiteSpace knowledge graph, systematically summarize recent research trends, clarify hotspots and future directions, and provide a reference for further development and application of C. spicatus. Methods Chinese and English literature on C. spicatus in traditional Chinese medicine was retrieved from China National Knowledge Infrastructure (CNKI) and the Web of Science (WOS) Core Collection from 1 January 1995 to 30 October 2025. Records from each database were imported into NoteExpress for automatic and manual de-duplication, and then screened according to predefined inclusion and exclusion criteria. Based on the final dataset, CiteSpace and Microsoft Excel were used to perform visual analyses of publication output, authors, institutions, country distribution and keywords. In addition, the top 10 most-cited Chinese and English articles were subjected to high-cited literature analysis. Results A total of 626 publications were included, comprising 274 Chinese and 352 English articles, with an overall fluctuating upward trend in annual output. The most productive authors were Song Liqun and Ismail Zhari. Co-authorship networks showed relatively close collaboration within research teams but limited cooperation between different teams. The most productive institutions were the Yunnan Branch of the Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences, and Universiti Sains Malaysia, with core institutions mainly being traditional Chinese medicine research institutes and universities. Malaysia was the most productive country. Keyword analysis indicated that Chinese studies mainly focused on medicinal resources, traditional uses and clinical research, whereas English literature emphasized phytochemical profiling, pharmacological activities, data mining and molecular mechanisms. High-cited literature analysis revealed that both Chinese and English classic studies chiefly concentrated on chemical constituents and material basis, pharmacological effects and mechanisms, preparation technology and modern methodological innovation, as well as research progress, collectively forming a “constituent-pharmacology-formulation” research framework. Conclusion Research on C. spicatus in the field of traditional Chinese medicine has shown sustained growth, with its pharmacological mechanisms and clinical applications in the prevention and treatment of kidney diseases, rheumatoid arthritis, gout, osteoarthritis, and cancer becoming major research hotspots. Future studies should integrate systems pharmacology and multi-omics technologies, strengthen mechanistic exploration, and establish scientific quality control and clinical evaluation systems, thereby promoting the modernization and innovative development of C. spicatus and providing a reference for research on this herb and other ethnic medicines.

Objective This study employed iPBS molecular marker technology to analyze the genetic diversity and genetic differentiation relationships among 32 Dendrobium linawianum germplasm resources from different regions. Methods A “0, 1” matrix was constructed using Excel 2016, and the genetic similarity coefficient (G_s) was calculated using NTSYS-pc 2.10e to construct a UPGMA cluster dendrogram. Additionally, various genetic diversity indices were calculated using Popgene 1.32. Results A total of 84 clear bands were amplified from the six selected iPBS primers, including 76 polymorphic bands, with a polymorphism rate of 90.48% and an average polymorphism information content (PIC) of 0.46, indicating high polymorphism of the iPBS primers. The genetic similarity coefficients ranged from 0.547 6 to 0.965 5, with an average of 0.766 8. At a genetic similarity coefficient of 0.732, the 32 germplasm samples were classified into four populations. The average observed allele number (N_a), average effective allele number (N_e), Nei's genetic diversity index (h), and Shannon diversity information index (I) were 1.631 0, 1.381 6, 0.219 0, and 0.326 7, respectively. Genetic structure analysis revealed that the genetic diversity (H_t), gene differentiation coefficients (H_s) among populations, and genetic differentiation coefficients (G_st) were 0.281 3, 0.219 0, and 0.221 3, respectively. Genetic variation among populations accounted for 22.13% of total variation, while genetic variation within populations accounted for 77.87%, indicating that genetic variation was primarily derived from within populations. Furthermore, DNA fingerprinting of 32 germplasm resources was successfully constructed based on 19 polymorphic loci with six primers, which provided a reference for the classification and identification of D. linawianum. Conclusion The genetic diversity of D. linawianum was significantly lower between populations than within populations, and the genetic variation was mainly within populations. The genetic differentiation characteristics of germplasm resources are significantly correlated with their geographical distribution patterns. This study not only elucidates the genetic relationships among different germplasm of D. linawianum and the characteristics of population genetic diversity, but also provides an important theoretical basis for the subsequent scientific conservation and sustainable utilization of D. linawianum germplasm resources.

