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.

To analyze the changing pattern of blood stasis degree in different TCM syndromes of non-traumatic osteonecrosis of femoral head (NONFH), the present study adopted a retrospective cross-sectional design. The clinical and digital subtraction angiography (DSA) data were collected from 60 NONFH patients with blood stasis syndrome treated at the Third Affiliated Hospital of Beijing University of Chinese Medicine from February 2022 to March 2025. According to Chinese Medicine Identification Criteria for Necrosis of the Femoral Head (2019 edition), 60 patients were allocated into three groups of phlegm-stasis obstruction of collaterals, meridian obstruction, and liver-kidney deficiency, with 20 patients in each group. The morphological features of blood vessels were observed by DSA, and the diameter of the profunda femoris artery, diameters of medial/lateral circumflex femoral arteries, and vessel count were recorded. The degrees of blood stasis in different syndrome groups were analyzed. The results showed that there were 38 males and 22 females among the 60 patients, with mean age of (41.18±14.56) years. Two, thirty-three, twenty-two, and one patients were in ARCO stages Ⅰ, Ⅱ, Ⅲ and Ⅳ, respectively. There was no difference in the baseline data (age, BMI, etc.) among the three groups. Regarding the vascular parameters, the meridian obstruction group had the profunda femoris artery diameter of (6.59±0.94) mm, medial circumflex femoral artery diameter of (2.99±0.42) mm, medial/lateral ratio of 0.50 (0.43, 0.58), and total number of arteries being 8.00 (7.00, 8.00), which were all lower than those in the phlegm-stasis obstruction group and the liver-kidney deficiency group (P<0.05, P<0.01). The results indicated that the DSA of NONFH patients with blood stasis presented abnormal vascular morphology, and their vascular images showed a certain pattern throughout the disease course, with lower vessel diameter and count in the middle stage. This result suggested that the degree of blood stasis in the patients varied from mild to severe and back again with the progression of the disease. This study not only provides quantifiable imaging indicators for the macroscopic diagnosis of blood stasis syndrome and verify its dynamic evolution law, but also suggests that in clinical practice, imaging techniques such as DSA can be used to assist in syndrome differentiation and classification. This provides objective references for staged treatment and the optimization of intervention timing and intensity of blood-activating and stasis-resolving regimens.

A long-term follow-up non-randomized controlled study was conducted to evaluate the medium- and long-term efficacy of the Jianpi Huogu (Kanggu) Formula in treating early-stage (ARCO Ⅰ-Ⅱ stage) femoral head necrosis, thereby providing evidence for the early prevention and treatment of the disease with TCM, improvement of symptoms, and delay of disease progression. Patients who received Jianpi Huogu (Kanggu) Formula (experimental group, n = 61) or formula combined with TCM infusion therapy (control group, n = 168) between January 2003 and June 2020, with a follow-up of ≥3 years, were included. Clinical and imaging data at baseline and annual follow-ups were collected, and 40 matched pairs (80 cases) were selected using propensity score matching. The primary endpoints were clinical efficacy (hip pain scores, hip flexion function scores, walking distance scores, and total scores of clinical efficacy) and the Beijing University of Chinese Medicine X-ray Evaluation (BUCMXE), including femoral head morphology scores, osteoarthritis scores, necrotic lesion density scores, and total scores of BUCMXE. The efficacy of the two groups was compared before treatment and at 3, 5, and 10 years post-treatment. Clinical efficacy showed that, compared with pre-treatment, both groups had significantly reduced hip pain scores and improved hip flexion function scores, walking distance scores, and total scores of clinical efficacy at 3, 5, and 10 years (P<0.001). Imaging evaluation showed that, compared with pre-treatment, femoral head morphology scores in experimental group increased at 3 (0.65±1.00, P<0.05), 5 (0.70±1.02, P<0.001) and 10 years (0.80±1.08, P<0.05); osteoarthritis scores increased at 3 (0.32±0.66, P<0.05) and 5 years (0.32±0.67, P<0.05); necrotic lesion density scores decreased at 3 (1.18±0.50, P<0.05) and 5 years (1.19±0.52, P<0.05); total scores of BUCMXE increased at 3 (2.15±1.29, P<0.05) and 5 years (2.22±1.32, P<0.05). Compared with control group, the femoral head morphology scores increased in experimental group (P<0.05). These results confirm that the Jianpi Huogu (Kanggu) Formula can significantly improve clinical symptoms (pain and joint function) in patients with ARCO Ⅰ-Ⅱ stage femoral head necrosis. Medium- and long-term imaging data suggest slow disease progression (scores not reaching grade Ⅰ pathological criteria). For patients with ARCO Ⅰ stage or mild disease, monotherapy with this formula may be considered.

