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08 July 2025, Volume 29 Issue 19 Previous Issue   

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Programmed death receptor 1 inhibits osteogenic differentiation of rat bone marrow mesenchymal stem cells in a high glucose environment Han Nianrong, Huang Yifei, Akram · Osman, Liu Yanlu, Hu Wei 2025, 29 (19):  3961-3967.  doi: 10.12307/2025.075 Abstract ( 152 )   PDF (2623KB) ( 108 )   Save BACKGROUND: The mechanism of programmed death receptor-1 (PD-1) effect on osteogenic differentiation of bone marrow mesenchymal stem cells in high glucose environment remains unclear.  OBJECTIVE: To explore the effect of PD-1 on osteogenic differentiation of rat bone marrow mesenchymal stem cells in high glucose environment and its regulatory mechanism. METHODS: Rat bone marrow mesenchymal stem cells were randomly divided into normal glucose group (5.6 mmol/L), high glucose group (30 mmol/L), PD-1 overexpression group, PD-1 overexpression no-load group, PD-1 knockdown group, PD-1 knockdown no-load group, and PI3K/AKT pathway inhibitor group (PD-1 knockdown+5 μmol/L LY294002). Rat bone marrow mesenchymal stem cells were cultured in high glucose to simulate the diabetic environment in vitro. The mRNA expression of PD-1 and ligand PD-L1 and the mRNA expression of osteogenic markers Runx2 and OSX in rat bone marrow mesenchymal stem cells were detected by qRT-PCR. The osteogenic differentiation ability was observed by alkaline phosphatase staining and alizarin red staining. Cell proliferation was detected by CCK-8 assay. The protein expressions of PD-1, PD-L1, p-PI3K, and p-AKT were detected by western blot assay.  RESULTS AND CONCLUSION: (1) The levels of PD-1 and PD-L1 were significantly increased in the high glucose environment in vitro, and the osteogenic differentiation ability of bone marrow mesenchymal stem cells was inhibited in the high glucose environment. (2) Knockdown of PD-1 expression could promote osteogenic differentiation of bone marrow mesenchymal stem cells, increase cell proliferation activity, and activate the PI3K/AKT pathway. (3) After addition of PI3K/AKT pathway inhibitor LY294002, the ability of bone marrow mesenchymal stem cells to differentiate into osteoblasts decreased. The results show that PD-1 is dependent on the PI3K/AKT signaling pathway to inhibit osteogenic differentiation of rat bone marrow mesenchymal stem cells under high glucose environment. Figures and Tables | References | Related Articles | Metrics

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Mogroside V promotes osteogenic differentiation of bone marrow mesenchymal stem cells by modulating M1 polarization of macrophages under high glucose condition Ye Zhimao, Hui Jiuying, Zhong Xiaoxia, Mai Yuying, Li Hao 2025, 29 (19):  3968-3975.  doi: 10.12307/2025.062 Abstract ( 156 )   PDF (2198KB) ( 211 )   Save BACKGROUND: The diabetic microenvironment can cause excessive M1 polarization of macrophages, and this hyperglycemic inflammatory state can inhibit osteogenic differentiation of bone marrow mesenchymal stem cells, thus affecting the healing of diabetic bone defects. Studies have indicated that mogroside V possesses anti-inflammatory, antioxidant, and hypoglycemic properties. However, its potential to modulate M1 polarization of macrophages and osteogenic differentiation of bone marrow mesenchymal stem cells under high glucose and inflammatory condition remains unclear. OBJECTIVE: To explore the effect of mogroside V on regulating M1 macrophage polarization and its effect on osteogenic differentiation of bone marrow mesenchymal stem cells under high glucose and inflammatory condition.  METHODS: Murine diabetic models were established using C57BL/6 mice. Bone marrow-derived macrophages were isolated from tibia and fibula of normal and diabetic mice, and cultured in low-glucose and high-glucose media. Then M1 polarization of bone marrow-derived macrophages was induced using lipopolysaccharide and interferon-γ. Bone marrow-derived macrophages were treated with 160, 320, and 640 μmol/L mogroside V. Flow cytometry was employed to determine the proportion of F4/80+CD86+ cells. qRT-PCR was utilized to assess mRNA expression levels of inducible nitric oxide synthase, interleukin 1β, and interleukin 6. ELISA was employed to evaluate tumor necrosis factor-α secretion in bone marrow-derived macrophage supernatants. Bone marrow mesenchymal stem cells were isolated from tibia and fibula of C57BL/6 suckling mice, and induced osteogenic differentiation using low- or high-glucose osteogenic induction medium. Bone marrow mesenchymal stem cells were treated with M1 macrophage-conditioned mediums with or without 320 μmol/L mogroside V in osteogenic differentiation process. qRT-PCR was employed to assess the mRNA expression of alkaline phosphatase, Runt-related factor 2, osteocalcin, and osteopontin on day 14 after osteogenic induction. Alizarin red staining and quantitative analysis were conducted to evaluate calcium deposition on day 21 after osteogenic induction.  RESULTS AND CONCLUSION: (1) Flow cytometry results showed that with the treatment of 320 and 640 μmol/L mogroside V, the proportion of F4/80+CD86+ bone marrow-derived macrophages was significantly lower than that in the high-glucose control group (P < 0.05). (2) qRT-PCR results showed that with the treatment of 160, 320, and 640 μmol/L mogroside V, the mRNA expression levels of inducible nitric oxide synthase and interleukin 6 were significantly lower than that in the high-glucose control group (P < 0.05). With the treatment of 320 and 640 μmol/L mogroside V, the mRNA expression level of interleukin 1β was significantly lower than that in the high-glucose control group (P < 0.05). (3) ELISA results exhibited that with the treatment of 160, 320, and 640 μmol/L mogroside V, the tumor necrosis factor-α secretion level was significantly lower than that in the high-glucose control group (P < 0.05). (4) With the treatment of 320 μmol/L mogroside V, calcium salt deposition was increased in bone marrow mesenchymal stem cells under high glucose and inflammatory conditions (P < 0.05), and the mRNA relative expression levels of alkaline phosphatase, Runt-related factor 2, and osteopontin were increased (P < 0.05). These findings indicate that mogroside V can promote osteogenic differentiation of bone marrow mesenchymal stem cells by inhibiting the M1 polarization of bone marrow-derived macrophages under high glucose and inflammatory conditions and reducing the generation of inflammatory factors.  Figures and Tables | References | Related Articles | Metrics

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Treadmill training activates endogenous neural stem cells to promote spinal cord injury repair in mice Chen Chanjuan, Shangguan Zeyu, Li Qizhe, Tan Wei, Li Qing 2025, 29 (19):  3976-3982.  doi: 10.12307/2025.043 Abstract ( 81 )   PDF (1987KB) ( 116 )   Save BACKGROUND: Treadmill training is one of the effective ways to promote the recovery of motor function after spinal cord injury. Treadmill training can promote neurogenesis, but the effect of different intensities of treadmill training on the activation of endogenous stem cells is still unclear. OBJECTIVE: To analyze the activation effect of different intensities of treadmill training on endogenous neural stem cells in the spinal cord of mice after spinal cord injury. METHODS: Fifty female C57BL/6J mice were divided into control group, spinal cord injury group, low-, moderate-, and high-intensity exercise groups with 10 mice in each group by random number table method. T10 segment spinal cord injury model was constructed by the clamp method in spinal cord injury group, low-, moderate-, and high-intensity exercise groups. On day 7 after spinal cord injury, mice in the low-, moderate-, and high-intensity exercise groups were respectively trained on the treadmill with corresponding intensity, 3 times /d, 10 min/ times, 6 times a week for 28 consecutive days. At 3, 7, 14, 21, and 28 days after treadmill training, the hind limb motor function was evaluated by BMS score. At 28 days after treadmill training, the spinal cord tissue of the injured area was obtained, and the expression of epidermal growth factor receptor, glial fibrillary acidic protein, and 5-Ethynyl-2'-deoxyuridine (EdU), a proliferative marker, was detected. Hematoxylin-eosin staining was used to observe the morphology of spinal cord.  RESULTS AND CONCLUSION: (1) The BMS score of mice in the spinal cord injury group was lower than that in the control group (P < 0.05). With the extension of treadmill training time, the BMS scores of mice with spinal cord injury gradually increased, and the BMS scores of mice in moderate-intensity exercise group on days 14 and 21 after treadmill training were higher than those in spinal cord injury group and low- and high-intensity exercise groups (P < 0.05). The BMS score of mice in moderate- and high-intensity exercise group was higher than that in spinal cord injury group and low-intensity exercise group at 28 days after treadmill training (P < 0.05). (2) Compared with the control group, the proportion of epidermal growth factor receptor and EdU positive cells was increased in spinal cord injury group (P < 0.05). Compared with spinal cord injury group, the proportion of epidermal growth factor receptor and EdU positive cells was increased in low-, moderate-, and high-intensity exercise groups (P < 0.05), and the highest was found in moderate-intensity exercise group. Compared with control group, the proportion of glial fibrillary acidic protein positive cells was increased in spinal cord injury group (P < 0.05). Compared with spinal cord injury group, the proportion of glial fibrillary acidic protein positive cells was lower in low-, moderate-, and high-intensity exercise groups (P < 0.05), and the moderate-intensity exercise group was the lowest. (3) Hematoxylin-eosin staining showed that a large cavity was formed in the injured area of mice with spinal cord injury, and the cavity in the injured area of mice with spinal cord injury decreased after different intensities of treadmill training, and the decrease was most obvious in the moderate-intensity exercise group. (4) These results indicate that low-, moderate-, and high-intensity treadmill training can promote the recovery of motor function of mice with spinal cord injury by activating endogenous neural stem cells, and the effect of moderate-intensity exercise training is the most obvious. Figures and Tables | References | Related Articles | Metrics

