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28 December 2026, Volume 30 Issue 36 Previous Issue   

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Aucubin promotes osteogenic differentiation of mouse cranial pre-osteoblasts MC3T3-E1 Wang Houyuan, Xiao Jiacong, Shao Shoujia, He Paian, Yang Nan, Jiang Ziwei 2026, 30 (36):  9381-9392.  doi: 10.12307/2026.908 Abstract ( 95 )   PDF (16786KB) ( 27 )   Save BACKGROUND: Oxidative stress is a major contributor to osteoporosis and other bone metabolic disorders. Aucubin, a natural compound with antioxidant and osteogenic effects, shows therapeutic potential, but its mechanisms remain unclear. OBJECTIVE: To investigate the potential targets and pathways through which aucubin promotes osteogenic differentiation using network pharmacology combined with in vitro validation. METHODS: Potential aucubin targets related to osteogenesis were identified from TCMSP and GeneCards databases, and a protein-protein interaction network with enrichment analysis was constructed to predict target pathways. An oxidative stress model of MC3T3-E1 cells was established using hydrogen peroxide, and the cells were assigned to control, model, low- and high-dose aucubin intervention, and Aucubin+JAK2/STAT3 inhibitor groups. Cell viability and malondialdehyde level were detected, and alkaline phosphatase staining and alizarin red staining were performed. Western blot and RT-qPCR were used to detect the expression of Janus kinase 2/signal transducer and activator of transcription 3 pathway-related proteins and osteogenic proteins and genes. RESULTS AND CONCLUSION: (1) Network pharmacology predicted that aucubin functioned mainly via the Janus kinase 2/signal transducer and activator of transcription 3 pathway, involving key targets such as Janus kinase 2, signal transducer and activator of transcription 3, Janus kinase 1, interleukin 2, and matrix metalloproteinase 9. (2) In vitro experiments demonstrated that aucubin reduced oxidative stress, decreased malondialdehyde levels, enhanced alkaline phosphatase activity and mineralization, and upregulated the expression of pathway proteins as well as the gene and protein expression of osteogenic markers (Runt-related transcription factor 2, osteocalcin, osteopontin). These effects were partially reversed by the pathway inhibitor AG490. Taken together, these findings indicate that aucubin alleviates oxidative stress and promotes osteogenic differentiation of MC3T3-E1 cells through activation of the Janus kinase 2/signal transducer and activator of transcription 3 pathway. Figures and Tables | References | Related Articles | Metrics

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Mechanism of high glucose-induced osteoblast cuproptosis Lian Yong, Xie Zhixing, Lu Jiaxin, Pan Zhaofeng, Chen Qingzhen, Shao Min 2026, 30 (36):  9393-9401.  doi: 10.12307/2026.381 Abstract ( 87 )   PDF (2164KB) ( 29 )   Save BACKGROUND: Cuproptosis is a newly identified copper-dependent cell death. It has attracted more attention to various diseases in recent years. Cuproptosis has been studied in cancer, but its role in osteoblasts remains unclear. OBJECTIVE: To investigate the mechanism of high glucose-induced cuproptosis in osteoblasts. METHODS: (1) MC3T3-E1 cells were cultured in four groups: 20 μmol/L CuCl2 group, 40 μmol/L CuCl2 group, 20 μmol/L CuCl2+high glucose group, and 40 μmol/L CuCl2+high glucose group. The high glucose environment was simulated by adding 25 mmol/L glucose and 200 mmol/L sodium palmitate. After 48 hours of osteogenic induction, western blot assay was used to detect the protein expression of copper ion transporters (copper ion influx transporter SLC31A1 and copper ion efflux transporter ATP7B). (2) MC3T3-E1 cells were cultured in four groups: the normal group without any treatment, the high glucose group with the addition of 25 mmol/L glucose and 200 mmol/L sodium palmitate, the CuCl2 group with the addition of 20 μmol/L CuCl2, and the high glucose + CuCl2 group with the simultaneous addition of 25 mmol/L glucose, 200 mmol/L sodium palmitate, and 20 μmol/L CuCl2. After osteogenic induction, mitochondrial alterations were observed via transmission electron microscopy. The osteoblast differentiation and mineralization were evaluated using alkaline phosphatase staining and alizarin red staining. qRT-PCR was used to detect osteoblast-specific gene expression, and western blot was used to detect osteoblast- and cuproptosis-related protein expression. RESULTS AND CONCLUSION: (1) Western blot analysis showed that the expression of SLC31A1 protein was higher in the 20 μmol/L CuCl2+high glucose group than the 20 μmol/L CuCl2 group (P < 0.05), while the expression of ATP7B protein was lower (P < 0.05). In the 40 μmol/L CuCl2+high glucose group, the expression of SLC31A1 protein was higher than in the 40 μmol/L CuCl2 group (P < 0.05), and the expression of ATP7B protein was lower (P < 0.05). (2) Under the transmission electron microscopy, CuCl2 induced changes in the mitochondrial structure of MC3T3-E1 cells, primarily characterized by the disappearance of mitochondrial cristae, mitochondrial shrinkage, reduced volume, and disruption of mitochondrial membrane structure. The combined exposure to high glucose exacerbated these structural alterations. Alkaline phosphatase and alizarin red staining results indicated that CuCl2 inhibited osteogenic differentiation and mineralization of MC3T3-E1 cells, and the inhibitory effect of CuCl2+high glucose environment on osteogenic differentiation and mineralization was stronger than that of CuCl2 alone. qRT-PCR and western blot analysis revealed that the CuCl2+high glucose environment significantly inhibited the expression of osteogenic-specific gene at mRNA and protein levels, inducing osteoblast cuproptosis. These findings suggest that high glucose may induce and exacerbate osteoblast cuproptosis by disrupting copper transport and cuproptosis-related protein expression.  Figures and Tables | References | Related Articles | Metrics

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Mechanisms of Liuwei Dihuang Pills in a rat model of postmenopausal osteoporosis Xu Pengchao, Xie Lihua, Huang Jingwen, Chen Juan, Chen Xuan, Li Shengqiang, Ge Jirong 2026, 30 (36):  9402-9412.  doi: 10.12307/2026.867 Abstract ( 85 )   PDF (6310KB) ( 18 )   Save BACKGROUND: Liuwei Dihuang Pill has been widely used in the treatment of osteoporosis with liver and kidney yin deficiency syndrome, but its therapeutic targets are still unclear and need further research. OBJECTIVE: To analyze the mechanism of Liuwei Dihuang Pills in the treatment of postmenopausal osteoporosis using network pharmacology and single-cell sequencing. METHODS: (1) The active ingredients and targets of each traditional Chinese medicine component of Liuwei Dihuang Pills were analyzed using the TCMSP and ETCM databases. The genes related to osteoporosis were retrieved from the GeneCards, OMIM, Disgenet, PharmGkb, and TTD databases. The drug-disease-target network was constructed and the core genes were screened by using analytical tools such as Cytoscape and STRING. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were performed by core genes. (2) Nine Sprague-Dawley rats were randomly divided into sham group (n=3), model group (n=3) and Liuwei Dihuang Pill group (n=3). Sham group did not undergo modeling, while the model and Liuwei Dihuang Pill group were modeled by bilateral oophorectomy. Four weeks after modeling, the Liuwei Dihuang Pill group was administered by gavage with a suspension prepared by Liuwei Dihuang Pills, while the sham group and the model group were administered by gavage with the same amount of normal saline. Administration in each group was given twice daily, for 12 consecutive weeks. After the final administration, peripheral blood was collected for single-cell sequencing and network pharmacology analyses.  RESULTS AND CONCLUSION: (1) There were a total of 116 intersection targets between Liuwei Dihuang Pills and osteoporosis. The "drug-disease-target" network diagram and protein-protein interaction analysis indicated that 15 genes such as B-cell lymphoma 2, Fos proto-oncogene 1, estrogen receptor 1, and interleukin-10 might be its core targets. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses indicated that the therapeutic mechanism of Liuwei Dihuang Pills might be related to osteoclast differentiation, cell senescence and apoptosis, steroid hormone regulation and inflammatory response. (2) Single-cell sequencing indicated that monocytes might be the key cell population in the treatment of postmenopausal osteoporosis with Liuwei Dihuang Pills. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses indicated that mononuclear macrophage populations were closely related to immunity, apoptosis, reactive oxygen species metabolism, etc. The combined analysis of single-cell sequencing and network pharmacology indicated that Liuwei Dihuang Pills for postmenopausal osteoporosis was mainly achieved through osteoclast and apoptosis-related pathways. To conclude, Liuwei Dihuang Pills treat postmenopausal osteoporosis through multiple targets, multiple effects and multiple mechanisms, and its main mechanism is related to the differentiation and apoptosis of osteoclasts. The CD16+ monocyte subset is the key cell subset responsible for this effect. Figures and Tables | References | Related Articles | Metrics

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Identification of antimicrobial peptides as key therapeutic targets for necrosis by sodium overload in osteoarthritis using multiple machine learning approaches: cytological validation Wang Huaijing, Guo Jinrong, Wan Dongping, Mei Qijie, Yuan Jingzhao, Xu Wenfei, Zeng Chao, Zheng Haijun, Yuan Changshen, Duan Kan 2026, 30 (36):  9413-9422.  doi: 10.12307/2026.910 Abstract ( 95 )   PDF (7517KB) ( 17 )   Save BACKGROUND: Necrosis by sodium overload (NECSO) plays a significant role in cardiovascular diseases, but its mechanism in osteoarthritis remains unclear. OBJECTIVE: To screen key genes related to NECSO in osteoarthritis through bioinformatics and to explore their mechanisms in osteoarthritis. METHODS: Osteoarthritis microarray datasets GSE117999 and GSE169077 were obtained from the GEO database. Differentially expressed genes were intersected with NECSO-related genes to identify osteoarthritis-NECSO differentially expressed genes. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses and protein-protein interaction network analysis were performed to obtain Hub genes. Immune cell infiltration analysis and weighted gene co-expression network analysis were conducted to further screen NECSO-related and immune-related differentially expressed genes in osteoarthritis. Key genes were screened using least absolute shrinkage and selection operator, extreme gradient boosting, and random forest methods to construct and validate a diagnostic model. The correlation between key NECSO genes and diagnostic genes was analyzed using the Pearson’s correlation analysis, and potential drugs targeting diagnostic genes were predicted. Finally, cell experiments were conducted for validation. RESULTS AND CONCLUSION: (1) Nine osteoarthritis-NECSO differentially expressed genes were identified, mainly enriched in biological processes such as cellular serine hydrolase activity, serine-type peptidase activity, and serine-type endopeptidase activity, as well as the interleukin-17 signaling pathway. (2) Immune infiltration analysis revealed differential expression of plasma cells and resting dendritic cells, suggesting these two immune cells play certain roles in osteoarthritis pathogenesis. (3) Weighted gene co-expression network analysis identified three genes: lactoferrin, cathelicidin antimicrobial peptide, and S100 calcium-binding protein A8. Combined with immune infiltration analysis, one gene, alpha-S1-casein, was identified. (4) Protein-protein interaction analysis identified five Hub genes: matrix metalloproteinase 3, apolipoprotein D, cathelicidin antimicrobial peptide, S100 calcium-binding protein A8, and lactoferrin. (5) Machine learning screened out one diagnostic gene, cathelicidin antimicrobial peptide, which was correlated with Hub genes. (6) Five drugs targeting the diagnostic gene were predicted: PEG-conjugated Toll-like receptor 7/8 agonist NKTR-262, ropocamptide, clobetasol 17-butyrate, recombinant vesicular stomatitis virus expressing interferon-beta and tyrosinase-related protein 1, and giloralimab. (7) Cell experiments showed significantly higher expression of cathelicidin antimicrobial peptide protein in chondrocytes treated with interleukin-1β compared with the blank group. These findings suggest that the cathelicidin antimicrobial peptide gene has diagnostic value for osteoarthritis, and NECSO-related genes play a role in its pathogenesis. Figures and Tables | References | Related Articles | Metrics

