Based on Wuyun Liuqi (five movements and six qi) theory recorded in Huangdi Neijing (Yellow Emperor’s Inner Canon) and the Qiantang Medical School’s qi-transformation materia medica thought of “observing manifestations to differentiate qi and inspecting quality according to growth season”, this study integrates the “Sanyou” theory of genuine regional medicinal materials (Dao-di herbs) to propose a novel concept of “Yunqi Spatiotemporal Materia Medica”, constructing an integrated framework of “spatiotemporal qi-transformation-macroscopic properties-quality evaluation”. Taking Lurong (Cervi Cornu Pantotrichum), a genuine regional medicinal material (Dao-di herb) from Northeast China, as a typical example, systematic correspondences between its five stages of generation, growth, transformation, convergence, and storage and the governing periods of the six qi were established: antler casting and bud sprouting correspond to “generative transformation” of Jueyin wind-wood; the rapid growth period aligns with the “luxuriant transformation” of Shaoyang minister fire, during which amino acid and growth factor contents reach their peaks; The osteogenic transition responds to the pivot of the “moistening transformation” of Taiyin damp-earth; complete ossification manifests the astringency of “hardening transformation” of Yangming dryness-metal; And winter dormancy accords with the latency of the “storing transformation” of Taiyang cold-water. The proposal of Yunqi Spatiotemporal Materia Medica aims to refine the spatiotemporally integrated quality evaluation system for medicinal materials, thereby facilitating the regional planning of genuine medicinal materials, season-appropriate harvesting, and dynamic quality control.

Objective To discover anti-atherosclerotic compounds from Ganoderma cochlear. Methods Silica gel column chromatography, reversed-phase C₁₈ column chromatography, and semi-preparative HPLC were used for isolation and purification, and the structures of the isolates were elucidated by various spectroscopic techniques (1D and 2D NMR). The in vitro anti-atherosclerotic activity of compound was evaluated using an ox-LDL-induced macrophage lipid deposition model, and the expression of phenotype-related protein was detected by Western blotting. Results A previously undescribed meroterpenoid, designated as cochlemeroid A (1), was isolated and identified from G. cochlear. BODIPY and Oil Red O staining showed that compound 1 significantly inhibited ox-LDL-induced macrophage lipid deposition in a dose-dependent manner, accompanied by reduced TC and TG levels. Furthermore, Western blotting results demonstrated that compound 1 downregulated the expression of cluster of differentiation 36 (CD36) and class A scavenger receptor (SR-A), while upregulating the expression of the ATP-binding cassette transporter A1 (ABCA1). Conclusion In this study, a new meroterpenoid was successfully isolated from G. cochlear. The bioactivity results indicated that compound 1 possessed significant antiatherosclerotic potential.

Objective To investigate the C₂₁ steroids from Gymnema latifolium and their glucose uptake-promoting activity. Methods The compounds were isolated and purified by various chromatographic techniques including silica gel, ODS, Sephadex LH-20 and semi-preparative HPLC, and their structures were elucidated on the basis of spectroscopic data. The glucose uptake-promoting activity of the isolates was evaluated using a HepG2 cellular glucose uptake model. Results A total of 18 C₂₁ steroids were isolated and identified as gymlatifoside E (1), 20-O-benzoyl-12-O-cinnamoyl-3β,5α,8β,12β,14β,17β,20-heptahydroxy-(20S)-pregn-6-ene (2), gymnepregoside H (3), prosapogenin (4), sylvepregoside E (5), gymnepregoside G (6), penupogenin (7), gymnepregoside I (8), cissasteroid A (9), cynsaccatol L (10), gymnemarsgenin (11), gnetumoside A (12), cynanchin F (13), gagamine (14), 12-O-benzoylsarcostin (15), cissasteroid B (16), 12,20-O-dibenzoylsarcostin (17), hemoside A (18). Among them, compound 1 is a new C₂₁ steroid, and compounds 2—18 were isolated from G. latifolium for the first time. At non-cytotoxic concentrations, treatment with compounds 1—18 resulted in glucose uptake levels ranging from 0.70- to 1.73-fold of the control group in HepG2 cells, among which compounds 4, 8–10, 12–14, 16, and 17 significantly promoted glucose uptake, reaching 1.38- to 1.73-fold of the control group. Conclusion G. latifolium is rich in C₂₁ steroids, and some of these compounds exhibit promising potential anti-diabetic activity, warranting further investigation.

Objective To screen acetylcholinesterase (AChE) inhibitors from the specific medicinal herbs Peganum multisectum by affinity ultra-filtration-liquid chromatography-mass spectrometry (AUF-LC-MS), and to explore their inhibition mechanisms. Methods The AChE inhibitory activities of various polar fractions of P. multisectum were evaluated by Ellman’s test, and the active components were precisely screened by AUF-LC-MS. The action mechanisms of the active components on AChE were investigated by enzyme inhibition kinetics, molecular docking and molecular dynamics simulations. Results A total of seven active components, including one flavonoid and six alkaloids, were identified from the dichloromethane and n-butanol fractions. Enzyme inhibition kinetics suggested that these components belonged to three inhibition types: reversible competitive inhibition (linarin), non-competitive (vasicine, vasicinone and harmol), and reversible mixed inhibition (deoxyvasicine, deoxyvasicinone and harmaline). Molecular docking results revealed that all compounds exhibited good affinities to AChE. And the binding sites and interaction forces of these inhibitors with AChE differed. Molecular dynamics simulation indicated that the system had reached a stable state, and the inhibitors had strong and stable interactions with AChE. Conclusions This study provided a scientific basis for elucidating the potential of P. multisectum in the prevention and treatment of Alzheimer’s disease and offered a new reference for the discovery of high-efficiency AChE inhibitors.

Objective To investigate the chemical constituents and biological activities of Wikstroemia micrantha(Thymelaeaceae). Methods The chemical constituents were isolated and purified via multiple chromatographic techniques including normal-phase silica gel column chromatography, ODS reversed-phase column chromatography and Sephadex LH-20 gel permeation chromatography. The structures of isolated compounds were elucidated by physicochemical properties, 1D NMR, and ESI-MS spectroscopic analyses. The cytotoxic activities of all compounds against five human tumor cell lines (human breast cancer MCF-7 cells, human ovarian cancer SKOV3 cells, human colon cancer HCT-116 cells, human promyelocytic leukemia HL-60 cells, and human hepatocellular carcinoma HepG2 cells) were screened by the CCK-8 assay. Enzyme inhibition assays were performed to evaluate the inhibitory effects of the compounds on α-glucosidase, butyrylcholinesterase (BChE) and acetylcholinesterase (AChE). Results A total of ten compounds were isolated from the ethanol extract of the dried branches and leaves of W. micrantha, and their structures were identified as syzalterin (1), 2-(p-hydroxyphenoxy)-6,8-dimethyl-5,7-dihydroxychromone (2), genkwanin (3), luteolin (4), 7-hydroxycoumarin (5), daphnoretin (6), daphnetin (7), 2,2′-oxybis(1,4-di-tert-butylbenzene) (8), (+)-3-hhydroxy-1,5-diphenylpentan-1-one (9), 1,5-diphenylpentan-1-one (10). In vitro antitumor activity screening revealed that compounds 2 and4 exhibited weak cytotoxicity against various tumor cell lines, with their median inhibitory concentration (IC₅₀) values ranging from 18 to 50 μmol/L. In α-glucosidase inhibitory assay, compounds 1, 2 and 7 displayed moderate inhibitory activity, with IC₅₀ values of (15.51 ± 0.49), (15.99 ± 1.15) and (19.65 ± 1.71) μmol/L, respectively. Results from the in vitro BChE inhibitory assay indicated that compound7 exerted potent inhibitory activity with an IC₅₀ value of (5.63 ± 0.20) μmol/L, whereas compound 9 exhibited moderate inhibitory activity [IC₅₀ = (13.22 ± 0.84) μmol/L]. None of the isolated compounds 1—10 showed notable inhibitory activity against AChE. Conclusion Compounds 1—10 were isolated from W. micrantha for the first time. Bioactivity screening demonstrated that several isolates possessed favorable antitumor activity and selective BChE inhibitory effects. This study provides chemical material basis and experimental data support for further research and development of the medicinal material W. micrantha.

