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.