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.