Based on extensive research into the current status of understory medicinal herb cultivation and an in-depth analysis of existing problems, this study proposes policy and technical recommendations to promote sustainable and high-quality development. The research team conducted field investigations in advantageous cultivation regions across seven provinces (autonomous regions), including Guangxi, Yunnan, Fujian, and Jilin, supplemented by policy reviews and literature analysis. The findings reveal that although understory medicinal herb cultivation benefits from ecological advantages and policy support, it still faces a series of problems, such as an underdeveloped service system, insufficient regulatory capacity, low market alignment, and immature technical systems. It should be particularly noted that understory cultivation commonly encounters specific challenges, including the economic dilemma of high input and low output, mismatches between forest canopy density and the light requirements of medicinal plants, ambiguous boundaries between ecological protection and utilization in policy implementation, and weak research support, leading to high trial-and-error costs in technology adoption. The following development recommendations are proposed: ① establish a coordination mechanism between forestry and traditional Chinese medicine authorities to integrate planning and policy synergy; ② enhance the alignment of understory medicinal herb cultivation with market demand; ③ develop an independent technical system and standards for understory medicinal herb cultivation, such as establishing a variety suitability framework based on canopy density classification and ecological cultivation protocols; ④ strengthen research focused on industrial development, with priority given to large-scale plantations (e. g., pine and fir forests), dual-purpose food-medicine species (Polygonatum, Gastrodia elata), vine species, and intercropping effects; ⑤ establish a quality certification system and corresponding regulatory framework for understory-cultivated medicinal materials to promote premium pricing; ⑥ improve land-use policies and supporting infrastructure, clarifying utilization rights for different forest types and simplifying approval procedures for temporary facilities.

Based on integrated analysis of multi-source heterogeneous biomedical data combined with animal experimental validation, this study systematically explored the advantageous therapeutic pathways and molecular mechanisms of Jianpi Huogu Formula (JPHGF) in treating steroid-induced osteonecrosis of the femoral head (SONFH). First, the candidate active components and targets of JPHGF were obtained from the Encyclopedia of Traditional Chinese Medicine (ETCM v 2.0). Meanwhile, the Human Phenotype Ontology (HPO) database was used to identify potential genes associated with the corresponding syndrome pattern. Finally, clinical transcriptomic data were analyzed to obtain relevant targets for the phlegm-blood stasis blocking collateral syndrome of SONFH. The intersection of these three types of targets was used to construct a multidimensional "drug-ingredient-disease-syndrome" network. The STRING database was employed for protein-protein interaction (PPI) network analysis, and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of core targets was performed via the DAVID platform to predict key biological processes and signaling pathways. Pharmacodynamic and mechanistic validation was subsequently conducted using a rat model of SONFH with phlegm-blood stasis obstructing collateral syndrome. Data integration and mining yielded a "disease and syndrome geneformula and drug target" network containing 146 core targets. Pathway enrichment analysis indicated that JPHGF primarily regulates lipid-coupled intervention in the SONFH phlegm-blood stasis blocking collateral phenotype through the purinergic receptor P2Y12 (P2RY12) -phosphoinositide 3-kinase (PI3K) -serine/threonine kinase (Akt) -fibrinogen gamma chain (FGG) signaling pathway. Hematoxylin-eosin (HE) staining revealed that rats with SONFH of the phlegm-blood stasis blocking collateral type exhibited reduced calcification in femoral head tissues, loosely arranged trabeculae, hypoplastic osteocytes, significantly increased lacunae, and elevated numbers of intramedullary adipocytes. Lipid profile analysis showed elevated triglyceride, total cholesterol, and low-density lipoprotein (LDL) levels, along with decreased high-density lipoprotein (HDL). Coagulation assays demonstrated shortened activated partial thromboplastin time (APTT) and prothrombin time (PT), as well as prolonged thrombin time (TT) and increased plasma fibrinogen levels. Immunohistochemical staining showed that compared with the normal group, the model group exhibited significantly increased positive expression areas of P2RY12, phosphorylated PI3K (p-PI3K), phosphorylated Akt (p-Akt), and FGG proteins in femoral head tissues (P<0.05). In contrast, all drug-treated groups showed significantly reduced expression of these proteins (P<0.05), with the most pronounced inhibitory effect observed in the high-dose group. These results indicate that JPHGF modulates the P2RY12-PI3K-Akt-FGG signaling axis in a dose-dependent manner. In conclusion, JPHGF exerts a syndrome-specific therapeutic effect through multi-target and multi-pathway synergistic actions. By particularly suppressing the excessive activation of the P2RY12-PI3K-Akt-FGG signaling axis, it improves lipid and bone metabolism abnormalities in SONFH with phlegm-blood stasis blocking collateral syndrome.