Objective To prepare berberine hydrochloride self-assembled nanoparticles (BBH-SAN) and realize stable self-assembly of BBH without carrier materials, and to systematically evaluate its physicochemical properties and in vitro transdermal performance. Methods BBH-SAN were prepared using a heat-assisted pH-driven method, with average particle size and polydispersity index (PDI) as evaluation indicators for process optimization. The nanoparticles were characterized by scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FT-IR), thermogravimetric analysis (TG), and X-ray diffraction (XRD). Molecular dynamics simulations were employed to elucidate the self-assembly mechanism of BBH-SAN, and in vitro transdermal experiments were conducted to investigate the drug’s transdermal performance. Results The prepared BBH-SAN exhibited lamellar sheet structures under SEM, with an average particle size of (385.67 ± 13.50) nm, a PDI of 0.06 ± 0.04, the ζ potential was determined to be (−34.03 ± 1.01) mV, and an encapsulation efficiency of (87.57 ± 0.05)%. XRD results indicated peak reconstruction and reduced crystallinity, while FT-IR revealed broadening and attenuation of characteristic peaks, suggesting a transition of BBH from a regular crystalline state to a low-crystalline aggregated form. Molecular dynamics simulations revealed that the formation of BBH-SAN is a self-assembly process dominated by van der Waals interactions, regulated by electrostatic interactions, and stabilized synergistically by hydrogen-bond networks. In the in vitro transdermal experiments, the cumulative permeation amount of BBH-SAN within 24 h was (61.25 ± 1.78) μg/cm², and the skin retention (Q_s), dermal retention (Q_d), and stratum corneum retention (Q_sc) of BBH-SAN were all significantly higher than those of BBH (P < 0.001). Conclusion This study achieved the stable self-assembly of berberine hydrochloride without the use of any carrier through a heat-assisted pH-driven method. The resulting nanoparticles exhibit uniform particle size, good dispersibility, excellent thermal stability, and significantly improved transdermal performance, providing a new strategy for the self-assembly delivery of alkaloid drugs.

Objective To achieve quality control during the production of Zhisou Qingguo Wan (ZQW, 止嗽青果丸), the pixel unmixing method from remote sensing was introduced into the uniformity evaluation of traditional Chinese medicine preparations, so as to visually analyze the mixing uniformity of medicinal powder and honey in ZQW. Methods Multiple preprocessing methods, including Savitzky-Golay (SG) smoothing, first derivative (1D), second derivative (2D), standard normal variate (SNV) and multiplicative scatter correction (MSC) were adopted to optimize spectral data and eliminate interferences such as background noise and spectral scattering. Based on pure substance spectra combined with the mixture tuned matched filtering (MTMF) algorithm, the abundance scores and mixture feasibility scores of target endmembers were integrated. Material abundance estimation and binarization at the pixel level of mixed samples were then performed, enabling the visualization of the spatial distribution of medicinal powders. Meanwhile, a residual network (ResNet) model was constructed for the prediction and pixel-level visualization of honey content. Results Compared with traditional quantitative models such as partial least squares regression (PLSR), multiple linear regression (MLR), ridge regression (RR), least absolute shrinkage and selection operator (Lasso), random forest (RF) and ElasticNet, the R² of the prediction set was significantly improved from 0.837 6 to 0.931 1 with the proposed model. Conclusion The method combining pixel unmixing and deep learning intuitively presents the spatial distribution of medicinal raw materials and excipients in the preparation. A novel insight for the refined production and quality control of traditional Chinese medicine preparations is provided.

Objective To isolate and prepare extracellular vesicle-like nanoparticles from Tiepishihu (Dendrobium officinale) flowers (DF-EVLPs), and evaluate their specific targeting ability for colon cancer cells, as well as investigate their in vitro targeting and anti-tumor effects against colon cancer cells, in order to provide experimental evidence for the development of targeted delivery systems based on medicinal and edible plants. Methods DF-EVLPs were isolated from D. officinale flowers and their morphology and particle size were characterized. The uptake of DF-EVLPs by CT26 colon cancer cells was observed using pKH26 fluorescence labeling. The effects of DF-EVLPs on the proliferation, migration and apoptosis of CT26 cells and their mechanism were evaluated using CCK-8 assay, scratch wound healing experiment, flow cytometry and Western blotting. Results The obtained DF-EVLPs showed a uniform cup-shaped morphology with an average particle size of (156.9 ± 7.2) nm. The cell uptake experiment demonstrated that DF-EVLPs could be efficiently internalized by CT26 colon cancer cells, and the uptake efficiency of DF-EVLPs for colon cancer cells was significantly higher than that of normal colonic epithelial cells, indicating their good specific targeting ability. The in vitro functional experiments showed that DF-EVLPs inhibited the proliferation of CT26 cells in a concentration-dependent manner, significantly reduced cell migration ability, and induced cell apoptosis. Conclusion This study successfully prepared extracellular vesicle-like nanoparticles from D. officinale flowers. As a new type of food-derived nanotherapeutic agent, DF-EVLPs can specifically target colon cancer cells and exert in vitro anti-tumor activity. This study reveals a new biological activity function of D. officinale flowers and provides experimental evidence for dietary intervention for colorectal cancer based on the “medicinal and edible” concept.

Objective To prepare diosgenin silk fibroin nanoparticles (Doi-SF-NPs) and evaluate their physicochemical properties, tumor targeting and in vivo anti-tumor effects . Methods The main influencing factors of Doi-SF-NPs were investigated by single factor experiments, and Box-Behnken design-response surface methodology (BBD-RSM) was employed to optimize prescription process of Doi-SF-NPs. Doi-SF-NPs was characterized by transmission electron microscopy (TEM) and Fourier transform infrared spectroscopy (FT-IR). Drug release behavior of Doi-SF-NPs in phosphate buffer saline solution with different pH values was investigated. MTT method and Annexin V/PI double staining method were used to investigate the inhibitory and proapoptotic effects of Doi-SF-NPs on PC3 cells, respectively. The nude mouse of prostate cancer was established, compared the concentrations of diosgenin and Doi-SF-NPs in tumor tissues, and then evaluated the tumor targeting ability of Doi-SF-NPs using the relative uptake rate and ratio of peak concentration. The nude mouse model of prostate cancer was divided into model (normal saline) group, positive control (cisplatin, 2.0 mg/kg) group, diosgenin (10 mg/kg) group, low-dose Doi-SF-NPs (5 mg/kg) group and high-dose Doi-SF-NPs (10 mg/kg) group, the in vivo anti-tumor effects of Doi-SF-NPs was compared with diosgenin. Safety evaluation of Doi-SF-NPs was also performed. Results Optimal prescription process of Doi-SF-NPs nanoparticles as follows: The mass ratio of silk fibroin to diosgenin was 11.94:1, the volume ratio of the organic phase to aqueous phase was 5.99:1, and the preparation time was 2.75 h. Average envelopment efficiency, drug loading, particle size and ζ potential were (84.07 ± 0.70)%, (6.41 ± 0.05)%, (164.29 ± 4.43) nm and (−23.01 ± 1.17) mV, respectively. Diosgenin formed Doi-SF-NPs with silk fibroin through hydrogen bonding. The appearance of Doi-SF-NPs was spherical nanoparticles. Doi-SF-NPs existed higher cumulative release in phosphate buffer solution of pH 5.5 and 6.0, which demonstrated that Doi-SF-NPs had obvious pH-sensitivity. MTT method results showed that IC₅₀ of diosgenin and Doi-SF-NPs on PC3 cells were 72.44 and 55.68 μmol/L, respectively. Doi-SF-NPs more effectively induced apoptosis of PC3 cells than that of diosgenin. The relative uptake rate of Doi-SF-NPs in in tumor tissue was 3.23, and the peak concentration ratio was 2.60, which demonstrating that Doi-SF-NPs could significantly increase the accumulation in tumor tissue. Doi-SF-NPs (10 mg/kg) increased the tumor inhibition rate of diosgenin (10 mg/kg) from 29.47% to 66.18%. Doi-SF-NPs did not affect the major organs of tumor-bearing nude mouse, which showed that the safety of Doi-SF-NPs was high. Conclusion Doi-SF-NPs could improve the tumor targeting of diosgenin and significantly enhanced anti-tumor efficacy. Therefore, Doi-SF-NPs had the potential to be developed into an antitumor drug.

