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18 July 2024, Volume 28 Issue 20 Previous Issue   

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Effects of dry swallowing and swallowing tasks of varying consistencies and volumes on the hyoid muscles in healthy adults Li Zhenzhen, Zhou Zhipeng 2024, 28 (20):  3117-3122.  doi: 10.12307/2024.386 Abstract ( 329 )   PDF (1178KB) ( 573 )   Save BACKGROUND: Modification of food consistency and volume is a commonly used method of swallowing compensation in clinical practice. Dry swallowing is a commonly used method of evaluation. The hyoid muscles are very important in swallowing. The effects of dry swallowing and swallowing tasks of different consistencies and volumes on hyoid muscle activation levels are still unclear. OBJECTIVE: To explore the effects of simple dry swallowing and swallowing tasks of different consistencies and volumes on the hyoid muscles in healthy adults. METHODS: A total of 44 healthy adults were included from April to August 2019, including 19 males and 25 females, with an average age of (21.7±2.8) years. They randomly performed dry swallowing and swallowing tasks of different consistencies (the International Dysphagia Diet Standardisation Initiative (IDDSI) frame levels 0-4) and volumes (5, 10, 20 mL), and the surface electromyogram signals of the hyoid muscles during each swallowing task were recorded. After processing the raw surface electromyogram signals, the activation levels of the hyoid muscles were compared between dry swallowing and swallowing tasks of different consistency and volume. RESULTS AND CONCLUSION: The mean amplitude values of the suprahyoid muscles corresponding to swallowing tasks of 20 mL for levels 0-4, 10 mL for level 3, and 5 mL for level 4 were higher than those of dry swallowing (P < 0.05). The mean amplitude values of the suprahyoid muscles corresponding to the 20-mL swallowing tasks of different consistencies were higher than those of the 5-mL swallowing tasks of the corresponding consistencies, except for level 3 (P < 0.05). The mean amplitude values of the suprahyoid muscles corresponding to the 20-mL swallowing tasks of different consistencies were higher than those of the 10-mL swallowing tasks of the corresponding consistencies, except for levels 2 and 3 (P < 0.05). The mean amplitude values of the infrahyoid muscles corresponding to all swallowing tasks were higher than that of dry swallowing (P < 0.05). The mean amplitude values of the infrahyoid muscles corresponding to the 20-mL swallowing tasks of different consistencies were higher than that of the 5- and 10-mL swallowing tasks of the corresponding consistencies (P < 0.05). The mean amplitude values of the infrahyoid muscles corresponding to the 10-mL swallowing tasks of different consistencies were higher than those of the 5-mL swallowing tasks of the corresponding consistencies, except for level 3 (P < 0.05). To conclude, in healthy adults performing swallowing tasks of different volumes and consistencies, the level of activation of the hyoid muscles is less susceptible to IDDSI frame levels 0-4 consistency and more susceptible to volume. The higher volume indicates the higher activation level of the hyoid muscles. Figures and Tables | References | Related Articles | Metrics

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Exploration of signaling pathways with unclear action status and possible effects on related diseases or functions after knockdown of silencing information regulator 1 gene in chondrocytes Ye Haiming, Zeng Hui, Yang Qi, Zhang Geng, Weng Jian, Yu Fei 2024, 28 (20):  3123-3129.  doi: 10.12307/2024.352 Abstract ( 339 )   PDF (1192KB) ( 2455 )   Save BACKGROUND: silencing information regulatory 1 (SIRT1) regulates the function of related proteins in chondrocytes in a deacetylated manner and participates in chondrocyte proliferation and differentiation, thereby promoting cartilage defect repair. OBJECTIVE: To screen for signaling pathways with unclear action status after SIRT1 gene knockdown in chondrocytes, as well as diseases or functions that produce changes using high-throughput technology. METHODS: ATDC5 chondrocytes from mice in logarithmic growth phase were divided into two groups: the cells were transfected with SIRT1 gene knockdown negative control lentivirus in control group and SIRT1 gene knockdown lentivirus in experimental group. GeneChip® Mouse Genome 430 2.0 Array was used to detect the mRNA expression at 72 hours after transfection. Applied bioinformatics technology was also used to screen for unclear activation or inhibition signaling pathways and their related factors. Moreover, enrichment of disease or function modules was analyzed. RESULTS AND CONCLUSION: After knocking down the SIRT1 gene, there were 245 signaling pathways with unclear activation or inhibition status in the mouse ATDC5 chondrocytes. According to the ranking of -Log (P-value), we reported the factors in the top 20 signaling pathways with unclear activation or inhibition status, including IGFBP4, TGFBR1, CTGF, COL4A5, LHX2, IL1RL1, and KLF6. According to the ranking of -Log (P-value), there were significant changes in 14 disease or function modules, including cellular growth and proliferation, organism survival, cell death and survival. According to the number of differentially expressed genes, there were significant changes in three disease or function modules, including organismal injury and abnormalities, cancer, and cell death and survival. According to the comprehensive ranking of -Log (P-value) and the number of differentially expressed genes, the disease or function module related to intrinsic immune response was significantly activated. Figures and Tables | References | Related Articles | Metrics

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Effect of moderate-intensity exercise on the level of autophagy in bone tissue of ovariectomized rats Li Xun, Zhang Weichao, Li Yingjie, Liu Rong, Tian Xuewen, Zhang Pengyi, Wang Xiaoqiang 2024, 28 (20):  3130-3136.  doi: 10.12307/2024.399 Abstract ( 308 )   PDF (1280KB) ( 141 )   Save BACKGROUND: Exercise is an effective method for preventing and treating osteoporosis, but it is unclear whether its effect on postmenopausal osteoporosis is related to changes in bone autophagy levels. OBJECTIVE: To observe the effects of exercise via cellular autophagy on the morphology and mechanical properties of bone tissue in ovariectomized rats, and to explore the mechanism of exercise on bone mass in ovariectomized rats from the perspective of autophagy. METHODS: A rat model of postmenopausal osteoporosis was established, and a 24-week moderate-intensity exercise was used for intervention. After the experiment, serum estradiol levels were measured by ELISA, and bone mineral density and bone microstructure of the cortical and trabecular bone were detected by micro-CT. The biomechanical indicators of the tibia were tested by a three-point bending test. Autophagosomes were observed by transmission electron microscopy. The expression of LC3 and ATG7 proteins was analyzed by western blot. RESULTS AND CONCLUSION: The serum estradiol level in the ovariectomized group was significantly lower than that of the sham-operation group and ovariectomized+exercise group (P < 0.01). The body mass of rats in each group increased, and the order was the ovariectomized group > the ovariectomized+exercise group > the sham-operation group > the sham-operation+exercise group. The bone mineral density and bone mass of rats in all groups significantly increased (P < 0.01), but the increase in the ovariectomized group was significantly lower than that of the other groups, and the increase in the ovariectomized+exercise group was significantly higher than that of the ovariectomized group. Compared with the sham-operation group, the bone mineral density of the tibial cancellous bone in the sham-operation+exercise group was significantly increased (P < 0.01), while the bone mineral density in the ovariectomized and ovariectomized+exercise groups was significantly decreased (P < 0.01). Compared with the ovariectomized+exercise group, the ovariectomized group showed significantly lower bone volume fraction, number of trabeculae, and bone mineral density of cancellous bone (P < 0.05), extremely significantly lower trabecular thickness (P < 0.01), and significantly higher mean trabecular pattern factor, trabecular separation, and structural model index (P < 0.01). Compared with the ovariectomized group, the LC3-II/LC3-I ratio and the relative expression of ATG7 protein significantly increased in the ovariectomized+exercise group (P < 0.05). Compared with the sham-operation and ovariectomized groups, the number of autophagosomes increased in the sham-operation+exercise and ovariectomized+exercise groups, respectively. To conclude, moderate-intensity treadmill exercise can improve the bone microstructure and biomechanical properties of the tibial cancellous bone and increase bone mass in ovariectomized rats by increasing serum estradiol levels and bone autophagy levels. Figures and Tables | References | Related Articles | Metrics

