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    18 February 2023, Volume 27 Issue 5 Previous Issue    Next Issue
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    Effects of exercise training on bone mass and bone microstructure in aged osteoporotic rats
    Wang Jinling, Huang Xiarong, Qu Mengjian, Huang Fujin, Yin Lingwei, Zhong Peirui, Liu Jin, Sun Guanghua, Liao Yang, Zhou Jun
    2023, 27 (5):  676-682.  doi: 10.12307/2022.1020
    Abstract ( 402 )   PDF (1204KB) ( 51 )   Save
    BACKGROUND: Exercise training can maintain bone mass and improve bone mineral density. However, its effect on senile osteoporosis and its possible mechanism are not clear.
    OBJECTIVE: To study the effects of exercise training on senile osteoporosis and to explore its possible molecular mechanism.
    METHODS: Sixteen Sprague-Dawley rats aged 24 months old were regarded as the model of senile osteoporosis and randomly divided into elderly model group (n=8) and exercise training group (n=8), while eight young Sprague-Dawley rats of 3 months old were randomly selected as young control group. All rats moved freely in their cages and those in the exercise training group underwent exercise training every day for 8 weeks. After 8 weeks of intervention, serum bone-specific alkaline phosphatase, type I procollagen amino-terminal peptide, tartrate-resistant acid phosphatase 5b and type I collagen amino-terminal peptide were detected using ELISA assay. Bone mineral density of the femur and the fourth lumbar vertebra were measured by dual energy X-ray absorptiometry, while bone microstructural parameters of the tibia and the fifth lumbar vertebra were determined by Micro-CT in each group. mRNA and protein expressions of bone morphogenetic protein 2, Runt-related transcription factor 2, Osterix, and peroxisome proliferator-activated receptor γ in bone marrow of the femur were detected by reverse transcriptase polymerase chain reaction and western blot.
    RESULTS AND CONCLUSION: Compared with the elderly model group, the serum levels of bone-specific alkaline phosphatase and type I procollagen amino-terminal peptide (P < 0.05), bone mineral density of the femur (P < 0.05) but not that of the fourth lumbar vertebra (P > 0.05), bone volume fraction and trabecular number of the tibia (P < 0.05), and the mRNA and protein expression of bone morphogenetic protein 2, Runt-related transcription factor 2, and Osterix in the femur (P < 0.05) were significantly increased in the elderly model group, while the serum levels of tartrate-resistant acid phosphatase 5b and type I collagen amino-terminal peptide (P < 0.05), trabecular separation of the tibia and the fifth lumbar vertebra (P < 0.05), and the mRNA and protein expression of peroxisome proliferator-activated receptor γ were significantly decreased (P < 0.05). In addition, exercise training could improve the bone volume fraction, trabecular number and trabecular thickness of the fifth lumbar vertebra in rats with senile osteoporosis, although the differences in changes were not always statistically significant 
    (P > 0.05). The trabecular thickness of the tibia had no significant changes after exercise training (P > 0.05). To conclude, exercise training can significantly improve bone mineral density and bone microstructure, promote osteogenic differentiation, and inhibit adipogenic differentiation in aged osteoporotic rats. The mechanism may be related to the bone morphogenetic protein 2/Runt-related transcription factor 2/Osterix signaling pathway.
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    Screening and verification of key genes of necroptosis in osteoarthritis
    Yuan Changshen, Guan Yanbing, Li Zhe, Rong Weiming, Liao Shuning, Chen Lewei, Mei Qijie, Duan Kan
    2023, 27 (5):  695-700.  doi: 10.12307/2023.141
    Abstract ( 611 )   PDF (1691KB) ( 37 )   Save
    BACKGROUND: Osteoarthritis is closely related to chronic and low-grade inflammation. Necroptosis is a new cell death pathway with extensive inflammatory characteristics, which can affect the progression of osteoarthritis, but the related targets of necroptosis in osteoarthritis have not been found.