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Effects of long non-coding RNA KIAA0125 on proliferation and apoptosis of acute myeloid leukemia U937 cells Hu Huali, Deng Fahua, Liu Yuancheng, Wang Siqi, Zhang Jingxin, Lu Tingting, Huang Hai, Wei Sixi 2025, 29 (19):  3983-3991.  doi: 10.12307/2025.510 Abstract ( 101 )   PDF (3173KB) ( 103 )   Save BACKGROUND: U937 cells can be used as a cell model for studying the biological characteristics, signaling pathways, and therapeutic targets of acute myeloid leukemia. Although it has been reported that long non-coding RNA KIAA0125 is highly expressed in acute myeloid leukemia, its biological function in U937 cells remains unclear, and its mechanism of action in the occurrence and development of acute myeloid leukemia needs to be further clarified.  OBJECTIVE: To investigate the expression level of long non-coding RNA KIAA0125 in peripheral blood of patients with acute myeloid leukemia and its effect on the proliferation and apoptosis of U937 cells.  METHODS: RNA-sequencing was used to analyze the bone marrow monocyte samples from acute myeloid leukemia patients, and the differentially expressed gene long non-coding RNA KIAA0125 was screened. The expression of long non-coding RNA KIAA0125 in peripheral blood of patients with acute myeloid leukemia was detected by qRT-PCR. The relationship between long non-coding RNA KIAA0125 mRNA expression and prognosis in bone marrow cells of 173 acute myeloid leukemia patients and 70 healthy people was statistically analyzed by GEPIA database. Subsequently, recombinant lentivirus technology and CRISPR/Cas9-SAM technology were used to construct U937 cell lines with knockdown/overexpression of long non-coding RNA KIAA0125. qRT-PCR was used to detect the knockdown/overexpression efficiency of long non-coding RNA KIAA0125. Next, CCK-8 assay, flow cytometry, and western blot assay were used to detect the effects of knockdown/overexpression of long non-coding RNA KIAA0125 on the proliferation and apoptosis of U937 cells. Finally, western blot assay was used to detect the effect of knockdown/overexpressed long non-coding RNA KIAA0125 on Wnt/β-catenin signaling pathway-related proteins.  RESULTS AND CONCLUSION: (1) The results of qRT-PCR showed that long non-coding RNA KIAA0125 was highly expressed in peripheral blood of acute myeloid leukemia patients. The results of GEPIA database showed that long non-coding RNA KIAA0125 was highly expressed in bone marrow cells of acute myeloid leukemia patients, and the high expression group had worse overall survival. (2) The knockdown efficiency of long non-coding RNA KIAA0125 in knockdown group was 70%, and the U937 cells that stably down-regulated long non-coding RNA KIAA0125 expression were successfully constructed. The expression of long non-coding RNA KIAA0125 in overexpression group was four times that of vector group, and stable U937 cells were successfully constructed. (3) Knockdown of long non-coding RNA KIAA0125 inhibited the proliferation of U937 cells and promoted their apoptosis. Overexpression of long non-coding RNA KIAA0125 promoted the proliferation of U937 cells but had no significant effect on the apoptosis of U937 cells. (4) Knockdown of long non-coding RNA KIAA0125 inhibited the activity of Wnt/β-catenin signaling pathway, while overexpression of long non-coding RNA KIAA0125 activated Wnt/β-catenin signaling pathway. These results confirm that long non-coding RNA KIAA0125 is highly expressed in acute myeloid leukemia peripheral blood. Long non-coding RNA KIAA0125 may affect the proliferation and apoptosis of U937 cells by regulating the Wnt/β-catenin signaling pathway, and may be a potential prognostic marker for acute myeloid leukemia.  Figures and Tables | References | Related Articles | Metrics

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Effect of oxymatrine on expression of stem markers and osteogenic differentiation of periodontal ligament stem cells Luo Jing, Yong Min, Chen Qi, Yang Changyi, Zhao Tian, Ma Jing, Mei Donglan, Hu Jinpeng, Yang Zhaojun, Wang Yuran, Liu Bo 2025, 29 (19):  3992-3999.  doi: 10.12307/2025.056 Abstract ( 141 )   PDF (1600KB) ( 181 )   Save BACKGROUND: Human periodontal ligament stem cells are potential functional cells for periodontal tissue engineering. However, long-term in vitro culture may lead to reduced stemness and replicative senescence of periodontal ligament stem cells, which may impair the therapeutic effect of human periodontal ligament stem cells.  OBJECTIVE: To investigate the effect of oxymatrine on the stemness maintenance and osteogenic differentiation of periodontal ligament stem cells in vitro, and to explore the potential mechanism.   METHODS: Periodontal ligament stem cells were isolated from human periodontal ligament tissues by tissue explant enzyme digestion and cultured. The surface markers of mesenchymal cells were identified by flow cytometry. Periodontal ligament stem cells were incubated with 0, 2.5, 5, and 10 μg/mL oxymatrine. The effect of oxymatrine on the proliferation activity of periodontal ligament stem cells was detected by CCK8 assay. The appropriate drug concentration for subsequent experiments was screened. Western blot assay was used to detect the expression of stem cell non-specific proteins SOX2 and OCT4 in periodontal ligament stem cells. qRT-PCR and western blot assay were used to detect the expression levels of related osteogenic genes and proteins in periodontal ligament stem cells.  RESULTS AND CONCLUSION: (1) The results of CCK8 assay showed that 2.5 μg/mL oxymatrine significantly enhanced the proliferative activity of periodontal stem cells, and the subsequent experiment selected 2.5 μg/mL oxymatrine to intervene. (2) Compared with the blank control group, the protein expression level of SOX2, a stem marker of periodontal ligament stem cells in the oxymatrine group did not change significantly (P > 0.05), and the expression of OCT4 was significantly up-regulated (P < 0.05). (3) Compared with the osteogenic induction group, the osteogenic genes ALP, RUNX2 mRNA expression and their osteogenic associated protein ALP protein expression of periodontal ligament stem cells were significantly down-regulated in the oxymatrine + osteogenic induction group (P < 0.05). (4) The oxymatrine up-regulated the expression of stemness markers of periodontal ligament stem cells and inhibited the bone differentiation of periodontal ligament stem cells, and the results of high-throughput sequencing showed that it may be associated with WNT2, WNT16, COMP, and BMP6.  Figures and Tables | References | Related Articles | Metrics