Objective To optimize the preparation process of Bajitian Formula (复方巴戟天) nano-suspension and evaluate its pharmacological efficacy in prophylactic treatment for mice with memory-impaired Alzheimer’s disease after performance characterization. Methods Taking Bajitian Formula as the study subject, and based on single-factor analysis, the formulation process of Bajitian Formula nano-suspension was optimized via high-pressure homogenization with stabilizer dosage, homogenization pressure, and number of homogenization cycles as influencing factors, followed by performance characterization. The paddle method was employed to identify the optimal in vitro release medium, and the in vitro release behavior of polysaccharides in the Bajitian Formula nano-suspension was investigated. Pharmacodynamic experiments were performed to evaluate the preventive effect of the nano-suspension on memory impairment in Alzheimer’s disease model mice. Results The optimized formulation process for Bajitian Formula nano-suspension was determined as follows: the mass ratio of crude herbs to soybean lecithin was 1:0.6, high-pressure homogenization pressure was 40.0 MPa (400 bar), and homogenization cycles were 7. The particle size of the nanoparticle suspension was measured as (203.46 ± 2.67) nm, with a PDI of 0.200 ± 0.002. The optimal in vitro release medium was a 10 mg/mL SDS aqueous solution, achieving a cumulative total polysaccharide release rate of (72.10 ± 1.19)% after 24 h. After 30 d of oral administration of the optimized nanoparticle suspension, compared with the model group, the latency period of escape behavior was significantly shortened in both the positive control group and the medium-and high-dose groups of Bajitian Formula nano-suspension in the spatial navigation experiment (P < 0.05, 0.01). In the spatial exploration experiment, the number of target quadrant entries was significantly increased in the positive control group, decoction group, and low-, medium-, and high-dose groups of Bajitian Formula nano-suspension. The swimming distance to target quadrants was significantly increased in the positive control group and low-, medium-, and high-dose groups of Bajitian Formula nano-suspension (P < 0.05, 0.01). In the positive control group and low-, medium-, and high-dose groups of Bajitian Formula nano-suspension, the acetylcholine (ACh) content in the hippocampus and prefrontal cortex was significantly increased. In the positive control group and medium-and high-dose groups of Bajitian Formula nano-suspension, choline acetyltransferase (ChAT) activity in the hippocampus and prefrontal cortex was significantly elevated, while acetylcholinesterase (AChE) activity in these regions was significantly reduced (P < 0.05, 0.01). Conclusion The formulation optimization process of Bajitian Formula nano-suspension is rational and feasible, with preparation properties meeting nanoscale requirements, demonstrating uniform particle dispersion and good stability. Characterized by slow and sustained release, with excellent in vitro release behavior. It exhibits significant preventive effects against memory decline induced by Alzheimer’s disease, surpassing the efficacy of optimized decoction formulations. This study provides a beneficial reference for the modernization and technical upgrading of traditional dosage forms.

Objective To establish an ultrasound-assisted nanonization process for realgar and elucidate the scientific connotation of achieving toxicity reduction and efficacy enhancement within safety thresholds through nanonization. Methods The parameters of zirconium ball water-grinding were optimized using central composite design-response surface methodology (CCD-RSM), and nano-realgar was prepared by combining acid washing with ultrasonic shearing. Four processed products (dry-ground Realgar, traditional water-floating Realgar, SDS-assisted water-floating Realgar and nano-Realgar) were characterized by laser particle size analyzer, scanning electron microscope (SEM), specific surface area and porosity analyzer based on the Brunauer-Emmett-Teller (BET) method, and liquid chromatography-atomic fluorescence spectrometry (LC-AFS). Toxicity was evaluated using a Caenorhabditis elegans model, and efficacy was assessed using a mouse wound model. Results The optimal process parameters of zirconium ball water-grinding were determined as milling for 3.9 h, three cycles of levigation, and the addition of 190 g of zirconia balls (the mass ratio of zirconium balls with diameters of 1, 2, 3, 4 mm is 1∶1∶2∶4); The particle size of Realgar sample can be reduced to 300 nm 30 min of sonication at 250 W. BET analysis showed that nano-realgar had well-developed mesopores. LC-AFS results indicated that, compared with the dry-milled product, all processing methods reduced the As³⁺ content, with nano-Realgar being slightly higher but still below the safety limit. In the C. elegans assay, nano-Realgar exhibited the lowest toxicity (P < 0.001). In the mouse wound model, it achieved the highest healing rate, significantly reducing inflammatory factors and upregulating growth factor expression (P < 0.05). Conclusion The zirconium ball water-grinding combined with ultrasound-assisted nanonization process successfully prepares nano-Realgar, which possesses dual advantages of toxicity reduction and efficacy enhancement within safety thresholds, providing a theoretical basis for upgrading the processing technology of Realgar.

Objective To elucidate the molecular mechanism by which Lobelia chinensis-derived extracellular vesicle-like nanoparticles (LC-EVLP) induce lethal autophagy in hepatocellular carcinoma (HCC) by remodeling the glutamine metabolic network, and to systematically evaluate their antitumor efficacy and biosafety. Methods LC-EVLP were isolated by differential ultracentrifugation and physicochemically characterized. By integrating network pharmacology, molecular docking and multidimensional cell-based assays, the effects of LC-EVLP on the proliferation, migration and invasion of human HCC Mahlavu cells were assessed. Rescue experiments using an autophagic-flux inhibitor and the metabolic intermediate α-ketoglutarate (α-KG) were conducted to interrogate the “metabolism-autophagy” regulatory axis. A nude mouse subcutaneous xenograft model was established to evaluate the in vivo anti-tumor effects of LC-EVLP and histopathological changes in major organs. Results LC-EVLP was successfully isolated and characterized as typical vesicular structures with particle sizes mainly distributed between 126.0 and 151.7 nm, and were efficiently internalized by Mahlavu cells. Functional assays demonstrated that LC-EVLP significantly reduced the viability of Mahlavu cells at concentrations ≥ 12.5 μg/mL (P < 0.001), while exerting no significant cytotoxicity toward LO2 cells. Further analyses revealed that LC-EVLP significantly inhibited colony formation and DNA synthesis (P < 0.05, 0.01, 0.001), and significantly suppressed cell migration and invasion (P < 0.001). The autophagy inhibitor bafilomycin A1 (BafA1) significantly rescued cell viability (P < 0.001), indicating that autophagic cell death was the dominant mechanism underlying the anti-HCC activity of LC-EVLP. LC-EVLP significantly upregulated Beclin 1 expression (P < 0.05, 0.01, 0.001), increased microtubule associated protein 1 light chain 3-II/I (LC3-II/I) value (P < 0.05, 0.01), and reduced p62 protein expression (P < 0.01, 0.001). Moreover, BafA1 further enhanced LC3-II/I accumulation and reversed the decrease in p62 expression (P < 0.01, 0.001), confirming the promotion of a complete autophagic flux. In terms of metabolic regulation, LC-EVLP significantly downregulated the expressions of pyruvate dehydrogenase kinase 1 (PDK1), pyruvate kinase M (PKM) and glutaminase 1 (GLS1) (P < 0.01, 0.001), while α-KG supplementation reversed the abnormal autophagic phenotype (P < 0.01). The in vivo experimental results showed that LC-EVLP significantly inhibited tumor growth (P < 0.05, 0.001), downregulated Ki67 and PCNA expressions (P < 0.001), and induced changes in autophagy related proteins (P < 0.05, 0.01, 0.001), with no significant toxicity observed. Conclusion LC-EVLP exerts anti-tumor effects by regulating PDK1/PKM axis, disrupting cellular metabolic balance and inducing enhanced autophagy. Its low toxicity characteristics suggest its potential for further research as a natural nanomaterial-based antitumor agent.