    OBJECTIVE: To explore the key genes of necroptosis in osteoarthritis based on bioinformatics and verify them with animal experiments, so as to provide new targets for the prevention and treatment of osteoarthritis from the perspective of necroptosis. 
    METHODS: Osteoarthritis- and necroptosis-related genes were screened from GEO database and GeneCard database respectively, and then the intersection of them was taken to obtain osteoarthritic necroptosis genes. Gene ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were further carried out to construct a protein-protein interaction network. The key genes of osteoarthritic necroptosis (Hub genes) were identified through five calculation methods using Cytoscape software. Finally, male  Sprague-Dawley rats were selected to metaphase models of osteoarthritis. The expression of Hub genes in the metaphase models of osteoarthritis of male Sprague-Dawley rats detected by RT-PCR. 
    RESULTS AND CONCLUSION: A total of 38 osteoarthritic necroptosis genes were identified. Gene ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses showed that they were significantly enriched in leukocyte activity, T cell receptor and interleukin-17 signal pathway. Five Hub genes, CYBB, FADD, IL1B, MYC and EGFR, were screened by the five different algorithms of Cytoscape. RT-PCR showed that EGFR and IL1B were highly expressed in the metaphase models of osteoarthritis of male  Sprague-Dawley rats (P < 0.05), while CYBB and MYC were lowly expressed (P < 0.05), but FADD had no significant changes (P > 0.05). To conclude, CYBB, IL1B, MYC, and EGFR, as the key genes of osteoarthritic necroptosis, are expected to become new targets for the prevention and treatment of osteoarthritis.
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    Anatomic risk factors for medial meniscus posterior root tears combined with anterior cruciate ligament injuries
    Liu Guangluan, Guo Zonglei, Ge Jin, Huang Dong, Wang Yehua
    2023, 27 (5):  663-668.  doi: 10.12307/2023.119
    Abstract ( 400 )   PDF (943KB) ( 66 )   Save
    BACKGROUND: Numerous studies have shown that the increased posterior inclination of the tibial plateau is an important risk factor for anterior cruciate ligament injury, which may promote the occurrence of medial meniscus posterior root tear. However, there are few studies on the anatomical risk factors of meniscus injury.
    OBJECTIVE: To compare the anatomic risk factors for medial meniscus posterior root tear at the proximal tibia in the case of anterior cruciate ligament injury and to improve the vigilance of clinicians to specific patients with anterior cruciate ligament injury.
    METHODS: Clinical data of 107 patients with anterior cruciate ligament injury who were admitted to the Department of Orthopedics of Xuzhou Medical University Hospital from January 2018 to December 2021 were retrospectively analyzed. They were divided into two groups according to whether there was medial meniscus posterior root tear confirmed by clinical and imaging diagnosis. Fifty-one patients with anterior cruciate ligament injury combined with medial meniscus posterior root tear were included in observation group, while fifty-six patients with simple anterior cruciate ligament injury were included in control group. Preoperative MRI was used to measure the medial and lateral posterior tibial slope (MPTS, LPTS), the medial and lateral posterior meniscal slope (MMS, LMS), the difference between the MPTS and LPTS (Δ-TS) and between the MMS and LMS (Δ-MS), and the medial tibial plateau depth (MTPD) of the two groups in a blind way.
    RESULTS AND CONCLUSION: There was no significant difference between the two groups in LPTS, LMS, Δ-TS, and MTPD (P > 0.05). The MTPS, MMS and Δ-MS in the observation group were significantly higher than those in the control group (P < 0.05). Multivariate Logistic regression analysis showed that the MPTS was an independent risk factor for anterior cruciate ligament injury with medial meniscus posterior root tear (P < 0.05). Receiver operating characteristic curve showed that the MPTS had a certain predictive value for anterior cruciate ligament injury with medial meniscus posterior root tear. All the results indicate that increased MPTS, increased MMS, and increased asymmetry of Δ-MS are important risk factors for anterior cruciate ligament injury with medial meniscus posterior root tear. This study provides important information for the diagnosis of medial meniscus posterior root tear, thus avoiding missed diagnosis and providing a theoretical basis for early ligament repair to reduce the risk of meniscus tear in clinical practice.