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Differences in visual deprivation and visual interference for postural control on unstable surfaces in individuals with chronic ankle instability Liu Mengyu, Shen Yurui, Zhao Yuting, Yang Kexin, Zhou Zhipeng, Zheng Liangliang 2026, 30 (36):  9423-9428.  doi: 10.12307/2026.915 Abstract ( 67 )   PDF (1596KB) ( 29 )   Save BACKGROUND: Postural control ability is significantly decreased in individuals with chronic ankle instability, and this decline is closely related to vision, vestibular sensation, and proprioception.   OBJECTIVE: To investigate the effects of visual conditions and support conditions on static postural control ability in individuals with chronic ankle instability.   METHODS: Twenty-six participants with chronic ankle instability were recruited. Each participant completed a single-leg stance test under three visual conditions (eyes open, stroboscopic vision, and eyes closed) and two support conditions (firm surface and foam pad). Center of pressure data were collected using a three-dimensional force platform under different test conditions. Two-way repeated measures analysis of variance was used to examine the effects of visual and support conditions on postural control ability in individuals with chronic ankle instability.   RESULTS AND CONCLUSION: (1) For the mean velocity of center of pressure displacement in the mediolateral direction, there was a significant interaction between visual and support conditions (P < 0.01). Under the same support condition, the mean velocity of displacement in the stroboscopic vision group was higher than that in the eyes-open group (P < 0.05), and the mean velocity of displacement in the eyes-closed group was higher than that in the stroboscopic vision group (P < 0.05). Under the same visual condition, the mean velocity of displacement in the foam pad support group was higher than that in the firm surface support group (P < 0.05). For the mean velocity of center of pressure displacement in the anteroposterior direction, there was also a significant interaction between visual and support conditions (P < 0.05). Under the same support condition, the mean velocity of displacement in the stroboscopic vision group was greater than that in the eyes-open group (P < 0.05), and the mean velocity of displacement in the eyes-closed group was greater than that in the stroboscopic vision group (P < 0.05). Under the eyes-closed condition, the mean velocity of displacement in the foam pad support group was greater than that in the firm surface support group (P < 0.05). (2) For the maximum displacement of the center of pressure in the mediolateral direction, there was no significant interaction between support and visual conditions (P > 0.05). On the firm surface, the maximum displacement in the stroboscopic vision group and eyes-closed group was longer than that in the eyes-open group (P < 0.05). On the foam pad, the maximum displacement in the stroboscopic vision group was longer than that in the eyes-open group (P < 0.05), and the maximum displacement in the eyes-closed group was longer than that in the stroboscopic vision group (P < 0.05). Under stroboscopic vision or eyes-closed conditions, the maximum displacement in the foam pad support group was longer than that in the firm surface support group (P < 0.05). (3) For the 95% ellipse area of center of pressure displacement, there was no significant interaction between support and visual conditions (P > 0.05). Under the same support condition, the 95% ellipse area in the stroboscopic vision group and eyes-closed group was greater than that in the eyes-open group (P < 0.05). To conclude, these findings indicate that visual interference and unstable surfaces significantly impair postural control in individuals with chronic ankle instability. Therefore, future rehabilitation programs can utilize unstable surfaces combined with stroboscopic glasses to design progressive functional rehabilitation training.   Figures and Tables | References | Related Articles | Metrics

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Effects of icariin-containing serum active ingredients on inflammatory chondrocytes: a metabolomics analysis Chen Weinan, Li Yusheng, Li Linzhen, Jiao Hongzhuo, Zhang Mingzhe, Wang Jianlong, Zhang Juntao 2026, 30 (36):  9429-9436.  doi: 10.12307/2026.907 Abstract ( 62 )   PDF (8376KB) ( 18 )   Save BACKGROUND:  Icariin, the main active component of the herb Epimedium, has been confirmed to possess cartilage-protective effects, but its in vivo metabolic transformation and mechanisms remain incompletely elucidated. In recent years, metabolomics technology has provided new tools for analyzing the complex in vivo metabolic processes of traditional Chinese medicine (TCM), while studies on medicated serum offer a more realistic reflection of the pharmacodynamic material basis of TCM.  OBJECTIVE: To analyze the pharmacodynamic material basis of icariin-containing serum using untargeted metabolomics. METHODS: Untargeted metabolomics was employed to compare differential metabolites and related signaling pathways between icariin-containing serum and blank serum. The cell counting kit-8 assay was used to evaluate the effect of gentisic acid on the viability of C57BL/6J mouse chondrocytes and determine the optimal concentration. A lipopolysaccharide-induced inflammatory chondrocyte model was established. After 24 hours of intervention with 75 μmol/L gentisic acid, mitochondrial membrane potential and mRNA expression of ATP5F1A and ATP5F1B mRNA were detected. RESULTS AND CONCLUSION: (1) Untargeted metabolomics revealed that the effects of icariin-containing serum might be associated with taurine and hypotaurine metabolism pathways, with the key metabolite gentisic acid influencing chondrocyte energy metabolism by regulating mitochondrial ATP synthase (ATP5F1A/ATP5F1B). (2) Gentisic acid promoted chondrocyte proliferation and restored mitochondrial membrane potential in inflammation-induced chondrocytes. (3) Gentisic acid significantly increased the expression levels of ATP5F1A and ATP5F1B in lipopolysaccharide-treated chondrocytes. These findings indicate that icariin-containing serum has a defined metabolic material basis for promoting cartilage repair, with the taurine and hypotaurine metabolism pathways serving as its core mechanism. This may involve promoting L-cysteine expression and modulating these metabolic pathways to exert anti-inflammatory effects, thereby maintaining cartilage homeostasis and facilitating repair. Gentisic acid may interact with ATP5F1A/ATP5F1B proteins to regulate chondrocyte energy metabolism, contributing to cartilage repair, and holds promise as a potential therapeutic agent for knee osteoarthritis. Figures and Tables | References | Related Articles | Metrics

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Association between mitochondrial dynamics imbalance and cartilage degeneration in osteoarthritis Fang Fengchao, Yang Wenwu, Wu Hongtao, Peng Peng, Liang Zhiye, Jiang Tao, Chen Weijian, Liu Wengang, Zhao Chuanxi 2026, 30 (36):  9437-9444.  doi: 10.12307/2026.918 Abstract ( 36 )   PDF (2213KB) ( 5 )   Save BACKGROUND: During the pathogenesis of osteoarthritis, chondrocytes exhibit insufficient energy supply. As mitochondria serve as the energy generators of cells, it remains unclear how changes in mitochondrial dynamic homeostasis affect chondrocyte function and cartilage degeneration. OBJECTIVE: To investigate the effect of mitochondrial dynamics homeostasis in articular chondrocytes on cartilage degeneration in knee osteoarthritis. METHODS: (1) Sprague-Dawley rats were randomly divided into a control group and a model group. The model group underwent anterior cruciate ligament transection to establish a rat model of knee osteoarthritis, and all rats were normally fed for 4 weeks. Articular cartilage changes were observed using hematoxylin-eosin and safranin O-fast green staining. The protein and mRNA expression levels of optic atrophy 1 and dynamin-related protein 1 in cartilage, as well as mitochondrial DNA expression, were detected using immunohistochemistry, immunofluorescence, western blot, and RT-PCR. Adenosine triphosphate expression was measured using an ATP colorimetric assay. (2) Additionally, 10 tibial plateau cartilage samples were collected. Five samples from patients without osteoarthritis undergoing amputation or trauma surgery served as the normal group, and five samples from patients with knee osteoarthritis undergoing knee replacement surgery served as the lesion group. Western blot and RT-PCR were used to examine the above indicators, along with the expression of mitofusin 1 and mitochondrial fission protein 1. RESULTS AND CONCLUSION: (1) Cartilage observation indicators in rat models: Compared with the control group, the model group showed thinning of the cartilage layer, destruction of subchondral bone structure, disorganized chondrocyte arrangement, and reduced cell number. The levels of the mitochondrial fusion protein (optic atrophy 1) and its mRNA expression, as well as mitochondrial DNA expression, were decreased in chondrocytes, while the levels of the mitochondrial fission protein (dynamin-related protein 1) and its mRNA expression were increased, and adenosine triphosphate content was decreased. (2) Human tibial plateau cartilage observation indicators: Compared with the normal group, the lesion group showed decreased protein expression of mitochondrial fusion proteins (optic atrophy 1 and mitofusin 1) and their mRNA expression, as well as decreased mitochondrial DNA expression, while the protein expression of mitochondrial fission proteins (dynamin-related protein 1 and mitochondrial fission protein 1) and their mRNA expression were increased, and adenosine triphosphate content was decreased. In conclusion, the imbalance of mitochondrial dynamics homeostasis in articular chondrocytes is manifested by downregulation of mitochondrial fusion protein expression, upregulation of fission protein expression, and impaired adenosine triphosphate synthesis, which is significantly positively correlated with cartilage damage. Figures and Tables | References | Related Articles | Metrics

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Warm acupuncture inhibits pyroptosis and improves cartilage degeneration in rabbit knee joints Liu Di, Wang Minglei, Li Chun, Wang Duo, Li Long, Wu Yongli 2026, 30 (36):  9445-9452.  doi: 10.12307/2026.898 Abstract ( 69 )   PDF (2854KB) ( 9 )   Save BACKGROUND: Warm acupuncture has a significant therapeutic effect on knee osteoarthritis. It can improve articular cartilage degeneration by inhibiting periarticular inflammation. However, whether this effect is related to pyroptosis mediated by the Toll-like receptor 4/nuclear factor-κB (TLR4/NF-κB) signaling pathway remains unclear. OBJECTIVE: To investigate the mechanism of warm acupuncture in the treatment of knee osteoarthritis from the perspective of pyroptosis. METHODS: Forty New Zealand rabbits were randomly divided into blank, model, western medicine, and warm acupuncture groups using a random number table, with 10 rabbits in each group. The blank group received no intervention, while knee osteoarthritis models were established in the other three groups using the plaster cast fixation method. Four weeks after modeling, the model group was immobilized in a fixator for 15 minutes once daily for 2 consecutive weeks. The warm acupuncture group received warm acupuncture at “Heding” (EX-LE2), “Neixiyan” (EX-LE4), and “Waixiyan” (ST35) for 15 minutes once daily for 2 weeks. The western medicine group received celecoxib by gavage once daily for 2 weeks. Samples were collected on the second day after treatment. Interleukin-1β and interleukin-18 levels in synovial fluid were detected by ELISA. Morphological changes in knee cartilage were observed by hematoxylin-eosin staining. The expression of TLR4, NF-κB p65, Caspase-1, and gasdermin D in knee cartilage was detected by qPCR, immunohistochemistry, and western blot. RESULTS AND CONCLUSION: (1) Interleukin-1β and interleukin-18 levels were significantly higher in the model group than in the blank group (P < 0.05), and significantly lower in the warm acupuncture and western medicine groups than in the model group (P < 0.05). (2) Hematoxylin-eosin staining results showed that the cartilage surface in the model group was rough and significantly damaged, with irregular and disorganized chondrocyte distribution. In the warm acupuncture and western medicine groups, the cartilage surface was relatively smooth, and chondrocytes were more uniformly and regularly distributed. (3) Results from qPCR, immunohistochemistry, and western blot showed that compared with the blank group, the mRNA and protein expression levels of TLR4, NF-κB p65, Caspase-1, and gasdermin D were significantly increased in the model group (P < 0.05). Compared with the model group, the mRNA and protein expression levels of TLR4, NF-κB p65, Caspase-1, and gasdermin D were significantly decreased in the warm acupuncture and western medicine groups (P < 0.05). These findings indicate that warm acupuncture can alleviate articular cartilage degeneration in rabbits with knee osteoarthritis by regulating the TLR4/NF-κB signaling pathway to downregulate the expression of pyroptosis-related proteins Caspase-1 and gasdermin D, as well as downstream inflammatory factors interleukin-1β and interleukin-18, thereby inhibiting pyroptosis. Figures and Tables | References | Related Articles | Metrics