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Effect and mechanisms of highly active umbilical cord mesenchymal stem cells on aging spleen in elderly tree shrews Ye Li, Tian Chuan, Zhao Xiaojuan, Chen Mengdie, Ye Qianqian, Li Qiang, Liao Zhuyin, Li Ye, Zhu Xiangqing, Ruan Guangping, He Zhixu, Shu Liping, Pan Xinghua 2025, 29 (19):  4000-4010.  doi: 10.12307/2025.047 Abstract ( 119 )   PDF (4326KB) ( 159 )   Save BACKGROUND: Spleen has the functions of blood storage, hematopoiesis, and immunity. With the increase of age, the structural degeneration and functional decline of spleen lead to the impairment of immune system function, thus accelerating the aging process of the body. The treatment of spleen aging in tree shrews with highly active umbilical cord mesenchymal stem cells has not been reported.  OBJECTIVE: To explore the intervention effect and mechanism of highly active umbilical cord mesenchymal stem cells on spleen aging in tree shrews.  METHODS: Highly active umbilical cord mesenchymal stem cells were isolated, cultured, and obtained from the umbilical cord tissue of newborn tree shrews by caesarean section. The differentiation abilities of adipogenesis, osteogenesis, and chondrogenesis were detected by three-line differentiation kit. Cell cycle and surface markers were detected by flow cytometry. The second generation of highly active umbilical cord mesenchymal stem cells were transfected with Genechem Green Fluorescent Protein with infection complex values of 100, 120, 140, 160, 180, and 200, respectively, to screen the best transfection conditions. After transfection, the fourth generation of highly active umbilical cord mesenchymal stem cells was injected into the tail vein of tree shrews in the elderly treatment group. The young control group and the aged model group were not given special treatment. After 4 months of treatment, the spleen tissue was taken and the structure of the spleen was observed by hematoxylin-eosin staining. β-Galactosidase staining was used to detect the activity of aging-related galactosidase. Immunohistochemical staining was used to detect the expression levels of p21 and p53 proteins. Ki67 and PCNA immunofluorescence staining was used to detect cell proliferation activity. Immunofluorescence staining was used to detect the expression levels of spleen autophagy protein molecules Beclin 1 and APG5L/ATG5. Reactive oxygen species fluorescence staining was used to detect the content of reactive oxygen species in spleen tissue. CD3 immunofluorescence staining was used to detect the change of the proportion of total T lymphocytes. The secretion levels of interleukin 1β and transforming growth factor β1 in spleen were detected by enzyme linked immunosorbent assay. The distribution of highly active umbilical cord mesenchymal stem cells labeled with green fluorescent protein in spleen tissue was observed by DAPI double staining of nucleus.  RESULTS AND CONCLUSION: (1) Highly active umbilical cord mesenchymal stem cells grew in a short spindle shape with fish-like growth, with a large proportion of G0/G1 phase, and had the potential to differentiate into adipogenesis, osteogenesis, and chondrogenesis. (2) Multiplicity of infection=140 and transfection for 72 hours were the best conditions for labeling tree shrews highly active umbilical cord mesenchymal stem cells with Genechem Green Fluorescent Protein. (3) Compared with the aged model group, in the aged treatment group, the spleen tissue cells of tree shrews were arranged closely, and the area of white pulp was increased (P < 0.01); the boundary between red pulp and white pulp was clear; the proportion of germinal centers did not show statistically significant difference (P > 0.05). The activity level of galactosidase related to spleen tissue aging was decreased (P < 0.001), and the expression levels of aging protein molecules p21 and p53 were down-regulated (P < 0.001). The expression levels of proliferation-related molecules Ki67 and PCNA were up-regulated (P < 0.001, P < 0.05); expression levels of autophagy-related molecules Beclin 1 and APG5L/ATG5 were up-regulated (P < 0.001), and the content of reactive oxygen species decreased (P < 0.001), and the proportion of CD3+ T cells increased (P < 0.05). The secretion level of interleukin 1β in the aging-related secretion phenotype decreased (P < 0.001); no significant difference was found in transforming growth factor β1 level (P > 0.05). Compared with the young control group, the above indexes were significantly different in the elderly treatment group (P < 0.05). (4) Green fluorescent cells labeled with green fluorescent protein were observed in spleen tissue of tree shrews the elderly treatment group by frozen tissue section observation. The results show that intravenous infusion of highly active umbilical cord mesenchymal stem cells can migrate to spleen tissue, inhibit the production of reactive oxygen species, down-regulate the expression of aging-related proteins, induce autophagy, promote cell proliferation, reduce chronic inflammation, and then improve the structure and function of spleen tissue.  Figures and Tables | References | Related Articles | Metrics

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Panax notoginseng saponins regulate differential miRNA expression in osteoclast exosomes and inhibit ferroptosis in osteoblasts Tao Hongcheng, Zeng Ping, Liu Jinfu, Tian Zhao, Ding Qiang, Li Chaohui, Wei Jianjie, Li Hao 2025, 29 (19):  4011-4021.  doi: 10.12307/2025.064 Abstract ( 122 )   PDF (3242KB) ( 200 )   Save BACKGROUND: Steroid-induced femoral head necrosis is mostly caused by long-term and extensive use of hormones, but its specific pathogenesis is not yet clear and needs further study. OBJECTIVE: To screen out the differential miRNAs in osteoclast exosomes after the intervention of Panax notoginseng saponins, and on this basis, to further construct an osteogenic-related ferroptosis regulatory network to explore the potential mechanism and research direction of steroid-induced osteonecrosis of the femoral head. METHODS: MTT assay was used to detect the toxic effects of different concentrations of dexamethasone and different mass concentrations of Panax notoginseng saponins on Raw264.7 cell line. Tartrate resistant acid phosphatase staining and TUNEL assay were used to detect the effects of Panax notoginseng saponins on osteoclast inhibition and apoptosis. Exosomes were extracted from cultured osteoclasts with Panax notoginseng saponins intervention. Exosomes from different groups were sequenced to identify differentially expressed miRNAs. CytoScape 3.9.1 was used to construct and visualize the regulatory network between differentially expressed miRNAs and mRNAs. Candidate mRNAs were screened by GO analysis and KEGG analysis. Finally, the differential genes related to ferroptosis were screened out, and the regulatory network of ferroptosis-related genes was constructed. RESULTS AND CONCLUSION: (1) The concentration of dexamethasone (0.1 μmol/L) and Panax notoginseng saponins (1 736.85 μg/mL) suitable for intervention of Raw264.7 cells was determined by MTT assay. (2) Panax notoginseng saponins had an inhibitory effect on osteoclasts and could promote their apoptosis. (3) Totally 20 differentially expressed miRNAs were identified from osteoclast-derived exosome samples, and 11 differentially expressed miRNAs related to osteogenesis were predicted by target mRNAs. The regulatory networks of 4 up-regulated differentially expressed miRNAs corresponding to 155 down-regulated candidate mRNAs and 7 down-regulated differentially expressed miRNAs corresponding to 238 up-regulated candidate mRNAs were constructed. (4) Twenty-four genes related to ferroptosis were screened out from the differential genes. Finally, 12 networks were constructed (miR-98-5p/PTGS2, miR-23b-3p/PTGS2, miR-425-5p/TFRC, miR-133a-3p/TFRC, miR-185-5p/TFRC, miR-23b-3p/NFE2L2, miR-23b-3p/LAMP2, miR-98-5p/LAMP2, miR-182-5p/LAMP2, miR-182-5p/TLR4, miR-23b-3p/ZFP36, and miR-182-5p/ZFP36). These results indicate that Panax notoginseng saponins may regulate osteoblast ferroptosis by regulating the expression of miRNAs derived from osteoclast exosomes, thus providing a new idea for the study of the mechanism of steroid-induced femoral head necrosis.  Figures and Tables | References | Related Articles | Metrics

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Osteogenic/odontogenic differentiation ability of human dental pulp stem cells under photocrosslinked composite hydrogel scaffold Yang Dujuan, Cheng Mengke, Liu Jia 2025, 29 (19):  4022-4028.  doi: 10.12307/2025.071 Abstract ( 115 )   PDF (2402KB) ( 250 )   Save BACKGROUND: The composite hydrogel scaffold formed by crosslinking of gelatin-methacryloyl (Gel-MA) and treated dentin matrix (TDM) under a certain proportion of ultraviolet light has good porosity, mechanical properties, swelling properties, and biodegradation rate, which provides a new idea and method for clinical pulp regeneration of young permanent teeth. OBJECTIVE: To explore the effect of Gel-MA/TDM composite hydrogel scaffold with 1:2 mass ratio on the proliferation ability and osteogenic/odontoblast differentiation ability of human dental pulp stem cells.  METHODS: The passage 3 dental pulp stem cells were inoculated into the Gel-MA/TDM composite hydrogel scaffold with a mass ratio of 1:2. The proliferation ability of human dental pulp stem cells in the composite hydrogel scaffold was detected by CCK-8 assay. Dental pulp stem cells at passage 3 were cultured in Gel-MA/TDM composite hydrogel scaffold with a mass ratio of 1:2 for osteogenic induction. The formation of mineralized nodules was observed by alkaline phosphatase and alizarin red staining. The gene expression levels of odontogenic factors (dentin matrix protein 1, dentin sialophosphoprotein), and osteogenic factors (osteocalcin, Runt-related transcription factor 2) were detected by RT-PCR. RESULTS AND CONCLUSION: (1) The results of CCK-8 assay showed that the proliferation ability of dental pulp stem cells increased significantly in the first 7 days, and slowed down on day 10. (2) The results of alkaline phosphatase staining and alizarin red staining showed that the alkaline phosphatase activity and the formation of mineralized nodules of dental pulp stem cells in the Gel-MA/TDM composite hydrogel group were stronger than those in Gel-MA hydrogel group (P < 0.05). (3) RT-PCR results showed that the gene expression levels of dentin matrix protein 1, dentin sialophosphoprotein, osteocalcin, and Runt-related transcription factor 2 in dental pulp stem cells in Gel-MA/TDM composite hydrogel group were significantly higher than those in Gel-MA hydrogel group (P < 0.05). The gene expression level at 14 days was significantly higher than that at 7 days (P < 0.05). The results conclude that the dental pulp stem cells cultured on Gel-MA/TDM composite hydrogel scaffolds with a mass ratio of 1:2 exhibit a good proliferation ability, which can strengthen the osteogenic and odontogenic differentiation abilities of dental pulp stem cells. Figures and Tables | References | Related Articles | Metrics