Objective To study the effect of gamabufotalin (CS-6) on the proliferation of nasopharyngeal carcinoma cells, and explore the mechanism by which CS-6 inhibits nasopharyngeal carcinoma cell proliferation through cell cycle arrest and autophagy. Methods The inhibitory effect of CS-6 on the proliferation of HK-1 and 5-8F cells was detected through CCK-8 assay and colony formation assay. Flow cytometry was used to detect the effect of CS-6 on the cell cycle distribution and apoptosis of HK-1 and 5-8F cells. Western blotting was used to detect the expressions of Cyclin B1 and cyclin dependent protein kinase 1 (CDK1). Transcriptome sequencing was used to screen for differentially expressed genes after CS-6 treatment, and IPA software was used to analyze the enrichment pathways of differentially expressed genes. The ultrastructure of autophagosomes after CS-6 acts on cells was observed using transmission electron microscopy. Western blotting was used to detect the expressions of microtubule associated protein light chain 3 (LC3), p62, and glycogen synthase kinase-3β (GSK-3β)/mammalian target of rapamycin (mTOR) pathway related proteins. After transient transfection of mCherry-EGFP-LC3 plasmid, the changes in autophagy flux of cells treated with CS-6 were observed using laser confocal microscopy. In order to investigate the effect of PFKFB4 on CS-6-induced proliferation, cell cycle arrest and autophagy of nasopharyngeal carcinoma cells, HK-1 and 5-8F cells were transfected with PFKFB4 plasmid, qRT-PCR and Western blotting were used to detect the expression of 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase-4 (PFKFB4), as well as changes in cell survival rate, colony formation, autophagy flux, cell cycle and autophagy related protein expressions. A mouse model of nasopharyngeal carcinoma transplantation in vivo was established, after CS-6 administration, tumor volume and weight were measured, pathological changes in tumor, liver and kidney tissues were observed using hematoxylin-eosin staining, Ki67 expression in tumor tissues was detected using immunohistochemistry, LC3, p62, Cyclin B1, CDK1, p-mTOR, PFKFB4 and p-GSK-3β protein expressions in tumor tissues were detected using Western blotting. Results After 48 h of treatment, CS-6 significantly inhibited the proliferation of HK-1 and 5-8F cells (P < 0.05, 0.001), with half-maximal inhibitory concentrations of 8.684 and 14.320 nmol/L, respectively. CS-6 caused significant G₂/M phase arrest in HK-1 and 5-8F cells (P < 0.05, 0.01), while increasing protein expressions of Cyclin B1 and CDK1 (P < 0.05, 0.01, 0.001). CS-6 did not induce apoptosis in HK-1 and 5-8F cells under treatment conditions of 48 h. CS-6 significantly reduced the mRNA and protein expressions of PFKFB4 (P < 0.05, 0.01, 0.001), typical double-membrane autophagosomes and autolysosomes were observed in the cells, CS-6 upregulated LC3-II/I and downregulated p62 protein expressions (P < 0.05, 0.01, 0.001), promoted the fusion of autophagosomes and lysosomes. Overexpression of PFKFB4 could partially reverse the inhibitory effect of CS-6 on cell proliferation and regulate the expressions of cell cycle and autophagy related proteins (P < 0.05, 0.01, 0.001), and inhibit the fusion of autophagosomes and lysosomes. CS-6 significantly increased the expression of p-GSK-3β and inhibited the expression of p-mTOR (P < 0.05, 0.01, 0.001). Overexpression of PFKFB4 could partially reverse the regulatory effects of CS-6 on p-GSK-3β and p-mTOR (P < 0.05, 0.01). In vivo, CS-6 significantly inhibited tumor growth in tumor bearing mice (P < 0.001), induced pathological damage to tumor tissue, inhibited tumor cell proliferation (P < 0.001), upregulated LC3-II/I, p-GSK-3β expressions and downregulated p62 expression in tumor tissue (P < 0.001), downregulated protein expressions of p-mTOR, PFKFB4, Cyclin B1 and CDK1 (P < 0.01, 0.001), and had no effect on liver and kidney tissues. Conclusion CS-6 exerts an anti-nasopharyngeal carcinoma effect by downregulating PFKFB4, thereby regulating GSK-3β/mTOR signaling pathway and inducing autophagy and cell-cycle arrest.

Objective To elucidate the material basis of Quzhou Chenpi (Citri Reticulatae Pericarpium) extract (CPE) and to investigate its mechanism in improving functional dyspepsia (FD) by repairing the intestinal barrier. Methods CPE was prepared by water decoction combined with freeze-drying. Qualitative and quantitative analyses of its chemical components were performed using ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) and high-performance liquid chromatography (HPLC), followed by network pharmacology to predict potential targets and pathways. In vivo, an FD rat model was established by iodoacetamide combined with irregular feeding. After drug intervention, gastric emptying rate and small intestinal propulsion rate were measured. Histopathological changes in the stomach and duodenum were examined by hematoxylin-eosin staining. Levels of lipopolysaccharide (LPS), diamine oxidase (DAO) in serum as well as tumor necrosis factor-α (TNF-α) in duodenal tissue were determined by ELISA. The expressions of zonula occludens-1 (ZO-1) and Occludin in duodenal tissue was assessed by immunohistochemistry. In vitro, a TNF-α-induced Caco-2 intestinal barrier injury model was used. Transepithelial electrical resistance (TEER) value and fluorescein isothiocyanate-dextran with relative molecular mass 4 × 10⁴ (FD-4) permeability were measured. The distribution of ZO-1 and Occludin proteins was observed by immunofluorescence staining. The mRNA expressions of ZO-1, Occludin, interleukin-6 (IL-6) and TNF-α were detected by qRT-PCR. IL-6 secretion was measured by ELISA. The phosphorylation levels of cellular Src (c-Src) and protein kinase B (Akt) were detected by Western blotting. Results A total of 84 chemical components were identified from CPE, eight of which were reported in Citri Reticulatae Pericarpium for the first time. The contents of the four major flavonoids were ranked from high to low as hesperidin > narirutin > nobiletin > tangeretin. Network pharmacology predicted that the potential core targets of CPE for improving FD involved SRC, AKT1, etc., with enrichment in phosphoinositide 3-kinase (PI3K)/Akt signaling pathway and tight junction regulation. In vivo, CPE significantly ameliorated gastrointestinal motility disorders in FD rats (P < 0.05, 0.01, 0.001), alleviated duodenal mucosal injury, reduced levels of LPS and DAO in serum (P < 0.01, 0.001), suppressed TNF-α protein and mRNA expressions in duodenal tissue (P < 0.01, 0.001), and up-regulated Occludin protein expression (P < 0.05, 0.001). In vitro, the actual hesperidin content in CPE was only 10.48% of that in the hesperidin monomer group, yet CPE was equivalent to hesperidin monomer in stabilizing TEER and reducing FD-4 permeability (P < 0.01, 0.001). CPE significantly up-regulated the mRNA expressions of ZO-1 and Occludin (P < 0.001), maintained their continuous network-like distribution, significantly inhibited IL-6 transcription and secretion (P < 0.01, 0.001), and significantly reduced the values of p-c-Src/c-Src and p-Akt/Akt (P < 0.05). Conclusion CPE significantly ameliorates gastrointestinal motility disorders and duodenal mucosal barrier injury in FD rats, and alleviates barrier dysfunction and inflammation in TNF-α-induced Caco-2 cells. Its effect depends on the inhibition of over-activation of c-Src/Akt signaling pathway. The overall efficacy demonstrates the multi-component synergistic characteristics of traditional Chinese medicine, providing an experimental basis for the application of Quzhou Citri Reticulatae Pericarpium in gastrointestinal diseases.

Objective To investigate whether Yiqi Fumai Lyophilized Injection [注射用益气复脉（冻干）, YQFM] ameliorates myocardial hypertrophy and pressure overload-induced heart failure (HF) by upregulating the cardiac endothelial mechanosensitive ion channel Piezo1 and activating the nitric oxide (NO) signaling pathway. Methods A mouse HF model was prepared via transverse aortic constriction (TAC) surgery. Two weeks post-operation, mice received intraperitoneal injections of different doses of YQFM (0.13, 0.26, 0.53 g/kg) or captopril (20 mg/kg) for four weeks. Cardiac function was assessed by echocardiography. Pathological injury and fibrosis were evaluated by hematoxylin and eosin (HE) and Masson staining. Cardiac remodeling and cardiomyocyte hypertrophy were assessed by heart index, heart weight-to-tibia length ratio and wheat germ agglutinin (WGA) staining. Serum biochemical assays, qRT-PCR, Western blotting and immunofluorescence were employed to detect HF-related biomarkers and the expressions of key proteins in Piezo1/NO pathway. Results YQFM significantly improved TAC-induced cardiac dysfunction and remodeling (P < 0.05, 0.01), and alleviated histopathological disorganization, cardiomyocyte hypertrophy and myocardial fibrosis (P < 0.01). Mechanistic studies showed that YQFM reduced levels of cardiac injury markers cardiac troponin T (cTn-T) and N-terminal pro-B-type natriuretic peptide (NT-proBNP) in serum (P < 0.01), and significantly downregulated the expressions of hypertrophy-related genes natriuretic peptide A (Nppa), natriuretic peptide B (Nppb), myosin heavy chain 7 (Myh7) and fibrosis-related genes collagen type I alpha 1 chain (Col1a1), collagen type III alpha 1 chain (Col3a1) in cardiac tissue (P < 0.05, 0.01). YQFM significantly decreased the levels of follistatin-like 1 (Fstl1) and endothelin-1 (ET-1) (P < 0.05, 0.01), while elevating the levels of NO in serum and cardiac tissue and cyclic guanosine monophosphate (cGMP) in cardiac tissue (P < 0.01). Furthermore, YQFM significantly upregulated the expressions of Piezo1 protein and phosphorylated endothelial nitric oxide synthase (p-eNOS) in cardiac endothelial cells (P < 0.01). Conclusion YQFM improves cardiac function and remodeling, and inhibits cardiomyocyte hypertrophy in HF by upregulating endothelial Piezo1 expression and activating eNOS/NO signaling pathway.