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Protective effect of salidroside on angiotensin II-induced fibrosis in cardiac fibroblasts Hai Zhen, Ning Zhongping 2024, 28 (20):  3137-3142.  doi: 10.12307/2024.335 Abstract ( 348 )   PDF (1790KB) ( 224 )   Save BACKGROUND: Previous studies have shown that salidroside has an ameliorative effect on multi-organ fibrosis. However, the protective effect of salidroside on angiotensin ii-induced fibrosis in cardiac fibroblasts is unclear. OBJECTIVE: To investigate the protective effects of salidroside on angiotensin ii-induced oxidative stress and extracellular matrix deposition in cardiac fibroblasts of Sprague-Dawley rats and its mechanism of action. METHODS: Angiotensin II was used to induce fibrosis in cardiac fibroblasts, and there were five experimental groups: normal control group, model group (final concentration of angiotensin II in culture medium was 1 μmol/L), salidroside low and high dose groups (treatment with salidroside 50, 100 μmol/L for 2 hours, followed by co-incubation with angiotensin II for 48 hours), SIRT1 inhibitor group (treatment with SIRT1 inhibitor EX527 10 μmol/L for 2 hours, followed by high dose of salidroside for 2 hours and then co-incubation with angiotensin II for 48 hours). The cell viability was detected using the cell counting kit-8 method, the cell migration rate was detected by Transwell, the intracellular reactive oxygen species level was detected by DCFH-DA fluorescent probe, and the intracellular malondialdehyde content, superoxide dismutase and catalase activities were detected by relevant kits. The protein and mRNA expression levels of SIRT1, LOXL2, α-SMA, type I collagen and type III collagen were detected by western blot and qRT-PCR, respectively. RESULTS AND CONCLUSION: The cells were identified as cardiac fibroblasts by Vimentin fluorescence. Compared with the normal control group, cell viability, cell migration rate, reactive oxygen species level, and malondialdehyde content were significantly increased, superoxide dismutase and catalase activities were significantly decreased, LOXL2, α-SMA, type I collagen, type III collagen mRNA and protein expression were significantly increased, and SIRT1 protein expression level was significantly decreased in the model group (all P < 0.01). Compared with the model group, the above indexes showed opposite changes in the salidroside low and high dose groups (all P < 0.05). Moreover, salidroside showed dose-dependent regulation. Compared with salidroside groups, cell migration rate and α-SMA protein expression level were significantly increased in the SIRT1 inhibitor group (both P < 0.001). To conclude, salidroside has a protective effect on angiotensin II-induced cardiac fibroblasts and can dose-dependently inhibit oxidative stress and extracellular matrix deposition. Figures and Tables | References | Related Articles | Metrics

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Deferoxamine mesylate improves the repair of jaw bone defects in an ovariectomized rat model of osteoporosis Tian Ai, Li Li, Xiao Tianjiao, Kang Jiabing, Zhan Jifan, Wei Yan, Chen Helin 2024, 28 (20):  3143-3149.  doi: 10.12307/2024.359 Abstract ( 335 )   PDF (2607KB) ( 1420 )   Save BACKGROUND: Deferoxamine mesylate is a potential anti-osteoporosis drug with iron chelation, vascular regeneration, and antioxidant effects. Recent studies have shown that the application of deferoxamine mesylate can be extended to the field of tissue regeneration engineering. OBJECTIVE: To investigate whether deferoxamine mesylate can promote the repair effect of iron overload osteoporotic rats after bone grafting for mandibular bone defects by simulating the state of iron accumulation in patients with postmenopausal osteoporosis with high iron intervention in osteoporotic rats. METHODS: An iron accumulation ovariectomized osteoporosis model was firstly constructed. The model group underwent bilateral ovariectomy, and the intraperitoneal injection of ferric ammonium citrate (90 mg/kg, twice a week, for 11 weeks) was started in the 2nd week, while the sham-operated group had some fat around the ovaries removed and was given an equal amount of saline for 11 weeks. After the successful modeling, the experimental rats were divided into sham-operated group (n=6), high iron ovariectomtized group (n=6) and high iron ovariectomized deferoxamine mesylate treatment group (deferoxamine mesylate group, n=6). Bone defects of 5 mm in diameter were established in the rat’s bilateral mandibles and implanted with Bio-Oss bone powder. Intraperitoneal injection of deferoxamine mesylate (100 mg/kg, 3 times a week) was started on postoperative day 4 in the deferoxamine mesylate group, and equal volume of saline was given in the sham-operated and high iron ovariectomized groups. The bone samples of the mandible, liver and blood were taken at 2 and 12 weeks after bone grafting for Prussian blue staining of the jaw and liver and ELISA detection of serum ferritin to detect iron levels in various body tissues; hematoxylin-eosin staining and Masson staining were performed to observe inflammatory cell infiltration and early osteogenesis in the bone defect area; tartrate resistant acid phosphatase staining was performed to observe osteoclast differentiation; ELISA was performed to detect serum calcitonin and type I collagen C-terminal peptide levels; and Micro-CT and hematoxylin-eosin staining were performed to observe osteogenesis in the middle and late stages. RESULTS AND CONCLUSION: The number of tibial trabeculae was reduced and the trabeculae were sparsely arranged in the high iron ovariectomized group. Iron levels in the liver, jaw bone and serum were significantly higher in the high iron ovariectomized group than the sham-operated group at 2 weeks after bone grafting, while the iron levels were significantly decreased after deferoxamine mesylate intervention (P < 0.05). In the early stage of bone defect repair, more inflammatory cell infiltration, less new bone matrix and less type I collagen fiber production were observed in the high iron ovariectomized group than in the sham-operated group (P < 0.05); after deferoxamine mesylate treatment, inflammatory cell infiltration was reduced, a small amount of new bone matrix was produced and collagen fibers increased significantly (P < 0.05). In the middle and late stages of bone defect repair, Micro-CT results showed a reduction in new bone production in the high iron ovariectomized group compared with the sham-operated group and increased new bone matrix after deferoxamine mesylate treatment (P < 0.05). Compared with the sham-operated group, the osteoclast number, serum calcitonin level, and serum type I collagen C-terminal peptide level were increased in the high-iron ovariectomized group, while the osteoclast number was decreased and bone metabolic indexes were improved after treatment with deferoxamine mesylate. To conclude, in ovariectomized rats with high iron intervention, elevated iron levels can be seen in multiple tissues, accompanied by reduced new bone production in the mandibular bone defect area. Deferoxamine mesylate can improve bone metabolism and inhibit osteoclast activity by removing iron deposits in tissues, improve bone formation in iron-accumulated osteoporotic rats, and promote bone healing in the mandibular bone defect area. Figures and Tables | References | Related Articles | Metrics

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Mechanism by which strength training improves bone injury in ovariectomized rats Yang Mengxiao, Fu Changxi 2024, 28 (20):  3150-3156.  doi: 10.12307/2024.342 Abstract ( 259 )   PDF (1312KB) ( 1031 )   Save BACKGROUND: Postmenopausal osteoporosis significantly increases the risk of fracture, which seriously affects the quality of life of patients. Exercise therapy is an important non-drug means and prevention and treatment strategy for patients with osteoporosis, in which strength training is the best mode, but its specific biological mechanism has not been determined.  OBJECTIVE: To investigate the effects of strength training on bone morphology, materials and biomechanics in ovariectomized rats and to explore the mechanism of extracellular matrix remodeling.  METHODS: Forty-eight female Sprague-Dawley rats were divided into sham operation group, sham operation exercise group, ovariectomized group and ovariectomized exercise group according to the random number table method. The menopausal animal model was established by bilateral ovariectomy in the ovariectomized group and ovariectomized exercise group, while sham operation was performed in the sham operation group and sham operation exercise group. Four weeks after operation, the sham operation exercise group and the ovariectomized exercise group underwent 12-week tail weight-bearing ladder training, and the sham operation group and the ovariectomized group were raised quietly in the cage. The bilateral femur and tibia were separated after training. The right tibia was used for dual-energy X-ray densitometry and biomechanical, biophysical and biochemical analyses, the left tibia was detected using micro-computed tomography for bone microstructural examination, the right femur was subjected to hematoxylin-eosin staining for histological observation, and the left femur was used for western blot and gelatin zymography detection of protein expression and enzyme activity of extracellular matrix metabolism-related factors, respectively.  RESULTS AND CONCLUSION: Compared with the sham operation group, the maximal load and stiffness decreased (P < 0.05), bone density, bone mineral density, bone inorganic matter content, bone calcium content decreased (P < 0.05), bone water content increased (P < 0.05), trabecular bone volume fraction, trabecular connectivity density, and trabecular number decreased (P < 0.05), trabecular separation, structural model index increased (P < 0.05), bone adipocyte number and cross-sectional area increased (P < 0.05), matrix metalloproteinase-2 activity decreased (P < 0.05), and protein expression of tissue inhibitor of metalloproteinase-1 and osteoprotegerin increased (P < 0.05) in the ovariectomized group. Compared with the ovariectomized group, the maximal load, stiffness, fracture load and resilience increased (P < 0.05), bone mineral density, bone mineral content, bone mineral density, bone inorganic matter content, and bone calcium content increased (P < 0.05), bone water content decreased (P < 0.05), trabecular separation and bone marrow area decreased (P < 0.05), trabecular bone thickness, cortical bone volume fraction, cortical bone area fraction, cortical bone thickness, and cortical bone porosity increased (P < 0.05), bone adipocyte number and cross-sectional area reduced (P < 0.05), matrix metalloproteinase-2 activity increased (P < 0.05), and protein expression of tissue inhibitor of metalloproteinase-1, Runt-related transcription factor 2 and osteoprotegerin decreased (P < 0.05) in the ovariectomized exercise group. To conclude, strength training can protect against bone injury caused by estrogen deficiency, which is characterized by improvement of bone biomechanical properties, bone tissue composition and bone microstructure, and its mechanism is related to the regulation of extracellular matrix remodeling. Figures and Tables | References | Related Articles | Metrics