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Material basis and action mechanism of drug-containing serum of Modified Erxian Pill inhibiting macrophage pyroptosis Li Siyuan, Wang Yuru, Xu Ye, Guo Di, Nan Nan, Liu Yang, Zhao Jie, Hao Huiqin 2025, 29 (19):  4029-4037.  doi: 10.12307/2025.077 Abstract ( 77 )   PDF (1875KB) ( 197 )   Save BACKGROUND: Our previous study found that Modified Erxian Pill could alleviate inflammation in collagen-induced arthritis rats, but its mechanism needs to be further verified. OBJECTIVE: To analyze the components absorbed in the blood of Modified Erxian Pill, and observe the effect of the drug-containing serum of Modified Erxian Pill on pyroptosis of J774A.1 macrophages.  METHODS: (1) Analysis of components absorbed in the blood of Modified Erxian Pill: Ultra-high performance liquid chromatography-high resolution mass spectrometry was used to detect and identify Modified Erxian Pill and its components absorbed in the blood. (2) Effect of the drug-containing serum of Modified Erxian Pill on pyroptosis of J774A.1 macrophages: Molecular docking technology was used to initially verify the sesquiterpenoids and NLRP3 in components absorbed in the blood of Modified Erxian Pill. J774A.1 macrophages were randomly divided into blank control group, lipopolysaccharide+adenosine triphosphate group, and lipopolysaccharide+adenosine triphosphate+Modified Erxian Pill with low (2.5%), medium (5%), and high (10%) dose groups. The release of lactate dehydrogenase in the cell supernatant of each group was detected according to the kit instructions. The levels of interleukin-1β and interleukin-18 in cell supernatant were detected in each group by ELISA. The cell membrane damage was detected by Hoechst/PI staining. The expression levels of NLRP3, Caspase-1, GSDMD, and GSDMD-N protein in the cells of each group were detected by western blot assay. RESULTS AND CONCLUSION: (1) A total of 32 active components of Modified Erxian Pill were identified, and 21 components entered the blood. The main components into blood included a variety of sesquiterpenoids. (2) Molecular docking results showed that 3-O-Acetyl-13-deoxyphomenone, Incensol oxide, Atractylenolide III, Rupestonic acid, and 3,7-Dihydroxy-9,11-eremophiladien-8-one had good binding activity with NLRP3. (3) Compared with the blank control group, lactate dehydrogenase activity and the expression levels of interleukin-1β and interleukin-18 were significantly increased in cell supernatant of lipopolysaccharide+adenosine triphosphate group (P < 0.001). Hoechst/PI staining showed that the number of PI-positive cells was significantly increased. After the intervention of lipopolysaccharide+adenosine triphosphate+Modified Erxian Pill group, all of them showed different degrees of reduction. (4) Compared with the blank control group, NLRP3, Caspase-1, GSDMD, and GSDMD-N protein expression levels were significantly increased in the lipopolysaccharide+adenosine triphosphate group (P < 0.05). Compared with lipopolysaccharide+adenosine triphosphate group, the protein expressions of NLRP3, Caspase-1, GSDMD, and GSDMD-N were significantly decreased in the lipopolysaccharide+adenosine triphosphate+Modified Erxian Pill group (P < 0.05), and had a certain dose dependence. These findings verify that the drug-containing serum of Modified Erxian Pill may inhibit the pyroptosis of J774A.1 macrophages by regulating the NLRP3/Caspase-1/GSDMD pathway. Figures and Tables | References | Related Articles | Metrics

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Electrical stimulation induces miR-741-3p to regulate Radil and promote Schwann cell migration Liu Qing, Gao Bo, Yang Xiao, Jiang Yu, Wang Pei 2025, 29 (19):  4038-4043.  doi: 10.12307/2025.080 Abstract ( 136 )   PDF (1985KB) ( 74 )   Save BACKGROUND: More and more animal experiments and clinical studies have confirmed that electrical stimulation can promote the repair of peripheral nerve injury, but the specific mechanism is not yet fully understood. OBJECTIVE: To investigate the effect of electrical stimulation-induced miR-741-3p regulating Radil on Schwann cell migration. METHODS: (1) Twelve male SD rats were randomly divided into electrical stimulation group and control group. The electrical stimulation group received continuous electrical stimulation for 7 days after sciatic nerve compression injury, while the control group was not treated after sciatic nerve compression. The injured nerves were taken on day 7 after operation. The expression difference of miR-741-3p between the two groups was verified by fluorescence in situ hybridization. (2) The target genes of miR-741-3p were predicted by miRDB, TargetScan, and miRWalk databases. (3) Schwann cells were transfected with miR-741-3p mimetic and its control, miR-741-3p inhibitor and its control, Radil siRNA and its control, miR-741-3p inhibitor+Radil siRNA and miR-741-3p inhibitor+siRNA control. The transfection efficiency was detected by RT-PCR. The migration ability of Schwann cells was detected by Transwell chamber.  RESULTS AND CONCLUSION: (1) The fluorescence intensity of miR-741-3p in the electrical stimulation group was lower than that in the control group. (2) The results of database prediction showed that 69 genes might be the target genes of miR-741-3p. Radil was one of the predicted target genes, which was mainly involved in cell adhesion and migration. (3) Compared with the miR-741-3p inhibitor control group, the number of Schwann cell migration increased in the miR-741-3p inhibitor group (P < 0.05). Compared with the miR-741-3p mimic control group, the number of Schwann cell migration in the miR-741-3p mimic group decreased (P < 0.05). Compared with the siRNA control group, the number of Schwann cell migration was decreased in the Radil siRNA group (P < 0.05). (4) Compared with miR-741-3p inhibitor control group, the expression level of Radil was increased in miR-741-3p inhibitor group. Compared with miR-741-3p mimic control group, the expression level of Radil was decreased in miR-741-3p mimic group. (5) Compared with miR-741-3p inhibitor+siRNA control group, the number of Schwann cell migration was reduced in miR-741-3p inhibitor+Radil siRNA group (P < 0.05). The results showed that electrical stimulation promoted the migration of Schwann cells by down-regulating miR-741-3p and targeting Radil gene. Figures and Tables | References | Related Articles | Metrics

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Inhibitory effect of hydroxy safflower yellow A on neuronal pyroptosis after glucose-oxygen deprivation/reglucose-reoxygenation treatment Wang Zeqian, Duan Yanzhe, Wu Yige, Ma Dong, Huang Jianjun, Yan Yuqing, Song Lijuan 2025, 29 (19):  4044-4051.  doi: 10.12307/2025.057 Abstract ( 92 )   PDF (1800KB) ( 79 )   Save BACKGROUND: Hydroxy safflower yellow A has anti-ischemia, anti-oxidation, anti-thrombotic and anti-inflammatory effects. Whether it affects neuronal pyroptosis after glucose-oxygen deprivation/reglucose-reoxygenation is still unclear. OBJECTIVE: To investigate the protective effect of hydroxy safflower yellow A on neuronal pyroptosis and its mechanism.  METHODS: HT22 cells in logarithmic growth phase were randomly divided into five groups: normal group, model group, hydroxy safflower yellow A group, colivelin group, and colivelin+hydroxy safflower yellow A group. HT22 cells were treated with glucose-oxygen deprivation/reglucose-reoxygenation to establish neuronal pyroptosis model, and then treated with STAT3 agonist Colivelin and hydroxy safflower yellow A. JC-1 probe was employed to assess changes in mitochondrial membrane potential. Reactive oxygen species kit was used to determine the content of reactive oxygen species in cells. GSDMD/TUNEL staining was conducted to observe cell pyroptosis. Immunofluorescence analysis was performed to detect STAT3 and GSDMD protein expression. RT-PCR was utilized for assessing mRNA expression levels of STAT3, NLRP3, and Caspase-1. Western blot assay was utilized to measure the protein expression levels of p-STAT3, NLRP3, GSDMD, Cleaved-caspase-1, and interleukin-1β.  RESULTS AND CONCLUSION: (1) Compared with the normal group, the number of pyroptotic cells increased in HT22 cells in the model group along with a significant increase in protein expression levels of p-STAT3, NLRP3, Cleaved-caspase-1, GSDMD, and interleukin-1β. Compared with the model group, the number of pyroptotic cells reduced, and the expression of pyroptosis-related proteins significantly decreased in the hydroxy safflower yellow A group. (2) In comparison with the model group, pyroptosis worsened in the colivelin group where mitochondrial membrane potential decreased along with elevated reactive oxygen species content and increased mRNA expression levels of STAT3, NLRP3, and Caspase-1, as well as increased protein expression levels of p-STAT3, NLRP3, GSDMD, Cleaved-caspase-1, and interleukin-1β. Compared with the Colivelin group, above indexes were improved in the colivelin+hydroxy safflower yellow A group. These results suggest that hydroxy safflower yellow A plays a neuroprotective role through STAT3 signaling pathway to inhibit HT22 pyroptosis after glucose-oxygen deprivation/reglucose-reoxygenation treatment. Figures and Tables | References | Related Articles | Metrics