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    Correlation between autophagy and the Hippo-YAP protein pathway in periodental ligament cells on the pressure side of a mouse model of orthodontic tooth movement
    Wu Yujie, Wan Xiaofang, Wei Mianxing, Peng Shiyuan, Xu Xiaomei
    2023, 27 (5):  683-689.  doi: 10.12307/2023.086
    Abstract ( 381 )   PDF (2711KB) ( 38 )   Save
    BACKGROUND: Autophagy is involved in the response of periodontal ligament cells to orthodontic forces, but how mechanical stimulation induces autophagy remains unclear. The Hippo-Yes-associated protein (YAP) pathway is sensitive to mechanical stimulation and is involved in the regulation of autophagy. 
    OBJECTIVE: To observe changes of the Hippo-YAP pathway and autophagy-related factors in the pressure side of orthodontic teeth and to explore whether the Hippo-YAP pathway is involved in the response of periodontal ligament cells to mechanical stimulation and its relationship with autophagy.
    METHODS: Fifty-four C57BL/6 mice were randomly divided into a blank group and eight experimental groups (orthodontic force applied for 30 minutes, 1 hour, 2 hours, 4 hours, 12 hours, 1 day, 3 days, 7 days). Mice in the experimental groups were modeled and killed at the set time after modeling to take their upper right first molars. Hematoxylin-eosin staining was used to observe the periodontal ligament tissue morphology on the pressure side. Tartrate-resistant acid phosphatase staining was used to count osteoclasts. Expressions of Beclin-1, LC3B, and active-YAP were detected by immunohistochemical staining.
    RESULTS AND CONCLUSION: Hematoxylin-eosin staining showed that with the application of orthodontic force, the periodontal ligament on the pressure side was first compressed and then slightly restored. Osteoclasts began to increase in number after 12 hours of orthodontic force applied (P < 0.01), and then the number of cells gradually increased and peaked at 7 days (P < 0.01). In the experimental groups, the expression of Beclin-1 and LC3B in periodontal ligament cells on the pressure side was correlated with the active-YAP expression during tooth movement and they showed similar trends. Correlation analysis showed that Beclin-1 and LC3B were both highly correlated with active-YAP (P < 0.01). These findings indicate that the Hippo-YAP pathway is involved in the response of periodontal ligament cells to mechanical stimulation on the pressure side, which may be an upstream regulation of autophagy. However, further experimental investigations are required.
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    Effects of enriched environment combined with melatonin on learning and memory function and brain neuron apoptosis in SAMP8 mice
    Zhao Siqi, Du Juan, Qu Haifeng, Li Jianmin, Zhang Yuxin, Liu Junjie
    2023, 27 (5):  701-706.  doi: 10.12307/2023.093
    Abstract ( 510 )   PDF (1507KB) ( 44 )   Save
    BACKGROUND: Neuronal apoptosis increases after brain aging and the mechanism is not clear. Recent studies have shown that melatonin can regulate the expression of a variety of apoptosis factors and enriched environments have been used as an effective treatment for Alzheimer’s disease. However, whether the combination of the two has better efficacy and their respective mechanisms of action remain unclear.
    OBJECTIVE: To investigate the effects of enriched environment combined with melatonin intervention on learning and memory function and brain neuron apoptosis in SAMP8 mice.