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Du Huo Ji Sheng Decoction delays cartilage degeneration in mice with knee osteoarthritis Ma Zhiguo, Zhou Zhengxin, Xue Song, Xiang Xinhong, Zhu Lei 2026, 30 (36):  9453-9461.  doi: 10.12307/2026.905 Abstract ( 56 )   PDF (2142KB) ( 14 )   Save BACKGROUND: Modified Du Huo Ji Sheng Decoction has clinical advantages in the treatment of knee osteoarthritis. However, the specific mechanism by which it ameliorates cartilage degeneration through regulating the Hedgehog signaling pathway remains unclear. OBJECTIVE: To investigate the protective effects of modified Du Huo Ji Sheng Decoction on cartilage degeneration in a mouse model of knee osteoarthritis. METHODS: Fifty male C57BL/6J mice were randomly divided into five groups: sham-operated group, model group, low-dose modified Du Huo Ji Sheng Decoction group, high-dose modified Du Huo Ji Sheng Decoction group, and celecoxib group (positive control). Mice in the sham-operated group underwent only skin incision and joint cavity exposure as a simulated surgical procedure, while mice in the other groups underwent medial meniscus destabilization surgery to establish a knee osteoarthritis model. Fourteen days after modeling, intragastric administration was initiated. The low-dose and high-dose modified Du Huo Ji Sheng Decoction groups received doses of 1.34 and 4.02 g/kg, respectively, and the celecoxib group received 26 mg/(kg·d) celecoxib. All treatments were administered once daily for 8 weeks. Histopathological staining, Micro-CT scanning, immunohistochemistry, real-time quantitative PCR, and western blot assay were employed to assess the degree of cartilage degeneration, subchondral bone microstructure, cartilage matrix metabolism-related proteins (type II collagen and matrix metalloproteinase 13), and the expression levels of key Hedgehog signaling pathway genes and proteins (Patched 1, Indian hedgehog, and Smoothened). RESULTS AND CONCLUSION: (1) Toluidine blue staining results showed that, compared with the sham-operated group, the model group exhibited erosive damage on the cartilage surface, disordered tissue structure, and an elevated Osteoarthritis Research Society International score. Compared with the model group, each dosage group of modified Du Huo Ji Sheng Decoction showed restored cartilage surface smoothness, progressively regular cartilage arrangement, and reduced Osteoarthritis Research Society International scores. (2) Micro-CT three-dimensional reconstruction of the knee joint subchondral bone showed that, compared with the sham-operated group, the model group exhibited significantly reduced medullary cavity area in the subchondral bone region, presenting a hardened and dense subchondral bone structure, with increased bone volume fraction and trabecular thickness, and decreased trabecular number and trabecular separation. Compared with the model group, each dosage group of modified Du Huo Ji Sheng Decoction showed increased medullary cavity area in the subchondral bone, reduced subchondral bone sclerosis, decreased bone volume fraction and trabecular thickness, and restoration of trabecular number and trabecular separation to near-normal levels. (3) Immunohistochemical results showed that type II collagen expression was decreased and matrix metalloproteinase 13 expression was increased in the cartilage of the model group. After intervention with modified Du Huo Ji Sheng Decoction, their expression was restored. (4) TUNEL staining results showed that the number of positive cells in the cartilage of the model group was increased, and after intervention with modified Du Huo Ji Sheng Decoction, the number of positive cells was significantly reduced. (5) Real-time quantitative PCR results showed that the expression of Hedgehog signaling pathway-related genes Patched 1, Indian hedgehog, and Smoothened was increased in the cartilage of the model group. After intervention with modified Du Huo Ji Sheng Decoction, these showed a dose-dependent decrease. (6) Western blot results showed that the expression of Patched 1, Indian hedgehog, Smoothened, and matrix metalloproteinase 13 was increased, and type II collagen expression was decreased in the cartilage of the model group. After intervention with modified Du Huo Ji Sheng Decoction, this trend was significantly reversed. These findings indicate that modified Du Huo Ji Sheng Decoction can restore the balance between synthesis and catabolism of the cartilage matrix, inhibit chondrocyte apoptosis, and delay cartilage degeneration in knee osteoarthritis by regulating the Hedgehog signaling pathway. Figures and Tables | References | Related Articles | Metrics

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Shaoyao Gancao Decoction combined with shock wave therapy for treating gluteus medius syndrome: a randomized controlled trial on structural improvement of the tendon Shi Tengbo, Zhang Mengyu, Wang Xingfei, Fu Zhuo, Li Chenyang, Meng Xiangxi, Liu Hao, Guo Chaowei, He Zike, Tang Yanfeng 2026, 30 (36):  9462-9468.  doi: 10.12307/2026.901 Abstract ( 57 )   PDF (4454KB) ( 13 )   Save BACKGROUND: Gluteus medius syndrome is a chronic painful disorder characterized by tendon degeneration, aseptic inflammation, or myofascial injury of the gluteus medius muscle. Current Western treatments primarily include extracorporeal shock wave therapy and nonsteroidal anti-inflammatory drugs, but long-term efficacy remains limited. Shaoyao Gancao Decoction, a classical formula, is known for its blood-activating and pain-relieving effects. However, the clinical efficacy of its combination with extracorporeal shock wave remains unclear. OBJECTIVE: To evaluate the clinical efficacy of Shaoyao Gancao Decoction combined with extracorporeal shock wave therapy in treating gluteus medius syndrome (qi stagnation and blood stasis syndrome) and assess its synergistic effects on pain relief, functional recovery, and inflammation modulation. METHODS: A prospective randomized controlled trial was conducted, enrolling 165 patients with gluteus medius syndrome meeting both Western and traditional Chinese medicine diagnostic criteria. Participants were randomly divided into three groups: the extracorporeal shock wave therapy group (extracorporeal shock wave therapy alone, every 3 days for a total of 4 consecutive sessions), the Shaoyao Gancao Decoction group (Shaoyao Gancao Decoction alone, once daily for 15 days), and the combination group (Shaoyao Gancao Decoction + extracorporeal shock wave therapy), with 55 patients in each group. Patients’ visual analog scores, Harris hip scores, and ultrasound echo intensity were compared before and after treatment. Serum levels of inflammatory cytokines (prostaglandin E2, substance P, interleukin-6, tumor necrosis factor-α) were also detected to evaluate the efficacy and adverse reactions of traditional Chinese medicine. RESULTS AND CONCLUSION: (1) The combination group exhibited significantly lower visual analog scale scores at 3 day, 1 week, and 3 months after treatment compared with monotherapy groups (P < 0.05). At 3 months post-treatment, the Harris Hip Score in the combination group (86.4) surpassed those of the extracorporeal shock wave therapy group (79.6) and the Shaoyao Gancao Decoction group (76.8). (2) The repair of gluteus medius tendon structure in the combination group was more significant, and the echo intensity was 112.6, which was better than that of the extracorporeal shock wave therapy group (138.7) and the Shaoyao Gancao Decoction group (158.4). (3) Inflammatory markers (prostaglandin E2, substance P, interleukin-6, tumor necrosis factor-α) decreased more markedly in the combination group (P < 0.05). (4) The total effective rate was 80% in the combination group, significantly higher than that in the extracorporeal shock wave therapy group (64%) and the Shaoyao Gancao Decoction group (59.2%), with a lower incidence of adverse events. To conclude, the combination of Shaoyao Gancao Decoction and extracorporeal shock wave therapy synergistically alleviates pain, promotes functional recovery, and suppresses inflammatory responses in gluteus medius syndrome, demonstrating superior efficacy to monotherapies with a favorable safety profile. Figures and Tables | References | Related Articles | Metrics

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Effect of shock wave therapy combined with facilitated stretching on the electromyographic characteristics of the upper trapezius muscle in patients with varying degrees of neck and shoulder pain Gong Xun, Zhou Yong, Ji Lina 2026, 30 (36):  9469-9478.  doi: 10.12307/2026.704 Abstract ( 79 )   PDF (1641KB) ( 11 )   Save BACKGROUND: Traditional monotherapy (such as medication and physical therapy) for neck and shoulder myofascial pain syndrome has high recurrence rates and limited improvement in deep myofascial adhesions. Shock wave therapy combined with facilitated stretching may be more effective in restoring muscle function through mechanical and metabolic regulation, but there is currently a lack of dynamic assessment evidence on neuromuscular activation characteristics in patients with different pain levels. OBJECTIVE: To observe the effect of shockwave therapy combined with facilitated stretching on the electromyographic characteristics of the upper trapezius muscle in patients with varying degrees of neck and shoulder pain.  METHODS: A total of 120 patients with neck and shoulder pain admitted to the Second Affiliated Hospital of Hainan Medical University from January 2019 to December 2023 were selected as the research subjects and randomly divided into a control group (60 cases) and an observation group (60 cases). The observation group was treated with shock wave therapy combined with facilitated stretching, while the control group was treated with facilitated stretching alone. All treatments were given thrice a week for 6 consecutive weeks. Both groups underwent tests for neck pain and disability index, visual analogue scale for subjective pain assessment, and surface electromyography before and after treatment.  RESULTS AND CONCLUSION: (1) The clinical efficacy rate of patients with different pain levels in the observation group was higher than that in the control group (P < 0.05). (2) Compared with before treatment, both groups showed a downward trend in neck pain and disability index and visual analogue scale scores after treatment, with the observation group being lower than the control group (P < 0.05); the shoulder joint function scores (Constant-Murley scores) in both groups showed an upward trend after treatment, with the observation group being higher than the control group (P < 0.05). (3) The resting absolute and relative values of surface electromyography showed a downward trend after treatment, with the observation group being lower than the control group (P < 0.05); the frequency domain indicators of muscle fatigue, median frequency and mean power frequency, increased after treatment, with the observation group being higher than the control group (P < 0.05). (4) The analysis of electromyographic indicators using the generalized estimating equation model at different time points, in different groups, and with different levels of pain showed that the therapeutical effect of the observation group was superior to that of the control group for patients with different pain levels. (5) There was a negative correlation between the resting absolute and relative values of surface electromyography and visual analogue scale (P < 0.05), and between mean power frequency and median frequency with visual analogue scale score (P < 0.05). In conclusion, shock wave therapy combined with facilitated stretching can effectively alleviate muscle tension in the upper trapezius of patients with varying degrees of neck and shoulder pain, and thereby relieve shoulder pain. Figures and Tables | References | Related Articles | Metrics

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Effects of low-frequency pulsed electromagnetic fields on skeletal muscle structure in post-stroke hemiplegic patients and the potential mechanism for health promotion Li Wenhao, Sun Tai, Li Boyang, Sun Shiqiang, Du Xinran, Wang Youpeng, Bai Shi, Li Zhongshan 2026, 30 (36):  9479-9487.  doi: 10.12307/2026.909 Abstract ( 59 )   PDF (2032KB) ( 14 )   Save BACKGROUND: For post-stroke hemiplegic patients, traditional resistance training, while effective, has application limitations in those unable to tolerate active exercise. As a passive intervention, the efficacy specificity, safety, and feasibility of low-frequency pulsed electromagnetic fields for this vulnerable population remain unclear, particularly lacking objective imaging assessments of morphological changes in post-stroke muscle atrophy. OBJECTIVE: To evaluate the improving effects of low-frequency pulsed electromagnetic fields on the muscle structure of the vastus lateralis muscle in the lower limbs of post-stroke hemiplegic patients. METHODS: Twenty-three patients with obvious symptoms of lower limb muscle atrophy after stroke hemiplegia were recruited and randomly divided into an experimental group receiving low-frequency pulsed electromagnetic field stimulation and a control group receiving sham treatment. There were 12 cases in the experimental group and 11 cases in the control group. The trial lasted for a total of 4 weeks. Patients in the experimental group received a 15-minute low-frequency pulsed magnetic field stimulation intervention every 48 hours, while the control group received sham treatment alongside routine medical care, with intervention time points consistent with the experimental group. After 4 weeks, B-mode ultrasound imaging was used as an assessment tool to observe changes in the structural indicators of the vastus lateralis muscle, including pennation angle, muscle thickness, and cross-sectional area, in both the unaffected and affected legs of patients in different groups, as well as changes in the difference values. RESULTS AND CONCLUSION: (1) After 4 weeks of low-frequency pulsed electromagnetic field stimulation, 18 patients completed the trial, with 9 cases in the experimental group and 9 cases in the control group. In the experimental group, the post -test values of the three indicators of muscle morphology in the affected leg, including muscle thickness, pennation angle, and cross-sectional area, were significantly higher than the pre-test values (P < 0.05). The difference values for muscle thickness, pennation angle, and cross-sectional area in the affected leg were significantly lower than those before the trial (P < 0.05). In the control group, muscle thickness and cross-sectional area of the affected leg were significantly higher than before the trial, and the cross-sectional area of the unaffected leg significantly increased; however, there were no significant changes in the difference values of the three muscle morphology indicators between the unaffected and affected legs. (2) Inter-group comparison revealed that the experimental group had significantly higher values than the control group for the three indicators of muscle morphology in the affected leg, including muscle thickness, pennation angle, and cross-sectional area (P < 0.05). The rate of change in the difference values for muscle thickness, pennation angle, and cross-sectional area between the unaffected and affected legs was significantly higher in the experimental group than in the control group (P < 0.05). These findings confirm that after 4 weeks of low-frequency pulsed electromagnetic field stimulation, indicators related to skeletal muscle structure and morphology in the affected vastus lateralis muscle of the lower limb were significantly improved in post-stroke hemiplegic patients, providing morphological evidence supporting the use of this technology as an exercise alternative and medical strategy for improving muscle atrophy symptoms in post-stroke hemiplegic patients. Figures and Tables | References | Related Articles | Metrics