This study aimed to elucidate the therapeutic effects and molecular mechanisms of Periplaneta americana extract (PAE) on ground-glass nodule (GGN) -type early-stage lung adenocarcinoma (LUAD) using patient-derived organoid (PDO) models. GGN-type patient-derived early-stage LUAD organoid (GGN-PDO) were successfully established. At the level of drug quality control and component analysis, the content of free amino acids and peptides in PAE were determined by high-performance liquid chromatography (HPLC), and the main components were further analyzed by liquid chromatography-mass spectrometry (LC-MS). Analysis of variance was used to evaluate the similarity between drugs of the same batch (Pearson correlation coefficient 0.87-0.99) and ensure quality control. At the histopathological level, hematoxylin-eosin (HE) staining and immunohistochemical (IHC) analysis of key molecular markers, including cytokeratin 7 (CK7), thyroid transcription factor-1 (TTF-1), and Ki-67, confirmed that the organoids recapitulated the tumor characteristics of parental tissues. Pharmacodynamic evaluation demonstrated that PAE significantly inhibited the proliferation and viability of GGN-PDOs in a dose-dependent manner, while immunofluorescence (IF) staining showed a marked reduction in the proliferation marker Ki-67. Flow cytometry further confirmed that PAE induced apoptosis in GGN-PDO, and Western blot analysis demonstrated that PAE downregulated the anti-apoptotic protein Bcl-2 and upregulated the pro-apoptotic protein Bax, thereby activating the apoptotic pathway. Notably, for the first time, the pro-apoptotic mechanism of PAE was shown to involve inhibition of the Janus kinase 1 (JAK1)/phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway, as evidenced by markedly decreased protein levels of JAK1, PI3K, p-PI3K, AKT, and p-AKT following PAE treatment. In summary, this study is the first to use a clinically relevant GGN-PDO models to demonstrate that PAE induces apoptosis in early-stage LUAD via JAK1/PI3K/AKT pathway inhibition, providing a solid theoretical and experimental basis for developing innovative pharmacotherapies and non-surgical interventions against GGNs.