Objective To clarify the dynamic accumulation pattern of secondary metabolites in the leaves of Acanthopanax senticosus during the growth season, providing theoretical support for precision-guided leaf harvesting tailored to different usage purposes. Methods Colorimetric method and untargeted metabolomics technology were used to analyze the dynamic accumulation characteristics of metabolites such as flavonoids, polyphenols and saponins in the leaves of A. senticosus. Results The growth season was divided into 13 stages (L1—L13) based on leaf morphodevelopmental traits. Total flavonoids, polyphenols, triterpenoids, and saponins exhibited bimodal accumulation peaks at L3 (the peak leaf expansion stage) and L11 (the end of July), with maximum contents at L11, reaching 8.02-, 1.68-, 1.61-, and 1.63-fold increases, respectively, compared to L1. KEGG enrichment analysis revealed that, relative to L3, L11 showed specific upregulation of phenylpropanoid and flavonoid biosynthesis pathways, coupled with synergistic accumulation of light signaling molecules (FAD, lumiflavin) and defense-related compounds (sinapine, quercetin glycosides), while nucleotide synthesis precursors significantly decreased. Additionally, the pharmacopoeial quality control indicator syringin increased by 2.51-fold in L11 relative to L3. Conclusion The secondary metabolism of A. senticosus leaves exhibits significant developmental stage-dependence, among which the L11 stage is a high-enrichment period, this provides metabolomic evidence for the precise utilization of A. senticosus leaves.

Objective This study aimed to investigate the correlation between bitterness and antioxidant activity in Coptis chinensis, systematically characterize its bitter-active compounds, and establish a corresponding quality control method, using an integration of electronic sensory techniques, UPLC-Q-TOF-MS/MS, and molecular docking. Methods The bitterness intensity and antioxidant capacity of C. chinensis. from three geographical origins were quantified via an electronic tongue and in vitro antioxidant assays, and a subsequent correlation analysis was conducted to explore their relationship. Chemical constituents were identified by UPLC-Q-TOF-MS/MS, and differential components and potential bitter compounds were screened by integrating plant metabolomics and correlation analysis between chemical components and bitterness. Molecular docking was performed to further screen components with strong binding affinity to the bitter taste receptors TAS2R1, TAS2R7, TAS2R8, and TAS2R14, and their binding modes were summarized. By integrating these results, the bitter-active compounds responsible for both taste and antioxidant effects were identified. Subsequently, a high-performance liquid chromatography (HPLC) method was developed for the simultaneous quantification of seven key bitter-active compounds (groenlandicine, jatrorrhizine, columbamine, epiberberine, coptisine, palmatine, and berberine) for quality control. Results Correlation analysis confirmed a positive relationship between bitterness intensity and antioxidant capacity across the three origins. UPLC-Q-TOF-MS/MS analysis led to the identification of 87 chemical constituents, from which origin-specific differential components and potential bitter-tasting substances were screened. Molecular docking identified 18 components with high binding affinity to the target bitter receptors, and their specific binding modes were elucidated. Integrated analysis pinpointed the bitter-active compounds in C. chinensis that contribute to both taste perception and antioxidant activity. A reliable HPLC method for the simultaneous quantification of the seven key bitter-active compounds was successfully established. Conclusion This study clarified the bitter-active compounds in C. chinensis and developed a quality control method based on these components. The findings provide a theoretical foundation and practical reference for elucidating the “bitter taste-efficacy” relationship and for optimizing the quality control system of C. chinensis.

Objective To elucidate the chemical composition differences among various parts of Pugongying (Taraxacum mongolicum), this study compared cultivated and wild T. mongolicum samples, dividing them into three parts: whole plant, above-ground parts, and roots. Methods A Venusil C₁₈chromatographic column was used with acetonitrile-0.2% phosphoric acid as the mobile phase for gradient elution at a flow rate of 1.0 mL/min, column temperature of 30 °C, and detection wavelength of 330 nm. A fingerprint spectrum of T. mongolicum medicinal materials was established, and the contents of six components (monocaffeoyl tartaric acid, chlorogenic acid, caffeic acid, ferulic acid, isochlorogenic acid C, and chicoric acid) were determined in different parts. The collected fingerprint spectrum data were analyzed using similarity evaluation, cluster analysis (CA), and orthogonal partial least squares discriminant analysis (OPLS-DA). Results The T. mongolicum fingerprint spectrum identified nine common peaks, with six major chemical components confirmed and five primary differential components screened. Quantitative analysis revealed significant differences in chemical composition content between T. mongolicum roots compared to whole plant and above-ground parts. Conclusion This method is simple, reliable, and highly reproducible. The comprehensive quality evaluation model provides objective analytical results suitable for holistic quality assessment of T. mongolicum medicinal materials. This study enriches research on quality analysis of T. mongolicum and offers evidence for future development, quality standardization, and clinical application.

