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    18 December 2025, Volume 29 Issue 35 Previous Issue    Next Issue
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    Asperosaponin VI promotes osteogenic differentiation of MC3T3-E1 cells under hypoxia environment #br#
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    Li Yunzhe, Niu Zefan, Wang Zirou, Ai Chongyi, Chen Gang, Wang Xinxing
    2025, 29 (35):  7481-7489.  doi: 10.12307/2025.991
    Abstract ( 117 )   PDF (3198KB) ( 77 )   Save
    BACKGROUND: Asperosaponin VI has good osteogenic effects, but its ability to promote cellular osteogenesis under hypoxia environment is not yet clear.
    OBJECTIVE: To investigate the effect and potential mechanism of Asperosaponin VI on osteogenic differentiation of MC3T3-E1 cells under hypoxia environment.
    METHODS: MC3T3-E1 cells were divided into three groups. Cells in the control group were cultured in a complete medium under normoxic conditions (volume fraction of 21% O2); cells in the hypoxia group were cultured in the complete medium under hypoxia conditions (volume fraction of 0.5% O2); and cells in the Asperosaponin VI group were cultured in the complete medium containing Asperosaponin VI under hypoxia conditions (volume fraction of 0.5% oxygen). After 24 hours of culture, cell counting kit-8 method and EdU staining were used to detect cell proliferation activity, TUNEL staining and western blot assay were performed to detect cell apoptosis, and flow cytometry was used to detect intracellular reactive oxygen species levels and cell cycle distribution. Each group of cells was cultured in an osteogenic induction medium containing 1×10-5, 1×10-6, and 1×10-7 mol/L Asperosaponin VI under hypoxia conditions for 7 days. The optimal concentration of Asperosaponin VI for intervention was identified using alkaline phosphatase staining under optical microscopy. Western blot was used to detect the expression of bone morphogenetic protein 2, osteopontin, and PI3K/AKT signaling axis-related proteins.
    RESULTS AND CONCLUSION: (1) Compared with the control group, the proliferation ability of MC3T3-E1 cells decreased under hypoxia conditions. 1×10-6 mol/L 
    Asperosaponin VI could significantly improve the cell proliferation ability under hypoxia conditions and reduce cell apoptosis. (2) Compared with the hypoxia group, the Asperosaponin VI group showed a decrease in intracellular reactive oxygen species and a significant increase in the proportion of cells in the S phase. (3) Compared with the hypoxia group, the cell morphology in the 1×10-6 mol/L Asperosaponin VI group was elongated, with more protrusions and darker alkaline phosphatase staining. (4) Compared with the hypoxia group, the expression of bone morphogenetic protein 2 and osteopontin increased in the Asperosaponin VI group, while the expression of p-PI3K and p-AKT proteins decreased. These findings indicate that under hypoxia conditions, Asperosaponin VI can promote osteogenic differentiation of MC3T3-E1 cells, possibly by regulating the PI3K/AKT pathway.

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    Differential effects of kartogenin on chondrogenic and osteogenic differentiation of rat and rabbit bone marrow mesenchymal stem cells
    Liu Chengyuan, Guo Qianping
    2025, 29 (35):  7490-7498.  doi: 10.12307/2025.568
    Abstract ( 131 )   PDF (2677KB) ( 62 )   Save
    BACKGROUND: Bone marrow mesenchymal stem cells possess multipotent differentiation potential and are an important source of cells for cartilage and bone tissue regeneration research. Kartogenin is a small-molecule drug that has been demonstrated to promote stem cell differentiation towards chondrogenesis. However, the ability to promote osteogenic differentiation is still controversial, and the specific effects of kartogenin on the chondrogenic and osteogenic differentiation of stem cells of different species have not been fully elucidated.
    OBJECTIVE: To investigate the effects of kartogenin on chondrogenic and osteogenic differentiation in rabbit-derived and rat-derived bone marrow mesenchymal stem cells.
    METHODS: Rabbit-derived and rat-derived bone marrow mesenchymal stem cells were obtained through whole bone marrow separation and adherence methods, and were treated with varying concentrations of kartogenin. Cell proliferation was detected using the CCK-8 assay. Chondrogenic and osteogenic differentiation was assessed via toluidine blue and alizarin red staining, respectively. After screening for the optimal concentration of kartogenin, alkaline phosphatase staining, qRT-PCR, and western blot assay were employed to analyze the expression of osteogenic and chondrogenic-related genes and proteins.
    RESULTS AND CONCLUSION: (1) Within the concentration range of 0 to 10 000 nmol/L, kartogenin did not significantly affect the proliferation of rabbit-derived or mouse-derived bone marrow mesenchymal stem cells. (2) The differentiation effect of kartogenin on rabbit-derived and rat-derived bone marrow mesenchymal stem cells showed significant differences. In rabbit-derived bone marrow mesenchymal stem cells, kartogenin predominantly promoted chondrogenic differentiation while inhibiting osteogenic differentiation. This was evident from positive toluidine blue staining, whereas alkaline phosphatase and alizarin red staining exhibited no significant changes. qRT-PCR analysis revealed upregulation of chondrogenic-related genes (Col2a1 and Sox9) and downregulation of genes associated with osteogenesis (Alpl, Col1a1, Runx2, and Bglap). Similar results were found in the western blot assay. (3) In contrast, kartogenin in mouse-derived bone marrow mesenchymal stem cells promoted both chondrogenic and osteogenic differentiation. While toluidine blue staining remained largely unchanged, alkaline phosphatase and alizarin red staining revealed increased positivity, indicative of enhanced osteogenesis. qRT-PCR analysis showed upregulation of not only chondrogenic-related genes (Col2a1 and Sox9) but also genes linked to osteogenesis (Alpl, Col1a1, Runx2, and Bglap). The western blot assay results showed similar results. These findings suggest that kartogenin exerts differential regulatory effects on the differentiation of rabbit and rat bone marrow mesenchymal stem cells, potentially stemming from variations in gene expression profiles and underlying signaling pathways. 
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    Comparison of six machine learning models suitable for use in medicine: support for osteoporosis screening and initial diagnosis
    Yang Lei, Liu Sanmao, Sun Huanwei, Che Chao, Tang Lin
    2025, 29 (35):  7499-7510.  doi: 10.12307/2025.947
    Abstract ( 137 )   PDF (2987KB) ( 270 )   Save
    BACKGROUND: With the increasing degree of population aging in China, the incidence of osteoporosis is rising annually. This growing demand for screening and diagnosis poses significant challenges to the healthcare system, increasing the time costs, financial burdens, and radiation exposure risks for patients.
    OBJECTIVE: To develop a novel interpretable prediction method based on traditional CT examination data and demographic data, aiming to reduce the number of patient examinations and enable multiple screenings from one examination.
    METHODS: A two-stage interpretable framework for osteoporosis prediction was designed. In the first stage, a human-computer collaborative method was used for annotating CT images, with an innovative vertebra 7-point CT value measurement technique. Patient’s sex and age were used as key demographic features to enrich the model’s input. In the second stage, the LightGBM model was enhanced by SHapley Additive exPlanations for quantitative analysis of feature importance, improving the interpretability of predictions and increasing clinical trust. Systematic experiments validated the effectiveness of the framework and the stability of the optimal feature set through the comparative analysis of different feature combinations with six machine learning models. To further assess the generalization ability of the model, the model was further tested on an external dataset.
    RESULTS AND CONCLUSION: The experiment compared six machine learning models suitable for medical applications, and the results showed that LightGBM model achieved an F1 score of 0.902 2 and an area under the curve of 0.938 7, outperforming the other models. In terms of interpretability, the clinical application credibility and operability of the model was increased by ranking and visualizing the contribution of input features to the results. Additionally, this study realized a prototype system, and testing results indicated that the system is user-friendly, capable of quickly processing data to provide prediction results, with visualized outcomes demonstrating good interpretability. This system effectively assists doctors in clinical decision-making and provides robust support for the screening and preliminary diagnosis of osteoporosis.
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    Improvements in automatic diagnosis methods for knee osteoarthritis based on deep learning
    Fang Ying, Zhang Yanwei, Li Xi, Yan Peidong, Bi Miao
    2025, 29 (35):  7511-7518.  doi: 10.12307/2026.533
    Abstract ( 146 )   PDF (3975KB) ( 430 )   Save
    BACKGROUND: Knee osteoarthritis is a common degenerative disease that significantly impacts patients' quality of life and increases the societal healthcare burden. Early and accurate diagnosis of knee osteoarthritis is crucial for the treatment and prognosis of patients. Traditional diagnostic methods are not only subjective and time-consuming but also do not guarantee consistently high accuracy.