Objective To explore the metabolic regulatory effects of different administration methods of brucine-loaded microneedles on rats with collagen-induced arthritis (CIA) through synovial fluid and serum metabolomics, and to provide experimental evidence for elucidating the mechanism of brucine-loaded microneedles in the treatment of rheumatoid arthritis (RA).Methods The non-targeted metabolomics method of ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was adopted. The control group, brucine-loaded hydrogel acupoint microneedles group (XWMN), brucine-loaded hydrogel non-acupoint microneedles group (FXWMN), blank microneedles group (KBMN) and traditional acupuncture group (ZC) were respectively compared with model group. The changes in the profiles of endogenous metabolites in the synovial fluid and serum of CIA rats after intervention were systematically analyzed.Results Based on the screening of synovial fluid metabolomics, differential metabolites were obtained for comparison between each treatment group and the model group. Analysis of the reversal trends of the differential metabolites showed that, compared with model group, three metabolites in XWMN group, five in FXWMN group, three in KBMN group, and three in ZC group were significantly restored toward normal levels. The metabolic regulation of XWMN group mainly involved the cyclic adenosine monophosphate (cAMP) signaling pathway, butyrate metabolism, nucleotide metabolism and other pathways, with a higher number of enriched pathways compared to other treatment groups. Based on serum metabolomics screening, differential metabolites were obtained for comparison between each treatment group and the model group. Through the analysis of the regression trend of differential metabolites, it was found that compared with model group, five metabolites in XWMN group showed significant regression, 15 metabolites in FXWMN group showed significant regression, 12 metabolites in KBMN group showed significant regression, and three metabolites in ZC group showed significant regression; The metabolic regulation of XWMN group mainly involved key pathogenic pathways such as purine metabolism and arachidonic acid metabolism in RA, while FXWMN group involved three pathways including purine metabolism and ABC transporter pathway, and KBMN group involves seven metabolic pathways including purine metabolism and linoleic acid metabolism. Conclusion Different administration methods of brucine-loaded microneedles can modulate the aberrant metabolism in CIA rats to varying degrees. Notably, acupoint administration of brucine-loaded microneedles exerts anti-inflammatory effects and regulates immune cell differentiation by modulating the expression of metabolites such as 3-hydroxybutyrate and deoxycytidine in synovial fluid, thereby influencing pathways like cAMP signaling, butanoate metabolism, and nucleotide metabolism. Concurrently, it exerts anti-inflammatory effects by modulating serum metabolic pathways including purine metabolism and arachidonic acid metabolism. Compared with non-acupoint administration, blank-microneedle treatment and traditional acupuncture, acupoint administration demonstrates a more prominent modulatory effect on key pathogenic pathways (e.g., purine metabolism, arachidonic acid metabolism).

Objective To explore the effect of total saponins of Platycodon grandiflorum on cartilage injury and Ras homolog gene family member A (RhoA)/Rho-associated coiled-coil kinase (ROCK) signaling pathway in knee osteoarthritis (KOA) rats.Methods Knee joint injection of papain was used to prepare a KOA model. The rats were randomly divided into model group, glucosamine sulfate (0.2 g/kg) group, total saponins of P. grandiflorum low- and high-dose (100, 200 mg/kg) groups and total saponins of P. grandiflorum (200 mg/kg) + RhoA activator LPA (1 mg/kg) group. A sham group was also included, with 15 rats in each group. After four weeks of continuous administration, micro CT scanning was used to observe changes in knee joints of rats in each group. Hematoxylin-eosin (HE) and Safranin O-fast green staining were used to observe the pathological changes of cartilage tissue. ELISA was used to detect the levels of matrix metalloproteinase-13 (MMP-13), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in joint fluid. TUNEL staining was used to observe chondrocyte apoptosis, and transmission electron microscopy was used to observe the microstructure of chondrocytes. Western blotting was used to detect expressions of aggrecan, type II collagen (Col II), B-cell lymphoma-2 (Bcl-2), Bcl-2 associated X protein (Bax), cleaved cystein-asparate protease-3 (cleaved Caspase-3), RhoA, ROCK1 and ROCK2 proteins in cartilage tissue. Results Compared with sham group, rats in model group showed severe pathological damage to the cartilage tissue, destruction of the ultrastructure of chondrocytes, Mankin score, osteoarthritis research society international (OARSI) score, chondrocyte apoptosis rate, levels of TNF-α, MMP-13, IL-6 in joint fluid, as well as protein expression levels of RhoA, ROCK1, ROCK2, Bax and cleaved Caspase-3 in cartilage tissue were significantly increased (P < 0.05), bone volume/total volume (BV/TV), trabecular thickness (Tb.Th) and protein expression levels of Bcl-2, aggrecan, Col-II in cartilage tissue were significantly reduced (P < 0.05). Compared with model group, glucosamine sulfate group and total saponins of P. grandiflorum groups showed reduced pathological damage to cartilage tissue, normal ultrastructure of chondrocytes, Mankin score, OARSI score, chondrocyte apoptosis rate, levels of TNF-α, MMP-13, IL-6 in joint fluid, as well as protein expression levels of RhoA, ROCK1, ROCK2, Bax and cleaved Caspase-3 in cartilage tissue were significantly decreased (P < 0.05). BV/TV, Tb.Th and protein expression levels of Bcl-2, aggrecan, Col-II in cartilage tissue were significantly increased (P < 0.05). After the combined use of RhoA activator LPA, the improvement effect of total saponins of P. grandiflorum on the above indicators was partially reversed (P < 0.05). Conclusion Total saponins of P. grandiflorum can improve cartilage damage in KOA rats by inhibiting RhoA/ROCK pathway.

Objective To construct HerbVLM, a vision-text fusion model for the identification of traditional Chinese medicinal materials, aiming to improve intelligent recognition performance in distinguishing visually similar categories and generalizing across scenarios. Methods Based on a vision-language pretrained dual-encoder framework, a traditional Chinese medicine knowledge-enhanced prompt module (TKPM) was introduced on the text side, while a hierarchical image adaptation and de-confusion module (HIA) was constructed on the image side. Dynamic fusion and joint loss optimization were further incorporated. Comparative and ablation experiments were conducted on the open-source Chinese-Medicine-163 dataset, and cross-domain and few-shot adaptation validation was performed on a self-collected external dataset of the “new Zhebawei”. Results On the test set, HerbVLM achieved top-1 accuracy (Top-1 Acc), macro precision (P_macro), macro recall (R_macro), and macro F1-score (F1_macro) of 98.69%, 98.70%, 98.71%, and 98.70%, respectively, outperforming comparative methods including residual network (ResNet), context optimization (CoOp), conditional context optimization (CoCoOp), training-free CLIP-Adapter (Tip-Adapter), multi-modal prompt learning (MaPLe), and contrastive vision-language pretraining in Chinese (Chinese-CLIP). Ablation experiments showed that text enhancement, image adaptation, de-confusion constraints, and dynamic fusion all brought stable performance gains. In external testing on the self-collected Zhejiang Eight Flavors dataset, the recognition rates for known categories Lingzhi (Ganoderma), Fupenzi (Rubi Fructus), Tiepishihu (Dendrobii Officinalis Caulis), and Qianhu (Peucedani Radix) were 96.00%, 100.00%, 100.00%, and 100.00%, respectively. For unknown categories, the average accuracies under 1-shot, 2-shot, and 5-shot settings were approximately 84.00%, 91.54%, and 98.20%, respectively, demonstrating favorable few-shot transfer capability. Conclusion HerbVLM significantly improves the fine-grained recognition accuracy and cross-domain generalization performance of traditional Chinese medicinal materials through multimodal collaborative modeling of image-text knowledge, and can provide methodological support for traditional Chinese medicinal material teaching, quality inspection assistance.