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Comparative finite element analysis of cervical intervertebral discs in adult humans and macaques Hao Yunteng, Shi Jun, Zhang Shaojie, Li Zhijun, Yang Yang, Wang Chaoqun, Ma Yuan, Zhao Hailong, Chen Jie, Dong Chongyang, Zhang Zhifeng, Li Kun, Wang Xing 2026, 30 (36):  9488-9496.  doi: 10.12307/2026.923 Abstract ( 50 )   PDF (10680KB) ( 16 )   Save BACKGROUND: The cervical vertebrae of macaques and humans exhibit remarkable homology with human cervical spines in terms of vertebral count, intervertebral disc structure, and the unique uncinate processes, rendering macaques an ideal biomechanical model for cervical spine research. However, comparative studies on their mechanical properties remain scarce. OBJECTIVE: To develop three-dimensional finite element models of adult human and macaque cervical spines and comparatively analyze the biomechanical characteristics of intervertebral discs under various loading conditions, thereby providing theoretical insights for clinical research. METHODS: Thin-slice CT scans of their cervical spine (C0-T1) were performed on one adult human and one adult macaque specimen to obtain DICOM data. The models were reconstructed using Mimics, refined with Pro/Engineer for optimized geometries (including intervertebral discs and endplates), meshed in Hypermesh, and subjected to finite element analysis using Abaqus. The study specifically compared and analyzed range of motion and stress distribution patterns on the intervertebral disc endplates between human and macaque cervical models. RESULTS AND CONCLUSION: (1) Under flexion-extension, lateral bending, and axial rotation conditions, both human and macaque models exhibited progressive increases in endplate stress and disc displacement values along the craniocaudal axis. Peak intervertebral disc stress and displacement occurred at the C5/6 disc level, while maximum endplate stresses and displacements localized to the C6 segment. (2) Significant lateral asymmetry in endplate stress distribution was observed under certain loading modes—with identical patterns emerging in both species (P < 0.05). Inter-species comparisons revealed no statistically significant differences in disc stresses or displacements. (3) These findings substantiate macaques as clinically relevant cervical spine models, offering mechanistic insights for disease pathophysiology and therapeutic optimization. Figures and Tables | References | Related Articles | Metrics

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Magnetic resonance imaging, virtual simulation, and motion capture analyses of biomechanical characteristics of the paracervical muscles in cervical spondylotic myelopathy Wei Haokai, Zuo Kuangshi, Liu Qiuli, Li Zhifei, Han Bin, Zhang Zhanming, Zhou Jinyan 2026, 30 (36):  9497-9504.  doi: 10.12307/2026.800 Abstract ( 53 )   PDF (4418KB) ( 15 )   Save BACKGROUND: Previous biomechanical studies on cervical spondylosis have mostly been limited to single data analysis, but dynamic analysis is now being used to prove the differences in movement patterns between healthy subjects and cervical spondylotic myelopathy patients with skeletal muscle disorders. OBJECTIVE: To provide more comprehensive evidence to support the biomechanical study of the cervical paraspinal muscles in spinal cord cervical spondylosis by analyzing the cross-sectional area of the cervical paraspinal muscles in spinal cord cervical spondylosis versus healthy populations and by simulating the differences in cervical paraspinal muscle force and coordinated activation of the cervical paraspinal muscles. METHODS: This prospective study included 30 participants, comprising 20 cases in the cervical spondylotic myelopathy group and 10 cases in the healthy control group. A Siemens MAGNETOM Verio 3.0 T MRI scanner was used to perform detailed scans on the participants. The scanning range extended from the first cervical vertebra to the second thoracic vertebra, covering all vertebrae, intervertebral discs, and paravertebral soft tissues. Spin-echo sequences and inversion recovery sequences were used for data processing. Imaging scan parameters were set with a slice thickness of 3 mm, a slice spacing of 1 mm, and an image matrix of 320×224, with the resulting image sequences exported. Subsequently, we used ImageJ software to select the anterior flexor muscles and posterior extensor muscles of the cervical spine for image analysis, measuring the cross-sectional area of the muscles. The polygon selection tool was used to outline the muscle boundaries, and the muscle area was calculated based on the original scale in the MRI images. Six inertial sensors were used to capture the head and neck movement trajectories, and the converted head and neck movement trajectory parameters were imported into the OpenSim virtual simulation system to establish a muscle simulation model. Based on the results of the detailed muscle scans, the muscle force parameters of the simulation muscle model were adjusted proportionally to fine-tune and train the muscle model. Finally, computer algorithms were used to statically optimize the cervical spine movement data captured by the inertial sensors, calculate muscle excitation, and analyze muscle coordination status. RESULTS AND CONCLUSION: (1) There were no significant differences in sex and age between the cervical spondylotic myelopathy and healthy subjects  (P > 0.05). (2) The results of MRI anchor measurement of cervical paraspinal muscles showed that the cross-sectional area of the sternocleidomastoid muscle in the healthy control group was significantly greater than that in the cervical spondylotic myelopathy group (t=-2.501, P=0.019). The cross-sectional area of the multifidus muscle in the healthy control group was significantly greater than that in the cervical spondylotic myelopathy group (t=-2.437, P=0.022). There were no significant differences in the other muscles between the two groups (P > 0.05). (3) Compared with the healthy population, patients with cervical spondylotic myelopathy exhibited more pronounced muscle atrophy in the anterior and lateral cervical muscles. (4) Parameters from OpenSim virtual muscle force simulation showed that muscle force was significantly reduced in the cervical spondylotic myelopathy group compared with the healthy population. (5) Muscle activation results showed that muscle synergistic activation was significantly lower in the cervical spondylotic myelopathy group than in the healthy control group, indicating that patients with cervical spondylotic myelopathy have impaired synergistic balance in the cervical muscle groups. (6) Differences in magnetic resonance imaging and head and neck movement were observed between patients with cervical spondylotic myelopathy and the healthy controls. Figures and Tables | References | Related Articles | Metrics

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Three-dimensional finite element analysis of the biomechanical effects on surrounding tissues following crown-by-crown treatment of impacted mandibular third molars Li Shenghao, Zhang Borui, Ma Yufeng, 2026, 30 (36):  9505-9518.  doi: 10.12307/2026.903 Abstract ( 56 )   PDF (17039KB) ( 18 )   Save BACKGROUND: Extraction of impacted mandibular third molars is one of the common procedures in oral and maxillofacial surgery. Traditional crown-splitting and bone-removing techniques are prone to complications such as adjacent tooth injury, inferior alveolar nerve damage, and jaw fracture. Although minimally invasive extraction techniques have significantly reduced intraoperative trauma, the parameters for sectioning operations (e.g., cutting depth, width, and angle) still rely on empirical judgment and lack biomechanical quantitative standards.   OBJECTIVE: To construct a three-dimensional finite element model of a horizontally and mesially impacted mandibular third molar and surrounding tissues, and to analyze the biomechanical effects of varying cutting depths, widths, and angles on crown fracture and surrounding tissues, thereby providing quantitative standards for precise crown sectioning.   METHODS: Cone-beam CT data of the mandibular third molar region from a healthy volunteer with complete dentition were extracted. The mandibular third molar region model, including the mandibular third molar, mandibular second molar, mandibular bone (cortical and cancellous bone), and inferior alveolar nerve canal, was generated using three-dimensional reconstruction software Mimics 21.0, reverse engineering software Geomagic 21.0, and three-dimensional computer-aided design software SolidWorks 2022. Finite element software Ansys 2021 R1 was used to simulate clinical crown sectioning by applying a 35 N vertical load to investigate the effects of cutting parameters (depth: 11 mm, 10.5 mm, 10 mm, 9.5 mm, 9 mm; width: 1.5 mm, 1 mm; angle: 0°, 10°, 20°, 30°) on crown fracture, stress distribution characteristics in surrounding tissues, and displacement response patterns.   RESULTS AND CONCLUSION: Three-dimensional finite element models of crown sectioning with different cutting parameters were successfully constructed. Through three-dimensional finite element analysis, the following conclusions were drawn: (1) When sectioning the crown using a 1.5 mm diameter bur, the optimal parameter combination is a cutting depth of 10-11 mm and a cutting angle of 0°-10°. This ensures complete crown fracture (maximum Von Mises stress value of the crown ≥ 250 MPa) while protecting adjacent teeth, the inferior alveolar nerve canal, and the mandibular bone, providing precise biomechanical guidance for crown sectioning in minimally invasive extractions. (2) In the extraction of horizontally and mesially impacted mandibular third molars, the choice of cutting depth and width should be based on ensuring complete crown fracture, fully eliminating resistance structures around the crown to provide ideal space for subsequent procedures and avoid damage to surrounding tissues. (3) In the extraction of horizontally and mesially impacted mandibular third molars, the cutting angle should be chosen as parallel as possible to the long axis of the second molar (0°-10°) to ensure crown fracture efficiency and protection of surrounding tissues.   Figures and Tables | References | Related Articles | Metrics

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Electroacupuncture regulates mitochondrial copper homeostasis to modulate neuroinflammatory injury in rats with ischemic stroke Yang Jiao, Wang Zhifeng, Shi Ting, Xu Shuangfeng, Tian Daman, Lan Junfeng, He Pengfen, Hao Zhihui, Pan Pan, Wang Jian 2026, 30 (36):  9519-9525.  doi: 10.12307/2026.904 Abstract ( 68 )   PDF (3590KB) ( 12 )   Save BACKGROUND: Electroacupuncture, a therapy that integrates traditional and modern elements, has been shown to improve the prognosis of stroke. However, whether it can intervene in neuroinflammatory injury by regulating mitochondrial copper homeostasis remains unclear.   OBJECTIVE: To observe the therapeutic effects of electroacupuncture stimulation at “Baihui” and “Dazhui” on cerebral ischemia-reperfusion injury, and to explore the possible mechanisms by which it improves copper homeostasis imbalance in cerebral ischemia-reperfusion injury.   METHODS: Male Sprague-Dawley rats were randomly divided into a sham-operated group, a model group, and an electroacupuncture group, with 12 rats in each group. The latter two groups were used to establish a rat model of cerebral ischemia-reperfusion injury using the modified middle cerebral artery occlusion method. The electroacupuncture group received acupuncture at “Baihui” and “Dazhui” acupoints, once daily for 30 minutes each time, for 14 consecutive days. Zea-Longa scores were used to assess the degree of neurological deficits. Triphenyltetrazolium chloride staining was used to measure cerebral infarct volume. Hematoxylin-eosin staining and Nissl staining were employed to observe pathological structural changes in brain tissue, and rhodanine staining was used to observe copper ion deposition. Transmission electron microscopy was used to observe the ultrastructure of rat cortical neurons. Colorimetric assays were used to detect the levels of reactive oxygen species, iron-sulfur clusters, and cuprous ions in the infarcted brain tissue. Immunohistochemical staining and western blot assay were used to observe the protein expression levels of Kelch-like ECH-associated protein 1 (Keap1), nuclear factor erythroid 2-related factor 2 (Nrf2), ATPase copper transporting beta (ATP7B), and dihydrolipoamide S-acetyltransferase (DLAT).   RESULTS AND CONCLUSION: (1) Compared with the sham-operated group, the model group showed significantly increased Zea-Longa scores (P < 0.001) and a significantly increased percentage of cerebral infarct volume (P < 0.001); in the infarcted area, the number of neurons was reduced, their arrangement was loose, neurons showed obvious pathological changes, and the number of organelles was decreased; the contents of reactive oxygen species and cuprous ions in brain tissue were significantly increased (P < 0.001), while the content of iron-sulfur clusters was significantly decreased (P < 0.001); in the cortex, the protein expressions of Nrf2, ATP7B, and DLAT were significantly decreased (P < 0.05, P < 0.001, P < 0.01, respectively), and the protein expression of Keap1 was significantly increased (P < 0.001). (2) Compared with the model group, the electroacupuncture group showed significantly decreased Zea-Longa scores (P < 0.001) and a decreased percentage of cerebral infarct volume (P < 0.05); in the infarcted area, the number of neurons was increased, their arrangement was relatively regular, the nuclear membrane of neurons was relatively intact, and the number of organelles such as mitochondria and endoplasmic reticulum was increased; the contents of reactive oxygen species and cuprous ions in brain tissue were significantly decreased (P < 0.05, P < 0.001), and the content of iron-sulfur clusters was increased (P < 0.01); in the cortex, the protein expressions of Nrf2, ATP7B, and DLAT were significantly increased (P < 0.05, P < 0.001, P < 0.05, respectively), and the protein expression of Keap1 was significantly decreased (P < 0.05). To conclude, these findings suggest that electroacupuncture can protect mitochondrial structural and functional integrity by regulating the Keap1-Nrf2 signaling pathway, promoting ATP7B-mediated mitochondrial copper transport, and maintaining mitochondrial copper homeostasis and normal energy metabolism, ultimately improving neurological function impairment after ischemia-reperfusion and promoting brain tissue repair.   Figures and Tables | References | Related Articles | Metrics