    METHODS: Twenty-four 3-month-old male SAMP8 mice were randomly divided into control group, enriched environment (EE) group, melatonin (MT) group, and enriched environment combined with melatonin (EE+MT) group, with 6 mice in each group. Mice in the control and MT groups were fed in a standard environment and those in the EE and EE+MT groups were fed in an enriched environment until they were 4 months old. At the age of 4 months, mice in the MT and EE+MT groups were subcutaneously injected with melatonin (8 mg/kg/d) for 30 days and those in the control and EE groups were given the same amount of normal saline for 30 days. After treatments, all the animals were kept in the original environment until 11 months of age, and their learning and memory abilities were evaluated by nest building test and Morris water maze test. After the completion of behavioral experiments, the apoptosis of hippocampal CAI cells was observed by immunofluorescence/TUNEL double standard staining and Nissl staining. The expression levels of apoptosis-related proteins Caspase-3, Bax, Bcl-2, and p53 were detected by western blot assay.
    RESULTS AND CONCLUSION: Compared with the control group, there was a significant increase in the score of the nest building test, cross-platform frequency, NeuN immunoreactivity in the hippocampal CA1 region, the number of Nissl-positive cells and the expression level of Bcl-2 protein in the hippocampus in the EE, MT, and EE+MT groups (P < 0.05). These indicators were significantly higher in the EE+MT group than the EE and MT groups (P < 0.05). Hematoxylin-eosin staining results indicated that compared with the control group, the EE, MT, and EE+MT groups showed better morphology and arrangement of nerve cells in the hippocampal CA1 region. Compared with the control group, Morris water maze escape latency, the number of immunofluorescence/TUNEL double positive cells in the hippocampal CA1 region and the expression levels of Caspase-3, Bax and p53 proteins in the hippocampus were significantly decreased in the EE, MT, and EE+MT groups (P < 0.05). These indicators in the EE+MT group were significantly lower than those in the EE and MT groups (P < 0.05). Overall, both enriched environment and melatonin can improve learning and memory function and brain neuron apoptosis in SAMP8 mice. The mechanisms may be related to the regulation of P53/P21 pathway in mouse hippocampal neurons to reduce apoptosis. Moreover, the combined application of enriched environment and melatonin is likely to be more effective than a single regimen treatment.
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    Changes in sensory neurons and astrocytes and the expression of interleukin 1beta and glial fibrillary acidic protein in the rat spinal cord after selective dorsal rhizotomy
    Chen Guodong, Zheng Meiyan, Zhang Peng, Wang Zhenchao, Jin Lixin
    2023, 27 (5):  726-731.  doi: 10.12307/2022.948
    Abstract ( 446 )   PDF (1216KB) ( 167 )   Save
    BACKGROUND: Excessive keratinization of astrocytes after spinal cord injury forms a mechanical barrier to repair the spinal cord and also plays an active role in spinal cord repair and regeneration. The regulation of keratinization to a suitable extent, to make it fully exert positive effects and decrease negative effects, is really a challenge to the research of spinal cord injury.
    OBJECTIVE: To observe the alteration of sensory neurons and astrocytes in the rat spinal dorsal horn, as well as the expression of interleukin-1β and glial fibrillary acidic protein in the abovementioned cells after a selective dorsal rhizotomy.
    METHODS: Thirty-three adult healthy Wistar rats were randomly divided into control group (n=3) and experimental group (n=30). On the operation side of the experimental group, the dorsal roots of the left lumbar 3 and 4 spinal nerves were exposed, separated, and cut off, while on the control side of the experimental group, the dorsal roots of the right lumbar 3 and 4 spinal nerves were only exposed and separated. The control group did not undergo any treatment. The rats were sacrificed under anesthesia at 1, 2, and 4 weeks after operation, and the segmental tissues of lumbar segments 3 and 4 were taken to make paraffin sections. The morphological changes of neurons and astrocytes in the rat spinal dorsal horn were observed by hematoxylin-eosin staining. The expression of interleukin-1β and glial fibrillary acidic protein in the spinal cord dorsal horn was observed by immunohistochemical staining.