    OBJECTIVE: To develop an automatic diagnostic method for knee osteoarthritis based on deep learning, utilizing deep learning networks to improve diagnostic accuracy and efficiency.
    METHODS: A new network model, YOLOV8-ViT, was proposed by replacing the backbone network of YOLOv8n with the Efficient-ViT network and incorporating attention mechanisms for the automatic identification and classification of X-ray images of knee osteoarthritis. The experimental dataset included 5 078 X-ray images of patients with knee osteoarthritis obtained from the Third Affiliated Hospital of Guangzhou University of Chinese Medicine. Three imaging physicians annotated the sites of knee osteoarthritis and classified them according to the Kellgren-Lawrence grading standard using Labelme software, and the results were combined. The evaluation indicators used in this study included Precision, F1 score, mean average precision (mAP), Recall, val/box_loss, val/cls_loss, and val/dfl_loss.
    RESULTS AND CONCLUSION: The experimental results showed that the YOLOV8-ViT model outperformed the YOLOv5n, YOLOv8n, and YOLOv9n models in terms of precision, mAP50, mAP50-95, F1 score, and Recall, while lowering val/box_loss, val/cls_loss, and val/dfl_loss by 0.496, 0.45, and 0.523; 1.037, 0.305, and 0.728; and 0.267, 0.654, and 0.854, respectively. These experimental data validate that this model has high detection accuracy.

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    Mechanism of Mongolian medicine Echinops sphaerocephalus L. in proliferation and angiogenesis of vascular endothelial cells
    Fang Yuan, Qian Zhiyong, He Yuanhada, Wang Haiyan, Sha Lirong, Li Xiaohe, Liu Jing, He Yachao, Zhang Kai, Temribagen
    2025, 29 (35):  7519-7528.  doi: 10.12307/2025.961
    Abstract ( 116 )   PDF (2777KB) ( 211 )   Save
    BACKGROUND: Mongolian medicine Echinops sphaerocephalus L. is a commonly used medicine for bone injury in Mongolian medicine. It is effective for tendon injury, fracture, bone nonunion, bone fever, tingling, sore and other diseases. Our previous studies have confirmed that Mongolian medicine Echinops sphaerocephalus L. can promote the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells, but its effect on angiogenesis in the process of bone defect repair is unknown.
    OBJECTIVE: To investigate the effect of Echinops sphaerocephalus L. on in vitro angiogenesis in human umbilical vein vascular endothelial cells and to explore the angiogenesis-promoting active ingredients and their mechanisms of action of Echinops sphaerocephalus L. using network pharmacology technology.
    METHODS: The ethanol extract of Echinops sphaerocephalus L. was prepared and preserved by freeze-drying. The proliferation, migration, chemotaxis and angiogenesis of human umbilical vein endothelial cells were observed after treatment with different concentrations (1 000, 100, and 10 μg/mL) of Echinops sphaerocephalus L. The active components and possible signaling pathways that promoted angiogenesis were enriched and analyzed by network pharmacology.
    RESULTS AND CONCLUSION: (1) The effect of Echinops sphaerocephalus L. on angiogenesis was regulated by its mass concentration: at low mass concentration (10 μg/mL), Echinops sphaerocephalus L. could promote the proliferation, migration, chemotaxis and angiogenesis of human umbilical vein vascular endothelial cells; on the contrary, Echinops sphaerocephalus L. inhibited the proliferation, migration, and chemotaxis of human umbilical vein endothelial cells at high mass concentration (1 000 μg/mL). However, the inhibitory effect of Echinops sphaerocephalus L. on angiogenesis was not significant at high mass concentration due to the limitation of experimental time. 10 μg/mL Echinops sphaerocephalus L. could up-regulate the mRNA expression of angiogenesis-associated factors, including kinase insert domain receptor, vascular endothelial growth factor A, and hypoxia-inducible factor α, and thereby influenced angiogenesis during bone repair. (2) Network pharmacological analyses indicated that Echinops sphaerocephalus L. may bind to eight core targets (TGFB1, TNF, IL-6, STAT3, CTNNB1, IL-1B, AKT1, and HIF-1A) through four core active components (apigenin, caffeic acid, quercetin, and chlorogenic acid) to exert an effect on angiogenesis, atherosclerosis, multiple viral infections, and tumor angiogenesis-related signaling pathways.
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    Simulation analysis of real-time continuous stiffness in muscle fibers and tendons of the triceps surae during multi-joint movement
    Li Chen, Liu Ye, Ni Xindi, Zhang Yuang
    2025, 29 (35):  7529-7536.  doi: 10.12307/2025.956
    Abstract ( 91 )   PDF (2672KB) ( 69 )   Save

    BACKGROUND: The stiffness of muscle fibers and tendons within skeletal muscles is regulated by the neuromuscular system and remains variable. However, observing the mechanical properties of muscle fibers and tendons during complex multi-joint movements is challenging, and the real-time variation patterns of their stiffness are not yet clear.
    OBJECTIVE: Taking the open-access simulation data of triceps surae at different running speeds and gait phases as an example, to explore the real-time stiffness change rules of muscle fiber stiffness and tendon stiffness.
    METHODS: OpenSim simulation results of muscle fiber activation, length, velocity parameters, and tendon length parameters of the triceps surae in five long-distance runners at different running speeds were collected from the Website of Stanford University. The instantaneous slope of the force-length relationship curve of muscle fibers and tendons in the Hill-Zajac muscle model used in the simulation was extracted as the real-time stiffness of the triceps surae muscle fibers and tendons. The temporal changes of stiffness indicators of muscle fibers and tendons during gait were analyzed.

    RESULTS AND CONCLUSION: The regulation of muscle fiber activation-length-velocity status and tendon strain resulted in the stiffness of muscle fibers and tendons changing in the same trend as the applied force. Compared with lower running speeds, the stiffness of the gastrocnemius muscle fibers was higher in the early support phase at higher running speeds (P ≤ 0.01), and the tendon stiffness of the medial head of the gastrocnemius was higher in the early support phase (P ≤ 0.02). The stiffness of the gastrocnemius muscle fibers and tendons was lower from the mid-support to the mid-swing phase (P ≤ 0.03), and the stiffness of the soleus muscle fibers was higher during the support phase (P ≤ 0.02). Under all running speeds, the stiffness of the triceps surae muscle fibers and tendons showed a trend of being higher during the support phase than during the pre-swing phase (P ≤ 0.03), and the stiffness of the gastrocnemius muscle fibers and tendons increased again in the late swing phase (P ≤ 0.05). These findings indicate that increasing running speed can increase the stiffness of triceps surae muscle fibers and tendons during the stance phase; when running speed and gait phase change, gastrocnemius and soleus muscles have different patterns of muscle fiber and tendon stiffness changes, whereas gastrocnemius can increase its muscle fiber stiffness and tendon stiffness in the late swing phase through pre-activation phenomenon.

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    Effect of Roujishuncuiyin on the improvement of skeletal muscle insulin resistance in a mouse model of type 2 diabetes mellitus
    Cai Zhixing, Xia Qiufang, Chen Lili, Zhu Danyang, Zhu Huiwen, Sun Yanan, Liang Wenyu, Zhao Heqian
    2025, 29 (35):  7537-7543.  doi: 10.12307/2025.945
    Abstract ( 128 )   PDF (1956KB) ( 74 )   Save

    BACKGROUND: Skeletal muscle insulin resistance is the key pathological link of type 2 diabetes. The traditional Chinese medicine compound Roujishuncuiyin can effectively improve skeletal muscle insulin resistance, but its mechanism has not been clarified.
    OBJECTIVE: To explore the mechanism of Roujishuncuiyin on skeletal muscle insulin resistance in type 2 diabetes mice.

    METHODS: Forty db/db mice with type 2 diabetes mellitus were randomized into a model group, a low-dose Roujishuncuiyin group, a high-dose Roujishuncuiyin group, and a positive drug group, with 10 mice in each group. The latter three administration groups were given 157.5 mg/g and 630 mg/g Roujishuncuiyin and 200 mg/g metformin hydrochloride aqueous solution by gavage once a day, respectively. In addition, 10 db/dm mice were selected as the blank control group. Mice in the model and blank control groups were given the same dose of 0.9% NaCl solution by gavage. After 12 weeks of intervention, fasting blood glucose was measured in each group of mice, and oral glucose tolerance test was performed to calculate the area under the blood glucose curve. ELISA was used to detect serum insulin level and calculate the resistance index. Mitochondrial structure of skeletal muscle tissue was observed under transmission electron microscopy. Western blot was used to detect the expression levels and phosphorylation levels of protein kinase B (AKT) and glycogen synthase kinase 3β (GSK-3β) proteins in skeletal muscle. 