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Mechanism and effect of aescin intervention against brain injury in a rat model of intracerebral hemorrhage Ji Lingshan, Qin Hewei, Cheng Shuaifang, Zhao Jing 2026, 30 (36):  9526-9537.  doi: 10.12307/2026.919 Abstract ( 44 )   PDF (2996KB) ( 9 )   Save BACKGROUND: Neuronal apoptosis induced by endoplasmic reticulum stress following intracerebral hemorrhage is a significant cause of secondary brain injury. Aescin has been demonstrated to possess anti-inflammatory, antioxidant, and anti-apoptotic effects in brain injury models. However, whether it exerts neuroprotective effects by regulating endoplasmic reticulum stress-related pathways remains to be clarified. OBJECTIVE: To investigate the effect of aescin on brain injury in rats with intracerebral hemorrhage and its potential mechanism of action. METHODS: Fifty-four rats were randomly divided into a sham operation group, an intracerebral hemorrhage group, and an intracerebral hemorrhage+aescin group using a random number table method. An intracerebral hemorrhage rat model was established in the latter two groups using autologous blood injection. Rats in the sham operation group underwent the same stereotaxic puncture procedure but without blood injection. After successful modeling, the intracerebral hemorrhage+aescin group received intraperitoneal injection of aescin (10 mg/kg), while the sham operation group and intracerebral hemorrhage group received an equal volume of normal saline. Neurological function of rats was assessed using Longa score, rotarod test, forelimb placing test, and corner turn test. Pathological damage of the brain and brain edema were evaluated using hematoxylin-eosin staining and brain water content measurement. A cell injury model was established in nerve growth factor-incubated PC12 cells under oxygen-glucose deprivation. An endoplasmic reticulum stress cell model was established using tunicamycin, with the endoplasmic reticulum stress inhibitor 4-PBA serving as a positive control; cells were also treated with IXA4 (IRE1 agonist) or CCT020312 (PERK agonist). Cell viability was detected using the cell counting kit-8 assay, and apoptosis was detected using flow cytometry. Expression of endoplasmic reticulum stress-related proteins was detected by western blot. The mechanism of pathway involvement was verified using inositol-requiring enzyme 1 (IRE1) or protein kinase R-like endoplasmic reticulum kinase (PERK) small interfering RNA (IRE1 siRNA or PERK siRNA) and pathway agonists. RESULTS AND CONCLUSION: (1) Compared with the sham operation group, rats in the intracerebral hemorrhage group showed significantly increased Longa scores (P < 0.05), significantly impaired sensorimotor function, obvious hemorrhagic foci, edema, and neuronal pyknosis in brain tissue, and significantly increased brain water content. In the aescin treatment group, these indicators were significantly improved from 12 hours to 5 days after surgery (P < 0.05). (2) Aescin intervention significantly reduced the expression levels of endoplasmic reticulum stress-related proteins (GRP78, CHOP, p-IRE1, p-PERK, etc.) in the brain tissue of intracerebral hemorrhage rats (P < 0.05). (3) Treatment with oxygen-glucose deprivation or the endoplasmic reticulum stress inducer tunicamycin significantly reduced the viability of nerve growth factor-incubated PC12 cells, increased the apoptosis rate (P < 0.05), and upregulated GRP78, CHOP, and the ratios of p-IRE1/IRE1 and p-PERK/PERK (P < 0.05). However, 10 and 20 μmol/L aescin significantly reversed these changes (P < 0.05). (4) Knockdown of IRE1/PERK produced effects consistent with aescin intervention, inhibiting the activation of the IRE1-XBP1 and PERK-eIF2α pathways induced by oxygen-glucose deprivation. Pathway agonists (IXA4 and CCT020312) reversed the protective effect of aescin (P < 0.05). (5) Correlation analysis showed that the expression of GRP78, p-IRE1, and p-PERK in rat brain tissue was positively correlated with brain edema volume and Longa score (P < 0.05). These findings suggest that aescin can alleviate endoplasmic reticulum stress by specifically blocking the IRE1/PERK signaling pathway, thereby ameliorating neurological impairment in rats with intracerebral hemorrhage and oxygen-glucose deprivation-induced injury in nerve growth factor-incubated PC12 cells. Figures and Tables | References | Related Articles | Metrics

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Mechanism of Huangpu Tongqiao Capsule on cognitive function in a rat model of Wilson’s disease Xu Wenlin, Wu Haixia, Li Jie, Zhang Manhua, Rao Zhihui, Zhu Qingxiong 2026, 30 (36):  9538-9545.  doi: 10.12307/2026.922 Abstract ( 53 )   PDF (1910KB) ( 12 )   Save BACKGROUND: Wilson’s disease not only affects liver function but can also cause oxidative damage to neurons in the brain through copper accumulation. Studies have found that the glucose-regulated protein 78/inositol-requiring enzyme 1α/tumor necrosis factor receptor-associated factor 2/c-Jun N-terminal kinase signaling pathway plays a key role in the neuropathological process of Wilson’s disease. Huangpu Tongqiao Capsule may reduce neuronal damage caused by copper overload and improve learning and memory function by scavenging oxygen free radicals, inhibiting neuroinflammation, and regulating endoplasmic reticulum stress-related pathways. OBJECTIVE: To further validate the effect of Huangpu Tongqiao Capsule on cognitive function in a rat model of Wilson’s disease and to explore its mechanism of action. METHODS: Forty SPF-grade male Sprague-Dawley rats were randomly divided into a normal group, a model group, a Huangpu Tongqiao Capsule group, and a positive control group, with 10 rats in each group. Rats in the latter three groups were fed a copper sulfate-loaded diet to establish a Wilson’s disease model. The normal and model groups received intragastric administration of normal saline. The Huangpu Tongqiao Capsule group received intragastric administration of 1.41 g/kg Huangpu Tongqiao Capsule decoction. The positive control group received intragastric administration of 0.09 g/kg penicillamine solution. All administrations were given continuously for 28 days. After administration, 24-hour urinary copper levels were measured. The Morris water maze test was used to evaluate learning and memory function. Hematoxylin-eosin staining was used to observe the structural integrity of neurons in the hippocampus. TUNEL staining was used to observe the number of positive neurons in the hippocampus. ELISA was used to detect the levels of superoxide dismutase and malondialdehyde in the rat hippocampus. Western blot was used to detect the expression of proteins related to the endoplasmic reticulum stress-associated glucose-regulated protein 78/inositol-requiring enzyme 1α/tumor necrosis factor receptor-associated factor 2/c-Jun N-terminal kinase signaling pathway in the hippocampus. RESULTS AND CONCLUSION: (1) Compared with the normal group, 24-hour urinary copper levels were increased in the model group (P < 0.05). Compared with the model group, 24-hour urinary copper levels were further increased in the Huangpu Tongqiao Capsule group and the positive control group (P < 0.05). (2) Compared with the normal group, rats in the model group showed increased escape latency and decreased number of crossings over the original platform (P <  0.05). Compared with the model group, rats in the Huangpu Tongqiao Capsule group and the positive control group showed shortened escape latency and increased number of crossings over the original platform (P < 0.05). (3) Compared with the normal group, the hippocampal tissue of rats in the model group showed disrupted neural structure, disorganized cell arrangement, and a significant increase in the number of TUNEL-positive apoptotic neurons. Compared with the model group, the structural integrity of hippocampal neurons in the Huangpu Tongqiao Capsule group and the positive control group was significantly improved, cell arrangement became more orderly, and the number of TUNEL-positive apoptotic neurons was also significantly reduced. (4) Compared with the normal group, the protein expression of glucose-regulated protein 78, inositol-requiring enzyme 1α, tumor necrosis factor receptor-associated factor 2, phosphorylated c-Jun N-terminal kinase, caspase-12, and Bax in hippocampal tissue was increased in the model group, while Bcl-2 protein expression in hippocampal tissue was decreased (P < 0.05). Compared with the model group, the Huangpu Tongqiao Capsule group and the positive control group showed opposite changes in the above indicators (P < 0.05). In conclusion, Huangpu Tongqiao Capsule can improve spatial learning and memory ability, reduce neuronal damage in the hippocampal tissue, and exert a neuroprotective effect in the rat model of Wilson’s disease. Its mechanism of action may be related to inhibiting the glucose-regulated protein 78/inositol-requiring enzyme 1α/tumor necrosis factor receptor-associated factor 2/c-Jun N-terminal kinase signaling pathway, thereby alleviating endoplasmic reticulum stress. Figures and Tables | References | Related Articles | Metrics

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Effects of Tuina on dorsal root ganglion neuron-satellite glial cell signaling and peripheral sensitization in rats with chronic sciatic nerve compression injury Huang Lanting, Chen Zuzhang, Huang Hongye, Wang Chenyu, He Luying, Cai Hengchang, Chen Shuijin, Lin Zhigang, Jiang Jingjing, Chen Lechun 2026, 30 (36):  9546-9552.  doi: 10.12307/2026.912 Abstract ( 46 )   PDF (5639KB) ( 8 )   Save BACKGROUND: The peripheral mechanism of Tuina analgesia remains unclear. This study aimed to investigate whether Tuina acupressure alleviates peripheral sensitization and neuropathic pain by regulating the abnormal signal interaction of the “neuron-satellite glial cell” functional unit in the dorsal root ganglion. OBJECTIVE: To observe the effects of Tuina on pain behavior and neuron-satellite glial cell signal interaction in the dorsal root ganglion of rats with chronic constriction injury of the sciatic nerve, and to explore the molecular mechanism of Tuina in alleviating neuropathic pain by regulating peripheral sensitization. METHODS: Twenty-four specific pathogen-free grade male Sprague-Dawley rats were randomly divided into blank control, model, and Tuina groups, with eight rats in each group. The latter two groups were established as chronic constriction injury models of the sciatic nerve. The Tuina group received acupressure intervention at the Weizhong acupoint starting from the 4th day post-surgery, once daily at a force of 5 N and a frequency of 120 times/min for 10 minutes each session, for a total of 14 days. Mechanical paw withdrawal threshold was measured using von Frey filaments, and thermal paw withdrawal latency was measured using a thermal pain instrument. Western blot was used to detect the expression of connexin 43, calcium/calmodulin-dependent serine protein kinase, glial fibrillary acidic protein, and pannexin 1 in the dorsal root ganglion. Enzyme-linked immunosorbent assay was used to detect the expression levels of inflammatory factors tumor necrosis factor α and interleukin 1β in the dorsal root ganglion. Hematoxylin-eosin staining was used to observe the pathological morphology of the gastrocnemius muscle. RESULTS AND CONCLUSION: (1) Compared with the blank control group, the mechanical paw withdrawal threshold and thermal paw withdrawal latency in the model and Tuina groups were significantly decreased on postoperative day 7 (all P < 0.001). On postoperative days 7, 10, 14, and 17, the mechanical paw withdrawal threshold was significantly higher and the thermal paw withdrawal latency was significantly longer in the Tuina group than in the model group (all P < 0.001). (2) The expression levels of connexin 43, calcium/calmodulin-dependent serine protein kinase, glial fibrillary acidic protein, pannexin 1, tumor necrosis factor α, and interleukin 1β in the dorsal root ganglion were significantly higher in the model group than in the blank control group (all P < 0.05), and these indicators were significantly lower in the Tuina group than in the model group (all P < 0.05). (3) Hematoxylin-eosin staining showed irregular shape and disordered nuclear arrangement of the gastrocnemius muscle in the model group, while the muscle fiber morphology was partially restored in the Tuina group. To conclude, these findings indicate that Tuina may improve peripheral sensitization and pain behavior in rats with chronic constriction injury of the sciatic nerve by inhibiting the expression of connexin 43 and pannexin 1 in neurons and satellite glial cells, attenuating signal interaction between neurons and satellite glial cells, reducing the release of pro-inflammatory factors, and decreasing neuronal excitability. This provides a theoretical basis from a molecular biology perspective for the peripheral analgesic mechanism of Tuina. Figures and Tables | References | Related Articles | Metrics