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Constructing a model of anterior cruciate ligament reconstruction with autologous Achilles tendon in southern Yunnan small-ear pigs Xiong Bohan, Yu Yang, Zheng Liling, Yang Tengyun, Lu Xiaojun, Wang Xu, Li Kaiwei, Yu Hong, Li Yajuan, Dong Kaiyan, Zhang Yaozhang, Liu Jinrui, Gu Ziming, Hu Bigeng, Li Yanlin 2024, 28 (20):  3157-3163.  doi: 10.12307/2024.331 Abstract ( 313 )   PDF (1609KB) ( 452 )   Save BACKGROUND: As a dominant breed pig in southwest China, the southern Yunnan small-ear pig has been widely used as an experimental animal in the basic research of other disciplines, but there are still no reports on its application in anterior cruciate ligament reconstruction. OBJECTIVE: To establish a large animal model of the southern Yunnan small-ear pig with anterior cruciate ligament with autologous Achilles tendon was established. METHODS: Twenty adult female Yunnan small-ear pigs were equally randomized into two groups. In the autologous Achilles tendon group, the right knee anterior cruciate ligament was reconstructed with autologous Achilles tendon as a graft, while in the sham-operated group, a similar operation was performed on the right knee without any treatment of the anterior cruciate ligament. General conditions of each pig were observed and recorded before and 12 months after surgery. Ligaments and grafts were taken for gross observation and MAS scoring. Hematoxylin-eosin staining was performed to observe morphological characteristics of ligaments. The staining and arrangement of type I and type III collagen were evaluated by immunohistochemistry. Transmission electron microscopy was used to observe the type, size, diameter, ratio, and distribution of collagen fibers in ligaments.  RESULTS AND CONCLUSION: All animals had normal diet and activity, good wound healing, no obvious inflammatory reaction, no local purulent infection, and no significant changes in mental and urinary conditions compared with those before surgery. The reconstructed cruciate ligament of the knee was intact, with no stiffness and normal range of motion. Both the anterior drawer and Lachman tests were negative. Gross observation of the graft: 12 months after surgery, the grafts was in good position, with good integrity, obvious tension, ligament color close to the original anterior cruciate ligament, and complete surface synovial coverage. Most of the intraarticular ligaments in the autologous Achilles tendon group were defined as MAS I type and a few were defined as MAS II type. In the sham-operated group, the intraarticular ligament was defined as MAS I type. Hematoxylin-eosin staining indicated that, 12 months after surgery, collagen fibers in the autologous Achilles tendon group began to appear bundled, isotropic, and uniformly arranged, with more obvious isotropic corrugations, and the nuclei were mainly linear or spindle-shaped, which were similar to those in normal anterior cruciate ligament tissue of the sham-operated group. Immunohistochemistry results indicated that, 12 months after surgery, there was a higher expression of type I collagen and significantly less expression of type III collagen in the reconstructed anterior cruciate ligament in the autologous Achilles tendon group. The degree of type I and type III staining was similar in the two groups. Under the transmission electron microscope, the diameter, arrangement and density of collagen fibers in the reconstructed anterior cruciate ligament of the autologous Achilles tendon group were similar to those of the original anterior cruciate ligament at 12 months after surgery, indicating that the ligament remodeling process had been basically completed in the autologous Achilles tendon group at 12 months after surgery. Through a comprehensive evaluation of animal general conditions, ligament general view, MAS score, hematoxylin-eosin staining, immunohistochemistry, and transmission electron microscopy observation, we successfully established a large animal model of anterior cruciate ligament reconstruction using autogenous Achilles tendon in southern Yunnan small-ear pigs, with good morphological, histological and ultrastructural results.  Figures and Tables | References | Related Articles | Metrics

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Protective effect and mechanism of 3-nitro-N-methyl salicylamide on the skeletal muscle of rats with limb ischemia-reperfusion injury Ji Weixiu, Bai Yi, Wang Shuo, Zhao Yungang 2024, 28 (20):  3164-3169.  doi: 10.12307/2024.358 Abstract ( 282 )   PDF (1278KB) ( 135 )   Save BACKGROUND: Mitochondrial reactive oxygen bursts have been shown to play a key role in skeletal muscle ischemia-reperfusion injury. 3-Nitro-N-methylsalicylamide (3-NNMS) can effectively reduce the electron transport rate and has a potential protective effect on limb ischemia-reperfusion injury, but there is no clear research and clinical application. OBJECTIVE: To investigate the protective effect of 3-NNMS on the skeletal muscle after limb ischemia-reperfusion injury in rats and its mechanism.  METHODS: Forty healthy 8-week-old Sprague-Dawley rats were randomly divided into control group, 0, 25 and 125 μg/mL 3-NNMS groups, with 10 rats in each group. Animal models of limb ischemia-reperfusion injury were prepared in the latter three groups. 3-NNMS was injected into the injury site 30 minutes before reperfusion. The animals were sacrificed 2 hours after reperfusion. Blood from the apical part of the heart, and the tissue of the rectus femoris muscle of the right lower limb were taken for testing. The pathological morphology of the rectus femoris muscle was detected by hematoxylin-eosin staining. Serum levels of creatine kinase found in the skeletal muscle (CK-MM), lactate dehydrogenase, and myeloperoxidase were detected using ELISA; the levels of nuclear factor κB, tumor necrosis factor α, interleukin 1β, cyclooxygenase 2, malondialdehyde, reactive oxygen species, superoxide dismutase, catalase and glutathione peroxidase in the rectus femoris muscle were measured; and adenosine triphosphate (ATP) level, ATPase activity, and mitochondrial respiratory control rate were tested. RESULTS AND CONCLUSION: Compared with the control group, the model rats with ischemia-reperfusion injury had increased serum levels of CK-MM, lactate dehydrogenase, and myeloperoxidase, increased levels of nuclear factor κB, tumor necrosis factor α, interleukin 1β, cyclooxygenase 2, malondialdehyde and reactive oxygen species in the rectus femoris muscle, decreased levels of catalase and glutathione peroxidase in the rectus femoris muscle, and reduced ATPase activity and mitochondrial respiratory control rate. Moreover, cell morphology was irregular, inflammatory cell infiltration was obvious, and the cells were swollen in rats after ischemia-reperfusion injury. Compared with the 0 μg/mL group, the serum CK-MM and lactate dehydrogenase levels decreased, the levels of nuclear factor κB and cyclooxygenase 2 in the rectus femoris muscle decreased, reactive oxygen species level decreased, and superoxide dismutase activity increased in the 25 μg/mL group; cell morphology was more regular, inflammatory cell infiltration was lighter, and cell swelling was alleviated. Compared with the 0 μg/mL group, the 125 μg/mL group had a reduction in the serum levels of CK-MM, lactate dehydrogenase, and myeloperoxidase and the levels of nuclear factor κB, tumor necrosis factor α, cyclooxygenase 2, malondialdehyde and reactive oxygen species in the rectus femoris muscle, as well as an increase in the levels of superoxide dismutase and glutathione peroxidase in the rectus femoris muscle, and mitochondrial respiratory control rate. Moreover, the cells were arranged neatly, the outline was clear and complete, and the inflammatory cell infiltration was light. To conclude, 3-NNMS can alleviate the functional impairment of the skeletal muscle caused by limb ischemia-reperfusion, and its mechanism of action may be through improving mitochondrial function, reducing reactive oxygen species production, decreasing oxidative stress and inflammatory response, and thus reducing tissue damage and repairing skeletal muscle function. Figures and Tables | References | Related Articles | Metrics