    RESULTS AND CONCLUSION: (1) Compared with the blank control group, fasting blood glucose, fasting insulin and insulin resistance index were significantly higher in the model group (P < 0.05), the area under the curve of the oral glucose tolerance test was significantly increased (P < 0.05), the expression of p-AKT and p-GSK3β proteins in tibialis anterior muscle was significantly decreased (P < 0.05), and there was a large amount of mitochondrial damage in tibialis anterior muscle and a large number of lipid droplets in the interstitium. (2) Compared with the model group, fasting blood glucose, fasting insulin, and insulin resistance index were significantly reduced in the low- and high-dose Roujishuncuiyin groups and the positive control group (P < 0.05), the area under the curve of the oral glucose tolerance test was reduced (P < 0.05), the expression of p-AKT and p-GSK3β proteins in the tibialis anterior muscle was significantly elevated (P < 0.05), and mitochondrial damage in the tibialis anterior muscle was significantly ameliorated, with decreased lipid droplets in the interstitium. (3) The above indexes were better in the high-dose Roujishuncuiyin group than the low-dose Roujishuncuiyin group (P < 0.05), while there was no significant difference between the high-dose Roujishuncuiyin group and positive control group (P > 0.05). To conclude, by upregulating the protein levels of p-AKT and p-GSK3β in skeletal muscle tissue, the traditional Chinese medicine compound Roujishuncuiyin can improve structural disorders and mitochondrial morphology in skeletal muscle tissue, reduce insulin resistance in the skeletal muscle and regulate glucose homeostasis in the body.  


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    Correlation of echocardiographic parameters with disability at 3 months after acute ischemic stroke
    Yang Bo, Pan Xinfang, Chang Liuhui, Ni Yong
    2025, 29 (35):  7544-7551.  doi: 10.12307/2025.955
    Abstract ( 97 )   PDF (1533KB) ( 95 )   Save
    BACKGROUND: Transthoracic echocardiography and speckle tracking echocardiography parameters are closely related to stroke occurrence and prognosis. However, the role of transthoracic echocardiography and three-dimensional speckle tracking echocardiography parameters in predicting functional disability following acute ischemic stroke remains unclear.
    OBJECTIVE: To explore the relationship between transthoracic echocardiography and three-dimensional speckle tracking echocardiography parameters and functional disability at 3 months post-stroke in patients with acute ischemic stroke.
    METHODS: A total of 299 patients with acute ischemic stroke admitted to the Second Affiliated Hospital of Soochow University between December 2020 and September 2022 were prospectively enrolled. Patients were divided into no disability (n=207) and disability (n=92) groups based on modified Rankin Scale (mRS) scores (≤ 1 as no disability; 1 < mRS score ≤ 5 as disability) at 3 months. Baseline characteristics and transthoracic echocardiography and three-dimensional speckle tracking echocardiography parameters were compared between groups. Independent risk factors for disability were identified using multivariate logistic regression. Predictive models were developed by incorporating variables associated with poor prognosis, and their performance was assessed using the receiver operating characteristic curve analysis. The optimal cutoff value for left ventricular ejection fraction was determined from the receiver operating characteristic curve, and patients were stratified into high and low left ventricular ejection fraction groups. Chi-square test was used to compare prognosis rate between two groups.
    RESULTS AND CONCLUSION: (1) The disability group had significantly lower left ventricular ejection fraction compared with the no disability group (P=0.011). Significant differences in the National Institute of Health Stroke Scale scores, Trial of ORG 10172 in Acute Stroke Treatment, and Oxfordshire Community Stroke Project classifications were observed (P < 0.05). Multivariate logistic regression showed that left ventricular ejection fraction, National Institute of Health Stroke Scale scores, and Oxfordshire Community Stroke Project classification were independent predictors of 3-month disability (P < 0.05). (2) The National Institute of Health Stroke Scale scores and Oxfordshire Community Stroke Project classification at admission were included in model 1, which added the left ventricular ejection fraction as model 2, and all univariate significant difference variables were included in model 3. Receiver operating characteristic analysis showed the area under the curve values of 0.769, 0.806, and 0.824 for the models used to predict the 3-month disability rate. (3) The optimal cutoff for left ventricular ejection fraction from the receiver operating characteristic curve was 60.25%, classifying patients into low left ventricular ejection fraction (≤ 60.25%) and high left ventricular ejection fraction (> 60.25%) groups. The 3-month disability rate was higher in the low left ventricular ejection fraction group than in the high left ventricular ejection fraction group (44% vs. 26%, P=0.002). These findings highlight that reduced left ventricular ejection fraction plays an important role in the assessment of functional disability 3 months after acute ischemic stroke. Left ventricular ejection fraction can be incorporated into routine prognostic assessments when clinically evaluating the prognosis of ischemic stroke.

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    A novel analysis and prediction method for potential mechanisms of traditional Chinese medicine based on artificial intelligence and omics data-driven approach
    Jiang Qiyu, Zeng Huiyan
    2025, 29 (35):  7552-7561.  doi: 10.12307/2025.968
    Abstract ( 150 )   PDF (16696KB) ( 12 )   Save
    BACKGROUND: The treatment of diseases with traditional Chinese medicine is a complex multi-target regulatory process. It is of great significance to explore the multi-target integration effect of traditional Chinese medicine by combining technologies from multiple fields such as artificial intelligence, single-cell transcriptomics, spatial transcriptomics, and bioinformatics. 
    OBJECTIVE: To propose a novel analytical prediction method for potential mechanisms of traditional Chinese medicines, which is different from network pharmacology, based on artificial intelligence and omics data driven, with an example of exploring the potential mechanisms of Dachaihu Decoction for treating hyperlipidemia and atherosclerosis.
    METHODS: (1) The pharmacodynamic protein targets of the constituent drugs of Dachaihu Decoction were collected through TCMSP database, and the disease targets of hyperlipidemia were obtained in Genecards, NCBI, and TTD. (2) The single-cell transcriptome samples of hyperlipidemia (the first set of single-cell data samples from aortic valves of wild-type, Apoe knockout, and Ldlr knockout mice; the second set of single-cell data samples from Ldlr knockout mice fed with high cholesterol versus normal feeding) and spatial transcriptome samples from human coronary atherosclerosis tissue sections were obtained from the GEO database. A deep neural network autoencoder model was developed to encode the transcriptome sequencing data, and the integrated coded values (MTIS) were mapped to the single-cell level and spatial organization level using single-cell transcriptome and spatial transcriptome technologies for comparative statistical analyses of the samples and identification of the main effector cells and effector genes. 
    RESULTS AND CONCLUSION: (1) There were significant differences in the data morphology and statistics of MTIS between wildtype and Apoe-knockout mice treated with Dachaihu Decoction (P < 0.000 1), as well as between wildtype and Ldlr-knockout mice treated with Dachaihu Decoction (P < 0.000 1). (2) The main effector cells of Dachaihu Decoction in Apoe-knockout mice were aortic valve stromal cells, while the main effector cells in Ldlr-knockout mice were white blood cells, fibroblasts, and vascular endothelial cells. Except for Ldlr and Apoe, the main effector genes are Vcam1, Fn1, and Mmp2. (3) There were statistically significant differences (P < 0.000 1) in MTIS between high cholesterol fed samples and normal fed samples of Ldlr-knockout mice treated with Dachaihu Decoction. The main effector cells were macrophages, and the main effector genes were Fn1, F7, Ptgs1, IL6 and App. (4) The spatial transcriptome comparisons of MTIS in human coronary artery slices showed that high MTIS value cells appeared to be distributed in both blood vessels and atherosclerotic plaque areas, while low MTIS value cells appeared to be mainly concentrated in the endothelial cells and atherosclerotic plaque areas. To conclude, this new analytical method achieves quantitative analysis of the multi-target integration effects of traditional Chinese medicine at the single-cell level and organ spatial tissue level, which is used to explore the potential mechanism of Dachaihu Decoction in treating hyperlipidemia and atherosclerosis.
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    Analysis of diagnostic biomarkers for ischemic stroke and experimental validation of targeted cuproptosis related genes
    Chen Ying, Guo Xiaojing, Mo Xueni, Ma Wei, Wu Shangzhi, Li Xiangling, Xie Tingting
    2025, 29 (35):  7562-7570.  doi: 10.12307/2025.957
    Abstract ( 133 )   PDF (4630KB) ( 179 )   Save
    BACKGROUND: Studies have shown that immune cells are involved in all processes of ischemic stroke, in which cuproptosis also plays a key role.
    OBJECTIVE: To screen diagnostic biomarkers related to the progression of ischemic stroke through bioinformatics, and analyze and validate cuproptosis-related genes closely related to the occurrence and development of ischemic stroke. 