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Effects of isorhamnetin on skin lesions in a rat model of auricular acne  Sun Xian, Hu Zhen, Liu Rui 2026, 30 (36):  9553-9559.  doi: 10.12307/2026.921 Abstract ( 50 )   PDF (2698KB) ( 11 )   Save BACKGROUND: Auricular acne is an infection of hair follicles or sebaceous glands, primarily manifesting as skin itching, redness, swelling, pain, and scarring. In severe cases, it can even lead to intracranial infection and facial paralysis. Isorhamnetin is a flavonoid compound with pharmacological activities including anti-inflammatory, antioxidant, cardioprotective, antitumor, and immunomodulatory effects, and may also play an important role in the treatment of skin inflammation-related diseases. OBJECTIVE: To investigate the effect of isorhamnetin on skin lesions and the transforming growth factor β1 (TGF-β1)/mothers against DPP homolog 3(Drosophila) (Smad3) signaling pathway in a rat model of auricular acne.  METHODS: A rat model of auricular acne was established. Successfully modeled rats were randomly divided into model group, low-dose isorhamnetin group, high-dose isorhamnetin group, and high-dose isorhamnetin+SRI-011381 (TGF-β1/Smad3 pathway activator) group. Another normal healthy rats served as the control group. After the intervention, skin lesion severity was scored, and the auricular swelling rate was calculated. Serum levels of inflammatory factors were detected by ELISA. Histopathological changes and fibrosis of auricular tissue were observed using hematoxylin-eosin and Masson staining. Expression of α-smooth muscle actin and type I collagen was detected by immunohistochemistry. Expression of TGF-β1/Smad3 pathway-related proteins was detected by western blot. RESULTS AND CONCLUSION: Compared with the control group, the model group showed local cartilage disruption, skin tissue thickening, epidermal spinous cell proliferation, sebaceous gland hyperplasia, enlarged hair follicle openings, loose dermal structure with visible edema and necrotic foci, marked inflammatory cell infiltration, increased blue collagen fiber deposition, and enlarged fibrotic area in the auricular tissue. Additionally, the model group exhibited significantly higher skin lesion severity scores, auricular swelling rates, levels of tumor necrosis factor α, interleukin 6, interleukin 1β, and interleukin 18, as well as increased expression of α-smooth muscle actin, type I collagen, TGF-β1, and phosphorylated Smad3/Smad3, while interleukin 10 levels were significantly decreased (P < 0.05). Compared with the model group, the low- and high-dose isorhamnetin groups showed varying degrees of improvement in auricular tissue pathological changes, reduced blue collagen fiber deposition, decreased fibrotic area, lower skin lesion severity scores, auricular swelling rates, levels of tumor necrosis factor α, interleukin 6, interleukin 1β, and interleukin 18, and reduced expression of α-smooth muscle actin, type I collagen, TGF-β1, and phosphorylated Smad3/Smad3, along with increased interleukin 10 levels (P < 0.05). The high-dose isorhamnetin+SRI-011381 group reversed the improvements in auricular tissue pathological damage, skin lesion severity scores, auricular swelling rates, inflammatory factors, and fibrosis observed with high-dose isorhamnetin alone. To conclude, these findings indicate that isorhamnetin can ameliorate skin lesions in a rat model of auricular acne, and this effect may be related to inhibition of the TGF-β1/Smad3 signaling pathway. Figures and Tables | References | Related Articles | Metrics

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Atractylenolide III ameliorates myocardial injury in a mouse model of myocardial infarction Huang Weiqing, Liao Weimin, Du Tingting, Tian Xin 2026, 30 (36):  9560-9565.  doi: 10.12307/2026.902 Abstract ( 59 )   PDF (1852KB) ( 22 )   Save BACKGROUND: Atractylenolide III can inhibit inflammatory response and cardiomyocyte apoptosis, exerting a cardioprotective effect, but its regulatory mechanism remains unclear. OBJECTIVE: To investigate the protective effect of atractylenolide III on myocardial injury in a mouse model of myocardial infarction based on the phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin signaling pathway. METHODS: Sixty 8-week-old male C57BL/6 mice were selected and randomly divided into sham operation group, model group, low-, medium-, and high-dose atractylenolide III groups, and aspirin group, with 10 mice in each group. Myocardial infarction was induced by ligation of the left anterior descending coronary artery. One week after modeling, the low-, medium-, and high-dose atractylenolide III groups were administered atractylenolide III 5, 30, 60 mg/kg by gavage, respectively, and the aspirin group was administered 25 mg/kg aspirin by gavage, once daily for 4 weeks. Echocardiography was used to detect ventricular function indicators. Hematoxylin-eosin staining was used to observe myocardial histopathological changes and determine the optimal dose of atractylenolide III. TTC staining was used to observe myocardial infarct size. ELISA was used to detect serum myocardial injury markers and inflammatory factor levels. TUNEL staining was used to detect cell apoptosis. Western blot and qPCR were used to observe the expression of phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin signaling pathway-related proteins and mRNAs. RESULTS AND CONCLUSION: (1) Compared with the model group, the ventricular function indicators improved significantly in the atractylenolide III groups and the aspirin group (P < 0.05). (2) Atractylenolide III ameliorated myocardial histopathological changes in mice, with the most significant effect observed in the high-dose atractylenolide III group. (3) Compared with the sham operation group, the model group showed increased percentage of myocardial infarct size (P < 0.05), higher levels of myocardial injury markers and inflammatory factors (P < 0.05), higher cardiomyocyte apoptosis rate (P < 0.05), and increased expression of phosphorylated phosphatidylinositol 3-kinase/phosphatidylinositol 3-kinase, phosphorylated protein kinase B/protein kinase B, phosphorylated mammalian target of rapamycin/mammalian target of rapamycin, as well as mRNA expression of phosphatidylinositol 3-kinase, protein kinase B, and mammalian target of rapamycin in myocardial tissue (P < 0.05). (4) Compared with the model group, the high-dose atractylenolide III group and the aspirin group showed decreased percentage of myocardial infarct size (P < 0.05), lower levels of myocardial injury markers and inflammatory factors (P < 0.05), lower cardiomyocyte apoptosis rate (P < 0.05), and decreased expression of the above-mentioned signaling pathway-related proteins and mRNAs in myocardial tissue (P < 0.05). These findings indicate that atractylenolide III can ameliorate myocardial histopathological damage and inflammatory response, and its mechanism is related to the regulation of the phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin signaling pathway. Figures and Tables | References | Related Articles | Metrics

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Ginsenoside Rb1 promotes wound healing in burn rats Jiang Xin, Han Zhixin, Ma Jiaqi, Yang Wen, Zhao Xiaojing 2026, 30 (36):  9566-9571.  doi: 10.12307/2026.917 Abstract ( 65 )   PDF (2858KB) ( 17 )   Save BACKGROUND: Ginsenoside Rb1 possesses anti-inflammatory and antioxidant effects and can be used to treat tissue cell damage caused by oxidative stress, inflammation, and other related conditions. OBJECTIVE: To investigate the effect of ginsenoside Rb1 on wound healing in burn rats. METHODS: Ninety-six Sprague-Dawley rats were randomly divided into control group, model group, low-dose ginsenoside Rb1 group, medium-dose ginsenoside Rb1 group, high-dose ginsenoside Rb1 group, and high-dose ginsenoside Rb1+activator group, with 16 rats in each group. The control group was not modeled, while the other five groups underwent establishment of a dorsal burn model (a 5 cm diameter metal weight heated in 95 °C water was applied vertically to the rat skin for 10 seconds). After modeling, the control and model groups received intraperitoneal injection of normal saline. The low-, medium-, and high-dose ginsenoside Rb1 groups received intraperitoneal injection of 10, 20, and 40 mg/kg ginsenoside Rb1, respectively. The high-dose ginsenoside Rb1+activator group received intraperitoneal injection of 40 mg/kg ginsenoside Rb1 and the Ras homolog gene family member A/Rho-associated coiled-coil kinase (RhoA/ROCK) signaling pathway activator lysophosphatidic acid. All injections were administered once daily for 2 consecutive weeks, and wound healing was observed. After the final administration, samples were collected. Hematoxylin-eosin staining was used to observe wound tissue morphology. TUNEL staining was used to detect the apoptosis rate of wound tissue cells. ELISA was used to measure serum levels of interleukin 6, tumor necrosis factor α, and interleukin 1β. Western blot was used to detect the protein expression of RhoA and ROCK in the wound tissue. RESULTS AND CONCLUSION: Compared with the control group, the model group showed extensive inflammatory cell infiltration in the wound, incomplete tissue structure, poor epithelial cell growth, and significantly increased apoptosis rate, levels of interleukin 6, tumor necrosis factor α, and interleukin 1β, as well as protein expression of RhoA and ROCK (P < 0.05). Compared with the model group, the low-, medium-, and high-dose ginsenoside Rb1 groups showed no obvious inflammatory cell infiltration in the wound, relatively intact tissue structure, good epithelial cell growth, significantly increased wound healing rate (P < 0.05), and significantly decreased apoptosis rate, levels of interleukin 6, tumor necrosis factor α, and interleukin 1β, as well as protein expression of RhoA and ROCK (P < 0.05), with the most significant changes observed in the high-dose ginsenoside Rb1 group. Compared with the high-dose ginsenoside Rb1 group, the high-dose ginsenoside Rb1+activator group showed obvious inflammatory cell infiltration in the wound, poor tissue structural integrity, poor epithelial cell growth, significantly decreased wound healing rate (P < 0.05), and significantly increased apoptosis rate, levels of interleukin 6, tumor necrosis factor α, and interleukin 1β, as well as protein expression of RhoA and ROCK (P < 0.05). In conclusion, ginsenoside Rb1 may promote wound healing in burned rats by inhibiting the RhoA/ROCK signaling pathway, thereby suppressing inflammatory response and apoptosis. Figures and Tables | References | Related Articles | Metrics

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Inducing cuproptosis in pancreatic β cells: regulating homocysteine via targeting SLC39A14 Li Shujuan, Ma Lingju, Li Xinru, Sun Rui, Ma Ben, Qi Ning, Zhang Qifan, Liu Li, Chai Yuee, Ma Shengchao 2026, 30 (36):  9572-9579.  doi: 10.12307/2026.920 Abstract ( 52 )   PDF (2147KB) ( 9 )   Save BACKGROUND: Hyperhomocysteinemia is closely associated with pancreatic β-cell dysfunction and insulin resistance, but its specific molecular mechanism remains to be elucidated. OBJECTIVE: To investigate the potential mechanism by which piR-000699 targets SLC39A14 to regulate homocysteine-induced cuproptosis in pancreatic β cells. METHODS: The mouse pancreatic β-cell line Min6 was cultured in vitro and divided into a control group (0 μmol/L homocysteine) and homocysteine groups (treated with 40, 80, 100, and 120 μmol/L homocysteine, respectively). A concentration of 100 μmol/L homocysteine was selected as the optimal dose for intervening in pancreatic β cells. qRT-PCR was used to detect the baseline expression of piR-000699 in homocysteine-treated cells. Western blot was used to detect the expression of cuproptosis-related proteins FDX1 and HSP70, as well as the piR-000699 downstream target gene SLC39A14, in homocysteine-induced pancreatic β cells. A copper ion assay kit was used to measure intracellular Cu²⁺ levels in pancreatic β cells. Cell transfection groups were as follows: (1) control group, homocysteine group (100 μmol/L), homocysteine+mimics-NC group, homocysteine+piR-000699 mimics group; (2) control group, homocysteine group (100 μmol/L homocysteine), homocysteine+inhibitor-NC group, homocysteine+piR-000699 inhibitor group. qRT-PCR was used to detect piR-000699 mRNA expression after transfection with piR-000699 inhibitor and piR-000699 mimics. Western blot was used to detect the protein expression of FDX1, HSP70, and SLC39A14 after transfection with piR-000699 inhibitor and piR-000699 mimics. A copper ion assay kit was used to measure Cu²⁺ expression levels after transfection. Wild-type and mutant dual-luciferase gene vectors for SLC39A14 were constructed to verify the targeted binding relationship between piR-000699 and SLC39A14. RESULTS AND CONCLUSION: (1) The optimal concentration of homocysteine for inducing pancreatic β-cell injury was 100 μmol/L. (2) Compared with the control group, piR-000699 mRNA expression was increased in the homocysteine group (P < 0.01), and the expression of cuproptosis-related proteins FDX1 and HSP70, as well as Cu²⁺ levels, were all increased (P < 0.001). (3) After interfering with piR-000699 in pancreatic β cells, homocysteine further downregulated the protein expression of FDX1, HSP70, and SLC39A14 (P < 0.01), while intracellular Cu²⁺ expression levels were decreased (P < 0.01). (4) After overexpressing piR-000699 in pancreatic β cells, homocysteine further upregulated the protein expression of FDX1, HSP70, and SLC39A14 in the cells (P < 0.01), while intracellular Cu²⁺ levels were significantly increased (P < 0.001). (5) Dual-luciferase reporter assay results indicated that, compared with mimics-NC, the relative luciferase activity was significantly increased in pancreatic β cells co-transfected with the SLC39A14 wild-type plasmid and piR-000699 mimics (P < 0.01). In conclusion, these findings indicate that piR-000699 can modulate the expression of cuproptosis-related proteins by targeting the SLC39A14 gene during homocysteine-induced cuproptosis in pancreatic β cells, providing new targets and strategies for the treatment and prevention of diabetes. Figures and Tables | References | Related Articles | Metrics