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Jiawei Xiaoyao San exerts anti-liver cancer effects via exosomal miRNA pathway Liu Xiaoming, Cheng Jinlai, Li Rushuang, Li Niuniu, Qin Qiuyun, Xia Meng, Yao Chun 2025, 29 (19):  4052-4062.  doi: 10.12307/2025.061 Abstract ( 115 )   PDF (2967KB) ( 296 )   Save BACKGROUND: Previous studies by our research group discovered that Jiawei Xiaoyao San has a significant anti-liver cancer effect, but the specific mechanism of action was unclear. OBJECTIVE: To investigate the regulatory effects of the traditional Chinese medicine formula Jiawei Xiaoyao San on the levels of miRNAs in plasma exosomes of rats with diethylnitrosamine chronically induced primary liver cancer, based on high-throughput sequencing combined with bioinformatics. METHODS: SD rats were randomly divided into a blank control group, a liver cancer model group, and a Jiawei Xiaoyao San treatment group. Liver cancer models were induced by continuous administration of diethylnitrosamine for 12 weeks. Starting from the 17th week, rats in the Jiawei Xiaoyao San treatment group were administered Jiawei Xiaoyao San once daily until the end of the 20th week, while rats in the blank control and liver cancer model groups were given an equivalent volume of saline. Anti-hepatocellular carcinoma effects were validated by assessing the morphological structure of rat liver tissues, along with the expression of the hepatocellular carcinoma markers, Glypican-3 protein and serum alpha-fetoprotein. Plasma exosomes from each group of rats were isolated using ultracentrifugation. High-throughput sequencing technology was used to screen for differentially expressed miRNAs in rat plasma exosomes. Bioinformatics was used to predict the potential biomarkers through which Jiawei Xiaoyao San exerts its anti-liver cancer effects via liver cancer-derived exosomal miRNAs, followed by functional analysis. RESULTS AND CONCLUSION: (1) Jiawei Xiaoyao San significantly improved the morphological structure of liver tissues in a rat model of liver cancer. Compared with the liver cancer model group, the expression of liver cancer markers Glypican-3 protein and serum alpha-fetoprotein was significantly reduced in the Jiawei Xiaoyao San treatment group. (2) Bioinformatics analysis showed that in the Jiawei Xiaoyao San group, upregulated miR-223-3p in the liver cancer model group had target binding sites with genes E2F1 and NCOA1, which were closely related to liver cancer survival and prognosis. Therefore, Jiawei Xiaoyao San has a therapeutic effect on liver cancer, possibly by targeting negative regulation of NCOA1/E2F1 through liver cancer plasma-derived exosomal miR-223-3p, thereby playing anti-liver cancer effect. Figures and Tables | References | Related Articles | Metrics

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Comparison of biological characteristics of adipose-derived stem cells in young and old mice Lin Meiyu, Yao Xiang, Gao Jing, Zhao Xilong, Pan Xinghua, Ruan Guangping 2025, 29 (19):  4063-4068.  doi: 10.12307/2025.076 Abstract ( 95 )   PDF (1962KB) ( 112 )   Save BACKGROUND: Adipose-derived stem cells have anti-aging effects, but whether adipose-derived stem cells from donors of different ages are different needs further study. OBJECTIVE: To compare the biological properties of adipose-derived stem cells in old and young mice.  METHODS: Adipose-derived stem cells were extracted from adipose tissue of C57BL mice aged 8 and 14 weeks, respectively. The differences of cell cycle, apoptosis, and proliferation of adipose-derived stem cells in old and young mice were compared. The expression levels of aging-related P21 and P27 genes and proteins of adipose-derived stem cells in old and young mice were detected by quantitative PCR and western blot assay.  RESULTS AND CONCLUSION: Compared with old mouse adipose-derived stem cells, young mouse adipose-derived stem cells were more active, more regular in morphology, less apoptosis, faster proliferation, and lower in expression of age-related P21 and P27 genes and proteins. It has been proven that adipose-derived stem cells from young mice have better anti-aging effects.  Figures and Tables | References | Related Articles | Metrics

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Analysis of oocyte granulosa cell transcriptome data in aged women Qin Lifeng, Wang Jiaqiang, Zhu Ling 2025, 29 (19):  4069-4075.  doi: 10.12307/2025.070 Abstract ( 125 )   PDF (1546KB) ( 144 )   Save BACKGROUND: The granulosa cell state greatly affects the quality of oocytes. As the increased proportion of aged women in assisted reproduction, granulosa cells from patients of different ages are examined at transcript levels in order to better assess oocyte quality. OBJECTIVE: To investigate the expression changes of mRNA and long non-coding RNA in granulosa cells from the young and aged patients. METHODS: Waste cumulus granulosa cells obtained during the assisted reproductive cycle. Young group was in 25-35 years old, and the aged group was in 38-45 years old. Granulosa cells collected from the young and aged females were analyzed using RNA sequencing, three replications for each group. RESULTS AND CONCLUSION: (1) The RNA sequencing analysis results showed that the average sequencing volume of samples was more than 14 G; the data quality Q30 after quality control was more than 93%; the average mapping rate of data was 98.4%, which indicates that the data had good quality. (2) The results of principal component analysis and correlation analysis showed that the samples of the aged group and the young group could be clearly distinguished. (3) The difference analysis results showed that compared with the young group, a total of 410 differentially expressed mRNAs were detected in the aged group (167 up-regulated and 243 down-regulated). GO analysis results showed that the down-regulated genes were mainly enriched in regulatedexocytosis and the emetabolicprocess. GSEA analysis results showed that secretion-related pathways in the aged group were down-regulated. (4) Compared with the young group, 662 differentially expressed long non-coding RNAs were detected in the aged group, and 1 772 protein-coding genes were directly regulated by these long non-coding RNAs, among which 59 genes overlaped with differentially expressed genes. The results showed that the expression of secretion-related genes and pathways in granulosa cells of the aged group was down-regulated, thus affecting oocyte quality. At the same time, these down-regulated genes were regulated by long non-coding RNA. Therefore, the expression of long non-coding RNA might be related to age.   Figures and Tables | References | Related Articles | Metrics

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Isolation, culture and differentiation of human urine-derived stem cells into smooth muscle cells Chen Jiahui, Dai Xiaoqi, Xu Yangang, Li Yuanchao, Huang Mei, Zhan Yifei, Du Yuxuan, Li Liuqiang, Guo Yaochuan, Bian Jun, Lai Dehui 2025, 29 (19):  4076-4082.  doi: 10.12307/2025.073 Abstract ( 164 )   PDF (1876KB) ( 142 )   Save BACKGROUND: Traditional methods of urinary tract reconstruction are limited by donor scarcity, high complication rates, and suboptimal functional recovery. Tissue engineering strategies offer new directions in this field. Since the urinary tract is mainly composed of muscle tissue, the key is to find suitable seed cells and efficiently induce them to differentiate into smooth muscle cells. Comparative studies on the efficacy of different smooth muscle cell induction regimens are still lacking. OBJECTIVE: To isolate, culture, and identify human urine-derived stem cells, and to compare the effects of two different induction protocols.  METHODS: Human urine-derived stem cells were isolated from urine samples of 11 healthy adult volunteers by multiple centrifugations. Surface markers were identified by flow cytometry. The multi-directional differentiation potential of human urine-derived stem cells was verified through osteogenic and adipogenic differentiation. Differentiation was induced by transforming growth factor-β1 or transforming growth factor-β1 combined with platelet derived growth factor for 14 days. Immunofluorescence staining and western blot assay were employed to compare the expression differences of smooth muscle-specific proteins (α-SMA and SM22).   RESULTS AND CONCLUSION: (1) Urine-derived stem cells were successfully isolated from the eight urine samples of healthy people. These cells exhibit a “rice grain”-like morphology and possess a robust proliferative capacity. (2) Urine-derived stem cells exhibited high expression of mesenchymal stem cell surface markers (CD73, CD90, and CD44) and extremely low expression of hematopoietic stem cell surface markers (CD34 and CD45). These cells did not express CD19, CD105, and HLA-DR. (3) After osteogenic and adipogenic differentiation, the formation of calcium nodules and lipid droplets was observed, with positive staining results from Alizarin Red S and Oil Red O staining. (4) After 14 days of smooth muscle induction culture, immunofluorescence staining revealed that the smooth muscle differentiation rate of urine-derived stem cells treated with a combination of transforming growth factor-β1 and platelet derived growth factor was significantly higher compared to those treated with transforming growth factor-β1 alone (P < 0.005). (5) After 14 days of smooth muscle induction culture, western blot assay further demonstrated that the expression levels of α-SMA and SM22 in the transforming growth factor-β1/platelet derived growth factor group were significantly elevated compared to those in the transforming growth factor-β1 only group (P < 0.005). These findings confirm that urine-derived stem cells can be non-invasively isolated using multiple rounds of centrifugation. Compared with transforming growth factor-β1 alone, the combination of transforming growth factor-β1 and platelet derived growth factor can improve the efficiency of inducing urine-derived stem cells to differentiate into smooth muscle cells. Figures and Tables | References | Related Articles | Metrics