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Cartilage protective effect of swimming exercise in aged mice with knee osteoarthritis Zhu Shijie, Yang Yiting, Cao Yuting, Zheng Liangdong, Lin Kaili, Zhu Rui 2024, 28 (20):  3170-3175.  doi: 10.12307/2024.353 Abstract ( 371 )   PDF (1268KB) ( 135 )   Save BACKGROUND: Swimming is an important non-pharmacological treatment for knee osteoarthritis, which can effectively alleviate the disease. However, the effect and mechanism of swimming on senile knee osteoarthritis are still unclear. OBJECTIVE: To investigate the effect of swimming exercise on the articular cartilage of aged mice with knee osteoarthritis. METHODS: Six 3-month-old male C57BL/6 mice were selected as the young group, and twelve 18-month-old male C57BL/6 mice were randomized into the aged group and the swimming group, with six mice in each group. Mice in the swimming group received adaptive swimming for 1 week and formal swimming for 8 weeks. After the intervention, stride length analysis and sampling were performed in each group. The total number of leukocytes and lymphocytes in peripheral blood was detected by blood routine examinations. The morphology of the articular cartilage was observed by hematoxylin-eosin and safranin O-fast green staining. Chondrocyte counts and the modified Mankin’s score were used to evaluate the degree of articular cartilage damage. The protein and mRNA expressions of type II collagen, aggrecan and matrix metalloproteinase 13 in articular cartilage were detected by immunohistochemical staining and RT-qPCR.  RESULTS AND CONCLUSION: Compared with the young group, the mice in the aged group showed significantly decreased stride length (P < 0.05), significantly increased numbers of peripheral leukocytes and lymphocytes (P < 0.05), significantly decreased count of chondrocytes (P < 0.05), significantly increased modified Mankin’s score (P < 0.05), significantly decreased protein and mRNA expression of type II collagen and aggreca (P < 0.05), and significantly increased matrix metalloproteinase 13 expression (P < 0.05). Moreover, hematoxylin-eosin and safranin O-fast green staining showed the uneven surface of the articular cartilage, abnormal chondrocytes, and proteoglycan loss in the aged group. Compared with the aged group, swimming exercise significantly improved the stride length of mice (P < 0.05), decreased the count of peripheral blood lymphocytes (P < 0.05), increased the count of chondrocytes (P < 0.05), decreased the modified Mankin’s score (P < 0.05), increased the protein and mRNA expression of type II collagen and aggrecan (P < 0.05), and decreased the expression of matrix metalloproteinase 13 (P < 0.05). Hematoxylin-eosin and safranin O-fast green staining showed that the articular surface of mice in the swimming group was smooth, chondrocytes were normal, and proteoglycan loss was less. All these findings indicate that swimming exercise can reduce the number of inflammatory cells in the blood of aged mice, improve articular chondrocytes, matrix composition and cartilage tissue morphology; thus, it has a protective effect on the cartilage of aged mice with knee osteoarthritis. Figures and Tables | References | Related Articles | Metrics

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Effect of electronic moxibustion on the volume of hippocampal subregion in patients with amnestic mild cognitive impairment Shi Jiao, Li Xingjie, Liu Qiqi, Liu Jun, Yuan Xu, Chen Shangjie 2024, 28 (20):  3176-3181.  doi: 10.12307/2024.327 Abstract ( 343 )   PDF (1167KB) ( 137 )   Save BACKGROUND: Current studies have shown that electronic moxibustion can improve memory function in amnestic mild cognitive impairment; however, its mechanism of action needs to be further investigated. The atrophy of hippocampal volume and impairment of functional connectivity are important imaging markers of amnestic mild cognitive impairment. Whether electronic moxibustion can regulate the volume of hippocampal subregion of partients with amnestic mild cognitive impairment is worth studying.  OBJECTIVE: To observe the effect of electronic moxibustion on the volume of hippocampal subregions in patients with amnestic mild cognitive impairment.  METHODS: Forty patients with amnestic mild cognitive impairment were recruited from April 1, 2018 to January 31, 2019 at the community service centers around the Second Affiliated Hospital of Shenzhen University (Baoan Hospital of Southern Medical University), Shenzhen, China. They were randomly divided into treatment group (n=20) and control group (n=20). The treatment group was treated with electronic moxibustion of regulating the mind and benefiting the intelligence, while the control group was treated with placebo moxibustion. Moxibustion was given at 45 oC, 20 minutes each time, once a day, 5 times a week, for 8 weeks in total. Memory evaluation using Rivermead behavioral memory test and magnetic resonance imaging scanning for detecting the hippocampal subregion volume were performed for each patient before and after treatment, and cognitive function of each patient was assessed using Montreal cognitive assessment and mini-mental state examination. Correlation of hippocampal subregion volumes with scores on each scale was analyzed. RESULTS AND CONCLUSION: After treatment, the volumes of the left parasubiculum and the left hippocampal-amygdala migrating area increased in the treatment group but decreased in the control group, and there was a significant difference between the two groups (P < 0.05). Compared with the pre-treatment data, the Rivermead behavioral memory test, Montreal cognitive assessment, and mini-mental state examination scores were significantly higher in the treatment group after treatment (P < 0.05), while there was no significant change in the three scale scores in the control group after treatment (P > 0.05). The three scale scores were higher in the treatment group than in the control group after treatment (P < 0.05). Pearson correlation analysis showed that the changes in the volume of the left parasubiculum was significantly and positively correlated with the Rivermead behavioral memory test scale score in the treatment group (r=0.418, P=0.014). To conclude, electronic moxibustion can improve memory in patients with amnestic mild cognitive impairment, and the mechanism may be the regulation of structural plasticity in hippocampal subregions. Figures and Tables | References | Related Articles | Metrics

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Sequencing, verification and functional analysis of differentially expressed genes in brain tissue of a rat model with acute intracerebral hemorrhage Gao Yuguang, Zhong Jie, Huang Deqing, Ma Yujuan, Liao Yuxiong, Liu Qiqi 2024, 28 (20):  3182-3189.  doi: 10.12307/2024.354 Abstract ( 406 )   PDF (2351KB) ( 652 )   Save BACKGROUND: There are differentially expressed genes in acute intracerebral hemorrhage, which are related to the occurrence and development of intracerebral hemorrhage.  OBJECTIVE: To screen differentially expressed genes and key genes in brain tissue of a rat model with acute intracerebral hemorrhage, to validate them through qPCR, and to analyze the relationships between key genes and the neurological function and brain tissue water content after intracerebral hemorrhage.  METHODS: Seventy-eight Sprague-Dawley rats were randomly divided into two groups: in intracerebral hemorrhage group, a rat model of acute intracerebral hemorrhage was made using collagenase injection at the right caudate nucleus; and in sham-operated group, rats were injected with equal amount of saline at the same site. RNA was extracted from rat brain tissues of both groups using the TRIzol method and transcriptome sequencing technology was used to identify differentially expressed genes in brain tissues of acute intracerebral hemorrhage, which were then verified by qPCR and analyzed for the relationships between the genes and neurological function and brain tissue water content after intracerebral hemorrhage. And the key genes were analyzed by GO and KEGG functional enrichment analysis in combination with bioinformatics.  RESULTS AND CONCLUSION: Ten key genes were identified, including CXCL8, SERPINE1, TFPI2, CXCR4, GDA, KCNQ5, ERICH3, SCN3B, CACNA1E, and CCL20. The contents of GDA, KCNQ5, ERICH3, SCN3B, and CACNA1E in the intracerebral hemorrhage group were lower than those in the sham-operated group (P < 0.05). The contents of CXCL8, SERPINE1, TFPI2, CXCR4 and CCL20 in the intracerebral hemorrhage group were higher than those in the sham-operated group (P < 0.05). The contents of GDA, KCNQ5, ERICH3, SCN3B, and CACNA1E were positively correlated with brain tissue water content and neurologic deficit score (P < 0.05), while the contents of CXCL8, SERPINE1, TFPI2, CXCR4 and CCL20 were negatively correlated with brain tissue water content and neurologic deficit score (P < 0.05). GO analysis indicated that differentially expressed genes were mainly enriched in two biological processes (leukocyte chemotaxis and chemokine-mediated signaling pathways), two cell components (cation channel complexes and ion channel complexes), and two molecular functions (gated channel activity and ion channel activity). KEGG analysis indicated that differentially expressed genes were concentrated in tumor necrosis factor signaling pathway, glutamatergic synapses and GABAergic synapses. To conclude, the differentially expressed genes in intracerebral hemorrhage include CXCL8, SERPINE1, TFPI2, CXCR4, GDA, KCNQ5, ERICH3, SCN3B, CACNA1E, and CCL20, and these genes are related to brain tissue water content and neurological function after intracerebral hemorrhage. These genes are mainly enriched in cell components, binding functions, cellular protrusions, and other related biological functions.  Figures and Tables | References | Related Articles | Metrics