Objective To address the strong coupling, long time delays, and nonlinear characteristics inherent in the traditional Chinese medicine (TCM) extraction process, as well as the issues of prediction lag and extreme-value smoothing commonly exhibited by traditional deep time-series models during operating condition transitions, a decoupled digital twin modeling method of key process parameters for the TCM extraction process, integrating Standard Operating Procedure (SOP) priors, named SOP-guided channel-independent patch time series Transformer (SGC-PatchTST), is proposed. Methods Based on the PatchTST architecture, the proposed model introduces three major enhancements: 1. An orthogonal decoupling late-fusion mechanism is employed to transform discrete SOP instructions into process-intention features, utilizing a soft-gating mechanism for fusion to suppress the lag caused by operating condition switches; 2. A decoupled structure comprising primary and auxiliary branches is constructed to independently model the reflux temperature, thereby mitigating the interference caused by multivariable coupling; 3. An extrema-sensitive hybrid loss function is designed, combining local statistics with extreme-value penalties to enhance the model's extrema-fitting and noise-resistance capabilities under the low-pass filtering conditions of Patch-based tokenization. All baseline models uniformly adopt the OneCycleLR learning rate schedule and an early-stopping strategy. Experimental data are independently partitioned into training, validation, and test sets on a batch-by-batch basis to strictly eliminate cross-batch data leakage. Actual production data from a TCM extraction workshop are utilized to construct the predictive model and evaluate it via autoregressive multi-step inference. Results Under a 50-batch stratified split, the single-evaluation Mean Absolute Errors (MAEs) of the upper and lower temperatures for SGC-PatchTST (optimized hyperparameters τ＝1.5σ，λ＝2.0) decrease to 0.36 ℃ and 0.35 ℃, respectively (which are 40.0% and 50.2% lower than the standard PatchTST model, respectively). Its capability to resist extreme deviations is significantly enhanced, with the 95% percentile errors (P95 errors) for the upper and lower temperatures controlled at 1.31 ℃ and 1.33 ℃, respectively. At the same time, benefiting from the decoupled primary and secondary architectures, the MAE of the reflux temperature, which is susceptible to external disturbances, is reduced to a lower value of 0.04 ℃. The prediction of minor fluctuations in the core safety parameter (in-tank pressure) is also maintained at a relatively high precision of 0.000 6 MPa. Furthermore, under three independent stratified splits, the average upper and lower temperature MAEs remain at 0.33 ℃ (coefficient of variation, CV = 6.8%) and 0.32 ℃ (CV = 6.4%), respectively, confirming its good statistical robustness. A six-step combination ablation study and a 25-group hyperparameter grid search validate the contribution of each module and the hyperparameter robustness, respectively. Conclusion The proposed SGC-PatchTST method achieves an ideal level of prediction accuracy and timeliness while demonstrating strong physical plausibility, providing reliable support for subsequent model-based reinforcement learning control.

Objective To explore the formulation rules of existing sleep-improving health foods, construct “data mining statistics-medicine and edible homology/classical formula screening-traditional Chinese medicine theoretical interpretation” new formula system and investigate its potential mechanisms of action. Methods The publicly available record information of sleep-improving health foods was collected from the State Administration for Market Regulation, and the raw materials and compatibility rules were analyzed. Based on the data mining results, a novel sleep-improving formula of medicine and edible homology was constructed. Network pharmacology was used to predict the potential action targets and signaling pathways, and molecular docking was performed to verify the binding affinity between active components and core targets. Results A total of 541 sleep-improving health foods meeting the inclusion criteria were included, involving 154 Chinese medicinal materials. The high-frequency materials included Suanzaoren (Ziziphi Spinosae Semen), Wuweizi (Schisandrae Chinensis Fructus), and Fuling (Poria). Their nature and taste are mainly sweet and neutral, and its meridians mainly focus on the heart and liver meridians. Association rule analysis revealed core drug pairs such as “Schisandrae Chinensis Fructus-Ziziphi Spinosae Semen” and “Poria-Ziziphi Spinosae Semen”. Capsules, tablets, and granules were the main dosage forms. The usage and dosage were mainly 1–2 times daily, and 99.8% of the products did not specify a course of administration. Besides the core function of sleep improvement, these products often also claimed enhancing immunity and relieving physical fatigue as combined functions. The suitable population was primarily individuals with poor sleep status, while the unsuitable population mainly included children, pregnant and lactating women. A novel formula consisting of Baihe (Lilii Bulbus), Dihuang (Rehmanniae Radix), Ziziphi Spinosae Semen, Tianma (Gastrodiae Rhizoma), Gegen (Puerariae Lobatae Radix) was constructed. Network pharmacology predicted 42 active components, 760 action targets, among which 189 were associated with sleep, 196 with immune enhancement, and 63 with gut microbiota regulation. The enrichment analysis indicated that the main pathways involved neuroactive ligand-receptor interaction, phosphatidylinositol-3-hydroxykinase (PI3K)-protein kinase B (Akt) signaling pathway, and endocrine resistance, which are closely related to sleep improvement, immune enhancement, and gut microbiota regulation. Molecular docking showed that the active components of the novel formula bound well with the core targets. Conclusion A new formula of health food for improving sleep, “Lilii Bulbus-Rehmanniae Radix-Ziziphi Spinosae Semen-Gastrodiae Rhizoma-Puerariae Lobatae Radix”, is screened out. Its sleep-improving effect is mainly closely related to enhancing immunity and regulating intestinal microbiota, which provides scientific theoretical support for the subsequent development of health products for sleep improvement..

Objective To analyze the current research status of Longyanrou (Longan Arillus) using bibliometric methods, and predict its future research hotspots and development trends. Methods Literature on Longan Arillus research was collected from the China National Knowledge Infrastructure (CNKI), Wanfang Data Knowledge Service Platform (WF), VIP Chinese Scientific Journals Database (VIP), and the Web of Science Core Collection (WOS). The retrieved literature was imported into NoteExpress, where duplicates and records not meeting the inclusion criteria were removed, and keywords were standardized. Excel, CiteSpace 6.4.R1 advanced, and VOSviewer 1.6.20 were used to statistically analyze publication volume of the literature, the countries of publication, institutions, authors, journals, literature with high citation frequency, keywords, etc. Results A total of 675 eligible articles were retrieved, including 605 Chinese articles and 70 English articles. The annual publication volume of Longan Arillus-related research showed an overall upward trend. The country with the highest publication volume is China. The institutions conducting research on Longan Arillus are also mainly Chinese institutions. The institutions with the most publications in both Chinese and English are Shandong University of Traditional Chinese Medicine (11 articles) and Guangdong Academy of Agricultural Science (19 articles), respectively. The authors with the most publications are Li Xuehua (nine articles) and Zhang Mingwei (14 articles), respectively. The journals with the highest publication volume are Shaanxi Journal of Traditional Chinese Medicine and Carbohydrate Polymers, respectively. The top 10 most cited Chinese and English articles mainly focused on the clinical application and pharmacological effects of Longan Arillus. The most frequent Chinese and English keywords were Guipi Decoction (144 times) and polysaccharides (15 times). Conclusion The dominant force in Longan Arillus research is concentrated in China, with domestic institutions serving as the core entities. With the strengthening of domestic and international collaboration, the growth of market demand, the improvement of economic support, the guidance of relevant policies, and advances in research technology, research on Longan Arillus has gradually developed from early traditional clinical application and experience summarization into a modern multidisciplinary research system at the microscopic molecular level. Current research hotspots are based on traditional Chinese medicine theory, exploring the compatibility mechanisms, clinical applications, chemical components, and pharmacological mechanisms such as antioxidation and antitumor effects of Longan Arillus itself and its classic formulas (such as Guipi Decoction). In the future, research will continue to take the promotion of traditional Chinese medicine theory as its foundation, strengthen international exchange and cultural dissemination, and integrate digital and other technological innovation concepts to address key technical challenges in modern traditional Chinese medicine research, deeply analyze the fine chemical composition and pharmacological mechanisms of Longan Arillus, and systematically reveal its pharmacodynamic material basis and pathways of action. At the same time, this study will deepen the collaborative cooperation between universities and enterprises, build an efficient transformation channel from research to industrialization, and promote the industrialization of related achievements, integrate traditional theories with modern technology to enhance innovation capabilities and international influence, and providing a reference for the modernization of traditional Chinese medicine theory on Longan Arillus and the industrialization transformation of related scientific and technological achievements.

Objective To investigate the regulatory mechanisms by which different cultivation substrates influence the accumulation of pharmacologically active components in near-wild Tiepishihu (Dendrobium officinale), and provide a basis for elucidating the molecular mechanisms underlying quality differences and for optimizing cultivation substrates. Methods D. officinale samples grown on stone-based and wood-based substrates were used as experimental materials. Non-targeted metabolomics was employed to identify and compare the secondary metabolite profiles under the two cultivation modes, while transcriptome sequencing was conducted to screen differentially expressed genes (DEGs). KEGG and other databases were used for pathway enrichment analysis, which was then integrated with metabolite variation data. Results A total of 2 878 metabolites were identified by metabolomics, of which 354 showed significant differences between the two cultivation modes. These metabolites were mainly involved in phenylpropanoids, flavonoids, alkaloids, lipids, and aromatic amino acids. Flavonoids, alkaloids, and certain amino acid-derived metabolites exhibited overall higher accumulation in stone-grown samples, whereas lipids and steroid-related compounds were relatively enriched in wood-grown samples. Transcriptome sequencing identified 2 966 DEGs. KEGG enrichment analysis indicated significant differentiation in pathways including phenylpropanoid biosynthesis (ko00940), flavonoid biosynthesis (ko00941), phenylalanine/tyrosine/tryptophan biosynthesis (ko00400), and tropane, piperidine and pyridine alkaloid biosynthesis (ko00960). Genes encoding key enzymes in flavonoid biosynthesis [such as phenylalanine ammonia lyase (PAL), chalcone synthase (CHS), flavanone 3-hydroxylase (F3H)] were upregulated in stone-grown samples, consistent with the accumulation patterns of the corresponding metabolites. Integrated analysis suggested that cultivation substrates may regulate the expression of genes involved in phenylpropanoid-flavonoid-alkaloid metabolism, thereby altering the direction of metabolic flux allocation and ultimately shaping distinct metabolic phenotypes. Conclusion Cultivation substrates influence the types and accumulation levels of active constituents in D. officinale by regulating the expression of genes in phenylpropanoid, flavonoid, and alkaloid biosynthetic pathways, resulting in quality differences between stone-grown and wood-grown materials. This study systematically elucidates the molecular mechanisms underlying quality variation in D. officinale and provides a theoretical basis for cultivation substrate optimization and graded utilization of medicinal materials.