Based on the adiponectin receptor/5′-AMP-activated protein kinase catalytic subunit α-1/carnitine palmitoyltransferase 1b (AdipoR/AMPKα/Cpt1b) signaling axis, this study investigated the therapeutic mechanisms of Jianpi Huogu Formula (JPHGF) in alcohol-induced osteonecrosis of the femoral head (AONFH). A total of 56 Sprague-Dawley rats were randomly divided into a normal group, a model group, and a JPHGF group (5.0 g·kg^-1). Except for the normal group, rats in the other two groups were administered 46% ethanol by gavage to establish the AONFH model, and JPHGF intervention was initiated in the treatment group at week 4 after model induction. After 4 and 8 weeks of intervention, samples were collected. Mechanical and cold pain thresholds were assessed using Von Frey filaments and the acetone drop test, respectively. Hindlimb muscle strength was evaluated by the inclined plane test. Micro-computed tomography (micro-CT) was used to determine bone mineral density (BMD), bone volume fraction (BV/TV), trabecular thickness (Tb. Th), and other bone morphometric parameters. Hematoxylin-eosin (HE) staining was performed to observe histopathological changes. Transcriptomic sequencing was conducted to identify differentially expressed genes (DEGs) regulated by JPHGF in AONFH, followed by Gene Ontology (GO) functional and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses. Western blot was used to detect the protein expression of AdipoR1, AdipoR2, phosphorylated AMPKα, and Cpt1b in femoral head tissues. The results showed that after 8 weeks of modeling, rats in the model group exhibited significantly decreased mechanical pain thresholds and increased cold sensitivity (P<0.05, P<0.01), indicating hyperalgesia. The maximum inclined angle was markedly reduced (P<0.01), suggesting weakened hindlimb muscle strength. The femoral head surface appeared rough, with enlarged dark-red cartilage areas and sparse, disrupted trabeculae; the proportion of empty lacunae and the number of adipocytes were significantly increased (P<0.01), while BMD, BV/TV, Tb. Th, and trabecular number (Tb. N) were significantly decreased (P<0.01). After 4 weeks of JPHGF intervention, mechanical and cold hyperalgesia were significantly alleviated (P<0.05), hindlimb strength was restored (P<0.01), and surface morphology of the femoral head was improved. After 8 weeks, trabecular structure was further restored, with decreased empty lacunae and reduced adipocyte numbers, and BMD, BV/TV, Tb. Th, and Tb. N significantly increased (P<0.05, P<0.01); cystic degeneration and collapse were markedly alleviated. Transcriptomic analysis revealed that JPHGF significantly regulated 231 key DEGs, which were mainly enriched in adipocytokine signaling pathways and closely associated with AdipoR1, AdipoR2, p-AMPKα, and Cpt1b. Western blot analysis showed that AdipoR1, AdipoR2, p-AMPKα, and Cpt1b expression levels were significantly downregulated in the AONFH model group compared with the normal group (P<0.01), but were markedly upregulated following JPHGF treatment (P<0.05, P<0.01). In summary, this study demonstrates that JPHGF exerts therapeutic effects against AONFH by activating the AdipoR/AMPKα/Cpt1b signaling axis, thereby improving bone marrow lipid homeostasis and restoring femoral head microarchitecture, effectively relieving pain and functional impairment. These findings provide new experimental evidence and molecular targets for the prevention and treatment of AONFH in clinical practice.

On the basis of significant achievements in static research on blood stasis syndrome (BSS) in non-traumatic osteonecrosis of the femoral head (NONFH), to further reveal the evolutionary law of the entire disease course, our team took the lead in proposing and systematically constructing the theoretical system of "dynamic evolution of BSS throughout the whole course of NONFH". The core viewpoint of this system is that BSS runs through the entire course of NONFH occurrence, development, and prognosis, and its severity shows a regular dynamic trajectory with disease progression: "initial appearance and gradual increase in the early stage (less→more)→significant peak in the middle stage (more) →gradual alleviation in the late stage (more→less)". "Stasis causing Bi (blockage)" is the core pathogenesis chain. Supported by the Key Program of the National Natural Science Foundation of China, the team has built a solid evidence chain through multi-dimensional studies: At the clinical syndrome level, large-scale data mining and syndrome element analysis revealed that the proportion of BSS-related syndrome types/manifestations presents a "low→high→low" evolution. At the molecular characterization level, network topological feature calculation and functional mining identified stage-specific core pathways (bone/lipid metabolism→immune inflammation/circulation→multi-system disorder), with animal models and candidate biomarkers verifying such dynamic correlation. At the imaging level, the BSS prediction model based on MRI and dynamic changes in DSA vascular indices consistently confirmed this evolutionary pattern. Based on this theory, an innovative "three stages (early, middle, late) and four types (Qi stagnation and blood stasis, phlegm-stasis blocking collaterals, meridian blockage, liver-kidney deficiency)" syndrome differentiation system was established, guiding the formation of stage-specific precise therapies: "strengthening spleen, resolving phlegm, activating blood and dredging collaterals (Jianpi Huogu Decoction) →tonifying Qi, activating blood, dredging meridians and relieving Bi (Huoxue Tongbi Decoction) →nourishing liver and kidney, replenishing essence and resolving stasis (Bushen Zhuanggu Decoction)", which showed significant clinical efficacy. This theoretical system represents a deepening and expansion of BSS research in NONFH, and provides important support for explaining the scientific connotation of BSS in chronic bone diseases, promoting the precise diagnosis and treatment of NONFH, and innovating the research paradigm of dynamic TCM syndromes.