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Stem cell therapy for amyotrophic lateral sclerosis: cell source, number, modification, and administration route Zhao Wen, Bi Yulin, Fu Xuyang, Duan Hongmei, Yang Zhaoyang, Li Xiaoguang 2025, 29 (19):  4083-4090.  doi: 10.12307/2025.054 Abstract ( 177 )   PDF (997KB) ( 239 )   Save BACKGROUND: With the continuous advancement of medical technology, stem cell therapy has been used to treat a variety of diseases, including amyotrophic lateral sclerosis. OBJECTIVE: To review the research progress of stem cell therapy for amyotrophic lateral sclerosis, and prospect the development trend of this field. METHODS: PubMed, China National Knowledge Infrastructure (CNKI), and WanFang Data were searched for articles published from 1995 to 2024 using the key words “amyotrophic lateral sclerosis, mesenchymal stem cells, neural stem/progenitor cells, pluripotent stem cells.” A total of more than 1 700 articles were retrieved, and 58 articles were finally included in this review. RESULTS AND CONCLUSION: Amyotrophic lateral sclerosis is a neurodegenerative disease that affects lower motor neurons in the brainstem and spinal cord and upper motor neurons in the motor cortex. The related research of stem cells in the treatment of amyotrophic lateral sclerosis has become a research hotspot. In this review, we summarize the application of different types of stem cells in amyotrophic lateral sclerosis research, including mesenchymal stem cells, neural stem progenitor cells, and induced pluripotent stem cells, and evaluate the key points of preclinical research such as stem cell source, cell volume, stem cell modification methods, and drug delivery routes, which lays the foundation for the future application of stem cell therapy.  Figures and Tables | References | Related Articles | Metrics

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Endothelial progenitor cell and mesenchymal stem cell therapy for vascular stent-associated diseases Li Qingyin, Li Linhua, Zhang Chunle, Fu Ping 2025, 29 (19):  4091-4101.  doi: 10.12307/2025.068 Abstract ( 133 )   PDF (949KB) ( 281 )   Save BACKGROUND: With advancements in stem cell research, the therapeutic efficacy of adult stem cells such as endothelial progenitor cells and mesenchymal stem cells in atherosclerosis and complications arising from atherosclerosis and vascular stent implantation is gradually being recognized. Due to the limitations of intravenous infusion of adult stem cells, including poor targeting and low treatment efficiency, recent research has focused on surface modification of vascular stents to achieve localized aggregation and functional modulation of endothelial progenitor cells or mesenchymal stem cells. OBJECTIVE: To discuss the therapeutic progress of endothelial progenitor cells and mesenchymal stem cells in vascular stent-related diseases, summarize the research status of the design of vascular stents based on endothelial progenitor cells and mesenchymal stem cells. METHODS: Relevant literature was retrieved on CNKI, WanFang, PubMed, and Web of Science databases since their inception. The Chinese search terms were “endothelial injury, stenting, thrombosis, intimal hyperplasia, atherosclerosis, endothelial repair, endothelial progenitor cell, mesenchymal stem cell, vascular stent.” English search terms were “endothelial injury, stenting, thrombosis, intimal hyperplasia, atherosclerosis, endothelial repair, endothelial regeneration, endothelial progenitor cell, mesenchymal stem cell, vascular stent, vascular scaffold.” According to inclusion and exclusion criteria, 127 articles were finally reviewed.  RESULTS AND CONCLUSION: Endothelial progenitor cells and mesenchymal stem cells can treat atherosclerosis and complications of stent implantation through differentiation and paracrine effects, mainly by protecting endothelial cells, regulating the expression of inflammatory cells and cytokines, and modulating smooth muscle cell proliferation and phenotype. Mesenchymal stem cells may have adverse reactions such as thrombosis and vascular calcification in therapeutic applications, and using extracellular vesicles and co-administration with heparin for surface modification is a feasible solution. Currently, there is more research on stents based on endothelial progenitor cells, mainly focusing on recruitment, capture, proliferation, differentiation, and activity. Research on stents based on mesenchymal stem cell capture in the vascular field is relatively scarce, but exosome-loaded stents derived from mesenchymal stem cells have been found to have highly effective therapeutic efficacy. Additionally, some underlying diseases such as diabetes may affect the activity of adult stem cells, leading to the loss of effectiveness in stem cell-based stent designs. Therefore, in future stent designs, consideration should be given to the background diseases. Figures and Tables | References | Related Articles | Metrics

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Adult stem cells from different germ layers applied in peripheral nerve injury repair Zheng Jiachen, Yang Entong, Zhu Yizhou, Liu Fang 2025, 29 (19):  4102-4110.  doi: 10.12307/2025.067 Abstract ( 96 )   PDF (942KB) ( 231 )   Save BACKGROUND: Adult stem cell therapy is one of the research hotspots in the field of peripheral nerve injury repair and regeneration. With excellent properties of mesenchymal stem cells such as high acquisition rate, wide source, and rapid proliferation, mesoderm have been regarded as the ideal source of adult stem cells, while ectoderm-derived adult stem cells, especially neural crest stem cells, have certain neurogenic properties and attract more and more attention from researchers. OBJECTIVE: To mainly review the role and mechanism of multifunctional adult stem cells from ectoderm and mesoderm in peripheral nerve injury repair and regeneration, so as to explore the research progress and application prospect of adult stem cells from different sources and discuss the potential application value of adult stem cell therapy and the problems to be solved in connection with clinical studies. METHODS: The first author searched the relevant articles published from December 2001 to February 2024 in PubMed and SinoMed by computer in February 2024. The Chinese and English search terms were “ectodermal stem cells, mesenchymal stem cells, peripheral nerve injury, repair, regeneration.” Finally, 69 articles were included and analyzed. RESULTS AND CONCLUSION: (1) Ectodermal adult stem cells have excellent differentiation and regeneration potential, especially epidermal neural crest stems cells, olfactory stem cells, and dental ectomesenchymal stem cells, which have certain neurogenic properties and can express neural specific markers in vitro. However, relevant clinical research needs to be accumulated. (2) There are many types of adult stem cells derived from mesoderm, which are easy to obtain and purify. Among them, the efficacy and safety of bone marrow mesenchymal stem cells and umbilical cord mesenchymal stem cells in the repair of peripheral nerve injury are supported by clinical trials; that is, they can improve sensory and motor nerve conduction and there are no complications and obvious adverse reactions in follow-up. The acquisition of bone marrow mesenchymal stem cells needs invasive surgery and requires the patient to match the donor bone marrow type, which limit the application to some extent. Although umbilical cord mesenchymal stem cells do not require invasive harvesting, their isolation is difficult and phenotypically unstable. (3) Adult stem cells derived from endoderm often fail to grow in vitro, so the possibility of clinical application is low. (4) In conclusion, bone marrow mesenchymal stem cells are still the first choice for adult stem cell therapy in the treatment of peripheral nerve injury, which is suitable for cases without surgical contraindications and meeting the matching requirements, followed by umbilical cord mesenchymal stem cells supplemented by improved isolation methods and advanced phenotypic stability strategies. (5) Dental ectomesenchymal stem cells and adipose-derived mesenchymal stem cells have high application potential and need to be further tested in clinical trials. Other adult stem cells derived from ectodermal and mesodermal layers have significant advantages in animal and cell experimental studies due to their excellent properties. Figures and Tables | References | Related Articles | Metrics

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Scientific basis for acupuncture combined with neural stem cells for repairing spinal cord injury Huang Xiaomeng, Zhang Zhilan, Shang Wenya, Huang Jing, Wei Huilin, Li Bing, Ren Yafeng 2025, 29 (19):  4111-4121.  doi: 10.12307/2025.072 Abstract ( 175 )   PDF (1631KB) ( 157 )   Save BACKGROUND: Spinal cord injury is a neurological disorder caused by traumatic or non-traumatic events, often leading to severe functional impairment below the injured segment. In recent years, neural stem cell transplantation has been considered to have significant therapeutic potential in regulating the inflammatory response after spinal cord injury, inhibiting excessive proliferation of glial scars, and promoting nerve regeneration. OBJECTIVE: To review and discuss the potential mechanism of action of acupuncture and neural stem cell transplantation therapy in inhibiting spinal cord injury-induced secondary injury, and to delve into the scientific basis for its treatment of spinal cord injury. METHODS: PubMed, Elsevier, WanFang, and CNKI databases were searched using “spinal cord injury, acupuncture, neural stem cells, SDF-1α/CXCR4 axis” as Chinese and English search terms. Totally 96 articles were finally included. The research findings of acupuncture combined with neural stem cells in the treatment of spinal cord injury were summarized and analyzed, and the mechanism of this combination therapy in the treatment of secondary injury after spinal cord injury was summarized. RESULTS AND CONCLUSION: (1) The stromal-derived factor 1α (SDF-1α)/chemokine receptor 4 (CXCR4) axis plays a crucial role in neural stem cell transplantation for spinal cord injury. This signaling mechanism not only affects neural stem cell migration, proliferation, and differentiation, but is also a key factor in determining the efficiency of stem cell homing to the injury site. Therefore, the regulation of targeting this axis is of great significance in enhancing the therapeutic effect of spinal cord injury. (2) Acupuncture, as a traditional Chinese medicine therapy, shows unique advantages in the regulation of secondary injury in spinal cord injury. It can effectively reduce secondary injury after spinal cord injury by regulating inflammatory response, inhibiting apoptosis, improving microcirculation, reducing glial scar formation, and counteracting oxidative stress. (3) Acupuncture was also able to influence the expression and function of the SDF-1α/CXCR4 axis, thereby enhancing the homing and survival ability of neural stem cells and promoting nerve regeneration and functional recovery. (4) The therapy combining acupuncture and stem cell transplantation is an innovative treatment strategy for spinal cord injury and suitable for repairing neural circuits. It combines the wisdom of traditional Chinese medicine with the advantages of modern biotechnology, providing a new treatment option for spinal cord injury patients. However, this combination therapy is still in the research and exploration stage, and its long-term efficacy and safety need to be further verified. (5) Taken together, acupuncture and neural stem cell transplantation for the treatment of spinal cord injury has great potential for clinical application, but in-depth research and optimization of treatment options are still needed. In the future, we look forward to further revealing the efficacy mechanism and optimal indications of this therapy through more clinical trials and mechanism studies, so as to bring better hope of recovery and more efficient therapeutic effects to spinal cord injury patients. Figures and Tables | References | Related Articles | Metrics