    METHODS: The GSE16561 microarray was obtained from the GEO database, containing data from 39 cases of ischemic stroke (ischemic stroke group) and 24 controls (control group). Differentially expressed genes from the ischemic stroke microarray data were analyzed. Gene ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were performed. By using LASSO and Random Forest methods, key genes affecting the occurrence and development of ischemic stroke were screened, and a diagnostic model was established and validated. Differential gene analysis was performed through immune cell infiltration and weighted gene co expression network. The differentially expressed immune-related genes were intersected with cuproptosis genes to obtain the hub genes related to cuproptosis immunity. In vitro cell experiments were conducted to divide rat hippocampal neurons into a normal group and an ischemic stroke group, and qPCR experiments were performed to verify the results. 
    RESULTS AND CONCLUSION: (1) 573 differentially expressed genes were obtained by differential analysis. Differentially expressed genes were mainly enriched in biological processes, such as positive regulation of immune response, and signaling pathways such as lipid and atherosclerosis. (2) Machine learning methods were used to screen diagnostic genes such as MFN2, PKM2, CREG1, and FOXO3A, which may have some diagnostic value for ischemic stroke. (3) Immune infiltration analysis revealed resting plasma cells, NK cells, macrophages, etc., indicating that immune cells play a certain role in the pathogenesis of ischemic stroke. (4) Weighted gene co-expression network analysis combined with immune infiltration analysis obtained 118 key module genes, which were intersected with cuproptosis genes to obtain 2 cuproptosis and immune characteristic genes. The correlation analysis between four diagnostic genes and Hub genes showed that the expression of FOXO3A and MFN2, PKM2 and BCL2L1, MTF1 and MFN2, ATP7B and BCL2L1 were correlated. (5) The qPCR results showed significant differences in the genes MTF1 and ATP7B between the ischemic stroke group and the blank group. To conclude, ATP7B and MTF1 can serve as characteristic genes for cuproptosis in ischemic stroke. It is possible to improve ischemic stroke by intervening in ATP7B and MTF1 to regulate cuproptosis. 

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    Chinese herbal prescription combined with head acupuncture exercise therapy improves limb spasticity in rats with ischemic stroke
    Zhu Chuanxi, Qiu Long, Li Lingxu, Ji Guangcheng
    2025, 29 (35):  7571-7577.  doi: 10.12307/2025.926
    Abstract ( 117 )   PDF (1743KB) ( 68 )   Save
    BACKGROUND: Chinese herbal prescription with head acupuncture exercise therapy has significant clinical efficacy in the treatment of limb spasticity after ischemic stroke; however, the exact mechanism is not clear.
    OBJECTIVE: To explore the possible mechanism of the combination of Chinese herbal prescription with head acupuncture exercise therapy in improving limb spasticity after stroke. 
    METHODS: Seventy Sprague-Dawley rats were divided into blank, model, exercise therapy, head-acupuncture, Chinese herbal prescription, Chinese herbal prescription with head acupuncture exercise therapy, and sham-operation groups using the random number method, with 10 rats in each group. Except for the blank group and the sham-operation group, a rat model of cerebral ischemia/reperfusion injury was made by using the middle cerebral artery cable embolism method in the rest of the groups. Interventions in each group started on the 3rd day after modeling. In the Chinese herbal prescription group, rats were given 1.5 g/kg Yi’nao Fujian herbal prescription by gavage; in the head acupuncture group, acupuncture was applied to Baihui and 2 mm on the left and right sides of Baihui, and was left in place for 30 minutes; in the exercise therapy group, rats were exercised on a treadmill for 30 minutes; in the Chinese herbal prescription with head acupuncture exercise therapy group, 1.5 g/kg Yi’nao Fujian herbal prescription was given by gavage followed by acupuncture treatment (identical to that in the head acupuncture group, and then the animals were exercised for 30 minutes on the treadmill while the acupuncture needles remained in place. Rats in the model and sham-operation groups were subjected to grasping under the same environmental conditions and placed on a rat immobilizer for 30 minutes, and the blank group did not have any intervention. The frequency of intervention in each group was once a day for 7 consecutive days. The neurological deficit score and muscle tone were evaluated in each group before and after treatment. Pathological changes in rat brain tissue were observed. Protein and mRNA expressions of glutamic acid decarboxylase 67, succinate semialdehyde dehydrogenase, and γ-aminobutyric acid transaminase in the cerebral cortex of the rats were detected. 
    RESULTS AND CONCLUSION: (1) Compared with the model group, the Chinese herbal prescription with head acupuncture exercise therapy group had lower neurological deficit scores and modified Ashworth spasticity scale (MAS) muscle tone scores (P < 0.01), decreased expression of succinate semialdehyde dehydrogenase andγ-aminobutyric acid transaminase at protein and mRNA levels, and increased expression of expression of glutamic acid decarboxylase 67 at protein and mRNA levels (P < 0.01); the neuronal cell structure was relatively intact, with only some irregular cavities visible and reduced nuclear consolidation. (2) Compared with the Chinese herbal prescription with head acupuncture exercise therapy group, the neurological deficit scores and MAS muscle tone scores were higher in the exercise therapy group (P < 0.05), and the MAS muscle tone scores were higher in the head-acupuncture group and the Chinese herbal prescription group (P < 0.05, P < 0.01), while the expressions of glutamic acid decarboxylase 67 protein and succinate semialdehyde dehydrogenase andγ-aminobutyric acid transaminase mRNAs were increased in all the other intervention groups (P < 0.05, P < 0.01). Neuronal cells in the Chinese herbal prescription with head acupuncture exercise therapy group were more orderly arranged and clearly outlined, with less nuclear consolidation and shape irregularity compared with the other intervention groups. (3) The neurological deficit score and MAS muscle tone score in the Chinese herbal prescription with head acupuncture exercise therapy group were significantly lower after treatment (P < 0.01). To conclude, Yi’nao Fujian herbal prescription combined with head-acupuncture exercise therapy can improve post-stroke spasticity by affecting the expression of γ-aminobutyric acid transaminase in rats, which is more effective than Chinese herbal prescription alone, exercise therapy alone, and head-acupuncture alone.
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    Exercise prevention and treatment of insulin resistance: role and molecular mechanism of Keap1/nuclear factor erythroid2-related factor 2 signaling pathway
    Liu Xuan, Ding Yuqing, Xia Ruohan, Wang Xianwang, Hu Shujuan
    2025, 29 (35):  7578-7588.  doi: 10.12307/2026.526
    Abstract ( 161 )   PDF (1682KB) ( 157 )   Save
    BACKGROUND: Exercise interventions play a key role in disease prevention and treatment, which can effectively activate the nuclear factor erythroid2-related factor 2 (Nrf2) signaling pathway to prevent the occurrence and development of insulin resistance. However, the potential mechanism of Nrf2-targeting exercise therapy strategies in alleviating insulin resistance remains unclear.
    OBJECTIVE: Based on the relationship between Nrf2, insulin resistance and exercise, to analyze the mechanism and influence of different exercise modes on the activation of Nrf2, thereby elucidating the potential mechanism of Nrf2-targeting exercise therapy strategies in the process of alleviating insulin resistance.
    METHODS: “Diabetes mellitus, insulin, insulin resistance, nuclear factor E2 related factor 2, oxidative stress, ferroptosis, autophagy, inflammatory response, exercise” were used as search terms in Chinese and English. WanFang Database, CNKI, Google Scholar and PubMed database were searched for relevant literature from inception to October 2024, and a total of 127 core related articles were obtained according to the inclusion and exclusion criteria.
    RESULTS AND CONCLUSION: (1) Keap1/Nrf2 signaling pathway, as an important endogenous antioxidant stress pathway, plays an important role in insulin resistance. Activation of Keap1/Nrf2 signaling pathway can enhance the antioxidant capacity of cells, reduce oxidative stress and inflammation, improve insulin signaling, and thus protect insulin resistance. (2) A variety of exercise methods (including aerobic exercise, resistance exercise and high-intensity intermittent exercise) can effectively activate Nrf2 signal, improve the activity of antioxidant kinase and enhance the antioxidant capacity of cells, so as to alleviate oxidative stress damage to a certain extent. (3) By activating Nrf2 signaling pathway, exercise can up-regulate the activity of antioxidant enzymes such as Heme oxygenase 1, superoxide dismutase, catalase, glutathione peroxidase, glutathione, and enhance the antioxidant capacity of cells. In addition, it can also regulate the key enzymes and proteins in the ferroptosis pathway, such as glutathione peroxidase 4, membrane iron transport protein 1, and ferritin heavy chain 1, inhibit the ferroptosis pathway and promote the balance of iron metabolism. In autophagy, the expression of autophagy related genes p62 and ATG5/7 can be enhanced, and the level of microtubule-associated protein 1 light chain 3 is increased, thus regulating autophagy process. At the same time, the activity of tumor necrosis factor-α, nuclear factor-κB, interleukin-6, interleukin-1β and other pro-inflammatory cytokines is decreased, and the inflammatory response can be effectively inhibited. These combined effects can reduce oxidative stress damage, increase insulin sensitivity and improve insulin signaling, and thus have a positive effect on improving insulin resistance. (4) Given that the Keap1/Nrf2 signaling pathway plays an important role in the pathogenesis and treatment of insulin resistance, exercise therapy strategies targeting the Keap1/Nrf2 signaling pathway will help promote the development of “exercise+drug” precision medicine for insulin resistance.