Objective Huanghuacai (Hemerocallis citrina) is a traditional vegetable with anti-inflammatory and anti-depressant effects. Its leaves are widely used in Taiwan and other regions to promote sleep, however, there are few reports on its chemical composition. This article aims to analyze the characteristic components of H. citrina leaves by fingerprint spectrum and to further determine the content of the index components, including chlorogenic acid, rutin, quercetin-3-O-glucoside, quercitrin, and quercetin, providing a basis for better utilization and quality evaluation of H. citrina leaves. Methods Firstly, the ethanol extraction method was used to prepare H. citrina leaves extract. Chromatographic separation was performed by DIKMA spursil C₁₈ column (250 mm × 4.6 mm, 5 mm) maintained at 30 ℃ and the mobile phase was acetonitrile-0.5% acetic acid water. The injection volume was 10 mL, the detection wavelength was 190—400 nm and the flow rate was 1 mL/min. Based on the principle of maximum absorption wavelength, the wavelengths of 258 nm was selected in two periods of 0—5.5 and 23—48 min, the wavelengths of 300 and 370 nm were selected in two periods of 5.5—23 and 48—80 min respectively, and a three-wavelength switching fingerprint was constructed. Secondly,a quantitative analysis of multi-components by single marker (QAMS) was used to determine the contents of chlorogenic acid, rutin, quercetin-3-O-glucoside, quercetin, and quercetin in ethanol extract of H. citrina leaves. Results The similarity of fingerprint spectrums of 10 batches of H. citrina leaves was above 0.90, and the three-wavelength switching fingerprint spectrums included 42 common peaks (accounting for more than 90% of the total peak area), of which 11, 25, and 26, 32 and 37 were identified as chlorogenic acid, rutin, quercetin-3-O-glucoside, quercitrin, and quercetin. With quercetin (S) as the internal reference, the relative correction factors of chlorogenic acid, rutin, quercetin-3-O-glucoside, and quercitrin were obtained by the QAMS as 1.402, 1.896, 1.268, 1.221. The average contents of the above five components in 10 batches of H. citrina leaves are 0.079, 0.086, 0.374, 0.325, 0.037 mg/g. There was no significant difference between the external standard method and QAMS. Conclusion The three-wavelength switching fingerprint spectrum constructed in this study has significantly increased the number of chromatographic peaks contained in any single wavelength fingerprint spectrum and can be used to characterize the chemical composition of H. citrina leaves. The QAMS further constructed can be used to determine the content of chlorogenic acid, rutin, quercetin-3-O-glucoside, quercitrin, and quercetin at one time. This study provides a methodological basis and reference for the comprehensive evaluation of H. citrina leaves quality.

Insomnia is a highly prevalent sleep disorder in clinical practice that severely impairs patients' daytime function and quality of life, and there are still unmet medical needs in its clinical treatment. Traditional Chinese medicine (TCM) has unique advantages in the treatment of insomnia, such as holistic regulation, mild adverse reactions, and long-lasting efficacy.In February 2026, the Center for Drug Evaluation of the National Medical Products Administration issued the Guiding Principles for Clinical Trials of Therapeutic Drugs for Chronic Insomnia (for Trial Implementation). However, it lacks targeted and specific regulatory guidance for the clinical trial design of innovative traditional Chinese medicines. Traditional Chinese medicines exert therapeutic effects on insomnia through multiple targets and pathways, which is significantly different from the clinical trial design of chemical drugs. Based on the clinical advantages and characteristics of TCM in the treatment of insomnia, combined with Chinese and Western medicine guidelines for insomnia, technical requirements for TCM clinical trials, and relevant research progress, this paper systematically analyzes key elements including trial background, diagnostic criteria, inclusion and exclusion criteria, run-in period and treatment course design, background therapy and concomitant medication, efficacy and safety indicators, control group selection, statistical analysis, and benefit-risk, and clarifies the design principles, implementation points, and precautions for each key element in the clinical trials of innovative TCM for insomnia, proposes scientific and rational clinical trial design strategies for innovative TCM in the treatment of insomnia, provides a reference for the standardized conduct of such clinical trials, and supports the establishment of high-quality TCM evidence-based evidence and the research, development and promotion of innovative traditional Chinese medicines.