This study aims to investigate the clinical efficacy and underlying mechanism of Huoxue Tongbi Formula in treating patients with steroid-induced osteonecrosis of the femoral head (SONFH) with meridian blockage syndrome. A total of 40 patients with SONFH and meridian blockage syndrome were retrospectively enrolled. Demographic data, clinical efficacy scores (pain and hip joint function), Harris scores, hip joint range of motion, imaging indices (ARCO stage, BUCMXE, bone marrow edema, and joint effusion), symptom/syndrome scores, and coagulation-related physiochemical indices (complete blood count, four coagulation tests, and four thrombosis tests) were collected and compared before treatment and after three months of treatment. Peripheral blood samples from three patients with significant efficacy were subjected to transcriptome sequencing before and after treatment to screen differentially expressed genes (DEGs). GO and KEGG enrichment and PPI network analysis were performed. Genes most relevant to the core phenotype of SONFH with meridian blockage syndrome were identified using Spearman correlation analysis. The results showed that Huoxue Tongbi Formula significantly improved the clinical efficacy scores (including pain and hip joint flexion function), Harris scores, and range of internal and external rotation of the hip (P<0.01). It also maintained stable imaging indices, alleviated various blood stasis-related clinical symptoms (hip pain, fixed pain, lower limb weakness) (P<0.05, P<0.001), and significantly reduced thrombin-antithrombin complex (TAT) levels (P< 0.01). Transcriptome analysis identified 368 DEGs (274 upregulated and 94 downregulated). GO functional enrichment revealed that the biological processes involved by DEGs mainly include positive regulation of tumor necrosis factor production, integrin-mediated signaling pathways, neutrophil chemotaxis, and regulation of tumor necrosis factor production. KEGG enrichment analysis revealed that these DEGs were primarily involved in pathways such as hematopoietic cell lineage, osteoclast differentiation, complement and coagulation cascades, and platelet activation. PPI network analysis combined with correlation analysis identified core genes UBA52 (r=0.83, P<0.05), RACK1 (r=0.88, P<0.05), and RPL19 (r=-0.87, P<0.05) were significantly correlated with blood stasis syndrome scores. CD74 was significantly correlated with osteonecrotic lesions (r=0.95, P<0.05) and TAT (r=0.88, P<0.05). These findings indicate that Huoxue Tongbi Formula ameliorates clinical symptoms and certain imaging indices in SONFH patients with meridian blockage syndrome by regulating hematopoiesis and bone metabolism (inhibiting osteoclast activity) and correcting hypercoagulability (inhibiting coagulation activation). Core genes such as UBA52, RACK1, and RPS3A may be key molecules linking blood stasis syndrome to the regulation of the coagulation-bone metabolism network.

This study systematically elucidated the scientific connotation of Jianpi Huogu Formula in treating phlegm-stasis blocking collaterals syndrome in steroid-induced osteonecrosis of the femoral head (SONFH) by integrating clinical transcriptomics, lipid metabolomics, and biomolecular network mining techniques. The invigorating the spleen and resolving phlegm group (Codonopsis Radix, Atractylodis Macrocephalae Rhizoma, Poria, and Citri Reticulatae Pericarpium) targeted effective target genes such as ALDH7A1 and ACOX1 through 115 chemical components, regulated the AMPK and sphingolipid signaling pathways, significantly reduced the levels of diacylglycerol (DAG) and cholesterol esters, and effectively corrected lipid metabolism disorders. The activating blood circulation and dredging collaterals group (Angelicae Sinensis Radix, Chuanxiong Rhizoma, Paeoniae Radix Rubra, Achyranthis Bidentatae Radix, and Cinnamomi Ramulus) exerts effects through 197 chemical components by intervening in the chemokine and NOD-like receptor pathways, inhibiting the formation of neutrophil extracellular traps (NETs), downregulating inflammatory factor expression, and promoting vascular function repair. The tonifying the kidney and strengthening bones group (Rehmanniae Radix Praeparata and Cervi Cornu) regulated the PI3K/Akt and Ras/MAPK signaling pathways, inhibited osteoclast differentiation genes CTSK and MMP9, and facilitated the reconstruction of bone homeostasis. Transcriptomic and lipid metabolomic analyses were performed on clinical blood samples collected before and after treatment with Jianpi Huogu Formula. Multi-omics integration revealed that the three efficacy groups synergistically constructed a multidimensional network involving "lipid metabolism-immunity-bone homeostasis". A total of 594 differentially expressed effector genes (367 downregulated and 227 upregulated) were identified via clinical transcriptomics, and 179 differential lipid metabolites (mainly triglycerides and DAG) associated with the formula's efficacy were identified via lipid metabolomics. Together with 2 657 disease/syndrome-related genes, a "disease-syndrome-symptom-formula" association network was constructed. Topological eigenvalue analysis and functional mining indicated that the core network targets of Jianpi Huogu Formula in intervening SONFH mainly regulated signaling pathways such as osteoclast differentiation, apoptosis, Ras, MAPK, PI3K/Akt, and p53, thereby inhibiting osteoclast activity, promoting osteoblast proliferation, balancing bone resorption and formation, and improving the bone marrow microenvironment. This study systematically reveals the scientific basis of Jianpi Huogu Formula in intervening in the pathological process of SONFH through the synergistic action of "lipid metabolism reprogramming-transcriptional regulatory network" from three key dimensions, i. e., lipid metabolism, inflammation, and bone remodeling. It fully demonstrates the holistic advantages of traditional Chinese medicine (TCM) formulas in multi-component, multi-target, and multi-pathway coordinated regulation, and provides an innovative perspective and data support for elucidating the mechanism of TCM in the prevention and treatment of SONFH.