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    Roles of SOX5 in bone metabolism and prevention of bone diseases and the relationship with exercise#br#
    #br#
    Li Zhipeng, Xing Rongxin, Hu Lianghong
    2025, 29 (35):  7589-7600.  doi: 10.12307/2025.752
    Abstract ( 133 )   PDF (1607KB) ( 201 )   Save
    BACKGROUND: SOX5 is an important transcription factor of the SRY-related HMG-box (SOX) family, which plays a key role in regulating the expression of genes related to bone development and remodeling, especially during osteoblast differentiation and chondrocyte maturation, through its unique HMG box DNA structural domains in concert with SOX6 and SOX9. In addition, the expression and activity of SOX5 and its family are regulated by a variety of diseases and different forms of exercise, among other factors, suggesting that SOX5 and its family have the potential to be effective as drugs and therapeutics to ameliorate related diseases in the future.
    OBJECTIVE: To provide new perspectives for future research on SOX5 and to provide scientific basis for the application of exercise intervention and drug therapy in the prevention and treatment of bone diseases.
    METHODS: CNKI and PubMed databases were searched for relevant literature published from 2001 to June 2024, and the search terms were “SRY-related HMG-box5, SOX5, Bone” in Chinese and English, respectively. After screening, analysis and summarization, 105 articles were included in the final review.
    RESULTS AND CONCLUSION: (1) Role of SOX5 in bone development: SOX5 is an important member of the SOX family, which plays a central role in the regulation of skeletal development, bone metabolism and cartilage formation. In synergy with SOX6 and SOX9, SOX5 activates gene expression in osteoblasts and chondrocytes by binding to specific DNA sequences to regulate bone formation and bone remodeling. (2) Abnormal expression of SOX5 is closely related to bone and joint diseases such as chondrodysplasia, osteoporosis and osteoarthritis, suggesting that it may be a key regulator of these diseases. Currently, a variety of drugs may be used to treat bone metabolism-related diseases by regulating SOX5 and its family, and upregulation of SOX5 in mesenchymal stem cells may be effective in improving the symptoms of bone metabolism disease patients in bone tissue engineering. (3) Exercise may effectively prevent osteoporosis and related bone diseases by enhancing bone metabolism and promoting osteoblast differentiation and bone density increase. This mechanism of action may be related to the specific regulatory mechanism of SOX5, especially in different types, intensities and durations of exercise need to be further explored and studied. In conclusion, SOX5 has an important regulatory role in bone development, chondrogenesis, and prevention of bone diseases, and its activity is regulated by a variety of factors, while exercise intervention provides a new scientific basis for the treatment of bone diseases.
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    Pathological processes, inflammatory responses, and related biomarkers of periodontitis: a multi-omics analysis
    Zhang Yixuan, Li Dongna, Liu Chunyan
    2025, 29 (35):  7601-7610.  doi: 10.12307/2025.988
    Abstract ( 201 )   PDF (1197KB) ( 387 )   Save
    BACKGROUND: In recent years, the rapid development of omics technologies has ushered oral medicine research into a new era. The extensive application of these technologies not only systematically reveals the complexity and dynamic changes within biological systems but also provides a comprehensive perspective for studying various diseases, including periodontitis.
    OBJECTIVE: To review the recent advancements in the application of omics technologies in periodontitis research, while also addressing existing challenges, thereby expanding the scope for the prevention and treatment of periodontitis and its applications in other clinical fields. 
    METHODS: A literature search was conducted on PubMed and CNKI databases, covering publications from June 1993 to August 2024. The search terms included “omics,” “periodontitis,” “transcriptomics and periodontitis,” “proteomics and periodontitis,” “genomics and periodontitis,” “metabolomics and periodontitis” and “multi-omics and periodontitis” in English and Chinese, respectively. A total of 72 relevant articles were included for further analysis.
    RESULTS AND CONCLUSION: (1) By using genomics, transcriptomics, proteomics, and metabolomics, researchers can conduct in-depth analyses of pathological processes, inflammatory responses, and associated biomarkers of periodontitis from multiple dimensions. (2) Recent studies have indicated that omics technologies can identify key genes and metabolites and other factors associated with periodontitis, facilitating a precise understanding and intervention of the disease. As these technologies continue to advance, omics analysis will be more widely applied in periodontitis research, driving the development of more effective preventive and therapeutic strategies, thereby improving patients’ quality of life.
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    Lipophagy, exercise intervention and prevention and treatment of nonalcoholic fatty liver disease
    Ji Long, Chen Ziyang, , Jin Pan, Kong Xiangkui, Pu Rui,
    2025, 29 (35):  7611-7619.  doi: 10.12307/2026.522
    Abstract ( 166 )   PDF (1504KB) ( 180 )   Save
    BACKGROUND: Lipophagy is closely related to a variety of chronic metabolic diseases. Exercise can delay the pathological process of non-alcoholic fatty liver by regulating lipophagy, which has become one of the most popular studies in the field of sports medicine.
    OBJECTIVE: To summarize the regulatory role of lipophagy in non-alcoholic fatty liver disease and the mechanism of exercise-mediated lipophagy in non-alcoholic fatty liver disease.
    METHODS: CNKI, PubMed, and Web of Science databases were searched for relevant literature published from 1980 to 2025, using the search terms of “lipophagy, lipid metabolism, non-alcoholic fatty liver disease, aerobic exercise, resistance training, combined aerobic resistance exercise” in Chinese and English. Totally 98 documents were included according to inclusion and exclusion criteria.
    RESULTS AND CONCLUSION: (1) Lipophagy plays an important regulatory role in the prevention and treatment of non-alcoholic fatty liver disease. (2) Different types of exercises affect lipophagy: Aerobic exercise can play a beneficial role can regulate the expression of lipophagy related factors, enhance autophagy flow, improve lysosome biosynthesis and promote the decomposition of lipase. Resistance training can increase the expression of autophagy related factors in adipose tissue. Combined aerobic resistance exercise can increase fat oxidation rate and improve insulin sensitivity. (3) Exercise can treat non-alcoholic fatty liver disease by improving liver inflammation, insulin resistance, and liver function. (4) Exercise plays a key role in the prevention and treatment of non-alcoholic fatty liver by regulating lipophagy, reducing lipid accumulation, inhibiting steatosis, alleviating liver inflammation and fibrosis and improving insulin resistance.
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    Influence and implications of basketball shoes' functional parameters on human biomechanics
    Yang Yicheng, Zheng Zhizhen, Liang Shuangxue, Wu Chengliang, Du Yunyun
    2025, 29 (35):  7620-7628.  doi: 10.12307/2025.962
    Abstract ( 206 )   PDF (1277KB) ( 394 )   Save
    BACKGROUND: Basketball shoes are one of the most important pieces of athletic equipment in basketball, helping to improve players’ performance and reduce the risk of injury. With the development of basketball, the demand for basketball shoe performance is getting higher and higher.
    OBJECTIVE: To systematically review and summarize the effects of different functional parameters of basketball shoes on the wearer’s biomechanical performance, and to investigate the effects of each functional parameter of basketball shoes on athletic performance and injury risk.
    METHODS: Relevant literature addressing the effect of basketball shoes’ functional parameters on human biomechanics from CNKI, WanFang, Web of Science, ScienceDirect and other databases was searched by computer, with the search terms of “basketball, basketball shoe, shoes, footwear, sports biomechanics, kinematics, kinetics” in Chinese and English. All the retrieved literature was evaluated and screened, and finally 61 articles were included for literature review.