Objective To screen species and identify DNA molecular markers based on the chloroplast whole genome data of four species of the genus Bupleurum for medicinal use. Methods Based on the published chloroplast genome sequences of Bupleurum plants, the chloroplast genome was analyzed using bioinformatics methods. A specific DNA barcode for accurately identifying the four original species of Bupleurum medicinal materials was developed based on the hypervariable regions of the chloroplast genome, and then verified. Results The chloroplast genomes of four species ranged in size from 155 478—178 668 bp. All the four genomes were represented as typical quadripartite structures. Two pairs of primers based on the hypervariable regions were designed to efficiently amplify 55 samples. The analysis of sequencing results confirmed that two hypervariable regions, rps16-psbK and petN-psbM, can be used as specific DNA barcodes for the identification of Bupleurum chinense, B. scorzonerifolium, B. smithii and B. marginatum var. stenophyllum. Conclusion The specific DNA barcodes rps16-psbK and petN-psbM, screened by comparative chloroplast genomics, could achieve precise classification and identification of the four original species of Bupleurum medicinal materials, providing important scientific support for ensuring the safety of clinical use and regulating the market order of Bupleurum medicinal materials.

Objective To screen high-quality germplasm resources of Dianchonglou (Paris polyphylla var. yunnanensis), reveal the associations between phenotypic traits and saponin content, and lay a foundation for the exploration of genetic resources and the breeding of improved varieties. Methods Through genetic diversity analysis, correlation analysis, cluster analysis (CA), principal component analysis (PCA), partial least squares-discriminant analysis (PLS-DA), and partial least squares-regression analysis (PLS-R), a comprehensive evaluation was conducted on 25 agronomic traits and 10 quality traits of 41 selected P. polyphylla var. yunnanensis germplasm resources. Results The RSD and genetic diversity index (H′) for agronomic traits ranged from 3.56% to 66.63% and from 1.80 to 2.25, respectively. For quality traits, RSD and H′ ranged from 34.26% to 130.18% and from 1.47 to 2.00, respectively. The variation in agronomic traits was mainly attributed to the length of the purple stem portion, pedicel length, and pedicel ratio, while the variation in quality traits was mainly attributed to the contents of polyphyllin V, polyphyllin H, and gracillin. Correlation analysis showed that the sum of polyphyllin I, II, and VII contents was positively correlated with plant height, stem diameter, leaf shape index, rhizome diameter, and powder color values L^*and E^*_ab. Cluster analysis classified the P. polyphylla var. yunnanensis germplasm into three major groups: Group I represented high-yield germplasm, Group II represented high-quality germplasm, and Group III exhibited overall poor growth, low active component contents, and low yield, indicating a generally poor comprehensive performance. PCA simplified the 35 traits into eight principal components, with a cumulative contribution rate of 85.602%. PLS-DA and PLS-R revealed that stem height, leaf length, rhizome diameter, powder color values L^* and E^*_ab, and the total content of polyphyllin I, II, and VII contributed substantially to the D value, and could serve as important indicators for selecting superior provenances. Conclusion The top three germplasm resources based on comprehensive ranking were SZ-16, SZ-11, and SZ-9, which exhibited high quality and high yield, making them potential candidates for cultivation as elite varieties. The stem height, leaf length, rhizome diameter, and powder color values L^*and E^*_ab among agronomic traits, together with the total content of polyphyllin I, II, and VII among quality traits, are important characteristics for distinguishing superior from inferior P. polyphylla var. yunnanensis germplasm.

Objectives Gentiana dahurica was selected as the focal species to assemble and sequence analyze its chloroplast genome, investigate the phylogeographic structure of wild populations across China based on chloroplast gene sequences, assess genetic diversity and population genetic structure, explore the causes and correlations between the genetic characteristics and geographical distribution patterns of the G. dahurica population, and infer its glacial refugia and the dynamic history of the population. Methods A total of 27 wild populations of G. dahurica were collected from six provinces, including Qinghai, Gansu, Shaanxi, Inner Mongolia, ect. All individuals were subjected to DNA resequencing. Following genome assembly and annotation, the chloroplast genomes of all individuals were obtained for comparative genomic analyses, phylogenetic analysis, population dynamic history, and phylogeographic analyses. Results In total, 81 complete chloroplast genomes were generated, all exhibiting the typical quadripartite circular structure with lengths ranging from 148 792 to 149 000 bp. A total of 134 genes were annotated, including 85 protein-coding genes. The IR/SC boundaries of the chloroplast genomes of G. dahurica were conserved across populations. Variation in non-coding regions was markedly higher than in coding regions. G. dahurica exhibited high within-population genetic diversity (h_S = 0.632) and total genetic diversity (h_T = 0.923). Phylogenetic analyses assigned all populations into two genetic clades, but no significant correlation was detected between genetic distance and geographic location. Population size remained relatively stable over historical periods, with no evidence of strong population expansion. Based on the consensus of non-coding sequences of G. dahurica, 27 haplotypes were identified, Based on the consensus of non-coding sequences of G. dahurica, 27 haplotypes were identified. No clear phylogeographic structure was observed among populations. Dispersal corridor analyses suggested that the northeastern Qinghai-Tibet plateau-Qilian mountains-Liupan mountains-Lvliang mountains-Taihang mountains constituted an important historical migration pathway for G. dahurica. Conclusions The chloroplast genome of G. dahurica is highly conserved in structure and stable in gene content. Two mutation hotspots identified here may serve as candidate markers for molecular identification of germplasm resources. Wild G. dahurica likely originated on the Qinghai-Tibet plateau, and mountain chains such as the Qilian, Liupan, Lvliang, and Taihang ranges may have acted as historical migration corridors that facilitated genetic connectivity among populations. This study provides a theoretical foundation for germplasm conservation and development and utilization of this medicinal plant.

Traditional Chinese medicine chemical biology (TCMCB) is an interdisciplinary discipline emerging in the modernization of traditional Chinese medicine (TCM). Guided by TCM theories, TCMCB employs bioactive components from Chinese medicines as molecular probes, focusing on the precise identification of action targets, regulation of signaling pathways, and intervention mechanisms of life processes induced by TCM bioactive ingredients. It provides critical technical support for the modern interpretation of the scientific connotation of TCM and its international promotion. This paper systematically reviews the disciplinary orientation, core technologies and methodologies of TCMCB, summarizes its applications and bottlenecks in the mechanistic elucidation of TCM bioactive components and innovative drug discovery and development, and highlights original research achievements over the past decade. TCMCB has gradually established an integrated research paradigm: “bioactive component screening-precise target identification-mechanism elucidation-clinical translational application” promoting the transformation of TCM theories from empirical description to mechanism-driven interpretation, and offering novel strategies for disease treatment in modern medicine and innovative drug development. Future efforts should focus on overcoming key challenges including the supply of TCM bioactive components, precise validation of targets, and international evaluation systems. Meanwhile, the construction of technology platforms with independent intellectual property rights and a global target database for traditional medicines will facilitate the advancement of TCM toward modern precision medicine.

Suxiao Jiuxin Pill (速效救心丸, SJP) is the first pure traditional Chinese medicine dripping pill officially recorded in the Pharmacopoeia of the People’s Republic of China. It is formulated with Chuanxiong (Chuanxiong Rhizoma) and Bingpian (Borneolum Syntheticum) as core components, and is clinically indicated for coronary heart disease and angina pectoris belonging to the qi stagnation and blood stasis syndrome, characterized by definite efficacy and rapid onset. Against the background that treatment of coronary heart disease emphasizes syndrome differentiation and individualized medication, the clinical demand for rapid-acting medicines for acute chest impediment has become increasingly prominent. This paper systematically reviews relevant domestic and international literatures, focusing on the key links of its “rapid onset” and the mechanism of “saving the heart”. From the perspectives of chemical constituents, formulation advantages, pharmacokinetic characteristics and molecular biological mechanisms, this review aims to explore the unique advantages of SJP and provide new ideas and theoretical references for its precise clinical application and further in-depth mechanistic research.