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 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.

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.

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.

Influenza viral pneumonia (IVP) is an infectious disease caused by influenza virus invading the lungs. With acute onset and rapid progression, it can lead to respiratory failure and even death in severe cases. Moreover, IVP is prone to cause large-scale incidence along with influenza epidemics, posing a serious threat to public health. The pathogenesis of IVP is complex, involving multiple pathological links such as viral replication, cytokine storm, immune imbalance, and oxidative stress. Studies have confirmed that the occurrence and development of IVP are closely related to signaling pathways including Toll-like receptors (TLRs)/nuclear factor-κB (NF-κB), Janus kinase (JAK)/signal transducer and activator of transcription (STAT), NOD-like receptor pyrin domain-containing protein 3 (NLRP3), mitogen-activated protein kinase (MAPK), phosphatidylinositol 3-kinase(PI3K)/protein kinase B (AKT), and nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase 1 (HO-1). TCM characterized by multi-pathway, multi-component, and multi-target actions shows unique potential in regulating the above pathways and ameliorating pathological damage in the lungs. Accordingly, this article clarifies the IVP pathogenesis mediated by these pathways and systematically reviews the recent research on TCM active ingredients, single herbs, and compound preparations in preventing and treating IVP via regulating these pathways, with focuses on the roles of TCM in inhibiting viral replication, regulating inflammation, restoring immune homeostasis, and alleviating oxidative stress damage. It aims to comprehensively reveal the potential mechanisms of TCM in treating IVP, providing new insights for clinical diagnosis and treatment of IVP.

TCM volatile oil exhibits excellent biological activities and wide application scenarios. However, its volatility and susceptibility to oxidation and deterioration severely restrict the quality of its preparations and clinical efficacy. Therefore, stabilizing TCM volatile oil is extremely necessary. Current research mainly focuses on the development of stabilization techniques and carrier materials, aiming to enhance its stability and endow it with sustained-release and controlled-release properties. In recent years, researchers have increasingly paid attention to the intrinsic mechanisms and potential interactions during the stabilization process, which is of great significance for the development of new stabilization technologies for volatile oil and the research on new dosage forms containing volatile oil. This article systematically reviews the mechanisms of action during the stabilization of TCM volatile oil and summarizes the key methods for characterizing interactions, with the aim of providing references for the development of new stabilization technologies for TCM volatile oil and the theoretical research on its stabilization.

Mitochondrial dysfunction is a key pathological mechanism in metabolic-associated fatty liver disease (MAFLD), and regulating mitochondrial function to restore normal activity is an important therapeutic strategy. The theory of "harmful hyperactivity and responding inhibition" is a classical TCM principle that explains cyclical patterns in nature, interprets physiological and pathological phenomena in the human body, and guides TCM clinical diagnosis and treatment. The mechanism of mitochondrial dysfunction closely aligns with the process described in this theory as when the responding element fails to control, the hyperactive element becomes harmful. Based on the relationship between mitochondrial function and the TCM pathogenesis of MAFLD, and applying the "harmful hyperactivity and responding inhibition" framework, this study proposes that MAFLD originates from spleen deficiency impairing nutrient transport and failing to disperse essence, kidney Yang deficiency leading to impaired Qi transformation, and insufficient mutual regulation of the two, resulting in pathological products such as excessive accumulation of phlegm-turbidity, dampness, and blood stasis. These pathological changes manifest as mitochondrial dysfunction, including mtDNA damage, hepatocyte injury, lipid peroxidation, and mitochondrial accumulation. Clinically, MAFLD treatment follows the principle of regulating the responding element and restraining the hyperactive element: reinforcing the spleen earth to restore regulation, strengthening kidney water to nourish the kidneys and harmonize the liver, and addressing phlegm-stasis obstruction and hyperactivity through blood-activating and phlegm-resolving therapies. By modulating mitochondrial structure and function, this approach improves hepatic lipid metabolism, reduces oxidative stress, and alleviates inflammatory responses, achieving multi-target therapeutic effects. This study provides new theoretical foundations and practical strategies for integrated TCM-western medicine treatment of MAFLD.

Chronic obstructive pulmonary disease (COPD) characterized by persistent airflow limitation is a heterogeneous lung condition prone to exacerbations and progression. Recent studies have revealed that gut microbiota dysbiosis can trigger metabolic disorders, immune abnormalities, and inflammatory responses, contributing to the pathogenesis and progression of COPD through the gut-lung axis. The spleen-tonifying and lung-replenishing therapy originates from the five elements theory of TCM, specifically the principle of tonifying the source in the case of deficiency. Enhancing the function of the spleen and stomach (earth element) indirectly nourishes the lungs (metal element), embodying the holistic concept of organ interdependence. The gut, as an extension of the transformation function of the spleen and stomach, reflects the modern interpretation of the spleen's role in transformation and transportation. When the spleen is deficient, gut microbiota dysbiosis leads to the accumulation of turbid toxins, which rise to affect the lungs, resulting in impaired flow of healthy Qi and the interplay of phlegm and stasis. The concept of tonifying the spleen not only involves tonifying the middle Qi but also encompasses multidimensional interventions such as improving gastrointestinal functions and regulating the gut microbiota to restore lung-spleen coordination. The spleen-tonifying and lung-replenishing therapy can influence the gut microbiota in COPD patients, effectively interrupting the pathological vicious cycle of spleen deficiency-microbiota dysbiosis-lung damage, slowing down disease progression, and reducing acute exacerbations. This method highlights the scientific value of the holistic thinking in TCM and provides a key pathway for integrating TCM and Western medicine in the prevention and treatment of COPD, underscoring the core clinical significance of tonifying the spleen and replenishing the lung.