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Role and mechanism of miR-155/leptin receptor/adenosine phosphate-dependent protein kinase axis in tuberculin-induced osteoclast formation Wang Zengshun, Suonan Angxiu, Liu Limin, Zhou Jingyuan 2024, 28 (20):  3190-3195.  doi: 10.12307/2024.333 Abstract ( 233 )   PDF (2227KB) ( 62 )   Save BACKGROUND: Abnormal activation of osteoclasts plays an important role in the bone destruction due to spinal tuberculosis. During the pathogenesis of osteoporosis, miR-155 knockdown activates adenosine phosphate-dependent protein kinase (AMPK) by increasing the expression of leptin receptors, thereby inhibiting osteoclast differentiation and bone resorption. However, the role of miR-155/leptin receptor(LEPR)/AMPK axis in the bone destruction due to spinal tuberculosis remains unclear. OBJECTIVE: To investigate the role and mechanism of miR-155/LEPR/AMPK axis in tuberculin-induced osteoclast formation.  METHODS: RAW264.7 cells were cultured and treated with different concentrations of purified protein derivative (PPD) (1.0, 2.5, 5.0, 10.0 IU/mL) and transfected with negative control (NC) sequence or miR-155 inhibitor, NC siRNA sequence or LEPR siRNA sequence. Tartrate resistant acid phosphatase staining was used to detect the number of osteoclasts. Fluorescence quantitative PCR was used to detect the expression of miR-155. Western blot was used to detect the expression of LEPR and p-AMPK. Double luciferase reporter gene was used to verify miR-155 targeting LEPR.  RESULTS AND CONCLUSION: Compared with the control group, the number of osteoclasts and the expression level of miR-155 significantly increased, while the expression level of LEPR and p-AMPK significantly decreased in 2.5, 5.0, and 10.0 IU/mL PPD groups (P < 0.05). Compared with NC+5.0 IU/mL PPD group, the number of osteoclasts and the expression level of miR-155 significantly decreased, while the expression level of LEPR and p-AMPK significantly increased in the miR-155 inhibitor+5.0 IU/mL PPD group (P < 0.05). Compared with the NC group, the fluorescence activity of LEPR wild-type double luciferase reporter gene was increased in the miR-155 inhibitor group, and decreased in the miR-155 mimic group (P < 0.05). Compared with si-NC+miR-155 inhibitor+5.0 IU/mL PPD group, the expression level of miR-155 had no significant change, the number of osteoclasts significantly increased, and the expression levels of LEPR and p-AMPL significantly decreased in si-LEPR+miR-155 inhibitor+5.0 IU/mL PPD group (P < 0.05). To conclude, tuberculin can induce osteoclast formation by increasing miR-155 expression and inhibiting downstream LEPR expression and AMPK activation. Figures and Tables | References | Related Articles | Metrics

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Machine learning combined with bioinformatics to identify and validate key genes for cellular senescence in osteoarthritis Yuan Changshen, Liao Shuning, Li Zhe, Wu Siping, Chen Lewei, Liu Jinyi, Li Yanhong, Duan Kan 2024, 28 (20):  3196-3202.  doi: 10.12307/2024.344 Abstract ( 384 )   PDF (1238KB) ( 525 )   Save BACKGROUND: Cellular senescence is closely related to the development and progression of osteoarthritis, but the specific targets and regulatory mechanisms are not yet clear.  OBJECTIVE: To mine key genes in cellular senescence-mediated osteoarthritis by integrating bioinformatics and machine learning approaches and validate them via experiments to explore the role of cellular senescence in osteoarthritis.  METHODS: The osteoarthritis gene expression profiles obtained from the GEO database were intersected with cellular senescence-related genes obtained from the CellAge database and the expression of the intersected genes was extracted for differential analysis, followed by GO and KEGG analysis of the differential genes. The key osteoarthritis cellular senescence genes were then screened by protein-protein interaction network analysis and machine learning, and in vitro cellular experiments were performed. Finally, the expression of the key genes was detected by qPCR.  RESULTS AND CONCLUSION: A total of 31 osteoarthritis cell senescence differential genes were identified. GO analysis showed that these genes were mainly involved in the biological processes, such as regulation of leukocyte differentiation, monocyte differentiation, regulation of T cell differentiation and exerted roles in DNA transcription factor binding, histone deacetylase binding, chromatin DNA binding, and chemokine binding. KEGG analysis showed that osteoarthritis cell senescence differential genes were mainly activated in the JAK/STAT signaling pathway, PI3K/Akt signaling pathway and FoxO signaling pathway. MYC, a key gene for osteoarthritis cellular senescence, was identified by protein-protein interaction network topology analysis and machine learning methods. The results of the in vitro cellular assay showed that the mRNA expression of MYC was significantly lower in the experimental group (osteoarthritis group) than the control group (normal group) (P < 0.05). To conclude, MYC can be a key gene in the senescence of osteoarthritic cells and may be a new target in the prevention and treatment of osteoarthritis by mediating immune response, inflammatory response and transcriptional regulation. Figures and Tables | References | Related Articles | Metrics

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The pathological progression of steroid-induced osteonecrosis of the femoral head caused by oxidative stress-induced osteoblast ferroptosis Zhang Jiahao, Liu Yuhao, Zhou Chi, Mo Liang, Fang Hanjun, Chen Zhenqiu 2024, 28 (20):  3202-3208.  doi: 10.12307/2024.334 Abstract ( 368 )   PDF (2335KB) ( 358 )   Save BACKGROUND: Studies have shown that imbalance of bone metabolism during glucocorticoid-induced osteonecrosis of the femoral head necrosis is closely related to oxidative stress. OBJECTIVE: To investigate the pathological mechanism by which oxidative stress-induced ferroptosis promote apoptosis in osteoblasts involved in steroid-induced osteonecrosis of the femoral head. METHODS: General data and serum specimens were collected from 47 patients with steroid-induced osteonecrosis of the femoral head. In addition, six femoral head specimens were collected from these patients. According to the Association Research Circulation Osseous (ARCO) staging system, serum specimens were grouped into ARCO II, III, and IV, while femoral head specimens were classified into ARCO III and IV. Serum levels of malondialdehyde and superoxide dismutase 1 were measured. The protein expression of superoxide dismutase 1, glutathione peroxidase 4, Bcl-2 in the femoral head was detected and verified by Data independent acquisition (DIA) for quantitative sequencing, western blot and alkaline phosphate detection. RESULTS AND CONCLUSION: The ARCO stage of patients with steroid-induced osteonecrosis of the femoral head was independent of age, sex and necrotic side. The serum levels of malondialdehyde and superoxide dismutase 1 were higher in patients with ARCO stage III compared with those with ARCO stage II and IV. The results of DIA protein quantification showed that the function of differential proteins was mainly related to redox. The levels of superoxide dismutase 1, glutathione peroxidase 4, and Bcl-2 in the necrotic region were lower than in the normal region, as well as lower in ARCO stage IV than in ARCO stage III. Western blot verified the results of DIA protein quantification. The alkaline phosphatase activity was lower in the necrotic region than in the normal region, as well as lower in ARCO stage IV than in ARCO stage III. In the necrotic and sclerotic regions, the function of differential proteins was also related to redox, and superoxide dismutase 1, glutathione peroxidase 4, Bcl-2 protein expression and alkaline phosphatase activity were lower in the necrotic area than in the sclerotic region, as well as lower in ARCO stage IV than in ARCO stage III. To conclude, glucocorticoids can influence the progression of steroid-induced osteonecrosis of the femoral head by upregulating oxidative stress levels, inducing osteoblast ferroptosis, and inhibiting osteogenic function. Figures and Tables | References | Related Articles | Metrics

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Correlations between brain function and olfactory function in patients with cerebral small vessel disease and Parkinson’s disease based on resting-state functional magnetic resonance imaging Huang Zhongxia, Wang Yu, Liu Yawen, Zhang Xiaoxu, Xu Dandan, Yang Yanping, Huang Mingming, Yu Hui 2024, 28 (20):  3209-3216.  doi: 10.12307/2024.345 Abstract ( 272 )   PDF (1233KB) ( 341 )   Save BACKGROUND: Olfactory dysfunction is an early biological marker of various diseases. However, the neuroimaging mechanism by which olfactory dysfunction occurs following cerebral small vessel disease is unclear. OBJECTIVE: To explore the different neuroimaging mechanisms of olfactory function regulation in patients with cerebral small vessel disease and Parkinson’s disease, and explore the potential application value of olfactory function assessment in patients with cerebral small vessel disease.  METHODS: Neuropsychological and olfactory tests, high-resolution structural magnetic resonance and resting-state functional magnetic resonance data were collected in 80 patients with cerebral small vessel disease, 44 healthy controls and 29 patients with Parkinson’s disease. DPABI, SPM12 and SPSS were used to analyze and compare the amplitude of low frequency fluctuation, regional homogeneity and functional connectivity values between the cerebral small vessel disease, control and Parkinson’s disease groups. Correlations between the significantly altered resting-state functional magnetic resonance imaging measures and olfactory and cognitive scores were evaluated.  RESULTS AND CONCLUSION: Compared with the control group, low-frequency fluctuation amplitude of the right dorsolateral superior frontal gyrus and the regional homogeneity of the left wedge leaf were significantly reduced in the cerebral small vessel disease and Parkinson’s disease groups. The right dorsolateral superior frontal gyrus and the left cuneiform lobe are the seed points. Compared with the Parkinson’s disease group, the functional connectivity values of the right anterior cunei, inferior temporal gyrus, anterior central gyrus and dorsolateral superior frontal gyrus, left posterior central gyrus and inferior temporal gyrus were significantly enhanced in the control and cerebral small vessel disease groups. The left cuneiform lobe was the seed point. Compared with the control group, the functional connectivity of the left lingual gyrus was significantly weakened in the cerebral small vessel disease and Parkinson’s disease groups. The functional connectivity values of the left middle temporal gyrus and the right posterior central gyrus were enhanced in the control group compared with the cerebral small vessel disease and Parkinson’s disease group, and that was enhanced in the cerebral small vessel disease group compared with the Parkinson’s disease group. Correlation analysis showed that the olfactory score and cognitive score were positively correlated in the cerebral small vessel disease group, and the regional homogeneity of the left wedge lobe was negatively correlated with the Montreal Cognitive Assessment Scale score, while the functional connectivity of left wedge lobe-left middle temporal gyrus in the Parkinson’s disease group was positively correlated with the olfactory recognition score, and the functional connectivity values of the left wedge lobe-left posterior central gyrus and left wedge lobe-left lingual gyrus were positively correlated with the olfactory identification score and the total olfactory score, respectively. The regulation of olfactory function in patients with cerebral small vessel disease has a different neuroimaging mechanism from that of olfactory dysfunction in patients with Parkinson’s disease. The olfactory function of patients with cerebral small vessel disease is related to cognitive function. It is speculated that the olfactory function following cerebral small vessel disease is a secondary change of brain dysfunction, while olfactory dysfunction following Parkinson’s disease is directly caused by abnormal function of olfactory-related brain areas. Olfactory function assessment in patients with cerebral small vessel disease has potential application in predicting cognitive function. Figures and Tables | References | Related Articles | Metrics