This study aimed to observe the promoting effect of Jianpi Huogu Formula (JPHGF) on angiogenesis-osteogenesis coupling in rats with steroid-induced osteonecrosis of the femoral head (SONFH), and to explore its mechanism of action through the vascular endothelial growth factor (VEGF) -Notch-Noggin signaling pathway. A total of 120 Sprague-Dawley rats were randomly divided into a normal group, a model group, low-, medium-, and high-dose JPHGF groups (2.5, 5, and 10 g·kg^-1), and a Jiangusheng Pills group (1.53 g·kg^-1). Except for the normal group, all rats were injected with lipopolysaccharide (20 μg·kg^-1) via the tail vein once daily on days 1 and 2, followed by intramuscular injection of methylprednisolone sodium succinate (40 mg·kg^-1, gluteal muscle) once daily from days 3 to 5. The normal group received an equivalent volume of normal saline. Four weeks after model establishment, the rats were administered the corresponding drugs by gavage. The therapeutic effects of JPHGF on SONFH rats were evaluated by measuring mechanical pain threshold, cold pain sensitivity, hemorheological parameters, four coagulation indices, and by pathological and imaging analyses of the femoral head. The microvascular distribution of the femoral head was observed by ink perfusion and Microfil angiography. Immunofluorescence was used to detect the expression of platelet endothelial cell adhesion molecule-1 (CD31), endothelial mucin (Emcn), and the osteoblast-specific transcription factor Osterix, as well as the co-expression of CD31 and Noggin proteins in the femoral head. Immunohistochemistry was employed to detect the expression of type Ⅰ collagen (collagen Ⅰ) and Notch-1 proteins, while Western blot was used to measure the expression levels of VEGF and Noggin proteins in the femoral head. The results showed that JPHGF significantly increased the mechanical pain threshold and reduced the number of foot lifts in response to cold stimulation in SONFH rats. It markedly increased trabecular bone area, reduced the number of adipocytes and the percentage of empty lacunae, improved imaging manifestations of femoral head necrosis, and increased the vascular area in ink perfusion and the vessel volume in Microfil perfusion within the medullary cavity of the femoral head. JPHGF also improved hemorheological parameters and coagulation function, upregulated the expression of CD31, Emcn, and Osterix proteins, enhanced the co-expression of CD31 and Noggin proteins, and promoted the expression of collagen Ⅰ, Notch-1, VEGF, and Noggin proteins. These findings indicate that JPHGF promotes angiogenesis-osteogenesis coupling by activating the VEGF-Notch-Noggin signaling pathway, thereby effectively treating SONFH in rats. This study elucidates the pharmacological connotation of JPHGF from the perspective of "angiogenesis-osteogenesis coupling" and provides an experimental basis for its clinical application.