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.

This study investigated the secondary metabolites of the endophytic fungus Penicillium sp. RM isolated from the toxic plant Rhododendron molle. Ten compounds were isolated and purified from the ethyl acetate extract of the fungal fermentation broth by silica gel column chromatography, gel column chromatography, and semi-preparative HPLC. The structures of these compounds were elucidated by comprehensive spectroscopic analyses, including NMR, MS, UV, IR, and CD. The identified compounds were as follows: penicidienone A (1), diversonol (2), alternariol (3), alternariol 9-methyl ether (4), methylated dechloromonilicin (5), norliquexantona (6), alternethanoxin A (7), α-diversonolic ester (8), (±) -asperlone A (9), and verruculotoxin (10). Compound 1 was identified as a new cyclopentenone derivative. Compound 9 showed inhibitory activity against the proliferation of HepG2 (IC₅₀=9.7 μmol·L^-1), HGC27 (IC₅₀ = 9.3 μmol·L^-1), and U251 (IC₅₀ = 8.4 μmol·L^-1) cell lines.

This study aims to investigate the antifungal secondary metabolites produced by the endophytic fungus Aspergillus sp. FH-1 isolated from Valeriana officinalis via a high-salt rice medium. Compounds were isolated and purified by silica gel and ODS column chromatography, semi-preparative HPLC, and other techniques. The chemical structures were elucidated through analysis of physicochemical properties and spectroscopic data of nuclear magnetic resonance (NMR), infrared (IR), and mass spectrometry (MS). The inhibitory effects of all the compounds against Colletotrichum gloeosporioides were evaluated via the mycelial growth rate method. Eight compounds were obtained from the ethyl acetate extract of Aspergillus sp. FH-1. They were identified as asperchoried A (1), butyrolactone Ⅱ (2), isobutyrolactone Ⅱ (3), aspernolide A (4), butyrolactone Ⅴ (5), clavatone (6), 1-hydroxy-6,8-dimethoxy-3-methylanthracene-9,10-dione (7), and (S) -7-methoxy-2, 5-dimethyl-2, 3-dihydrobenzofuran-6-ol (8). Among them, compound 1 is a novel chlorinated butanolide derivative and was designated asperchoried A. Compounds 1, 2, 6, and 7 possessed inhibitory effects on C. gloeosporioides, with EC₅₀ values of 15.08, 77.19, 89.94, and 62.02 μg·mL^-1, respectively.

Poria cocos is a commonly used bulk traditional Chinese medicinal material, with multiple parts used medicinally, such as Poriae Cutis, Rubra Poria, White Poria, and Poria cum Radix Pini. Polysaccharides are their main active components. Clarifying the composition and content differences of polysaccharides in different medicinal parts of P. cocos can provide a reference for processing and clinical application of P. cocos medicinal materials and decoction pieces. In this paper, high-performance gel permeation chromatography (HPGPC) and nuclear magnetic resonance (NMR) spectroscopy were employed to qualitatively compare the relative molecular mass distribution and chemical composition of alkali-soluble polysaccharides from different parts of crude P. cocos. The results showed that the composition of alkali-soluble polysaccharides in Rubra Poria and Poriae Cutis was consistent with that in White Poria, all of which were β-(1→3) -D-glucans. The relative molecular mass distribution was also relatively close, ranging approximately from 9.13×10⁴ to 1.04×10⁵. On the basis of the content determination method of β-(1→3) -D-glucan in P. cocos according to the 2025 edition of Chinese Pharmacopoeia, the preparation method of the test sample solution was optimized. Firstly, a desalination step was added, which can enable the test sample chromatogram to present only the chromatographic peak of glucose, thereby reducing the difficulty of separation. Secondly, this experiment discovered for the first time that under acidic conditions, the hydrolysis of β-(1→3) -D-glucan to glucose was accompanied by the generation of a small amount of 5-hydroxymethylfurfural (5-HMF) as a by-product. A more intense acid hydrolysis reaction condition indicated a greater conversion yield of 5-HMF. Therefore, according to the results of single-factor experiments and response surface methodology experiments, the acid hydrolysis time was shortened from 6 h to 1.5 h, which improved the analysis efficiency. After verification, the optimized method was simple and stable, and had good repeatability and accuracy. Then, the optimized quantitative method was used to evaluate the content of β-(1→3) -D-glucan in different parts of crude P. cocos and decoction pieces. The results demonstrated that the content of β-(1→3) -D-glucan in different medicinal parts of P. cocos was in the order of White Poria, Poria cum Radix Pini, Rubra Poria, and Poriae Cutis. Among the commercially available Poria decoction pieces, the content of β-(1→3) -D-glucan in the white and hard decoction pieces was higher than that in the dark and loose-textured decoction pieces.