Doxorubicin (DOX) is a widely used anthracycline anticancer agent; however, its dose-dependent and cumulative cardiotoxicity severely limits its long-term clinical use. Danshen (Salviae Miltiorrhizae Radix et Rhizoma) and Salviae Miltiorrhizae Radix et Rhizoma-based preparations have shown potential for the prevention and treatment of DOX-induced cardiotoxicity in both experimental and clinical studies. This review systematically summarizes the cardioprotective mechanisms of Salviae Miltiorrhizae Radix et Rhizoma extracts, active constituents (including tanshinone I, tanshinone II_A, and salvianolic acid B), Salviae Miltiorrhizae Radix et Rhizoma-containing herbal formulas, and Chinese patent medicines, with particular emphasis on their inhibitory effects on oxidative stress, apoptosis, pyroptosis, inflammation, and other pathological processes. Unlike previous reviews that have focused primarily on individual active constituents or the general cardiovascular protective effects of Salviae Miltiorrhizae Radix et Rhizoma, this review compares the pharmacological characteristics and mechanistic differences among different Salviae Miltiorrhizae Radix et Rhizoma-derived components in DOX-induced cardiotoxicity. Based on the current evidence, key barriers to clinical translation are further discussed to provide a reference for the clinical application and drug development of Salviae Miltiorrhizae Radix et Rhizoma-derived therapies.

Ulcerative colitis (UC) is a chronic relapsing inflammatory disease involving the colonic mucosa, clinically characterized by abdominal pain, diarrhea, and mucopurulent bloody stools. Rectal drug delivery has emerged as an important approach for UC treatment due to its advantages such as direct access to the lesion site, high local drug concentration, and avoidance of the first-pass effect. In recent years, the continuous development of traditional Chinese medicines, chemical drugs, and various rectal formulations have significantly improved the retention and controlled-release performance of rectal drug delivery systems. This review systematically summarizes the domestic and international development history, biological basis of rectal drug delivery, and the pathogenesis of UC, with a focus on analyzing the progress of formulation innovations and clinical studies in rectal drug delivery for UC treatment over the past five years, aiming to provide a reference for the individualized treatment strategies of UC.

Against the backdrop of accelerated innovation in the global biomedical sector and China’s transition toward innovation-driven development in the pharmaceutical industry, Jiangxi province—with its profound traditional Chinese medicine heritage and industrial foundation characterized by “one city, one capital, and multiple parks”—faces both opportunities and challenges. Prominent issues include the relatively small scale of emerging fields such as biologics and antibody drugs, insufficient clustering of high-end innovation factors, and low efficiency in translating research outcomes into applications. To study these challenges, this study employs methods such as literature review, case analysis, and comparative research to systematically assess the current status and core weaknesses of Jiangxi’s innovative drug industry. It further proposes a differentiated development pathway centered on “modernizing traditional Chinese medicine as the foundation while promoting differentiated and coordinated development of biologics and antibody drugs.” The study recommends that Jiangxi should focus on advancing the digital and intelligent transformation and standardization of the entire TCM industry chain, cultivate blockbuster pharmaceutical products with a market scale of RMB 10 billion, implement a gradient layout in key areas such as cell therapy and antibody-drug conjugate drugs, and build a synergistic innovation ecosystem integrating “talent, capital, platforms, and policies,” with strengthened clinical translation and financial support. These measures will enable the transition from a “major pharmaceutical province” to a “strong pharmaceutical province.”

Non-motor symptoms of Parkinson’s disease (PD) involve multidimensional manifestations such as hyposmia, rapid eye movement sleep behavior disorder, mood disorder, cognitive impairment and autonomic nerve dysfunction, which can occur in the prodromal and progressive stages of the disease. Its occurrence and development are closely related to abnormal folding, aggregation, clearance disorder of alpha-synuclein (α-syn) and prion-like transmission. Ciwujia (Acanthopanacis Senticosi Radix et Rhizoma seu Caulis)has the effects of invigorating qi and spleen, tonifying kidney and tranquilizing mind. Modern pharmacological studies have shown that its multi-components, multi-targets and multi-pathways regulation characteristics are highly compatible with the complexity of the pathological cascade of α-syn. Focusing on the pathological cascade of α-syn, by systematically combing the physiological conformation dynamics of α-syn, the basis of pathological transformation, the driving factors of aggregation, the imbalance of cell homeostasis and the mechanism of central nervous circuit damage, the potential role of Acanthopanacis Senticosi Radix et Rhizoma seu Caulisin indirectly affecting the abnormal accumulation of α-syn and the pathological process related to non-motor symptoms by improving α-syn pathological related microenvironment such as oxidative stress, neuroinflammation, protein clearance disorder, neurotransmitter imbalance and brain-gut axis abnormality was further analyzed, which provided a new research idea for the comprehensive intervention of non-motor symptoms of PD.

Traditional Chinese medicine (TCM) decoctions are not single true-solution systems, but complex heterogeneous and multiphase dispersed systems formed through the self-assembly of multiple constituents via non-covalent interactions. The supramolecular “imprinting template” theory of TCM provides an analytical perspective for interpreting multi-component recognition, non-covalent assembly, phase-state formation, and the material forms underlying pharmacological activity in decoctions. From this theoretical perspective, this review summarizes the formation mechanisms, physicochemical characterization, component distribution patterns, and pharmacological relevance of different phase states in TCM decoctions, including true solution, colloidal, suspension, and precipitate phases. Different phase states show marked physical differences in particle size, potential, and microscopic morphology, and chemical constituents exhibit phase-preferential distribution. Key active constituents such as alkaloids and flavonoids are often selectively enriched in colloidal or precipitate phases. These specific supramolecular phase-state structures may represent important material forms mediating pharmacological effects and are closely related to drug dissolution, stability, in vivo transport, and bioavailability. Among them, colloidal phases often show potential passive targeting characteristics and solubilization effects, whereas suspension and precipitate phases may play important roles in toxicity attenuation and sustained release through physical encapsulation, providing a phase-state-based explanation for understanding the compatibility of TCM formulas. In addition, this review further summarizes the main research strategies, including decoction kinetic evolution, phase separation based on particle size differences, physicochemical characterization and pharmacological evaluation of different phase states, and reverse assembly and mechanism verification based on characteristic constituents. The key challenges in applying phase-state characteristics to TCM quality evaluation are also discussed. Future studies may be deepened in terms of in situ characterization, in vivo transformation processes, quantitative analysis of phase-state fingerprints, and quality evaluation, providing references for elucidating the pharmacodynamic material basis of TCM decoctions and promoting the modern formulation transformation of classical prescriptions.

As a core ecological factor in the root zone of plants, rhizosphere microorganisms’ community structure and functional diversity directly affect medicinal plants’ nutrient utilization, stress response, and secondary metabolite synthesis. Systematically elucidating the mechanisms by which rhizosphere microorganisms regulate medicinal plant quality is of great significance for developing ecological cultivation techniques based on microecological regulation and stabilizing the genuine characteristics of medicinal herbs. This paper systematically reviews the core mechanisms by which rhizosphere microorganisms influence medicinal plant quality through improving nutrient absorption, regulating endogenous hormone balance, inducing systemic resistance (ISR), and directly participating in the synthesis of active compounds. It also summarizes application strategies such as rhizosphere microecological regulation mediated by root exudates, microbial inoculant improvement, and construction of synthetic microbial communities, while analyzing practical bottlenecks such as unstable field colonization of functional microbial inoculants and competition with native microorganisms, aiming to provide a theoretical reference for the rhizosphere microecology of medicinal plants and the green production of authentic medicinal herbs.

Ruxiang (Olibanum) is a commonly used traditional Chinese medicinal material with a long history of clinical application, and it is known for its effects of promoting blood circulation, relieving pain, reducing swelling, and promoting tissue regeneration. To date, more than 400 compounds have been isolated and identified from Olibanum, among which diterpenoids and triterpenoids are considered the major bioactive constituents. Olibanum and its active components exhibit a wide range of pharmacological activities, including anti-inflammatory, hepatoprotective, and antitumor effects. The underlying mechanisms involve multiple pathways, such as the inhibition of inflammatory mediator release and the regulation of oxidative stress. This review summarizes the recent advances in the chemical constituents and pharmacological activities of Olibanum, aiming to provide a reference for its further development and utilization, as well as for related basic and clinical research.