    RESULTS AND CONCLUSION: Existing studies on functional parameters of basketball shoes include shoe collar height and heel counter-stiffness, midsole hardness, midsole cushioning, midsole thickness, forefoot flexural stiffness, whole shoe mass, and outsole traction. The functional parameters of basketball shoes play a role in improving sports performance and reducing the risk of sports injuries: (1) Compared with low-top shoes, high shoe collars can reduce the risk of ankle sprains, but may affect sports performance, ankle proprioception, and increase the load on the knee. (2) Better midsole cushioning or softer midsoles can effectively reduce vertical impact, and the forefoot cushioning structure can effectively improve the athletic performance of lateral movement. (3) The midsole thickness of 11 mm in the forefoot and 20 mm in the rearfoot can obtain the best ankle stability and sprinting performance. (4) Improving the outsole traction and forefoot bending stiffness can significantly improve the performance of jumping, sprinting, and side-cutting. There are diminishing returns to the improvement of athletic performance by improving outsole traction. (5) Lighter basketball shoes can improve athletic performance, but this effect only occurs when the wearer is aware of the shoes’ weight.
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    All-inside versus traditional techniques of anterior cruciate ligament reconstruction: meta-analysis of therapeutic efficacy and radiological outcomes
    Wang Feng, Cao Chunfeng, He Chao, Zhang Tao, Zhou Zixian, Zhu Fengchen
    2025, 29 (35):  7629-7638.  doi: 10.12307/2025.938
    Abstract ( 99 )   PDF (2300KB) ( 66 )   Save
    OBJECTIVE:  To compare the clinical efficacy and radiological results of arthroscopic all-inside technique and traditional technique in anterior cruciate ligament reconstruction.
    METHODS: Databases such as CNKI, WanFang, VIP, PubMed, Web of Science, Embase, and Cochrane Library were searched for literature on all-inside technique and traditional technique in anterior cruciate ligament reconstruction. The search time was from the establishment of each database to September 2024. Meta-analysis was conducted on the included literature. 
    RESULTS: (1) A total of 17 articles were included in the meta-analysis, with 631 patients in the all-inside technique reconstruction group and 626 patients in the traditional technique reconstruction group. The earliest time for the last follow-up was half a year after surgery, and the latest was 5 years after surgery. Most follow-up period was 2 years. (2) Meta-analysis results showed that compared with traditional anterior cruciate ligament reconstruction, the all-inside technique had a thicker graft during anterior cruciate ligament reconstruction [mean difference (MD)=0.20, 95% confidence interval (CI) (0.09, 0.31), P=0.000 5], smaller postoperative maximum diameter [standardized mean difference (SMD)=-3.64, 95% CI (-6.00, -1.28), P=0.002] and volume [SMD=-3.69, 95% CI (-5.37, -2.00), P < 0.000 1] of the tibial tunnel, and higher International Knee Documentation Committee subjective scores [MD=2.41, 95% CI (0.49, 4.32), P=0.01] and Lysholm scores [MD=1.11, 95% CI (0.42, 1.8), P=0.002] 2 years after surgery. However, the operation time was relatively longer [MD=10.06, 95% CI (4.71, 15.4), P=0.000 2], and the knee stability was poorer after 2 years [SMD=0.3, 95% CI (0.04, 0.55), P=0.02]. No significant differences were found between the two groups in the following aspects: the subjective scores of the International Knee Documentation Committee at 6 months and 1 year postoperatively [MD= -0.05, 95% CI (-1.96, 1.83), P=0.96; MD=0.51, 95% CI (-1.17, 2.19), P=0.55]; the difference in anterior laxity of bilateral knees at 1 year postoperatively [SMD=-0.02, 95% CI (-0.3, 0.27), P=0.9]; the Lysholm score at 6 months postoperatively [MD=0.87, 95% CI (-0.15, 1.89), P=0.09]; the objective score of the International Knee Documentation Committee at the last follow-up [RR=0.95, 95% CI (0.86, 1.06), P=0.37]; the American Knee Society Score at the last follow-up [MD=0.33, 95% CI (-0.55, 1.21), P=0.47]; the Tegner score at the last follow-up [MD=0.05, 95% CI (-0.11, 0.22), P=0.53]; the negative rate of the pivot shift test at the last follow-up [RR=0.92, 95% CI (0.83, 1.01), P=0.09]; the postoperative revision rate at the last follow-up [RR=2.2, 95% CI (0.98, 4.92), P=0.05]; and the result of the single-leg hop test at the last follow-up [MD=-0.06, 95% CI (-4.99, 4.86), P=0.98]. 
    CONCLUSION: There were no significant differences in most functional outcome scores and the position of the tibial tunnel between the all-inside technique and the traditional technique after anterior cruciate ligament reconstruction. The all-inside technique was more favorable in terms of subjective International Knee Documentation Committee scores and Lysholm scores at 2 years postoperatively. Meanwhile, the knee joint was more stable 2 years after anterior cruciate ligament reconstruction using the traditional technique. In addition, it was found that the graft was thicker during the all-inside technique, while the diameter and volume of the tibial tunnel were smaller postoperatively, with more bone tissue preserved. Nevertheless, the operation time of the all-inside technique was longer.
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    Effects of different transcranial magnetic stimulation modes on refractory depression in adults: a network meta-analysis
    Tian Jinxin, Zhao Yuxin, Hu Tong, Cui Tiantian, Ma Lihong
    2025, 29 (35):  7639-7648.  doi: 10.12307/2025.966
    Abstract ( 162 )   PDF (1858KB) ( 237 )   Save
    OBJECTIVE: To assess the efficacy and safety of transcranial magnetic stimulation in the treatment of refractory depression and to compare the differences in efficacy between various transcranial magnetic stimulation treatment protocols in refractory depression, thereby providing a theoretical basis for the clinical selection of transcranial magnetic stimulation treatment protocols.
    METHODS: A comprehensive search was conducted across multiple databases, including PubMed, Embase, Cochrane Library, Web of Science, CNKI, WanFang Data, CBM and VIP. The search terms were “transcranial magnetic stimulation, treatment-resistant depression, randomized controlled trial” in Chinese, and “depressive disorder, treatment-resistant, transcranial magnetic stimulation, randomized controlled trial” in English. The objective was to identify randomized controlled trials on the treatment of patients with refractory depression published from the establishment of the databases to September 2024. The quality of the included studies was evaluated using the Cochrane risk of bias assessment tool, version 5.1.0, and the Physiotherapy Evidence Database scale. Meta-analysis of the outcome indicators was conducted using the Review Manager 5.4 and Stata 18.0 software.
    RESULTS: (1) Following a comprehensive review, 20 randomized controlled trials were included in the analysis. All of the trials were assessed to be of high or very high quality according to the Physiotherapy Evidence Database scale. (2) Meta-analysis results showed that, compared with the sham stimulation group, high-frequency repetitive transcranial magnetic stimulation could significantly reduce the scores of Hamilton Depression Rating Scale [mean difference (MD)=
    -3.89, 95% confidence interval (CI): -6.14 to -1.65, P < 0.05) or the Montgomery Depression Rating Scale (MD =-3.97, 95% CI: -6.57 to -1.36, P < 0.05). (3) The probability ranking results of the network Meta-analysis showed that, in terms of the Hamilton Depression Rating Scale score, the probability ranking results were as follows: high-frequency repetitive transcranial magnetic stimulation (69.9%) > intermittent theta burst stimulation (62.8%) > bilateral theta pulse stimulation (57.5%) > low-frequency repetitive transcranial magnetic stimulation (54.9%) > bilateral sequential transcranial magnetic stimulation (49.0%) > transcranial pulsed electromagnetic field (37.0%) > sham stimulation (18.9%). And in terms of the Montgomery Depression Rating Scale score, the probability ranking results were as follows: high-frequency repetitive transcranial magnetic stimulation (93.3%) > bilateral theta pulse stimulation (50.3%) > sham stimulation (45.9%) > low-frequency repetitive transcranial magnetic stimulation (32.1%) > bilateral sequential transcranial magnetic stimulation (28.4%).
    CONCLUSION: Transcranial magnetic stimulations can improve the depressive symptoms of patients with treatment-resistant depression. Among them, the high-frequency repetitive transcranial magnetic stimulation mode has the best effect on improving the depressive symptoms of patients with treatment-resistant depression, followed by the intermittent θ burst stimulation mode.
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    A systematic review of osteoporosis-specific quality-of-life scales
    Yan Jing, Qin Qiujun, Li Fen, Zhou Jun, Ding Yuanyuan, Jin Chunlin
    2025, 29 (35):  7649-7655.  doi: 10.12307/2025.975
    Abstract ( 157 )   PDF (1340KB) ( 127 )   Save
    OBJECTIVE: To review the development trend, target population, setting and construction process of osteoporosis-specific quality of life measures both domestically and internationally. 