This study is aimed to investigate the therapeutic effects of Junren Bufei Yixin Granules (JRBF) on hypoxic pulmonary hypertension (HPH) and explore the therapeutic mechanism through the polypyrimidine tract-binding protein 1 (PTBP1)/pyruvate kinase isozyme type M2 (PKM2) signaling pathway. Sixty SPF-grade C57BL/6 male mice were randomized into normal, model, sildenafil, and low/medium/high-dose JRBF groups. HPH was induced by hypoxic exposure (10% O₂, 8 h/day) for 4 weeks. The right ventricular systolic pressure (RVSP) was measured by right cardiac catheterization. Echocardiography was employed record the pulmonary artery acceleration time (PAAT), pulmonary ejection time (PET), tricuspid annular plane systolic excursion (TAPSE), and right ventricular anterior wall thickness at diastole (RVAWd). The histopathological changes were observed by hematoxylin-eosin (HE) and Masson staining. Western blot was employed to measure the expression levels of PTBP1, PKM2, and PKM1 in the lung tissue. The immunofluorescence assay was employed to detect the co-localization of PTBP1 with the marker alpha-smooth muscle actin (α-SMA) of pulmonary arterial smooth muscle cells (PASMCs). The lactate assay kit was used to measure the lactate levels in the serum and lung tissue. In the cell experiments, the hypoxia model was established by exposing human pulmonary artery smooth muscle cells (hPASMCs) to 1% O₂ for 48 h. The optimal (10%) JRBF concentration for intervention was determined by the cell-counting kit-8 (CCK-8). The cell proliferation (EdU), migration (Transwell), PTBP1/PKM2/PKM1, and lactate production were analyzed. The results showed that JRBF significantly reduced the RVSP, improved the PAAT/PET ratio and TAPSE, and attenuated pulmonary arteriolar remodeling and myocardial fibrosis. Furthermore, JRBF suppressed PTBP1 and PKM2 expression and reduced the lactate accumulation in the lung tissue and PASMCs. The cell experiment results showed that the 10% JRBF-containing serum inhibited the proliferation and migration of hPASMCs, while decreasing the PTBP1/PKM2 expression and lactate production. In conclusion, JRBF mitigates HPH progression by suppressing the PTBP1/PKM2 axis and restoring the glycolytic flux.

This study aimed to investigate the mechanisms by which Acanthopanax senticosus extract (ASH) exerts effects onα-synuclein (α-syn) overexpressing transgenic mouse model of Parkinson's disease (PD), with a focus on its regulation of brain lipid metabolism. Twenty PD model mice were randomly assigned to a model group or an ASH treatment group (45.5 mg·kg^-1 by gavage for 4 weeks), and 10 C57BL/6 mice served as a normal control group. Behavioral assessments revealed that, compared with controls, PD model mice showed prolonged pole test time, reduced spontaneous locomotor activity, shorter latency to fall in the rotarod test, and decreased total distance traveled in the open field test. Serum levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), caspase-9 were significantly elevated, B-cell lymphoma-2 (Bcl-2), and proliferating cell nuclear antigen (PCNA) expression was reduced, and marked neuronal damage was observed in brain tissue. ASH intervention significantly improved these behavioral and biochemical parameters and attenuated neuronal injury. Untargeted lipidomics analysis revealed significant alterations in sphingomyelin (SM), ceramide (Cer), phosphatidylcholine (PC), and phosphatidylserine (PS) across multiple brain regions (cortex, substantia nigra, cerebellum, and striatum) in PD mice, which were notably restored by ASH treatment. Pathway analysis indicated that these metabolites were predominantly involved in sphingolipid metabolism. Western blot further demonstrated that ASH downregulated the expression of key sphingolipid metabolic enzymes serine palmitoyltransferase long-chain base subunits 1 and 2 (SPTLC1 and SPTLC2) and upregulated UDP-glucose ceramide glucosyltransferase (UGCG), β-galactosylceramidase (GALC), and sphingosine kinase 2 (SPHK2), thereby suppressing abnormal SM and Cer accumulation in the substantia nigra and elevating PS and PC levels in the striatum. Spearman's correlation analysis supported the modulatory effect of ASH on brain lipid metabolic profiles. In conclusion, ASH improves behavioral deficits, exerts anti-inflammatory effects, and regulates sphingolipid metabolism to correct disordered lipid profiles, thereby providing neuroprotective effects in PD mice.