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Grape seed extract inhibits apoptosis in growth plate chondrocytes and promotes tibial growth in rats Ning Taoli, Xie Yan, Wang Na, Wang Qingfeng, Ji Jian, Zhang Dongna 2024, 28 (20):  3216-3222.  doi: 10.12307/2024.310 Abstract ( 395 )   PDF (1878KB) ( 1187 )   Save BACKGROUND: Grape seed extract has been shown to be effective in inhibiting the growth of androgen-dependent tumors (e.g., breast cancer), and thus grape seed extract could theoretically inhibit epiphyseal closure induced by estrogen in late adolescence. OBJECTIVE: To screen the effects of grape seed extract on apoptosis of growth plate chondrocytes and epiphyseal closure in rats.  METHODS: (1) In vitro experiment: Growth plate chondrocytes from rat large tibia and femur at logarithmic growth stage were obtained and cultured in groups: normal control group, model control group (adding 17β-estradiol to induce apoptosis), positive control group (adding letrozole and 17β-estradiol), grape seed extract group (adding 17β-estradiol and 10 µg/mL grape seed extract), Caspase-9 inhibitor group (adding 17β-estradiol and Caspase-9 inhibitor), Caspase-9 agonist group (adding 17β-estradiol and Caspase-9 agonist). Cell apoptosis was detected by flow cytometry after 48 hours of culture. (2) In vivo experiment: Thirty 3-month-old Sprague-Dawley rats were randomly divided into model control group, positive control group and low-, medium- and high-dose groups, with five rats in each group. All rats were injected subcutaneously with 17β-estradiol (3 times per week) to establish epiphyseal closure models, followed by intragastric administration of letrozole in positive control group and 0.05, 0.2 and 0.8 g/kg grape seed extract in low-, medium- and high-dose groups, respectively, once a day until over 2/3 of the epiphyseal plate in the model control group was closed. The length of the tibia was then observed. Another 18 Sprague-Dawley rats were randomly divided into model control group, positive control group, and medium-dose group, with 6 rats in each group, treated as above for 1.5 continuous months. The expression of Caspase-9 protein in rat growth plate cartilage was detected by western blot. RESULTS AND CONCLUSION: (1) In vitro experiment: 17β-estradiol could induce apoptosis in growth plate chondrocytes, and letrozole, grape seed extract, and caspase-9 inhibitors could all inhibit apoptosis in growth plate chondrocytes. (2) In vivo experiment: When more than 2/3 of the epiphyseal plate in the model control group was closed, the number of rats with epiphysis closure in the positive control and medium-dose groups was less than that in the model control group (P < 0.05), and the tibial length was longer than that in the model control group (P < 0.05), and the Caspase-9 protein expression in the tibial growth plate was lower than that in the model control group (P < 0.05). To conclude, the appropriate dose of grape seed extract can effectively inhibit the apoptosis of growth plate chondrocytes and delay epiphyseal closure, which has the potential to promote bone growth. Figures and Tables | References | Related Articles | Metrics

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Development of a three-dimensional digital children’s acupuncture point visualization system of Mongolian medicine Liu Yuhang, Sun Ruifen, Mu Rigen Jiya, Wang Xing, Li Zhijun, Liu Yanan, Hao Yunteng, Cai Yongqiang, Zhang Shaojie, Li Kun 2024, 28 (20):  3223-3228.  doi: 10.12307/2024.389 Abstract ( 404 )   PDF (1968KB) ( 1197 )   Save BACKGROUND: Nowadays, there are increasing reports on the digitization and visualization system of acupuncture points for adults in traditional Chinese medicine, and the digitization and visualization system of acupuncture points for children in traditional Chinese medicine and the simulation system of acupuncture manipulation for Mongolian medicine training have been reported. However, there are no reports on relevant systems for children in Mongolian medicine. OBJECTIVE: To develop a simulation system of acupuncture points for children in Mongolian medicine, in the hope that it can be used for clinical teaching, manipulation practice and research on acupuncture safety. METHODS: Based on the tomographic anatomical dataset of preschool boys, a three-dimensional (3D) digital virtual anatomical model of children with multiple internal organs and tissues was constructed by using PhotoShop.2021 and Digihuman Reconstruction System software. The relevant annotation information database of 27 acupoints such as Dinghui acupoint of Mongolian medicine was compiled by the Unity database language. The Mongolian gold needle and silver needle were selected to record the acupuncture point teaching video on the 3D printed head and neck resin model of children. In Unity3D software, children’s anatomical model, acupoint annotation information database and acupuncture operation video were integrated and coded, and a 3D digital children’s Mongolian acupuncture acupoint visualization system integrating simulation acupuncture training, clinical teaching and acupuncture safety research was successfully created. RESULTS AND CONCLUSION: This study was based on real children’s specimens. In order to reduce the error of two-dimensional segmentation, the manual layer-by-layer segmentation section image method was used to ensure the accuracy of the 3D model to the greatest extent. The Digihuman Reconstruction System was used to extract and save the individual segmentation data while maximizing the accuracy of the 3D model. PhotoShop.2021 software was used to complete the 3D reconstruction of the outer skin of the head and neck of children and the internal bony structure, cervical spinal cord, blood vessels and nerves, muscles and ligaments. After 3D reconstruction, the basic morphology of each independent structure and the integrity of the overall contour were verified in MeshLab software and the final fine adjustment and anatomical position confirmation were conducted using 3-matic research 13.0 software. The real anatomical morphology of the head and neck of preschool children was successfully simulated and restored. Unity3D software was used to integrate the 3D model of children, acupuncture operation video and acupoint annotation database, and the 3D digital Mongolian acupuncture acupoint visualization system for children was successfully constructed. Based on the real continuous fault ultra-thin dataset of preschool boys in China, China’s first 3D digitization and visualization system of acupuncture points in the head and neck of children in Mongolian medicine was developed. Compared with the previous acupuncture soft works, this system is more suitable for the anatomical morphological development characteristics of Asian children, and has a high value of application in the fields of research on the safety of acupuncture in Mongolian medicine, clinical teaching and acupuncture simulation training. Figures and Tables | References | Related Articles | Metrics

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Salvia miltiorrhiza attenuates white matter injury induced by hypoperfusion in neonatal rats Su Xuewen, Yuan Haifeng, Feng Wanyu, Song Ruixia, Chen Junlong, Yi Ruhan, Zhu Hua, Dou Zhongxia 2024, 28 (20):  3229-3234.  doi: 10.12307/2024.343 Abstract ( 248 )   PDF (2155KB) ( 504 )   Save BACKGROUND: Premature birth is a major global health problem associated with high mortality and morbidity. White matter injury is the most common brain injury in preterm infants. Salvia miltiorrhiza is a traditional herbal plant that is commonly used to treat cardiovascular and cerebrovascular diseases in Asian countries.  OBJECTIVE: To investigate the therapeutic effect of Salvia miltiorrhiza on white matter injury in preterm infants. METHODS: Eighteen neonatal male Sprague-Dawley rats at 3-day gestational age were selected and randomized into normal group, white matter injury group, and Salvia miltiorrhiza group. Animal models of preterm white matter injury were established by permanent ligation of the right common carotid artery in the latter two groups. Rats in the Salvia miltiorrhiza group were given intraperitoneal injection of Salvia miltiorrhiza (5 mg/kg•d) for 7 consecutive days. Normal group and white matter injury group were given the same volume of PBS for intervention. On the 14th day after modeling, the rats were sacrificed. Brains were pathologically observed by hematoxylin-eosin staining under microscope, and the expression levels of myelin basic protein and CC1 in brain tissue were visualized using immunofluorescence. Furthermore, liquid chromatography-tandem mass spectrometry was used to analyze possible pathways for the action of Salvia miltiorrhiza. RESULTS AND CONCLUSION: In the white matter injury group, the structure of the corpus callosum was irregular and the cells appeared swollen and necrotic. In addition, induction of white matter injury resulted in significantly reduced myelin formation, with irregular and loosely arranged nerve fibers and significantly decreased myelin sheaths. Interestingly, white matter injury rats treated with Salvia miltiorrhiza had reduced cellular swelling, reduced lesions, and increased myelin sheaths. The expression of myelin basic protein was closely related to myelin formation, and CC1 was a marker of myelin oligodendrocytes. Salvia miltiorrhiza significantly up-regulated the expressions of myelin basic protein and CC1 in white matter injury rats (P < 0.000 1), indicating that Salvia miltiorrhiza alleviated white matter injury. Liquid chromatography-tandem mass spectrometry analysis showed that the therapeutic effect of Salvia miltiorrhiza in the rat model of white matter injury was closely related to the regulation of complement and coagulation cascades. To conclude, Salvia miltiorrhiza may be a potential therapeutic agent for treating preterm white matter injury. Figures and Tables | References | Related Articles | Metrics