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Diagnosis and treatment of osteoarthritis with exosomes derived from different stem cells and carrying non-coding RNA Wang Zhe, Qi Yansong, Xu Yongsheng 2025, 29 (19):  4122-4131.  doi: 10.12307/2025.069 Abstract ( 117 )   PDF (1028KB) ( 112 )   Save BACKGROUND: Exosomes can be detected in synovial fluid and plasma at levels that vary with the progression of osteoarthritis in patients with osteoarthritis, and may play a relieving role in the local inflammation of osteoarthritis, cartilage calcification, and osteoarthritic joint degradation. OBJECTIVE: To comprehensively understand the function and mechanism of exosomes from different stem cells in the diagnosis and treatment of osteoarthritis, and to present the prospects and challenges of exosome therapy for osteoarthritis. METHODS: We searched the articles published from October 2003 to October 2023 included in PubMed and CNKI databases with the keywords of “exosomes, osteoarthritis, mesenchymal stem cells, stem cells” in Chinese and English, respectively. A total of 99 articles were finally included for review. RESULTS AND CONCLUSION: The appearance of exosomes brings hope to the diagnosis and treatment of osteoarthritis. The differences of RNA, protein, and lipid content in exosomes can be used as biomarkers for the diagnosis of osteoarthritis. At the same time, exosomes from various stem cells can effectively protect chondrocytes, relieve inflammation, maintain cartilage matrix metabolism, and regulate angiogenesis and subchondral bone remodeling, showing excellent potential in the treatment of osteoarthritis. The engineered exosomes break through the traditional limitations and enhance the specificity and efficiency of treatment by modulating the expression of specific non-coding RNA, providing a new strategy for the treatment of osteoarthritis. Figures and Tables | References | Related Articles | Metrics

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Mechanism of action and progress of mitophagy, ferroptosis, cuproptosis, and disulfidptosis in Alzheimer’s disease Liu Dandan, Qin Hewei 2025, 29 (19):  4132-4144.  doi: 10.12307/2025.066 Abstract ( 364 )   PDF (1887KB) ( 970 )   Save BACKGROUND: In recent years, with the in-depth study of programmed cell death, new models of programmed cell death (mitophagy, ferroptosis, cuproptosis, and disulfidptosis) involved in Alzheimer’s disease injury are gradually emerging and have large research space in the future. OBJECTIVE: To review the molecular mechanism of novel programmed cell death mode (mitophagy, ferroptosis, cuproptosis, and disulfidptosis), the crosstalk mechanism of novel cell death mode, and clinical transformation in Alzheimer’s disease, aiming to provide a new perspective for exploring the mechanism of action and drug targets in Alzheimer’s disease. METHODS: The first author used the computer to search the literature published between 1991 and 2024. 101 articles were finally included according to the inclusion criteria. RESULTS AND CONCLUSION: (1) Programmed cell death is a necessary regulatory pathway to maintain normal cell renewal and homeostasis. Among them, new types of programmed cell death such as mitophagy, ferroptosis, cuproptosis, and disulfidptosis are hot research fields in life science. (2) Mitophagy can clear damaged mitochondria in Alzheimer’s disease neurons, reduce intracellular reactive oxygen species, restore the energy metabolism and signal transduction of neurons in Alzheimer’s disease, and play a crucial role in regulating the health and function of neurons. (3) Studies on ferroptosis in Alzheimer’s disease have attracted much attention. It can regulate Alzheimer’s disease through various ways such as cystine/glutamate, iron metabolism, and polyunsaturated fatty acids, thus affecting Aβ deposition and Tau protein phosphorylation. Recent studies have shown that natural polyphenols, Suanzoren decoction, poria acid, and vitamin E can resist ferroptosis in Alzheimer’s disease. (4) Cuproptosis is a new and unique form of cell death involving copper dependence, accumulation of fatty acylated proteins, and reduction of iron-sulfur tufting proteins. Excessive copper exposure may directly interact with Aβ plaques and amyloid precursor proteins, exacerbating cognitive impairment in Alzheimer’s disease. Currently, the research field of cuproptosis is emerging, and the mechanism of action has not been fully clarified. (5) Disulfidptosis, as an emerging form of programmed cell death, is caused by disulfide stress due to excessive cystine accumulation and glucose starvation, resulting in damage to the actin skeleton associated with Alzheimer’s disease. (6) Various patterns of programmed cell death have a tandem mechanism in the pathogenesis of Alzheimer’s disease, forming an interaction network of various programmed cell death with autophagy as the core, providing great potential for multi-level and multi-target regulation of Alzheimer’s disease. The crosstalk network mechanism between autophagy, necroptosis, and pyroptosis/ferroptosis co-regulates Alzheimer’s disease. (7) Cuproptosis and disulfidptosis, as a new mode of programmed death, have not been reported deeply enough in Alzheimer’s disease, and further research and continuous attentions are still needed in the future. (8) Since most studies on mitophagy, ferroptosis, cuproptosis, and disulfidptosis are based on basic experiments or biogenic analysis, there is a lack of large-scale and long-term clinical research validation. Further in-depth studies are needed in the future to provide new ideas and effective strategies for the treatment of Alzheimer’s disease.  Figures and Tables | References | Related Articles | Metrics

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Analyzing current status of stem cell research based on National Natural Science Foundation of China's completed projects from 2018 to 2022 Jiang Jun, Zhang Bin 2025, 29 (19):  4145-4150.  doi: 10.12307/2025.045 Abstract ( 70 )   PDF (1648KB) ( 155 )   Save BACKGROUND: At present, most research focuses on in-depth analysis of a certain aspect of stem cells from the perspective of papers and project applications funded by the National Natural Science Foundation of China, or verifies certain evaluation methods, lacking a comprehensive analysis of the completed projects of the National Natural Science Foundation of China in the field of stem cells.  OBJECTIVE: To understand the funding status and research hotspots of National Natural Science Foundation of China projects in the field of stem cells from 2018 to 2022, providing a reference for subsequent research. METHODS: Using “stem cells” as the search terms, databases were searched such as Big Data Knowledge Management Service Portal of the National Natural Science Foundation of China to extract project information, clean and merge keywords, institutions, supporting units and regions. Software such as Excel, Echarts, and VOSviewer was used to perform word frequency statistics, cluster analysis, and visualization. RESULTS AND CONCLUSION: The National Natural Science Foundation of China’s Stem Cell Project has demonstrated significant characteristics of interdisciplinary integration, covering not only medicine and life sciences, but also multiple disciplines such as chemistry, information science, engineering and materials science, earth science, and mathematical science, bringing revolutionary progress to the development of the stem cell industry. Keyword clustering analysis shows that China’s funding for stem cells mainly focuses on areas such as mesenchymal stem cells, tumor stem cells, embryonic stem cells, neural stem cells, and hematopoietic stem cells. By comparison, the funding of China, the United States, and the United Kingdom each has their own emphasis, reflecting the different development directions in the field of stem cells in each country and providing broad space for international cooperation. In the past five years, although the number of completed projects has remained stable, the amount of completed projects has slightly decreased, indicating that China’s investment in the field of stem cells is relatively insufficient. Therefore, it is recommended to strengthen the investment of the National Natural Science Foundation in this field and guide more social capital to participate, forming a diversified investment pattern to promote the rapid development of the stem cell industry. Figures and Tables | References | Related Articles | Metrics