Impaired exercise capacity is an independent risk factor for poor prognosis in patients with cardiovascular disease (CVD). The vicious cycle triggered by CVD, centered on cardiac dysfunction, progressively compromises pulmonary ventilation/gas exchange, skeletal muscle metabolism, and skeletal function, collectively leading to exercise intolerance. The “cardio-pulmonary-muscular-skeletal” functional axis theory is proposed to elucidate this multisystem deterioration. Within the framework, this review systematically discusses the mechanisms by which traditional Chinese medicine (TCM) interventions synergistically reverse exercise intolerance. These mechanisms include improving cardiac and vascular function, enhancing pulmonary ventilation and oxygenation, increasing muscle mass and metabolic function, and maintaining bone quality and mechanical function, thereby leveraging the multi-targeted and holistic regulatory advantages of TCM. Although the overall quality of clinical evidence remains moderate, a multidimensional evidence system centered on exercise tolerance has been established. This evidence confirms the consistent benefits of TCM formulas on core indicators across the functional axis. The functional axis theory provides an innovative framework for understanding the multisystemic underlying mechanisms of exercise capacity decline in CVD patients and for guiding holistic TCM-based interventions, highlighting the unique value of TCM in cardiac rehabilitation.

As the precursor of sex hormone synthesis, depression is profoundly influenced by metabolic pathways, which also affect how patients respond to antidepressants. In addition, when sex hormones are dysregulated, it significantly increases both the risk of developing depression and the likelihood of its worsening over time, and the disorder of cholesterol metabolism can lead to depression by disrupting the homeostasis of sex hormone microenvironment in the brain. Therefore, drugs can exert antidepressant effects by mediating cholesterol metabolism, regulating the levels of sex hormones and their receptors, and restoring the homeostasis of brain sex hormone microenvironment. This article aims to review the mechanism of the imbalance of brain sex hormone homeostasis caused by cholesterol mediated metabolic disorders of neurosteroid deficiency, hypothalamic-pituitary-adrenal axis hyperactivation, and inflammatory pathway activation. Furthermore, stable isotope tracing, metabolomics, biological network regulation and other technologies were applied to reveal the internal mechanism of “cholesterol metabolism sex hormone homeostasis depression”, as well as the regulatory effect of drug intervention on depression. It provides novel research strategies for revealing the mechanism of depression and drug intervention from the perspective of homeostatic regulation of sex hormone microenvironment.

Micelles, formed through the self-assembly of amphiphilic molecules, hold significant promise as drug delivery systems for enhancing the solubility, stability, and targeting properties of active components in traditional Chinese medicine (TCM). Within the field of Chinese medicine formulation, numerous active constituents can spontaneously form micelles or be encapsulated within micellar carriers via specific preparation techniques, thereby effectively improving drug performance. This paper reviews the formation mechanisms, preparation techniques, and current applications of TCM micelles. In view of the hybrid nature of micelles within complex TCM systems, it combines structural characterisation techniques with key influencing factors in micelle formation. The study identifies existing issues such as disordered self-assembly and lack of controllability in current research, proposing corresponding suggestions and future directions. This aims to provide theoretical reference for modernisation of TCM and the development of innovative formulations.

Lung cancer is a malignant tumor with high incidence and mortality rates in the global. In recent years, traditional Chinese medicine (TCM) formulas have achieved breakthrough progress in regulating the tumor immune microenvironment (TME) and serving as adjuvant therapy for lung cancer. However, due to the multi-component, multi-target, and multi-pathway synergistic effects of TCM formulas, elucidating their mechanisms of regulating TME remains a significant challenge. This review systematically summarizes TCM’s understanding of the etiology and pathogenesis of lung cancer, as well as clinical treatment principles. It further explores the molecular mechanisms by which TCM regulates tumor immune microenvironments and provide insights into future research strategies for deciphering the mechanisms underlying the regulation of TME by TCM formulas. This review aims to provide a theoretical basis for understanding TCM-mediated TME regulation against lung cancer and promoting the precise application of TCM formulas in clinical lung cancer treatment.

Insomnia, as a common sleep disorder, exhibits a rising incidence rate annually, with its pathogenesis closely linked to gut microbiota dysbiosis. Currently, most of the clinical methods for the treatment of insomnia have limitations and adverse reactions. A review of recent domestic and international literature indicates that Lianzixin (Nelumbinis Plumula) possesses multi-component, multi-pathway, and multi-target characteristics. It exerts sedative and hypnotic effects by regulating the gut microbiota composition, influencing intestinal barrier function, microbial composition, and their metabolites, while also suppressing inflammatory responses and modulating immune homeostasis. However, its specific molecular mechanism network has not yet been fully elucidated. This review elaborates on mechanism of gut microbiota in occurrence and development of insomnia and the regulatory effects of Nelumbinis Plumula, establishes a systematic analysis of “Nelumbinis Plumula-gut microbiota-insomnia” axis, and provides theoretical support and reference for developing Nelumbinis Plumula-based insomnia treatments and microbiota-targeted drugs, advancing the collaborative prevention and treatment of insomnia through traditional Chinese medicine and Western medicine from basic research to precision clinical therapy.