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Silent information regulator 1: A potential target of semaglutide in the treatment of Alzheimer’s disease Chai Shifan, Li Xinru, Ye Yucai, Sun Junli, Cai Hongyan, Wang Zhaojun 2024, 28 (20):  3235-3239.  doi: 10.12307/2024.382 Abstract ( 371 )   PDF (1770KB) ( 2737 )   Save BACKGROUND: Studies have found that glucagon-like peptide-1 and its analogues have a significant neuroprotective effect, and some drugs have been applied to the clinical stage III study of Alzheimer’s disease. However, the mechanism of its neuroprotective effect is still unclear, which needs to be further explored and clarified.  OBJECTIVE: To screen out the genes related to the pathogenesis of Alzheimer’s disease and the related targets of semaglutide for the treatment of Alzheimer’s disease based on bioinformatics and network pharmacology analyses, to identify the potential target genes by comprehensive analysis of the two and to verify them at the cellular level.  METHODS: Using DisGeNET database, differentially expressed genes between Alzheimer’s disease patients and healthy population were screened out. The chemical structure formula and two-dimensional structure diagram of semaglutide were obtained using PubChem online database. GO/KEGG enrichment analysis was performed using DAVID online database. A protein-protein interaction network was constructed by using the STRING database. The HPA database was used to determine the distribution characteristics of the target proteins in various human tissues. Finally, western blot was used to detect relevant protein expression in HT22 cells after semaglutide intervention.  RESULTS AND CONCLUSION: With the dataset in DisGeNET database, 3 374 differentially expressed genes between Alzheimer’s disease patients and healthy people were obtained, and meanwhile, 101 target genes of semaglutide potential drugs were obtained. There were 23 intersection genes between them. Ten key genes were identified based on the protein-protein interaction network, which were silent information regulator 1 (SIRT1), CASP9, CCND1, CASP1, KEAP1, DLG4, CASP4, GRB2, GRIA1, and EDNRA. The results of GO gene functional annotation analysis of key genes showed that the positive regulatory activity of cysteine endopeptidase, the positive regulation of proteolysis, and the positive regulation of cysteine endopeptidase involved the cytoplasmic part of the apoptotic activity process; AMPA glutamate receptor complex, inflammatory complex, CARD domain binding, cysteine endopeptidase activity, and cysteine endopeptidase activity were involved in the apoptotic process. The results of KEGG signaling pathway analysis indicated that colorectal cancer, non-small cell carcinoma, and endometrial carcinoma were related to immune infiltration, inflammation and autophagic apoptosis. In addition, according to the association ranking of key genes and their distribution in different tissues of HPA online database, SIRT1 was identified as the most significant differential gene. The expression level of SIRT1 protein was significantly down-regulated in HT22 cells after β-amyloid protein 1-42 treatment, but it could be significantly increased after being treated with semaglutide. To conclude, SIRT1 may be a target gene for semaglutide in the treatment of Alzheimer’s disease.  Figures and Tables | References | Related Articles | Metrics

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The effective components of Chinese medicine combined with scaffold materials promote bone tissue regeneration Dong Xinyu, Dong Xinyue, Wang Wanting, Fan Haixia, Cheng Huanzhi 2024, 28 (20):  3240-3245.  doi: 10.12307/2024.385 Abstract ( 526 )   PDF (910KB) ( 1222 )   Save BACKGROUND: With the proven ability of traditional Chinese medicine such as icariin and berberine to promote bone regeneration by regulating various mechanisms and targets, researchers have combined active ingredients of traditional Chinese medicine with bone tissue engineering and found that they have unique advantages in treating bone defects. OBJECTIVE: Starting from the active ingredients of traditional Chinese medicines that promote bone formation, to screen cases of their effective combination with different drug-carrying scaffold materials, and summarize the active ingredients of traditional Chinese medicines that have the potential to be applied to bone tissue engineering. METHODS: CNKI, WanFang, PubMed, and Web of Science were searched for relevant literature published from 2000 to 2023, using the keywords of “bone tissue engineering, bone tissue-engineered scaffold materials, bone defect, bone repair, bone regeneration, traditional Chinese medicine” in Chinese and English. According to the inclusion and exclusion criteria, 87 papers were finally included for review. RESULTS AND CONCLUSION: There are various kinds of active ingredients of traditional Chinese medicine to promote bone regeneration, mainly including flavonoids, non-flavonoid polyphenols, alkaloids, glycosides. These active ingredients have anti-inflammatory and analgesic effects, promote osteoblasts, inhibit osteoclasts and promote early angiogenesis. The combination of active ingredients of traditional Chinese medicine with bone tissue engineering is effective in anti-inflammation, accelerating collagen and bone formation, and promoting the expression of osteogenic genes, which provides a theoretical basis for the application of traditional Chinese medicine in bone tissue regeneration, and at the same time provides a new idea for the repair of bone defects. Figures and Tables | References | Related Articles | Metrics

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The role of macrophage polarization in the pathogenesis and treatment of periodontitis Ge Ruiyang, Ni Can, Yang Kun, Yan Fuhua 2024, 28 (20):  3246-3251.  doi: 10.12307/2024.339 Abstract ( 510 )   PDF (987KB) ( 309 )   Save BACKGROUND: Host immune response triggered by plaque biofilm is an initiator of periodontitis progression and destruction. Macrophages are an important component of the innate immune response, which play an important role in inflammation occurrence and development.  OBJECTIVE: To review the relationship between macrophage polarization and periodontitis and the related progress in the treatment of periodontitis by regulating macrophage polarization. METHODS: PubMed and CNKI databases were searched for relevant literature published from 1990 to 2023, with “macrophage polarization, M1/M2 macrophage, periodontitis, periodontitis treatment, macrophage polarization and periodontitis, osteoimmunology, ferroptosis, macrophage polarization and ferroptosis, periodontitis and ferroptosis” as the English and Chinese search terms. After the initial screening, 96 articles were selected for review. RESULTS AND CONCLUSION: The switch between different phenotypes of macrophages is closely related to the tissue destruction caused by periodontitis, and various cytokines and inflammatory mediators secreted from macrophages are involved in the destruction and repair of periodontal tissue. Therefore,regulation of macrophage polarization and cytokine secretion in inflammatory state helps to alleviate periodontitis inflammation and improve the periodontal microenvironment, thereby reducing tissue destruction or promoting periodontal tissue regeneration. Many studies have been conducted to develop drugs or biomaterials to modulate macrophage function for the purpose of immunomodulatory treatment of periodontitis. However, macrophages act throughout the development of periodontitis and play an important role in the process of anti-infection, bone destruction and bone repair, and polarization is a complex and dynamic process influenced by many factors. Therefore, further exploration on possible mechanisms is still needed to clarify the interaction between materials or drugs and macrophages. Figures and Tables | References | Related Articles | Metrics

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Regulation of N6-methyladenosine on non-coding RNAs in pathological cardiac remodeling Yin Gonghua, Xu Ruoyao, Zhang Lijuan, Zhang Yifan, Qi Jie, Zhang Jun 2024, 28 (20):  3252-2358.  doi: 10.12307/2024.349 Abstract ( 313 )   PDF (1014KB) ( 264 )   Save BACKGROUND: N6-methyladenosine (m6A) is a hot research topic in the mechanism of pathological cardiac remodeling and plays an important role in the development of cardiovascular diseases. OBJECTIVE: To summarize the possible mechanism by which m6A modification in non-coding RNAs regulates the main processes of pathological cardiac remodeling, such as pathological cardiac hypertrophy, cardiomyocyte death, myocardial fibrosis and vascular remodeling. METHODS: “m6A, non-coding RNA, pathological cardiac hypertrophy, cardiomyocyte apoptosis, cardiomyocyte pyroptosis, cardiomyocyte ferroptosis, myocardial fibrosis, vascular remodeling” were used as search terms in Chinese and English. Relevant literature from CNKI, PubMed and Web of Science databases published from January 1974 to April 2023 was retrieved, and finally 86 eligible articles were reviewed. RESULTS AND CONCLUSION: m6A modification is a highly dynamic and reversible modification. Pathological cardiac remodeling mainly involves pathological cardiac hypertrophy, cardiomyocyte apoptosis, cardiomyocyte pyroptosis, cardiomyocyte ferroptosis, myocardial fibrosis and vascular remodeling. m6A-related enzymes can regulate pathological cardiac remodeling processes through various non-coding RNAs and different signaling pathways, which can be used as a new potential intervention for cardiovascular diseases. In pathological cardiac remodeling, research on the regulatory relationship between m6A modification and non-coding RNAs is still in its infancy. With the development of epigenetics, m6A modification in non-coding RNAs is expected to have a new development in the regulation of pathological cardiac remodeling. Figures and Tables | References | Related Articles | Metrics