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Ban’s Culuan Zhuyun Decoction alleviates oxidative stress injury in vitrified ovarian tissue Liu Yunjia, Yue Xiaolei, Wu Liming, Zhou Huanglei, Lan Xiang, Cui Jiayu, Bai Lin, Liao Yanbo, Liang Jing, Li Mingxing, He Guozhen, Yang Sufang 2026, 30 (36):  9580-9588.  doi: 10.12307/2026.913 Abstract ( 45 )   PDF (4915KB) ( 12 )   Save BACKGROUND: Vitrification causes cryoinjury to ovarian tissue. The protective effect and molecular mechanism of Ban’s Culuan Zhuyun Decoction on vitrified ovarian tissue urgently need to be further elucidated.   OBJECTIVE: To investigate the effect and molecular mechanism of Ban’s Culuan Zhuyun Decoction on vitrified mouse ovarian tissue.   METHODS: (1) Pre-vitrification experiment: Sixty female Kunming mice, 6-8 weeks old, were randomly divided into control group I, model group I, and low, medium-, and high-dose Ban’s Culuan Zhuyun Decoction groups. After continuous administration for 21 days, ovarian tissues were collected. Ovaries in the control group I were fresh, while those in the other groups were vitrified for 7 days. (2) Post-vitrification experiment: Ovarian tissues were collected from another 60 female Kunming mice, 6-8 weeks old. Ovaries in the control group II were fresh. The remaining ovaries, after vitrification for 7 days, were randomly divided into model group II and low-, medium-, and high-dose Ban’s Culuan Zhuyun Decoction-containing serum groups, and then cultured in vitro at 37 °C for 1 hour in medium containing 15% blank rat serum or different doses of Ban’s Culuan Zhuyun Decoction-containing serum. Ovarian morphology was observed by hematoxylin-eosin staining. Ovarian cell apoptosis was detected by TUNEL assay. The levels of ferrous ion glutathione, malondialdehyde, and glutathione peroxidase 4 were measured by colorimetric methods. The mRNA and protein expression levels of solute carrier family 7 member 11, tumor suppressor protein p53, and glutathione peroxidase 4 were detected by RT-qPCR and western blot, respectively.   RESULTS AND CONCLUSION: (1) Compared with control groups I and II, model groups I and II showed disorganized and partially absent ovarian granulosa cells, a significant increase in TUNEL-positive cell rate, ferrous ion content, malondialdehyde content, and tumor suppressor protein p53 mRNA and protein expression levels, as well as a significant reduction in glutathione content, glutathione peroxidase 4 content, and solute carrier family 7 member 11 and glutathione peroxidase 4 mRNA and protein expression levels (P < 0.05). (2) Compared with model groups I and II, Ban’s Culuan Zhuyun Decoction and its drug-containing serum reduced the number of atretic follicles in ovarian tissue, significantly decreased the TUNEL-positive cell rate, ferrous ion content, malondialdehyde content, and tumor suppressor protein p53 mRNA and protein expression levels, and significantly increased glutathione content, glutathione peroxidase 4 content, and solute carrier family 7 member 11 and glutathione peroxidase 4 mRNA and protein expression levels (P < 0.05). Therefore, Ban’s Culuan Zhuyun Decoction and its drug-containing serum can improve the morphological structure of ovarian tissue and follicles in vitrified mice, inhibit ovarian cell apoptosis and ferroptosis, thereby alleviating cryoinjury in ovarian tissue. Its mechanism may be related to inhibiting the abnormal activation of the tumor suppressor protein p53/solute carrier family 7 member 11/glutathione peroxidase 4 signaling pathway.   Figures and Tables | References | Related Articles | Metrics

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Diabetes prediction and analysis of influencing factors integrating genetic information Liu Yi, Lu Jiarong, Wu Jianyong 2026, 30 (36):  9589-9596.  doi: 10.12307/2026.899 Abstract ( 45 )   PDF (5068KB) ( 26 )   Save BACKGROUND: Early risk assessment and accurate diagnosis are crucial for the clinical prevention and management of diabetes mellitus. Genetic factors play a significant role in the pathogenesis of diabetes; however, most current studies insufficiently integrate genetic information into risk modeling. OBJECTIVE: To construct a comprehensive feature dataset integrating genetic factors, anthropometric indicators, and insulin metabolism metrics, and to develop an interpretable diabetes mellitus prediction model for early risk assessment and prediction.  METHODS: A publicly available diabetes prediction dataset from Kaggle was used, comprising 5 070 valid samples, including 1 936 diabetic and 3 134 non-diabetic individuals. The model was trained on the integrated feature dataset using the Histogram-based Gradient Boosting Decision Tree (HistGBDT) algorithm. Hyperparameters were optimized via grid search. Model performance was evaluated using accuracy, precision, recall, F1-score, and area under the receiver operating characteristic curve. The SHapley Additive exPlanations (SHAP) framework was applied to identify key features influencing diabetes risk and to enhance model interpretability. Ablation studies were conducted to validate the contribution of genetic factors. RESULTS AND CONCLUSION: The proposed interpretable diabetes prediction model outperformed existing mainstream models across all evaluation metrics, achieving an accuracy of 98.03%, precision of 97.66%, recall of 97.16%, F1-score of 97.41%, and the area under the receiver operating characteristic curve of 97.86%, representing an improvement of 1%–4% in overall performance. Ablation experiments demonstrated that integrating genetic factors enables more comprehensive and effective capture of diabetes risk characteristics. SHAP analysis identified triceps skinfold thickness, insulin release test, body mass index, oral glucose tolerance test, and diastolic blood pressure as the top five influential factors. The interpretability analysis provides a theoretical foundation and technical support for early diabetes identification and personalized health management. Figures and Tables | References | Related Articles | Metrics

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Protective effect of kaempferol against renal injury in lupus nephritis Zhang Bo, Zhao Jingjing, Li Wenting, Kong Jie 2026, 30 (36):  9597-9603.  doi: 10.12307/2026.916 Abstract ( 56 )   PDF (2334KB) ( 9 )   Save BACKGROUND: Studies have found that kaempferol may be a potential drug for improving immune and inflammatory mechanisms in advanced diabetic nephropathy by affecting ferroptosis. The Janus kinase 2/signal transducer and activator of transcription 3 (JAK2/STAT3) signaling pathway exhibits high transcriptional activity in autoimmune diseases and inflammation. OBJECTIVE: To investigate the effect of kaempferol against renal injury in mice with lupus nephritis. METHODS: MRL/lpr mice (systemic lupus erythematosus mice) were randomly divided into model group (n=12), low-dose kaempferol group (n=12), medium-dose kaempferol group (n=12), high-dose kaempferol group (n=12), high-dose kaempferol+STAT3 activator group (n=12), and positive drug group (n=12). Twelve C57BL/6 mice were selected as the control group. The control and model groups received intragastric administration of normal saline. The low-, medium-, and high-dose kaempferol groups received intragastric administration of 25, 50, and 100 mg/kg kaempferol, respectively. The high-dose kaempferol+STAT3 activator group received intragastric administration of 100 mg/kg kaempferol combined with intraperitoneal injection of the STAT3 activator colivelin. The positive drug group received intraperitoneal injection of prednisone acetate. All administrations were performed once daily for 4 consecutive weeks. At 2 weeks after the final administration, the following parameters were measured: 24-hour urinary protein level, renal function indicators, serum inflammatory factor levels, renal histopathological morphology, IgG deposition in renal tissue, and protein expression of phosphorylated JAK2, JAK2, phosphorylated STAT3, and STAT3 in renal tissue. RESULTS AND CONCLUSION: (1) Compared with the control group, the model group showed glomerular proliferation and sclerosis, peripheral inflammatory cell infiltration, increased IgG deposition, and significantly elevated levels of 24-hour urinary protein, serum creatinine, blood urea nitrogen, anti-double-stranded DNA, interleukin 6, tumor necrosis factor α, interleukin 1β, interleukin 17, and interleukin 23, along with upregulated protein expression of phosphorylated JAK2 and phosphorylated STAT3 (P < 0.05). (2) Compared with the model group, the low-, medium-, and high-dose kaempferol groups and the positive drug group showed gradually alleviated periglomerular inflammation, reduced IgG deposition, and significantly decreased levels of 24-hour urinary protein, serum creatinine, blood urea nitrogen, anti-double-stranded DNA, interleukin 6, tumor necrosis factor α, interleukin 1β, interleukin 17, and interleukin 23, along with downregulated protein expression of phosphorylated JAK2 and phosphorylated STAT3 (P < 0.05). (3) Compared with the high-dose kaempferol group, the high-dose kaempferol+STAT3 activator group showed increased periglomerular inflammatory cell infiltration, more IgG deposition, and significantly elevated levels of 24-hour urinary protein, serum creatinine, blood urea nitrogen, anti-double-stranded DNA, interleukin 6, tumor necrosis factor α, interleukin 1β, interleukin 17, and interleukin 23, along with upregulated protein expression of phosphorylated JAK2 and phosphorylated STAT3 (P < 0.05). Overall, these findings indicate that kaempferol may alleviate renal injury in mice with lupus nephritis by inhibiting the JAK2/STAT3 signaling pathway. Figures and Tables | References | Related Articles | Metrics

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Development and evaluation of a prediction model for functional language communication outcomes in post-stroke aphasia patients Huang Yunshi, Chai Linsong, Ni Jinglei, Zuo Shuang, Lin Bingbing, Huang Jia 2026, 30 (36):  9604-9612.  doi: 10.12307/2026.911 Abstract ( 67 )   PDF (7265KB) ( 32 )   Save BACKGROUND: Most patients with post-stroke aphasia still have basic communication deficits 1 year after onset, highlighting an urgent need for accurate prognostic prediction tools to guide clinical rehabilitation decisions. OBJECTIVE: To construct a machine learning-based model for predicting language function prognosis at discharge in patients with post-stroke aphasia, aiming to improve prediction accuracy. METHODS: Clinical data were collected from 245 patients with post-stroke aphasia admitted to the Rehabilitation Hospital Affiliated to Fujian University of Traditional Chinese Medicine from July 1, 2022 to July 1, 2025, with an aphasia quotient change ≥ 6 points as the outcome indicator. The study cohort was randomly divided into training (n=171) and test (n=74) sets in a 7:3 ratio. Predictive factors were screened using recursive feature elimination. Six machine learning algorithms (logistic regression, random forest, decision tree, support vector machine, Gaussian naïve Bayes, and extreme gradient boosting classifier) were used to construct models. Internal validation was performed using the bootstrap method. Model performance was evaluated using receiver operating characteristic curves, calibration curves, and Shapley additive explanations analysis. RESULTS AND CONCLUSION: The language function improvement rate among the 245 patients with post-stroke aphasia was 69.80%. Ten factors, including age, female sex, education level, anomic aphasia, non-fluent aphasia, global aphasia, and baseline total score of the Chinese functional communication profile, were selected as predictive factors. The Gaussian naïve Bayes model performed best in the test set, with an area under the curve of 0.71 and an F1 score of 0.83. The calibration curve showed good consistency between predicted probabilities and actual outcomes (Brier score=0.19). Shapley additive explanations analysis identified anomic aphasia (0.24), advanced age (0.17), female sex (0.08), baseline total score of the Chinese Functional Communication Profile (0.08), and non-fluent aphasia (0.07) as key risk factors. These findings indicate that the Gaussian naïve Bayes prediction model based on machine learning can effectively identify language functional prognosis risks at discharge in patients with post-stroke aphasia, providing decision support for individualized rehabilitation interventions. Figures and Tables | References | Related Articles | Metrics