    RESULTS AND CONCLUSION: (1) Hematoxylin-eosin staining: 1 and 2 weeks after selective dorsal rhizotomy, there were changes in the nuclei and cytoplasm of sensory neurons on the surgical side of the experimental group and at 4 weeks after operation, apoptosis in neurons appeared. Astrocytes were activated and the number of the cells increased accompanied by enlarged cell body and increased processes and reached a peak at 2 weeks after operation. The number of sensory nerve fibers on the surgical side of the experimental group was less than that on the control side of the experimental group and in the control group at 2 and 4 weeks after operation (P < 0.05), while the number of astrocytes was higher than that on the control side of the experimental group and in the control group at 1, 2, and 4 weeks after operation (P < 0.05). (2) Immunohistochemical staining: The positive expressions of interleukin-1β and glial fibrillary acidic protein on the surgical side of the experimental group were enhanced and peaked at 2 weeks after selective dorsal rhizotomy. The average absorbance values of glial fibrillary acidic protein and interleukin-1β positive cells on the surgical side of the experimental group were higher than those on the control side of the experimental group and in the control group (P < 0.05). (3) To conclude, selective dorsal rhizotomy can lead to a decrease in the number of sensory neurons in the dorsal horn of the spinal cord, reactive proliferation of astrocytes, and increased expression of interleukin-1β and glial fibrillary acidic protein.
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    Imiquimod combined with photodynamic therapy for hypertrophic scars: immune response and prognosis
    Lu Huixiu, Cao Haiyu, Lou Dan, Li Jianying, Liu Hongyuan, Sun Jing
    2023, 27 (5):  690-694.  doi: 10.12307/2023.095
    Abstract ( 574 )   PDF (830KB) ( 48 )   Save
    BACKGROUND: Hyperplasia scar is caused by dermal fibrosis due to fibroblast proliferation, uncontrolled growth and excessive collagen deposition. Both photodynamic therapy and imiquimod can inhibit fibroplasia. Imiquimod can further increase cell apoptosis and reduce the proliferation activity of fibroblasts by activating the immune response system.
    OBJECTIVE: To study the effect of imiquimod combined with photodynamic therapy on the immune response and prognosis of patients with hypertrophic scars.
    METHODS: A total of 110 patients with hypertrophic scars admitted to Shijiazhuang People’s Hospital from December 2017 to March 2020 were enrolled and randomly divided into a control group and a study group, with 55 patients in each group. Patients in the control group were given photodynamic therapy (every 10 days, four times in total) and those in the study group were treated with imiquimod cream (once a day for continuous 3 months) based on photodynamic therapy. The Vancouver Scar Scale was used to evaluate scar changes on day 2 after treatment. Clinical efficacy was evaluated and compared between the two groups of patients. The serum levels of interferon-γ, tumor necrosis factor-α, interleukin-2 and interleukin-6 were determined by enzyme-linked immunosorbent assay before and after treatment. The immunoturbidimetric method was used to determine the serum levels of IgA, IgG, and IgM in the two groups of patients. The occurrence of adverse reactions was recorded in the two groups.
    RESULTS AND CONCLUSION: (1) Before treatment, there was no significant difference in softness, thickness, blood vessel distribution, and color score between the two groups (P > 0.05). After treatment, the softness, thickness, blood vessel distribution and color scores of the two groups were significantly reduced, and the above-mentioned scores in the study group were significantly lower than those in the control group (P < 0.001). (2) The clinical response rate of the study group was 96%, which was significantly higher than that of the control group (78%; P < 0.05). (3) Before treatment, there were no significant differences in the serum levels of interferon-γ, tumor necrosis factor-α, interleukin-2 and interleukin-6 between the two groups (P > 0.05). After treatment, the serum levels of interferon-γ, tumor necrosis factor-α, interleukin-2 and interleukin-6 were significantly reduced in the two groups, and the levels of the above-mentioned indicators in the study group were significantly lower than those in the control group (P < 0.001). (4) Before treatment, there was no significant difference in serum IgA, IgG and IgM levels between the two groups (P > 0.05). After treatment, the serum levels of IgA, IgG and IgM increased significantly in the two groups, and the serum levels of IgA, IgG and IgM in the study group were significantly higher than those in the control group (P < 0.05). (5) The total adverse reaction rate in the control group was 13%, which was significantly higher than that of the study group (2%; P < 0.05). (6) To conclude, imiquimod combined with photodynamic therapy has a good effect in the treatment of hypertrophic scars, which can significantly reduce inflammatory responses, improve immune function, and have fewer adverse reactions in such patients.