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Analysis of regulation of prognosis, immune infiltration, and ferroptosis in sarcoma based on stemness index model Wei Jingxian, Meng Lian, Sun Hao, Zhang Tiantian, Liu Chunxia 2025, 29 (19):  4151-4160.  doi: 10.12307/2025.055 Abstract ( 89 )   PDF (3871KB) ( 205 )   Save BACKGROUND: The stemness index may be associated with the prognosis and immune infiltration of sarcoma, but the specific regulatory mechanism and characteristic genes have yet to be fully elucidated. OBJECTIVE: To investigate the correlation between stem cells and prognosis as well as immune infiltration in sarcoma employing the gene stemness index model and to identify the ferroptosis signature genes associated with sarcoma stem cells. METHODS: The sarcoma RNA sequencing data and related clinical information were obtained from the Cancer Genome Atlas (TCGA). The sarcoma RNA sequencing data were grouped using the sarcoma stemness index. Survival data were used to analyze prognosis between groups. Differentially expressed genes were obtained for pathway enrichment and immune infiltration analysis. Ferroptosis-related differential genes were used to construct a protein interaction network and analyze prognostic correlation. Rhabdomyosarcoma cell lines were cultured and divided into adherent cell group and stem cell group. The adherent cell group received no intervention, while the stem cell group was treated with serum-free culture to enrich stem cells in rhabdomyosarcoma cells. qRT-PCR was used to evaluate stemness markers, ferroptosis-related genes, and mRNA expression of ferroptosis-related markers in the cells. RESULTS AND CONCLUSION: (1) Patients were divided into high and low stemness index groups based on the median stemness index. The progression-free survival of patients in the high stemness index group was lower than that in the low stemness index group by disease risk prediction, suggesting poor prognosis. (2) According to GO and KEGG analysis, the groups with high and low stemness indices differed from one another. There were differences in immune infiltration between the high and low stemness index groups. Nine of the 23 ferroptosis-related genes in the differential genes have the potential to establish a highly correlated network of protein interactions. Patients with high expression of IDO1, IFNG, and AQP5 have a better prognosis, while those with high expression of CA9 have a poor prognosis. (3) The qRT-PCR results demonstrated a significant upregulation of stem cell-related markers NANOG, SOX2, and OCT4 mRNA expressions in the stem cell group compared to the adherent cell group (P < 0.05). Compared to the adherent cell group, the stem cell group exhibited decreased mRNA expression level of ferroptosis-related marker SLC7A11 (P < 0.05) while showing increased levels of ACSL4, GPX4, FTH1, and COX2 (P < 0.05). Compared to the adherent cell group, the stem cell group displayed decreased mRNA expression level of differentially expressed gene CA9 alongside elevated levels of IDO1, IFNG, and AQP5 (P < 0.05). Stem cells were strongly associated with sarcoma survival and ferroptosis by bioinformatics analysis and experimental verification. Sarcoma stem cells have aberrant expression of CA9, IDO1, IFNG, and AQP5, which may serve as new targets for sarcoma therapy as well as diagnostic indicators.  Figures and Tables | References | Related Articles | Metrics

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Action mechanism of Coptidis Rhizoma Alkaloids against cerebral ischemia based on transcriptome sequencing Tian Liangliang, Zhou Rui, Cao Guangzhao, Zhang Jingjing 2025, 29 (19):  4161-4171.  doi: 10.12307/2025.063 Abstract ( 77 )   PDF (2387KB) ( 201 )   Save BACKGROUND: Coptis chinensis can clear heat, dry dampness, relieve fire, and detoxify. Coptis chinensis and its components have a significant protective effect on cerebral ischemia. The action mechanism of anti-cerebral ischemia of Coptidis Rhizoma Alkaloids was explored based on network pharmacology and transcriptome sequencing. OBJECTIVE: Based on the study of the protective effects of Coptidis Rhizoma Alkaloids on cerebral ischemia of rats, the action mechanism of Coptidis Rhizoma Alkaloids intervention in cerebral ischemia was investigated by using network pharmacology and transcriptome sequencing technology.  METHODS: The SD rats were randomly divided into sham operation group, ischemia/reperfusion group, positive drug group, and Coptidis Rhizoma Alkaloids group. The ischemia/reperfusion model of middle cerebral artery occlusion was prepared by modified thread method in the latter three groups. No thread was inserted and the other operations were the same in the sham operation group. TTC staining, Longa 5 neurological deficient score, hematoxylin and eosin staining, and Nissl staining were used to evaluate the protective effect of Coptidis Rhizoma Alkaloids on ischemia/reperfusion model rats. Transcriptome sequencing was performed on the brain tissues of rats in sham operation group, ischemia/reperfusion group, and Coptidis Rhizoma Alkaloids group. Differentially expressed genes, gene Ontology analysis, Kyoto encyclopedia of genes and genomes analysis, and Correlation Analysis of Transcriptomics and Network Pharmacology were used to elucidate the effect of Coptidis Rhizoma Alkaloids on cerebral ischemia. Finally, ELISA and immunofluorescence staining were used to verify the key targets of Coptidis Rhizoma Alkaloids in the intervention of cerebral ischemia. RESULTS AND CONCLUSION: (1) Coptidis Rhizoma Alkaloids treatment decreased the Longa 5 neurological deficit scores and cerebral infarction area of ischemia/reperfusion model rats, increased the number of neurons and Nissl bodies. (2) Differentially expressed gene after Coptidis Rhizoma Alkaloids treatment analyzed by functional enrichment analysis of gene ontology includes biological processes such as inflammatory reaction and positive regulation of mitogen-activated protein kinase cascade. The enrichment analysis of Kyoto gene and genome encyclopedia analysis pathway mainly involves interleukin-17 signaling pathway, neuroactive ligand-receptor interaction, cyclic adenosine-3′,5′-mconophosphate signaling pathway and so on. (3) Analysis of transcriptomics showed that the main genes regulated by Coptidis Rhizoma Alkaloids were prostaglandin endoperoxide synthase 2, brain derived neurotrophic factor, and transient receptor potential A1. (4) Network pharmacology analysis revealed that nine components in Coptidis Rhizoma Alkaloids may exert their effects by associating with 87 targets related to prostaglandin endoperoxide synthase 2, brain derived neurotrophic factor, and transient receptor potential A1. (5) ELISA and immunofluorescence staining results further confirmed that Coptidis Rhizoma Alkaloids regulated the expression of prostaglandin endoperoxide synthase 2, brain derived neurotrophic factor, and transient receptor potential A1. (6) It is concluded that Coptidis Rhizoma Alkaloids treatment can significantly improve the injury in ischemia/reperfusion model rats, possibly by regulating prostaglandin endoperoxide synthase 2, brain derived neurotrophic factor, and transient receptor potential A1.  Figures and Tables | References | Related Articles | Metrics

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Cuproptosis-related genes in natural killer cells of Alzheimer’s disease Chen Guilin, Tang Qiqiang 2025, 29 (19):  4172-4180.  doi: 10.12307/2025.036 Abstract ( 161 )   PDF (2329KB) ( 708 )   Save BACKGROUND: The immune-related pathogenesis of Alzheimer’s disease is still unclear. Exploring the correlation between natural killer cells and cuproptosis mechanism in Alzheimer’s disease patients through bioinformatics can provide a new direction for the study of the occurrence and development of Alzheimer’s disease. OBJECTIVE: To screen the key genes related to cuproptosis of natural killer cells in peripheral blood of patients with Alzheimer’s disease by bioinformatics analysis and verify them in clinical specimens. METHODS: The GEO online database was used to screen the transcriptome differentially expressed genes and natural killer cell related genes in the peripheral blood of patients with Alzheimer’s disease, and intersected with the reported cuproptosis factors. Differentially expressed cuproptosis-related genes were obtained. Then RT-qPCR technology was used to verify the relative gene expression levels. The experimental samples were all from peripheral blood of hospitalized patients in the Department of Neurology of Anhui Provincial Hospital from 2021 to 2023, and 30 patients in the disease group and 20 in the control group were included according to the inclusion and exclusion criteria. The protein-protein interaction network was further constructed using the online GeneMANIA website. R language was used for immune infiltration analysis. Transcription factor prediction was conducted based on ENCODE database. RESULTS AND CONCLUSION: (1) The differential expression genes of peripheral blood transcriptome of Alzheimer’s disease patients in GSE63060 data set, natural killer cell related genes in GSE168522 data set, and reported cuproptosis genes were used to screen and obtain four differentially expressed cuproptosis-related genes by using online Venn diagram tool, LASSO algorithm, and random forest machine learning methods: ferredoxin 1 (FDX1), ATPase Cu2+ transporting alpha polypeptide (ATP7A), pyruvate dehydrogenase El subunit beta (PDHB), and dihydrolipoamide succinyltransferase (DLST). (2) Clinical sample experiments showed that FDX1 and ATP7A were up-regulated in peripheral blood of patients with Alzheimer’s disease (P < 0.001), and were differentially expressed in different genotypes of apolipoprotein E4 (P < 0.01, P < 0.001). The expression of PDHB and DLST in peripheral blood of patients with Alzheimer’s disease was down-regulated (P < 0.001), and there was no difference in apolipoprotein E4 genotypes (P > 0.05). (3) Protein-protein interaction network found that 20 functional proteins were associated with key genes, and immunoinfiltration analysis showed that key genes were significantly associated with 12 immune cells (P < 0.05 was considered to be relevant). (4) Bioinformatics analysis and experimental verification results suggest that FDX1, ATP7A, PDHB, and DLST are differentially expressed in Alzheimer’s disease, may participate in the occurrence and development of Alzheimer’s disease through the cuproptosis mechanism in peripheral blood natural killer cells, and also provide potential targets for the diagnosis and treatment of Alzheimer’s disease.  Figures and Tables | References | Related Articles | Metrics