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Molecular docking and network pharmacology analyses of Shalin Decoction in regulating crystal homeostasis: active components and multi-target mechanisms Su Gangling, Liu Chunying, Cui Yangqing, Zhou Shaohu, Liu Shiyong, Wu Zhimin 2026, 30 (36):  9613-9620.  doi: 10.12307/2026.906 Abstract ( 44 )   PDF (18141KB) ( 27 )   Save BACKGROUND:  Shalin Decoction, a traditional Chinese medicine formula, has been shown to significantly inhibit crystal formation. Although preliminary studies have revealed the multi-target regulatory potential of its compositions, the systemic regulatory network underlying its therapeutic effects remains to be fully elucidated.   OBJECTIVE: To identify the potential active components, target proteins, and their interaction networks in Shalin decoction using network pharmacology and molecular docking approaches, and to explore its potential molecular mechanisms in treating kidney stones.   METHODS: The active compounds of Shalin Decoction were retrieved from the TCMSP database (Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform), and potential drug-like molecules were screened. Candidate targets of Shalin Decoction were identified by intersecting with kidney stone-related genes from the GeneCard database. A protein-protein interaction network was then constructed using the STRING database, and core targets were extracted for Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses. Tissue-specific expression of core genes was analyzed using the BioGPS database. Molecular docking was further employed to visualize the binding affinities between potential targets and their corresponding compounds.   RESULTS AND CONCLUSION: A total of 15 potential active ingredients and 74 corresponding targets of Shalin Decoction were screened from the TCMSP database. Protein-protein interaction network analysis identified interleukin-6, caspase-3, tumor necrosis factor, proto-oncogene protein JUN, B-cell lymphoma 2, peroxisome proliferator-activated receptor gamma, transforming growth factor beta 1, and proto-oncogene protein FOS as the core targets with the highest degree centrality. Enrichment analysis indicated that these targets were significantly associated with kidney stone-related biological processes and Kyoto Encyclopedia of Genes and Genomes pathways. Molecular docking further confirmed that hydrogen bonding and hydrophobic interactions were key forces mediating the stable binding between active compounds and their target proteins.  Figures and Tables | References | Related Articles | Metrics

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Mechanism underlying Qu Shi Jie Du Formula for treating hyperuricemia in mice: an analysis integrating UHPLC-QE-MS with network pharmacology Lin Kun, He Xianshun, Zhang Junjiao, Han Mingli, He Mincong, Wei Qiushi 2026, 30 (36):  9621-9631.  doi: 10.12307/2026.900 Abstract ( 62 )   PDF (7299KB) ( 19 )   Save BACKGROUND: Qu Shi Jie Du Formula (QSJDF) is a clinical empirical prescription for preventing and treating hyperuricemia and gout based on the “dampness-toxin accumulation” theory. It exerts effects of eliminating dampness, detoxifying, and relaxing tendons and collaterals. However, its underlying mechanism remains unclear. OBJECTIVE: To investigate the therapeutic effects and bone protection mechanism of QSJDF in hyperuricemia mice. METHODS: (1) The chemical constituents of QSJDF were identified using ultra-high performance liquid chromatography – Q-Exactive mass spectrometry. Target genes of these components related to hyperuricemia were retrieved from relevant databases. Core targets for QSJDF in treating hyperuricemia were identified via network pharmacology and subjected to Kyoto Encyclopedia of Genes and Genomes pathway enrichment and Gene Ontology functional annotation. (2) Forty Balb/c mice were randomly divided into five groups (n=8 per group): blank control, model, allopurinol, QSJDF, and QSJDF + allopurinol combination. Except for the blank control group, hyperuricemia models were established. After 28 days of modeling, mice received daily intragastric administration for 4 weeks: blank control and model groups received sodium carboxymethyl cellulose; allopurinol group received allopurinol; QSJDF group received QSJDF; combination group received both QSJDF and allopurinol. Samples were collected 6 hours after the final dose. Serum levels of uric acid, creatinine, blood urea nitrogen, procollagen type I N-terminal propeptide, C-terminal telopeptide of type I collagen, and osteocalcin were measured. Renal pathology was observed using hematoxylin-eosin and Sirius Red staining. Bone mass of the knee joint was analyzed by Micro-CT. Osteoclast numbers were assessed via tartrate-resistant acid phosphatase staining. Immunofluorescence staining was performed to detect p53 and glucose transporter 9 in renal tissue and c-JUN expression in knee joints. Pearson correlation analysis was used to evaluate relationships between Micro-CT parameters and c-JUN fluorescence intensity. RESULTS AND CONCLUSION: (1) A total of 75 active components were identified in QSJDF, with the top ones being curcumin, quercetin, apigenin, soy isoflavones, and genistein. A total of 198 predicted targets were obtained, with core targets including TP53, tumor necrosis factor, interleukin-6, JUN, interleukin-1B, and nuclear factor-KB1, mainly enriched in p53, hypoxia-inducible factor-1, and nuclear factor-κB signaling pathways. (2) Serum assays showed that QSJDF reduced renal function indicators (uric acid, creatinine, blood urea nitrogen) and bone metabolism indicators (procollagen type I N-terminal propeptide and C-terminal telopeptide of type I collagen) in hyperuricemia mice. Hematoxylin-eosin and Sirius Red staining demonstrated renal protective effects. Micro-CT and tartrate-resistant acid phosphatase staining revealed increased tibial bone mass and reduced osteoclast numbers. Immunofluorescence showed decreased expression of p53 and glucose transporter 9 in the kidney and c-JUN in the knee joint. Pearson analysis indicated a significant positive correlation between trabecular separation and c-JUN fluorescence intensity (r > 0, P < 0.05), while bone volume fraction and trabecular number were significantly negatively correlated with c-JUN fluorescence intensity (r < 0, P < 0.05). To conclude, QSJDF exerts urate-lowering and renal protective effects by regulating the p53/glucose transporter 9 pathway and alleviates bone loss by modulating c-JUN/activating protein 1 signaling to influence osteoclast activity in hyperuricemia mice Figures and Tables | References | Related Articles | Metrics

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Visualization analysis of research hotspots and trends in the treatment of knee osteoarthritis Bei Chaoyong, Zhu Jialin, Yu Zhengze, Shi Niujin, Li Huarui, Peng Fenglin 2026, 30 (36):  9632-9639.  doi: 10.12307/2026.369 Abstract ( 52 )   PDF (3367KB) ( 20 )   Save BACKGROUND: Comparative studies have been conducted on different treatment methods for knee osteoarthritis, but there remains a relative paucity of visualized studies on global research trends in the treatment of knee osteoarthritis that span a long period of time and include a large number of literature sources. OBJECTIVE: To reveal global research trends over the past two decades in the treatment of knee osteoarthritis through bibliometric methods and to conduct visualized analyses of research entities and hotspots, thereby providing references for future studies. METHODS: This study selected relevant literature from the Web of Science Core Collection database. CiteSpace software was used for visual analysis of publication trends, main institutions, core authors, keywords, centrality, clustering and co-cited literature, to reveal the knowledge base and frontier hotspots in this field. RESULTS AND CONCLUSION: (1) A total of 5 806 publications from 2005 to 2024 were finally included; the overall number of publications from the selected articles showed an upward trend. China, the United States, and Australia are the three countries with the largest number of publications in this field. Harvard University is the most prolific research institution. (2) Keywords co-occurrence analysis displayed that platelet-rich plasma, arthroplasty, sodium hyaluronate and acupuncture have been attempted for the treatment of knee osteoarthritis in clinical practice. (3) Burstiness analysis of keywords identified emerging hotspots such as cells, unicompartmental knee arthroplasty, and stromal vascular fraction in recent years. (4) Co-citation analysis revealed that foundational literature focusing on pharmacological or non-pharmacological treatments and clinical guidelines constituted the basis of this field. (5) Cluster analysis revealed that there are controversies regarding the treatment of knee osteoarthritis with biological agents such as mesenchymal stem cells and platelet-rich plasma. Large sample randomized controlled trials are still needed to verify their efficacy, reflecting the current and future research hotspots in field.  Figures and Tables | References | Related Articles | Metrics

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Causal association study between resting-state brain network functional and structural connectivity and autoimmune diseases Wang Xiaoxuan, Xiao Lu, Miao Changhong, Yuan Weijie, Guo Dengzhou, Yuan Ziwei 2026, 30 (36):  9640-9648.  doi: 10.12307/2026.393 Abstract ( 54 )   PDF (1971KB) ( 14 )   Save BACKGROUND: Recent studies have shown a potential association between resting-state brain networks and autoimmune diseases. However, the underlying neuro-regulatory mechanisms remain unclear. OBJECTIVE: To explore the potential causal relationship between resting-state brain network functional/structural connectivity and autoimmune diseases using bidirectional two-sample Mendelian randomization. METHODS: The resting-state brain network genome-wide association study (GWAS) data from the UK Biobank database (a large biomedical database jointly constructed by the UK government, the Scottish government, and the Wellcome Trust), as well as GWAS data on autoimmune diseases integrated from the IEU Open GWAS database (a genome-wide association study database developed by the MRC Integrated Epidemiology Unit (IEU) at the University of Bristol in the UK), GWAS Catalog database (a genome wide association study database under the European Molecular Biology Laboratory-European Bioinformatics Institute (EMBL-EBI)), and FinnGen database (a large public-private partnership project initiated in Finland) were used to screen independent genetic variants significantly associated with exposure factors as instrumental variables. Bidirectional Mendelian randomization was conducted for seven resting-state brain networks (functional/structural connectivity) and nine autoimmune diseases (rheumatoid arthritis, systemic lupus erythematosus, multiple sclerosis, type 1 diabetes, Sjogren's syndrome, myasthenia gravis, ulcerative colitis, inflammatory bowel disease, and celiac disease). The inverse variance weighted method served as the primary analysis method to assess potential bidirectional causal relationships, supplemented by MR-Egger, weighted median, simple mode and weighted mode methods for complementary analysis. Multiple testing correction was performed using the false discovery rate (FDR) with q < 0.05. Cochran’s Q test was used to evaluate heterogeneity, Mendelian randomization Pleiotropy RESidual Sum and Outlier method was used to evaluate horizontal pleiotropy, leave-one-out method was used to test robustness of results, and cross-database consistency test was used to verify statistical significance of autoimmune disease data from different sources, ensuring reliability of research findings. RESULTS AND CONCLUSION: In forward Mendelian randomization analyses involving nine autoimmune diseases, increased functional connectivity in the default mode network was positively associated with a higher risk of rheumatoid arthritis [odds ratio (OR)=1.431, 95% confidence interval (CI)=1.108–1.850, P=0.006, q=0.048). Reduced functional connectivity in the dorsal attention network may correlate with decreased rheumatoid arthritis risk (OR=0.797, 95%CI=0.677–0.938, P=0.006, q=0.048). Decreased structural connectivity in the visual network was linked to reduced inflammatory bowel disease risk (OR=0.957, 95%CI=0.932–0.982, P < 0.001, q=0.014), while weakened structural connectivity in the ventral attention network was similarly correlated with diminished inflammatory bowel disease risk (OR=0.724, 95%CI=0.575–0.911, P=0.005, q=0.04). Reverse Mendelian randomization analyses revealed no significant causal effects of autoimmune diseases on resting-state brain network connectivity strength (q > 0.05). This Mendelian randomization study provides the first evidence of a bidirectional causal relationship between resting-state brain network connectivity abnormalities and autoimmune disease pathogenesis, thereby elucidating the pivotal role of brain network dysregulation in autoimmune pathology. These findings provide a theoretical basis for the development of neuroimaging biomarkers and neuroimmune targeted therapy strategies for autoimmune diseases. Figures and Tables | References | Related Articles | Metrics