    METHODS: All of the currently available publicly available osteoporosis-specific quality-of-life scales were found by searching the PubMed, Embase, CNKI, and WanFang Data. The search period included the time from database construction to August 2024. The basic information, development trend, target population, and the setting of the scale’s dimensions, entries, and levels were taken from the original research and development literature that included the scale. The content of the dimensions and the scale’s construction process were then compiled and examined. 
    RESULTS: A total of 20 osteoporosis-specific quality of life scales were included in the study, and the literature on the development of 34 scales was traced. The number of dimensions of the included scales ranged from 2-7 dimensions, all of which included physiological dimensions; the number of entries of the scales ranged from 10-84 entries, while the number of levels set under the entries was mostly focused on four or five levels. There are three main ways of constructing the scales, of which the construction of a pool of entries and the screening of the entries according to classical measurement theory and item response theory is an approach that has emerged in recent years.  
    CONCLUSION: According to the specific target population and diversified measurement objectives, the content of osteoporosis-specific quality of life scales has become increasingly streamlined, with fewer dimensions and entries overall; and its building procedure is gradually becoming more standardized.

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    Effect of insulin-like growth factor family member levels on inflammatory arthritis: a FinnGen biobank-based analysis
    Wang Xuepeng, , He Yong,
    2025, 29 (35):  7656-7662.  doi: 10.12307/2025.998
    Abstract ( 118 )   PDF (2925KB) ( 98 )   Save
    BACKGROUND: Studies have shown a significant association between members of the insulin-like growth factor family and the occurrence of inflammatory arthritis, but the causal relationship has not been accurately characterized. 
    OBJECTIVE: To explore the potential association between members of the insulin-like growth factor family and the occurrence and development of ankylosing spondylitis, rheumatoid arthritis, and psoriatic arthritis.
    METHODS: Genetic instrumental variables associated with 14 discrete members of the insulin-like growth factor family, primarily derived from the expansive genomic database of a genome-wide association study, were used. The pertinent summary statistics for ankylosing spondylitis, rheumatoid arthritis, and psoriatic arthritis were meticulously procured from the FinnGen Consortium’s extensive dataset. Our primary analytical methodology was anchored in the inverse-variance weighted approach, which is recognized for its robustness in Mendelian randomization studies. To augment the credibility and broader applicability of the findings, an array of complementary analyses were performed. These encompassed the weighted-median method, which mitigates the influence of potential outliers; the MR-Egger regression, a tool for assessing directional pleiotropy; the weighted mode and simple mode approaches, which provide alternative estimates of the causal effect; the MR pleiotropy residual sum and outlier test, designed to identify and correct for horizontal pleiotropy; Cochran’s Q statistic test, which evaluates the heterogeneity of the effect estimates; and the MR-Egger intercept analysis, a diagnostic for detecting and adjusting the impact of pleiotropic relationships. 
    RESULTS AND CONCLUSION: We identified four distinct causal associations: CYR61 protein was negatively correlated with ankylosing spondylitis (odd ratios [OR]: 0.919, 95% confidence interval [CI]: 0.848-0.997, P=0.042) and rheumatoid arthritis (OR: 0.946, 95% CI: 0.908-0.987, P=0.011); IGF-II receptor was negatively correlated with ankylosing spondylitis (OR: 0.909, 95% CI: 0.835-0.990, P=0.029); IGFBP-7 was positively correlated with psoriatic arthritis (OR: 1.104, 95% CI: 1.002-1.218, P=0.046). The results of sensitivity analyses were consistent (P < 0.05). This rigorous analytical approach has yielded evidence suggestive of a potential causal nexus between a constellation of insulin growth factor family members and the risk of inflammatory arthritis. These findings underscore the necessity for further research to delineate the precise mechanisms by which insulin-like growth factor family members influence the developmental trajectories of ankylosing spondylitis, rheumatoid arthritis, and psoriatic arthritis, thereby providing evidence for developing targeted interventions. In addition, by drawing on international research experience, future biomedical research in China should further focus on the genetic mechanisms of inflammatory and immune-related diseases, promote clinical translation and the development of precision medicine, and provide more efficient and personalized treatment plans for Chinese patients.
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    Causal relationship between gut microbiota and rheumatoid arthritis: data analysis in European populations based on GWAS data
    Wang Tao, Wang Shunpu, Min Youjiang, Wang Min, Li Le, Zhang Chen, Xiao Weiping
    2025, 29 (35):  7663-7668.  doi: 10.12307/2026.536
    Abstract ( 122 )   PDF (1507KB) ( 88 )   Save
    BACKGROUND: Studies have shown that gut microbiota may affect the progression of rheumatoid arthritis. However, the causal relationship between the two is unknown. Mendelian randomization analysis of the two using published Genome Wide Association Study (GWAS) data can explore the causal relationship between gut microbiota and rheumatoid arthritis, helping to develop targeted microbial therapies and provide methods and strategies for the prevention and treatment of rheumatoid arthritis.
    OBJECTIVE: To explore the potential causal relationship between gut microbiota and rheumatoid arthritis using two-sample two-way Mendelian randomization method. 
    METHODS: Gut microbiota GWAS data from the MiBio-Gen consortium and rheumatoid arthritis GWAS data from the IEU Open GWAS database (a large gene-phenotype association database developed at the MRC Integrative Epidemiology Unit (IEU) at the University of Bristol, UK) were used. Inverse variance weighting was used as the main analysis method, and MR-Egger regression method, weighted median method, weighted model and simple model method were used as supplements to study the causal relationship between gut microbiota and rheumatoid arthritis. Heterogeneity was assessed using Cochran’s Q test, horizontal pleiotropy was assessed using MR-PRESSO and MR-Egger intercept tests, robustness of results was tested using leave-one method, and reverse Mendelian randomization analysis was used to assess the presence or absence of reverse causality.   
    RESULTS AND CONCLUSION: (1) There was a causal relationship between five kinds of enteric bacteria and rheumatoid arthritis. Ruminococcus gauvreauii group (β=0.262, odds ratio [OR]=1.300, P=0.013) and Butyricimonas (β=0.001, OR=1.001, P=0.014) increased the risk of rheumatoid arthritis, while Anaerostipes (β=-0.225, OR=0.798, P=0.025), Lachnospiraceae-UCG010 (β=-0.177, OR=0.838, P=0.026) and Oxalobacter (β=-0.171, OR=0.843, P=0.001) reduced the risk of rheumatoid arthritis. Sensitivity analyses showed no significant heterogeneity or horizontal pleiotropy (all P > 0.05), and leave-one-out testing confirmed the robustness of the results, while the addition of the remaining four methods other than the inverse variance weighting method further validated the reliability and stability of the results. (2) Reverse Mendelian randomization analysis did not find a causal association between rheumatoid arthritis and the five kinds of enteric bacteria identified by Mendelian randomization analysis. These findings indicate that Ruminococcus gauvreauii group and Butyricimonas may be the risk factors of rheumatoid arthritis, while Anaerostipes, Lachnospiraceae-UCG010 and Oxalobacter may be the protective factors of rheumatoid arthritis. Gut microbiota may play an important role in the pathogenesis of rheumatoid arthritis, and provide new biomarkers for the prevention and treatment of rheumatoid arthritis. In addition, for the field of biomedical research in China, we can learn from international experience and gradually establish and improve a multi-center large-scale genetic database, so as to deeply explore the relationship between gut microbiota and disease risk, and promote the development of precision medicine and personalized treatment in China.
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    Action mechanism of Gegenmaqi prescription in treatment of periarthritis of shoulder combined with type 2 diabetes based on TCMSP database
    Wang Tong, Zheng Yu, Jia Chengming, Yang Hu, Zhang Guangfei, Ji Yaoyao
    2025, 29 (35):  7669-7678.  doi: 10.12307/2025.953
    Abstract ( 131 )   PDF (4720KB) ( 179 )   Save
    BACKGROUND: Gegenmaqi prescription has a good effect on periarthritis of shoulder combined with type 2 diabetes and has a good application prospect, but the specific mechanism is not clear. 
    OBJECTIVE: To explore the action mechanism of Gegenmaqi prescription on periarthritis of shoulder and type 2 diabetes by network pharmacology, molecular docking, and molecular dynamics. 
    METHODS: The active components and protein targets of Gegenmaqi prescription were retrieved from the Traditional Chinese Medicine System Pharmacology database and analysis platform, referred to as TCMSP jointly established by the Shanghai Institute of Materia Medica, Chinese Academy of Sciences and the Institute of Chinese Materia Medica, and China Academy of Chinese Medical Sciences in 2013. Genecards created by Professor Doron Lancet’s team at the Weizmann Institute of Science in Israel in 1997, Drugbank created by scientists at the University of Alberta in Canada in 2006, and the OMIM database established by Dr. Victor A. McKusick’s team at Johns Hopkins University in the United States in 1966 were used to search the disease protein targets of periarthritis of shoulder and type 2 diabetes, and the intersection targets were obtained based on the WeChat online tool. The protein-protein interaction network was constructed based on the STRING database created in 2000 by Peer Bork’s team at the European Bioinformatics Institute (EMBL), and the protein-protein interaction relationship was analyzed. The core targets were screened according to the degree value. The intersection targets were subjected to GO and KEGG enrichment analyses. Finally, molecular docking and molecular dynamics simulation were used to verify the binding of key components to key targets. 