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 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.

The therapeutic effects of Quzhou Aurantii Fructus aqueous extract (QAFA) and Aurantii Fructus aqueous extract (AFA) on functional dyspepsia (FD) in rats were evaluated and compared, and their underlying mechanisms were investigated. The FD rat models were constructed by a combined method of "pinching tail + irregular diet + drinking dilute acidic water". The rats were orally administered QAFA at a dose of 2.05 g·kg^-1(equivalent to 4 g·kg^-1 QAF) and AFA at a dose of 1.93 g·kg^-1(equivalent to 4 g·kg^-1 AF), respectively. After 14-day administration, the gastric emptying and intestinal propulsion were assessed. The pathological changes in the gastric antrum and duodenum were analyzed via histology, and the levels of serum gastrin (GAS) and vasoactive intestinal polypeptide (VIP) were measured by enzyme-linked immunosorbent assay. The underlying mechanisms were investigated by using 16S rRNA sequencing and serum metabolomics. The results showed that QAFA and AFA could increase gastrointestinal motility. Hematoxylin-eosin (HE) staining and immunofluorescence analysis showed that both QAFA and AFA repaired the damaged intestinal barrier, and ELISA analysis showed that they regulated GAS and VIP levels in serum. These results suggest that both can improve symptoms in FD rats. The intestinal microbiota analysis revealed that QAFA and AFA reduced the abundance of the harmful microbe Ruminococcus. AFA increased the abundance of the beneficial bacteria Prevotella and Lactobacillus, and QAFA increased the abundance of Clostridia UCG-014. Metabolomic analysis showed that both QAFA and AFA restored serum metabolites primarily through regulating phospholipid metabolism. Correlation analysis between metabolites and gut microbiota showed that both QAFA and AFA decreased the abundance of Ruminococcus, and AFA increased the abundance of Prevotella, thereby restoring serum metabolite levels. These findings suggest that QAFA and AFA may improve FD symptoms in rats by modulating specific gut microbiota to restore gastrointestinal hormones and phospholipid metabolism. However, there are some differences in the specific microbiotas they regulate.

This study aimed to elucidate the role of the G protein-coupled receptor 35 (GPR35)/AMP-activated protein kinase (AMPK)/mechanistic target of rapamycin (mTOR) signaling axis in mediating mitochondrial energy metabolism in radiation-induced myocardial fibrosis (RIMF) and to evaluate the interventional effect of astragaloside Ⅳ (AS-Ⅳ). Forty Wistar rats were randomly divided into four groups (n=10 per group). Except for the blank group, all rats were anesthetized and subjected to a single 40-Gy local thoracic X-ray irradiation to establish the RIMF model. After irradiation, AS-Ⅳ was administered intragastrically once daily at doses of 80 and 160 mg·kg^-1 for 30 consecutive days. After 30 days of continuous administration, the serum levels of N-terminal pro-B-type natriuretic peptide (NT-proBNP), cardiac troponinI (cTnI), and creatine kinase-MB (CK-MB) were quantified by ELISA. Hematoxylin-eosin (HE) and Masson staining were used to observe myocardial histopathological changes, and TUNEL staining was performed to detect cardiomyocyte apoptosis. The levels of adenosine triphosphate (ATP), adenosine diphosphate (ADP), and adenosine monophosphate (AMP), as well as the activities of mitochondrial respiratory chain complexes Ⅰ-Ⅳ, were measured colorimetrically. The fluorescence intensity of Ca²⁺ in myocardial tissue was detected using a fluorescent probe, and ultrastructural changes were observed by transmission electron microscopy. The fluorescence intensity and protein expression levels of GPR35, phosphorylated AMPK (p-AMPK), phosphorylated mTOR (p-mTOR), Caspase-3, Caspase-9, B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax), type Ⅰ collagen (COL-Ⅰ), and α-smooth muscle actin (α-SMA) in myocardial tissue were analyzed by immunofluorescence and Western blot. Results showed that compared with the blank group, serum NT-proBNP, cTnI, and CK-MB levels in the model group were significantly increased. Myocardial tissue exhibited focal myocyte necrosis and dissolution, connective tissue hyperplasia, and an increased area of collagen fiber positivity. The cardiomyocyte apoptosis rate was elevated. Myocardial ATP levels were significantly decreased, while ADP and AMP levels were significantly increased. The activities of mitochondrial respiratory chain complexes Ⅰ-Ⅳ were markedly reduced. The Ca²⁺ fluorescence intensity in myocardial cells was significantly enhanced. Severe mitochondrial damage was observed, showing flocculent changes, increased numbers of damaged mitochondria, swelling, rupture, disordered arrangement, disrupted cristae, and partial vacuolation. The fluorescence intensity and protein expression levels of GPR35, p-AMPK, Bax, Caspase-3, Caspase-9, COL-Ⅰ, and α-SMA in myocardial tissue were significantly increased, whereas those of p-mTOR and Bcl-2 were decreased. Compared with the model group, AS-Ⅳ significantly reduced serum NT-proBNP, cTnI, and CK-MB levels, markedly decreased the cardiomyocyte apoptosis rate, significantly increased ATP levels while reducing ADP and AMP content, enhanced the activities of mitochondrial respiratory chain complexes Ⅰ-Ⅳ, diminished Ca²⁺ fluorescence intensity, and alleviated mitochondrial damage, with mitochondria arranged more regularly. The fluorescence intensity and protein expression levels of GPR35, p-AMPK, Bax, Caspase-3, Caspase-9, COL-Ⅰ, and α-SMA were significantly decreased, while those of p-mTOR and Bcl-2 were increased. In conclusion, AS-Ⅳ exerts an inhibitory effect on RIMF, and its mechanism may be associated with the regulation of the GPR35/AMPK/mTOR signaling axis to improve mitochondrial energy metabolism.