This study aims to investigate the effects of glycyrrhizic acid and calcium ions from adjuvant solutions on the toxic lectin in Pinelliae Rhizoma Praeparatum and clarify the processing detoxification mechanism. Molecular docking and molecular dynamics simulations were employed to construct a 3D model of lectin and evaluate the interactions of glycyrrhizic acid and/or calcium ions with lectin. Wild-type and mutant lectin proteins were produced via a prokaryotic expression system in Escherichia coli. The interactions of glycyrrhizic acid and/or calcium ions with the lectin proteins were verified by Bio-Layer Interferometry (BLI), and the irritant toxicity was assessed via a rabbit model of conjunctival irritation. The findings revealed that the ternary complex formed by glycyrrhizic acid, calcium ions, and lectin exhibited greater stability. This complex showed significantly reduced root mean square deviation (RMSD), root mean square fluctuation (RMSF), solvent-accessible surface area (SASA), radius of gyration (Rg), Gibbs free energy of solvation (ΔGSOLV), and total binding free energy (ΔTOTAL), but significantly increased number of hydrogen bonds and gas-phase free energy (ΔGGAS). The combined action of glycyrrhizic acid and calcium ions led to a significant increase in the number of interactions in the amino acid residue region 51-56, 170-175, and 232-237 of lectin, with enhanced Coulomb interaction-short range (Coul-SR) and Lennard-Jones-short range (LJ-SR). When the wild-type lectin protein co-bound with glycyrrhizic acid and calcium ions, the affinity constant was 8.10×10^-6 mol·L^-1, which was significantly lower than that of the binary complex of wild-type lectin with glycyrrhizic acid or calcium ions and the ternary complex of mutant lectin with glycyrrhizic acid and calcium ions. Additionally, lectin soaked in a mixed solution of glycyrrhizic acid and calcium ions did not significantly exacerbate the irritation of the rabbit conjunctiva when being combined with toxic needle crystals. In conclusion, glycyrrhizic acid and calcium ions demonstrate a synergistic effect in forming a structurally stable and non-toxic supramolecular complex with lectin, thereby achieving the processing detoxification of Pinelliae Rhizoma Praeparatum.

Attention deficit hyperactivity disorder (ADHD), a common neurodevelopmental disorder in children, is characterized by inattention, hyperactivity, and impulsivity. Epidemiological surveys show that the prevalence of ADHD in children is gradually increasing worldwide, and it is the most common childhood mental disorder in China. Because of the complex clinical symptoms, multiple co-morbidities, and unknown etiology, ADHD has far-reaching negative impacts on individuals, families, and the society. Behavioral interventions, as a pillar in the management of ADHD, play a targeted role in improving children's social functioning, with significant benefits supported by evidence. However, they are constrained by uneven resources, poor compliance, and insufficient continuity, Western medicine has multiple adverse effects and unclear long-term effects in the treatment of ADHD despite the definite efficacy. Accordingly, there is an urgent need to find safe and effective therapies suitable for children. With a holistic view and treatment based on syndrome differentiation, traditional Chinese medicine (TCM) has significant advantages in treating ADHD via multiple targets, which involves dopamine (DA), norepinephrine (NE), 5-hydroxytryptamine (5-HT), cyclic adenosine monophosphate (cAMP), brain-derived neurotrophic factor (BDNF) and other signaling pathways. Through these pathways, TCM can treat ADHD through the regulation of neurotransmitters, enhancement of prefrontal and striatal functions, enhancement of neuronal protection, attenuation of neuroinflammation, and reduction of neuronal apoptosis. However, a systematic study remains to be conducted. This paper summarizes the signaling pathways related to the treatment of ADHD by TCM in the past two decades, aiming to provide reference for delving into the mechanism and exploring effective TCM prescriptions for ADHD in children and to give full play to the advantages of the efficacy and characteristics of TCM.