Ovarian cancer is often characterized as a “silent killer” due to its insidious onset, difficulty in early diagnosis, and high mortality rate. Furthermore, advanced stages of the disease exhibit high recurrence rates and formidable chemoresistance. In recent years, the synergistic and sensitizing value of traditional Chinese medicine (TCM) in the comprehensive management of ovarian cancer has become increasingly evident. This review systematically summarizes the pivotal mechanisms underlying the intervention of TCM monomers and compound formulas in ovarian cancer. It reveals that TCM constituents not only inhibit tumor invasion and metastasis by inducing cell cycle arrest, promoting apoptosis and ferroptosis, and modulating crucial signaling pathways—including the mitogen-activated protein kinase/extracellular signal-regulated kinase and Janus kinase/signal transducer and activator of transcription pathways—but also demonstrate unique advantages in reversing platinum resistance and alleviating chemotherapy-induced myelosuppression by interfering with glycolipid metabolic reprogramming and remodeling the immune microenvironment. Future research should integrate multi-omics technologies to comprehensively validate these complex molecular networks, thereby providing robust evidence-based medical foundations for clinical precision therapeutics and the development of novel antineoplastic agents.

Zingiberaceae plants are important medicinal and edible resources, widely used in traditional diets and medicine. They combine nutritional value with diverse physiological regulatory functions, holding significant potential for future applications. This study focused on seven Zingiberaceae species listed in the National Health Commission’s inventory of traditional food and medicinal materials: Zingiber officinale (ginger, including fresh and dried forms), Alpinia oxyphylla, Amomum tsaoko, Curcuma longa, A. villosum, Kaempferia galanga, and Alpinia officinarum. A systematic literature review and industrial analysis were conducted to summarize their core functional components, pharmacological activities, and industrialization status. The results showed that these plants are generally rich in volatile oils, gingerols, curcumin, flavonoids, and polysaccharides, exhibiting bioactivities such as anti-inflammatory, antioxidant, metabolic regulatory, neuroprotective, and potential antitumor effects. Furthermore, a comprehensive system for extracting and identifying these functional components and evaluating their bioactivities has been established. Analysis of the industrial chain revealed uneven development of Zingiberaceae resources, with common challenges including fragmented cultivation, extensive processing, lack of standardization, weak branding, and insufficient industry-academia-research collaboration. Corresponding recommendations were proposed: promoting whole-chain standardization, advancing deep processing and high-value product development, building collaborative innovation systems, and implementing branding and internationalization strategies. Future research should integrate advanced technologies like multi-omics to elucidate mechanisms of action and provide evidence-based support hrough high-quality clinical studies. Technological innovation will drive the development and layout of high-value products, facilitating the transformation of Zingiberaceae resources from a “resource advantage” into an “industrial advantage”, thereby contributing to the health industry and regional economic development.

Astragalus membranaceus, a traditional qi-tonifying herb, exhibits quality that is strongly influenced by ecological factors of the production region, cultivation practices, and processing techniques. Significant differences in morphology, chemical composition, and bioactivity are observed among different production areas: A. membranaceus from Hunyuan, Shanxi is characterized by high levels of calycosin-7-O-β-D-glucoside, that from Longxi, Gansu is enriched in astragaloside, while samples from Inner Mongolia contain higher concentrations of isoflavonoids. These variations arise from the intricate coupling between ecological factors (such as soil elemental composition, climate, and microbial communities) and genetic background, with associated plants and secondary metabolite regulatory networks serving as key mechanisms. Cultivation patterns exert considerable influence, as wild-simulated cultivation can approximate wild-type quality; appropriate planting density, water and nutrient management, and arbuscular mycorrhizal inoculation effectively enhance the accumulation of bioactive constituents. Harvesting and processing are equally critical: autumn harvest, hot air drying at 60 ℃, and honey-frying processing maximize retention of active compounds and improve immunomodulatory activity. Additionally, previous studies have examined chemical changes during processing and identified astragaloside and calycosin-7-O-β-D-glucoside as marker compounds, establishing quality control strategies from raw materials to final products. The 2025 edition of the Chinese Pharmacopoeia, which focus on single constituents, are insufficient to fully reflect the synergistic effects of constituent clusters. Expansion of cultivation areas exacerbates quality variability, and continuous cropping can disrupt soil microbial ecology. Future approaches should integrate multi-omics analyses and intelligent monitoring to develop a dynamic quality evaluation system based on bioactive constituents, and employ “soil–climate–genotype” matching models to optimize cultivation in non-authentic production regions, thereby promoting high-quality development of the A. membranaceus industry.