This study aimed to elucidate the mechanisms of diosgenin, a natural compound derived from traditional Chinese medicine, on the triple-negative breast cancer cell line MDA-MB-231, with a focus on its induction of cell cycle arrest and inhibition of cell migration. Multiple experimental approaches were employed, including RNA sequencing analysis, flow cytometry for cell cycle detection, wound healing assays to evaluate migration capacity, F-actin staining to observe cytoskeletal changes, transmission electron microscopy (TEM) to examine cellular morphology and filopodia alterations, as well as Western blot and immunofluorescence staining to detect the expression of related proteins. RNA sequencing revealed significant gene expression differences between the diosgenin-treated group and the control group, with KEGG and GO analyses showing enrichment of differentially expressed genes mainly in pathways related to cell cycle and migration. Flow cytometry demonstrated that diosgenin induced G₀/G₁ phase arrest in MDA-MB-231 cells in a dose-dependent manner. Further Western blot and immunofluorescence assays indicated that diosgenin regulated the cell cycle via the CDC25A-CDK2/4/6 axis. Wound healing assays showed that diosgenin significantly inhibited cell migration. F-actin staining indicated that diosgenin suppressed F-actin formation and disrupted its structure. TEM observations revealed a reduction in filopodia formation on the cell surface after diosgenin treatment. Moreover, Western blot and immunofluorescence results demonstrated that diosgenin inhibited migration by suppressing epithelial-mesenchymal transition (EMT) and the RhoA-ROCK1 signaling pathway. In conclusion, diosgenin induces G₀/G₁ phase arrest in MDA-MB-231 cells by regulating cell cycle-related proteins and significantly inhibits cell migration by suppressing F-actin formation, reducing filopodia on the cell surface, and inhibiting EMT and the RhoA-ROCK1 pathway.

Based on the holistic regulation and multi-target characteristics of TCM, this study investigated the mechanism of Panlongqi Tablets (PLQT) in ameliorating rheumatoid arthritis (RA) by regulating key proteins, including glycogen synthase kinase-3β (GSK-3β), phosphatidylinositol 3-kinase catalytic subunit alpha (PIK3CA), and protein kinase B3 (AKT3), within the PI3K-AKT signaling pathway. An adjuvant-induced arthritis (AIA) rat model was established by intradermal injection of type Ⅱ collagen-Freund's complete adjuvant into the right hind paw, and rats received different doses of PLQT for intervention. Pharmacodynamic effects were evaluated by monitoring changes in body weight, measuring paw swelling, observing ankle synovial tissue pathology via hematoxylin-eosin (HE) staining, and detecting serum levels of inflammatory cytokines, including tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β), using ELISA. Targets identified through prior network pharmacology screening were integrated with therapy-related biomarkers from untargeted metabolomics for key pathway enrichment and identification of critical targets. Western blot and RT-qPCR were subsequently used to validate the expression of these key targets. Animal experimental results showed that, compared with the model group, all PLQT dose groups significantly improved weight loss, reduced paw swelling, inhibited synovial tissue fibrosis hyperplasia and inflammatory cell infiltration, and decreased serum levels of pro-inflammatory cytokines such as TNF-α, IL-6, and IL-1β in RA rats, exhibiting a partial dose-dependent manner. Metabolomics analysis revealed that PLQT exerted regulatory effects on 179 of 276 RA-related biomarkers. Enrichment analysis revealed that GSK-3β, PIK3CA, and AKT3 proteins in the PI3K-AKT signaling pathway were the key targets for PLQT intervention in RA. Validation by Western blot and RT-qPCR demonstrated that PLQT significantly modulated the expression of these three proteins. In conclusion, the therapeutic effects of PLQT on RA are likely associated with regulation of GSK-3β, PIK3CA, and AKT3 within the PI3K-AKT signaling pathway.