    RESULTS AND CONCLUSION: (1) One hundred and forty-two active ingredients of Gegenmaqi prescription were obtained, including 65 intersections between component targets and disease targets, 5 key active ingredients (β-sitosterol, stigmasterol, kaempferol, quercetin, and formononetin), and 5 key targets (AKT1, tumor necrosis factor, interleukin-10, JUN, and TP53). (2) GO function enrichment included 508 items, 390 biological process items, 77 molecular function items and 41 cell component items. KEGG pathway analysis showed 146 pathways, mainly involving advanced glycation end products receptor signaling pathway, lipid and atherosclerosis signaling pathway, tumor necrosis factor signaling pathway, and interleukin-17 signaling pathway. (3) Molecular docking showed that the key components and key targets had good binding activity. Molecular dynamics simulation showed that β-sitosterol had stable interactions with AKT1, tumor necrosis factor and interleukin 10. (4) Gegenmaqi prescription has been comprehensively studied, and the material basis of its pharmacological effect has been primarily clarified. It is predicted that Gegenmaqi prescription can treat periarthritis of shoulder combined with type 2 diabetes through multi-components, multi-targets, and multi-pathways to exert anti-inflammatory and regulate insulin secretion. 
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    Machine learning combined with bioinformatics screening of key genes for pulmonary fibrosis associated with cellular autophagy and experimental validation
    Gong Yuehong, Wang Mengjun, Ren Hang, Zheng Hui, Sun Jiajia, Liu Junpeng, Zhang Fei, Yang Jianhua, Hu Junping
    2025, 29 (35):  7679-7689.  doi: 10.12307/2025.971
    Abstract ( 131 )   PDF (6590KB) ( 463 )   Save
    BACKGROUND: Early diagnosis of pulmonary fibrosis is the foundation for timely antifibrotic drug therapy. Therefore, exploring and discovering ideal biomarkers that can be effectively used for the early diagnosis of pulmonary fibrosis is crucial for the treatment of the disease.
    OBJECTIVE: To conduct an in-depth analysis of key autophagy-related genes involved in the process of pulmonary fibrosis by means of bioinformatics and machine learning techniques, in order to investigate whether autophagy-related core genes of pulmonary fibrosis can be used as reliable biomarkers in the assessment of the progression of pulmonary fibrosis.
    METHODS: Two datasets of pulmonary fibrosis, GSE24206 and GSE110147, were downloaded from the Gene Expression Omnibus (GEO) database (a public database developed and maintained by the U.S. National Center for Biotechnology Information to store and share bioinformatics data), and the gene expression matrices of these two datasets were normalized by using the “limma” package in R software. The autophagy-related genes were extracted from GeneCards database (a database created by the U.S. National Center for Biotechnology Information, which automatically integrates gene-centric data from about 200 Web sources, including genomic, transcriptomic, proteomic, genetic, clinical, and functional information). Differential gene analysis was performed on the pulmonary fibrosis dataset, and the common genes were extracted by cross-comparing the differential genes with the autophagy genes, so as to identify autophagy genes that may play a role in the process of pulmonary fibrosis. The intersecting genes were analyzed for functional enrichment and cellular immune infiltration by gene ontology and Kyoto Encyclopedia of Genes and Genomes. Core genes of pulmonary fibrosis associated with autophagy were screened by protein-protein interactions and machine learning, and core genes were subjected to the enrichment analysis. Diagnostic models were constructed from the identified core genes. Calibration curves were used to assess the predictive ability of the line graph model. An external dataset, GSE21369, was used to perform a receiver operating characteristic curve analysis to validate the expression profiles of pulmonary fibrosis genes associated with autophagy, as well as to predict Chinese herbs associated with the genes IL6 and COL1A2 via the Coremine database. Finally, human embryonic lung fibroblasts were cultured and modelled by transforming growth factor-β1 treatment, and the relative expression of genes in the model cells was verified using qRT-PCR.
    RESULTS AND CONCLUSION: (1) A total of 51 pulmonary fibrosis differential genes and 25 genes intersecting with autophagy genes were obtained. Gene ontology analysis showed that the 25 intersecting genes were related to extracellular matrix tissue, collagen metabolism, collagen pro-fibroblasts, and growth factor binding, etc. The results of Kyoto Encyclopedia of Genes and Genomes enrichment analysis indicated that they were mainly related to the Phosphatidylinositol 3-kinase/protein kinase B signaling pathway and the signaling pathway of the extracellular matrix-receptor interactions. (2) Immunoinfiltration analysis revealed that the expression of activated memory CD4+ T cells, M0 macrophages, and resting dendritic cells was significantly elevated in the pulmonary fibrosis group (P < 0.05), showing a strong correlation. (3) Two autophagy signature genes involved in the progression of pulmonary fibrosis were identified: COL1A2 and IL6. The column-line diagram model showed that the two core genes predicted the onset of pulmonary fibrosis more accurately, and the receiver operating characteristic curve analysis showed that the two characteristic genes had diagnostic significance. COL1A2 and IL6 were related to the cell-cycle pathway, mitogen-activated protein kinase signaling pathway, Janus kinase-signal transduction and activator of transcription signaling pathway and cytokine-cytokine receptor interactions. A total of 20 Chinese herbs were predicted to be related to COL1A2 and IL6 genes, and their efficacies were mainly to clear away heat and detoxify toxins and to invigorate blood and move qi. COL1A2 and IL6 were verified to be highly expressed in pulmonary fibrosis. To conclude, COL1A2 and IL6 may be potential diagnostic biomarkers for pulmonary fibrosis, but its specificity to pulmonary fibrosis needs to be further investigated.
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    Identification of immunodiagnostic biomarkers and drug screening for steroid-induced osteonecrosis of the femoral head
    Han Jie, Pan Chengzhen, Shang Yuzhi, Zhang Chi
    2025, 29 (35):  7690-7700.  doi: 10.12307/2025.973
    Abstract ( 126 )   PDF (10068KB) ( 101 )   Save
    BACKGROUND: The pathogenesis of steroid-induced osteonecrosis of the femoral head is not yet fully understood. A deep comprehension of the pathology and molecular mechanisms of steroid-induced osteonecrosis of the femoral head, as well as the search for diagnostic markers with high specificity and sensitivity, are crucial for the prevention and treatment of this condition.
    OBJECTIVE: To identify immune diagnostic markers for steroid-induced osteonecrosis of the femoral head and predict potential drug targets through drug enrichment analysis and molecular docking techniques.
    METHODS: The study utilized gene expression profile data (GSE123568 and GSE74089) from the GEO databases (a public gene expression database built by the U.S. National Center for Biotechnology Information). R software was used for data normalization and differential gene screening, followed by weighted gene co-expression network analysis (WGCNA) to identify disease-related genes. Immune-related genes were obtained from the GeneCards database and intersected with the differential genes and WGCNA gene sets to select immune-related genes for steroid-induced osteonecrosis of the femoral head. Mendelian randomization was used to validate the potential causal relationship between these immune-related genes and steroid-induced osteonecrosis of the femoral head. Gene Set Enrichment Analysis was conducted to analyze the immune-related pathways involved, and protein-protein interaction networks were used to assess functional associations. Finally, drug enrichment analysis and molecular docking were performed to predict potential drugs targeting these immune-related genes.
    RESULTS AND CONCLUSION: Three key immune-related genes - RNASEL, SECTM1, and HSPA6 - were identified. These genes were highly expressed in steroid-induced osteonecrosis of the femoral head and exhibited good diagnostic potential, which were involved in multiple immune-related signaling pathways. Mendelian randomization analysis confirmed their potential causal relationship with steroid-induced osteonecrosis of the femoral head. Drug enrichment analysis and molecular docking identified nine potential drugs, including β-ecdysterone, showing the possibility of intervening in the pathological process of steroid-induced osteonecrosis of the femoral head by regulating the HSPA6 protein. These findings provide new biomarkers and drug targets for the early diagnosis and personalized treatment of steroid-induced osteonecrosis of the femoral head. They also highlight the potential application of bioinformatics in Chinese biomedical research, facilitating the integration and translational use of international data in local disease studies.
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