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    High-intensity endurance exercise influences browning of white adipose tissue in a mouse model of high-fat diet induced obesity
    Li Yujiao, Su Kunxia
    2023, 27 (5):  707-713.  doi: 10.12307/2023.065
    Abstract ( 356 )   PDF (1033KB) ( 81 )   Save
    BACKGROUND: Studies have shown that uncoupling protein 1 (UCP-1), peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) and fibronectin type III domain containing protein 5 (FNDC5) are the key proteins that influence the browning of white adipocytes. Endurance exercise can effectively reduce the body mass of mice, but there is no conclusion on the impact of high-intensity endurance exercise on the expression of UCP-1, PGC-1α, and FNDC5 proteins in white adipose tissue of obese mice induced by high-fat diet.
    OBJECTIVE: To study the effects of high-intensity endurance exercise on the expression of UCP-1, PGC-1α and FNDC5 related to the browning of white adipocytes in mice with high-fat diet induced obesity.
    METHODS: Four-week-old male C57BL/6J mice were fed normal diet for 1 week and then fed 8 weeks of high-fat diet to establish a mouse model of nutritional obesity. The control mice were fed normal diet. After 1 week of adaptive treadmill exercise, obese mice were subjected to high-intensity endurance treadmill exercise, while the control mice did not have exercise. After 8 weeks, epididymal and groin white adipose tissue of mice was separated and the fat mass changes were compared between groups. A portion of adipose tissue was taken for hematoxylin-eosin staining, and the remaining adipose tissue was used to detect the expression of UCP-1, PGC-1α, and FNDC5 that were related to the browning of white adipocytes by western blot and the expression changes were compared between groups.
    RESULTS AND CONCLUSION: (1) High-fat diet could obviously increase the mass and cell volume of white adipose tissue in mice. (2) High-fat diet could remarkably reduce the expression of FNDC5 protein in white adipose tissue and slightly reduce the expression of UCP-1 and PGC-1α. However, there was no significant difference from the normal mice. (3) High-intensity endurance exercise could effectively reduce the mass and cell volume of white adipose tissue in mice with obesity induced by a high-fat diet. (4) High-intensity endurance exercise could considerably increase the expression of UCP-1, PGC-1α, and FNDC5 proteins in white adipose tissue, suggesting that high-intensity endurance exercise could remarkably promote the browning of white adipocytes in mice, thereby accelerating fat thermogenesis to achieve the purpose of reducing white adipose tissue.
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    Physiological function of the testis in rats undergoing different parts of vasectomy
    Yang Heng, Zheng Liying, Liu Zhili, Wang Qinzhang, Hao Zhiqiang, Wang Jingshen, Wang Yuan, Li Yongle, Tan Minghui, Zou Xiaofeng, Zhang Guoxi, Huang Ruohui, Jiang Bo, Qian Biao
    2023, 27 (5):  720-725.  doi: 10.12307/2023.060
    Abstract ( 464 )   PDF (1656KB) ( 113 )   Save
    BACKGROUND: Vasectomy is currently an effective method of male birth control, and the main concern is clinical complications after vasectomy.
    OBJECTIVE: To study the effects of different surgical methods of vasectomy on physiological function of the testis in rats.