Non-alcoholic fatty liver disease (NAFLD) is the most prevalent chronic liver disease worldwide, characterized primarily by hepatic inflammation and dysregulated lipid metabolism. As a classic representative formula of the "bitter-purging method", Modified Wushi Xiexin Decoction has demonstrated definite clinical efficacy; however, its anti-inflammatory effects and underlying mechanisms in the treatment of NAFLD remain unclear. This study combines network pharmacology with animal experimental validation to evaluate the therapeutic effect of Modified Wushi Xiexin Decoction on NAFLD and reveal its potential mechanisms of action. Drug targets were collected from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP). NAFLD-related disease targets were retrieved from the Therapeutic Target Database (TTD), GeneCards, Online Mendelian Inheritance in Man (OMIM), Pharmacogenomics Knowledge Base (PharmGKB), and Comparative Toxicogenomics Database (CTD). R software was used to screen the intersection targets of the two sets. Core target analysis and network visualization were performed by using the Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) database and Cytoscape 3.8.2 software. R software was applied for Gene Ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Core targets were analyzed based on the Gene Expression Omnibus (GEO) database, and receiver operating characteristic (ROC) curves were plotted to evaluate the diagnostic efficacy of the core targets. For animal experimental validation, a NAFLD mouse model was established by feeding a 60% high-fat diet combined with intraperitoneal injection of streptozotocin. Pathological improvements were assessed by hematoxylin-eosin (HE) staining and oil red O staining. Enzyme-linked immunosorbent assay (ELISA) was used to detect the level of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β) in liver tissue. Immunohistochemistry (IHC) and Western blot (WB) were performed to measure the expression of Toll-like receptor 4 (TLR4), nuclear factor-κB (NF-κB), and NOD-like receptor protein 3 (NLRP3) proteins in mouse liver tissue. Network pharmacology identified 97 active components, 108 intersection targets, and 10 core targets of Modified Wushi Xiexin Decoction. GO enrichment analysis indicated that anti-NAFLD effects were exerted by regulating biological processes such as oxidative stress and nutritional levels. KEGG enrichment analysis mainly involved signaling pathways, including TLR, NF-κB, and TNF. The GEO database indicates that there is a significant difference in TLR4 expression between healthy individuals and NAFLD patients (P<0.05), with the area under the ROC curve (AUC) being greater than 0.810, suggesting a high diagnostic efficacy. Animal experiments showed that Modified Wushi Xiexin Decoction could alleviate pathological damage in the liver tissue of NAFLD mice, reduce blood lipids, improve liver function, and downregulate the expression of inflammatory factors in liver tissue. IHC and WB confirmed that Modified Wushi Xiexin Decoction could downregulate the TLR4 expression, inhibit NF-κB activity, and suppress NLRP3 inflammasome activation. In conclusion, Modified Wushi Xiexin Decoction may alleviate inflammatory responses and improve liver function and blood lipids in NAFLD by regulating the TLR4/NF-κB signaling pathway and inhibiting NLRP3 inflammasome activation, thereby exerting therapeutic effects on NAFLD.

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 aims to investigate the effects of Naotaifang (NTF) on neuronal ferroptosis and pyroptosis in cerebral ischemia-reperfusion injury (CIRI), with a focus on nuclear factor erythroid 2-related factor 2 (Nrf2). A rat model of CIRI was established by middle cerebral artery occlusion/reperfusion (MCAO/R). Sprague-Dawley rats were randomly assigned to a sham group, a model group, low/medium/high-dose (4.5, 9, 18 g·kg^-1, respectively) NTF groups, and a butylphthalide (60 mg·kg^-1) group (n=15 per group). After modeling, the treatment groups received corresponding agents by gavage once daily for 7 days, and the sham and model groups received an equal volume of normal saline. After treatment, the neurological function was assessed based on the Zea-Longa score, the infarct volume by TTC staining, and cellular morphology by HE staining. The levels of Fe²⁺, interleukin-1β (IL-1β), and interleukin-18 (IL-18) were measured by ELISA, and those of malondialdehyde (MDA) and glutathione (GSH) by biochemical assays. Western blot was employed to examine the expression levels of Nrf2, glutathione peroxidase 4 (GPX4), NOD-like receptor family pyrin domain-containing 3 (NLRP3), and the N-terminal fragment of gasdermin D (GSDMD-N). In the cell experiment, oxygen-glucose deprivation/reperfusion (OGD/R) was used to model CIRI in SH-SY5Y cells. siRNA-mediated Nrf2 knockdown (si-Nrf2) was employed to probe into the role of Nrf2 in OGD/R-induced ferroptosis and pyroptosis and the intervention effect of NTF. Membrane damage was evaluated by Hoechst/propidium iodide staining and LDH release, and GPX4 and GSDMD immunofluorescence was employed to indicate ferroptosis and pyroptosis, respectively. Fe²⁺, MDA, and GSH were quantified biochemically, and the levels of IL-1β and IL-18 by ELISA. The expression of Nrf2, GPX4, NLRP3, and GSDMD-N was quantified by Western blot. The cellular ultrastructure was observed by transmission electron microscopy. The results of the animal experiment showed that NTF promoted the expression of Nrf2 to alleviate neuronal ferroptosis and pyroptosis, thus producing significant neuroprotection against CIRI in a dose-dependent manner. The results of the cell experiment showed that the neuroprotective effects of NTF after CIRI were mediated by Nrf2-dependent suppression of neuronal ferroptosis and pyroptosis. Taken together, NTF attenuates CIRI by suppressing neuronal ferroptosis and pyroptosis through Nrf2-mediated regulation.