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Measurement and evaluation of proprioception of foot and ankle complexes Feng Liang, Zhang Yafei, Huo Hongfeng 2024, 28 (20):  3259-3264.  doi: 10.12307/2024.356 Abstract ( 432 )   PDF (1161KB) ( 1678 )   Save BACKGROUND: Research on foot and ankle proprioception is crucial for the rehabilitation of chronic ankle instability and geriatric diseases as well as for the improvement of body posture control and motor performance. Previous studies have often studied the sensory evaluation of the foot and ankle joints separately, which has limitations for a comprehensive understanding of their sensory function. OBJECTIVE: The foot and ankle complex is the only part in direct contact with the support surface, and plays an important role in the collective sensory feedback and regulation and balance control. By combing the existing investigation and research of foot and ankle ontology, the measurement and evaluation methods of the sensation of the foot and ankle complex are combed, in order to pave the way and provide the theoretical basis for future related studies.  METHODS: Chinese terms “(foot OR foot ankle OR ankle) AND (sensation OR proprioception)” and English terms “(foot OR ankle) AND (feel OR proprioception)” were used as the keywords for retrieving relevant literature in the Web of Science, PubMed, and CNKI. We understood the basic concepts, current status and scope of research on the foot and ankle, summarized and evaluated the proprioceptive evaluation methods of the foot and ankle, and finally included 57 papers for further review.  RESULTS AND CONCLUSION: The evaluation of foot and ankle complex sensation was mainly divided into sensory evaluation of the foot and proprioceptive evaluation of the ankle joint. The sensory evaluation of the foot mainly describes the sensation of the skin and the sensory feedback under the intervention conditions. The methods mainly include the pressure sensory threshold test, the two-point discrimination test of the foot (planar and plantar), and the duration test of skin vibration sensation. Ankle joint proprioception evaluation focuses on the description of joint position, motion range, force value and functional performance. The methods are mainly divided into static joint angle reset test, motion minimum threshold test, force perception reproduction test and dynamic balance, speed and walking ability tests. The report of quantitative results is generally expressed by “an error,” which is generally divided into absolute error, relative error, constant error, etc. To conclude, the foot and ankle complex has specific sensory capabilities, including foot sensation and ankle proprioception, which affect the quality of life and athletic performance of humans. Weakness of both foot sensation and ankle proprioception is associated with reduced human balance, and the combined measurements of the two can comprehensively and effectively evaluate foot and ankle function. The combination of foot and ankle sensory measures is selected according to different research needs and various influencing factors such as environment, emotion and reporting style are fully considered, to improve the validity of measurement and evaluation. Figures and Tables | References | Related Articles | Metrics

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Mechanism of action and related signaling pathways of long non-coding RNAs in neuroimmuno-inflammatory response after ischemic stroke Wan Jun, Bai Yanjie, Wang Yan, Chen Shuying, Chen Limin, Xiao Yuqian, Sun Kexin 2024, 28 (20):  3265-3271.  doi: 10.12307/2024.350 Abstract ( 343 )   PDF (1184KB) ( 433 )   Save BACKGROUND: Long non-coding RNAs (lncRNAs), as important regulators of the inflammatory response, are involved in the immune-inflammation-brain crosstalk mechanism after ischemic stroke and have the potential to become a therapeutic agent for neurological dysfunction after ischemic stroke. OBJECTIVE: To analyze and summarize the molecular mechanism of lncRNA acting on glial cells involved in the neuroimmuno-inflammatory cascade response after ischemic stroke and the associated signaling pathways, pointing out that lncRNAs have the potential to regulate inflammation after ischemic stroke. METHODS: PubMed was searched using the search terms of “ischemic stroke, long non-coding RNA, neuroinflammation, immune function, signal pathway, microglia, astrocytes, oligodendrocyte, mechanism,” and 63 relevant documents were finally included for review. RESULTS AND CONCLUSION: In the early stage of ischemic stroke, the death of nerve cells due to ischemia and hypoxia activates the innate immune response of the brain, promoting the secretion of inflammatory factors and inducing blood-brain barrier damage and a series of inflammatory cascades responses. As an important pathogenesis factor in ischemic stroke, the neuroimmuno-inflammatory cascade has been proved to seriously affect the prognosis of patients with ischemic stroke, and it needs to be suppressed promptly in the early stage. Neuroinflammation after ischemic stroke usually induces abnormal expression of a large number of lncRNAs that mediate a series of neuro-immune-inflammatory crosstalk mechanisms through regulating the polarization of microglia, astrocytes and oligodendrocytes to exert post-stroke neuroprotective effects. LncRNAs, as important regulatory factors of the inflammatory response, inhibit the neuroimmuno-inflammatory cascade response after ischemic stroke through regulating nuclear factor-κB, lncRNA-miRNA-mRNA axis, Rho-ROCK, MAPK, AKT, ERK and other signaling pathways to effectively improve neurological impairment after ischemic stroke. Most of experimental studies on the interaction between lncRNAs and ischemic stroke are based on a middle cerebral artery occlusion model or a cerebral ischemia-reperfusion injury model, but no clinical trials have been conducted. Therefore, it remains to be further explored about whether lncRNAs can be safely applied in clinical practice. At present, there are many therapeutic drugs for the treatment of ischemic stroke, but there are relatively few studies on the application of lncRNAs, exosomes and other transplantation technologies for the treatment of ischemic stroke using tissue engineering technology, which need to be further explored. lncRNA has become an important target for the treatment of ischemic stroke with its relative stability and high specificity. In future studies, more types of inflammatory lncRNAs that function under ischemic-hypoxia conditions should continue to be explored, in order to provide new research directions for the treatment of neuroinflammation after ischemic stroke. Figures and Tables | References | Related Articles | Metrics

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Application of deferoxamine in bone tissue regeneration Wang Raokaijuan, Zhao Lixing 2024, 28 (20):  3272-3280.  doi: 10.12307/2024.291 Abstract ( 522 )   PDF (1327KB) ( 1615 )   Save BACKGROUND: Aside from iron chelating, deferoxamine is also considered as an effective hypoxia mimetic agent and hypoxia inducible factor-1α stabilizer. Deferoxamine has played a favorable effect on bone regeneration in both basic and clinical research recently. Deferoxamine solutions or deferoxamine loaded bio-scaffolds have been locally applied in bone tissue engineering, and their promotion of bone repair involves various functional properties and molecular mechanisms which have not been entirely clarified. Moreover, their advances in research of bone regeneration lack comprehensive summary as well.  OBJECTIVE: To review the functional properties, relative merits and advances in basic research and clinical practice of deferoxamine applied in bone regeneration, attempting to provide references and strategies for further studies. METHODS: Relevant articles were searched with the key words of “deferoxamine OR desferrioxamine OR desferal OR DFO,” “bone tissue engineering OR bone regeneration OR bone remodeling OR bone repair OR bone healing OR osteogenesis,” “angiogenesis OR vascularized bone regeneration OR angiogenic-osteogenic coupling” in English and Chinese by using PubMed, WanFang and CNKI databases. Eventually, 88 articles were selected for review. RESULTS AND CONCLUSION: Deferoxamine can recruit stem cells and regulate their function, activate relevant signaling pathways to advance hypoxia adaptation of the cells, exert anti-inflammatory and antioxidant properties to improve local inflammatory environment, and promote bone regeneration by coupling osteogenesis and angiogenesis as well as inhibiting bone resorption. Compared with growth factors or peptides loaded in conventional bone tissue engineering, deferoxamine has its unique advantages as a small molecule drug, while it also has toxic reactions and application limitations. Therefore, it is necessary to optimize its loading form and dosagey. The unique angiogenic-osteogenic coupling ability of deferoxamine can be used in different types of bone injuries including fractures, osteonecrosis, distraction osteogenesis, bone grafting, oral related osteogenesis, and bone defects. Due to the enhancement of angiogenesis, this ability enables deferoxamine to better adapt and solve the difficulties in bone repair caused by the complex and variable clinical situations and individual differences. However, it is also necessary to compare and optimize the application methods and safe dosage of deferoxamine to expand its application scope and enhance its clinical value. Figures and Tables | References | Related Articles | Metrics