Gardeniae Fructus (GF) is one of the most commonly used medicinal and edible resources in China, renowned for its anti-inflammatory and antipyretic effects. However, studies of the chemical characteristics, metabolic transformation and pharmacokinetic behavior of GF in vivo remain incomplete. This study utilized mass defect filtering (MDF) combined with UHPLC-ESI-Q-TOF-MS/MS to analyze the in vitro chemical constituents of GF aqueous extracts and its metabolites in vivo in rats, comprehensively characterizing 114 chemical constituents and 181 endogenous substances, including 18 prototype constituents and 163 metabolites (21 phase Ⅰ and 142 phase Ⅱ metabolites). Additionally, a quantitative method using UHPLC-QQQ-MS/MS was developed to analyze the in vivo dynamics of seven bioactive compounds, and the pharmacokinetic parameters of the main active components in the GF aqueous extracts, namely geniposidic acid, deacetyl asperulosidic acid methyl ester, gardenoside, genipin, genipin 1-β-D-gentiobioside, geniposide, and crocetin, were obtained. This study conducted a comprehensive and systematic investigation into the potential material basis and the in vivo biotransformation profile of GF aqueous extracts based on UHPLC-ESI-Q-TOF-MS/MS coupled with MDF technology and UHPLC-QQQ-MS/MS, providing crucial evidence for further elucidating its pharmacological mechanisms.

This study aims to systematically retrieve and sort out the clinical research evidence of oral Chinese patent medicines in the treatment of vascular dementia (VaD) and understand the distribution and quality of the existing evidence in this field. PubMed, Web of Science, Cochrane Library, EMbase, CNKI, VIP, Wanfang, and SinoMed databases were retrieved from their establishment to November 5, 2024. Evidence diagrams were used to comprehensively present the data distribution characteristics. A total of 211 pieces of literature were included, among which 176 were RCTs, 11 were non-RCTs, 3 were retrospective studies, 1 was cohort study, 17 were systematic reviews/Meta-analyses, and 3 were expert recommendations or consensuses. It involves 45 kinds of Chinese patent medicines. Among them, Ginkgo Leave Tablets/Capsules are the most widely used. Most of the research periods are 8-12 weeks, and the sample size is 51-100 cases. The main outcome indicators include clinical efficacy, cognitive function evaluation, quality of life evaluation, safety indicators, neurological function indicators, hemorheological indicators, etc. The literature quality of RCTs, non-RCTs, and systematic reviews/Meta-analyses are generally not high, and the quality level of evidence is relatively low. Most studies have shown that oral Chinese patent medicines have potential efficacy in the treatment of VaD, but there is a lack of high-quality clinical evidence. Chinese patent medicines have advantages in the treatment of VaD, but there is still a lack of high-quality studies. This suggests that in future clinical research, large-sample and multi-center RCTs need to be carried out. Meanwhile, clinical research design and reporting should be standardized in accordance with relevant guidelines or tools to explore the characteristic advantages of oral Chinese patent medicines in the treatment of VaD, select reasonable outcome indicators, and strive to improve the quality of clinical research in TCM. This will help standardize the research plan, produce advanced evidence-based medical evidence with rigor and scientificity, and provide strong support for the prevention and treatment of VaD with TCM.

Published in 1856, Catalogus Medicamentorum Sinensium stands as the earliest known modern pharmacological work in existence. Compiled by the Russian scholar Alexander Tatarinov during the 19th century, this groundbreaking study systematically documented and identified the botanical origins of 495 Chinese medicinal materials procured from Beijing, supplemented with plant specimens collected in China. It represents the first application of modern scientific nomenclature, combining Latin binomial names with specified medicinal parts, for the systematic classification of Chinese medicinal materials. This research conducted a comprehensive analysis of the cataloged substances through meticulous examination of relevant literature, archival records, and reference materials. By using methods such as materia medica verification, data analysis, etc., the information of the listed drugs was verified, classified, organized, and examined, and compared with the origins recorded in modern standards or specialized works. The compendium documented 495 medicinal entries (including 33 duplicate medicinal materials), comprising 396 botanical, 35 mineral, 40 zoological, and 24 other substances. This book identified a total of 190 Chinese medicinal materials whose origin was consistent with that recorded in the 2020 edition of the Pharmacopoeia of the People's Republic of China or Chinese Materia Medica. There were 76 cases with completely different origins. Another 189 cases had their origins not fully identified, but only partial information was marked. Lastly, there were 40 cases that were either untested or could not be matched with current Chinese medicinal materials. The research results indicate that this book is the first in modern times to use Linnaean binomial nomenclature to name the origin of Chinese medicinal materials. It laid the foundation for subsequent research on the origin of Chinese medicinal materials and established the methods and annotation formats for such research. It has been subsequently adopted and referenced by subsequent TCM researchers both in China and abroad. The publication of this book can be regarded as a historical turning point in the evolution of the definition of the origin of Chinese medicinal materials from traditional to modern times. It has advanced the description of the name and reality of Chinese medicinal materials in the form of text and pictures to the definition method using modern natural classification systems and labeled with biological Latin names, which is a pioneering work. This research can provide a reference for sorting out the development process of modern research on the origin of Chinese medicinal materials.

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.