    METHODS: After 1-week adaptive feeding, 12 male Sprague-Dawley rats were randomly selected and randomly divided into 4 groups with 3 rats in each group, including negative control group, sham-operation group, operation group and traditional group. The rats in the operation group were subjected to bilateral oblique vasectomy, the proximal end of the vas deferens was ligated and the distal end was separated but not ligated. The rats in the traditional group underwent bilateral oblique vasectomy, with both ends of the vas deferens ligated. In the sham-operation group, the both ends of the vas deferens were separated without ligation. The negative control group had no special treatment. Three months after the operation, methylene blue was injected and observed. After anesthesia, the rats were sacrificed and their testicles were collected for routine pathological examination using hematoxylin-eosin staining.  Testicular tissue was collected and the expressions of Bax, Bcl-2, 3βHSD and human epididymal protein 4 were detected by immunohistochemistry. Apoptosis in testicular tissue was detected by TUNEL assay. Serum interleukin-1 and testosterone levels were detected using ELISA/biochemical detection. 
    RESULTS AND CONCLUSION: Hematoxylin-eosin staining: compared with the sham operation group, the testis was swollen and hard in texture in the traditional and operation groups; the incidence of testicular aseptic inflammation in the traditional group was higher than that in the negative control group; no spermatogenic granuloma was found in the testis of the operation group and the histological structure of the testis in the traditional group was remarkably changed. TUNEL detection: Compared with the negative control and sham operation groups, apoptosis was increased in the operation and traditional groups; and there was less apoptosis in the operation group than the traditional group. Immunohistochemical detection: Compared with the negative control group, the expression of Bax changed insignificantly in the sham operation group and increased in the operation and traditional groups, especially in the traditional group. Compared with the negative control group, the expression of Bcl-2 was decreased in the operation and traditional groups, especially in the traditional group. Compared with the negative control and sham operation group, the protein expression of 3βHSD and epididymal protein 4 in the rat testis was lower in the operation and traditional groups and there was no significant difference between the operation and traditional groups. ELISA/biochemical detection: Compared with the negative control group, the levels of serum interleukin-1 and testosterone were significantly increased in the operation group (P < 0.05) but significantly decreased in the traditional group (P < 0.05). These findings indicate that pelvic vasectomy has little effect on the physiological function of the testis in rats.
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    Differential expression profiles of microRNAs in muscle tissue of denervated skeletal muscle atrophy rats
    Liu Hongwen, Li Jiao, Xu Wenhao, Nie Hua, Liu Shaojiang, Xu Jie, Yin Li
    2023, 27 (5):  732-737.  doi: 10.12307/2023.123
    Abstract ( 390 )   PDF (1922KB) ( 184 )   Save
    BACKGROUND: There is no effective treatment for denervated skeletal muscle atrophy with poor prognosis, and moreover, the expression profiles of microRNA in vivo are unclear. 
    OBJECTIVE: To analyze the microRNA expression profiles and the changes of transforming growth factor-β1/Smad3 signaling pathway in denervated skeletal muscle atrophy rats using high-throughput sequencing technology and to explore the role and mechanism of microRNA in skeletal muscle atrophy. 
    METHODS: Twelve Sprague-Dawley rats were randomized into two groups: experimental group and control group. Animal models of denervated muscle atrophy were established using truncation of the sciatic nerve in the experimental group. In the control group, only the sciatic nerve was exposed followed by suturing. The bilateral gastrocnemius muscles were completely removed to calculate the muscle wet weight ratio. Atrophy and fibrosis were observed by hematoxylin-eosin staining and the cross-sectional area of the muscle was determined. Illumina HiSeq 2500 sequencing technology was used to identify the microRNA expression profiles in skeletal muscle tissue. In combination with volcano plot, hierarchical clustering map, Gene Ontology, Kyoto Encyclopedia of Genes and Genomes pathway, differentially expressed genes involved in skeletal muscle metabolism were analyzed. Real-time fluorescent quantitative PCR was used to verify the differential expression of candidate genes in muscle tissue and western blot was used to detect the expression of transforming growth factor-β1 and p-Smad3 proteins. 
    RESULTS AND CONCLUSION: Compared with the control group, the muscle wet weight ratio and cross-sectional area were significantly lower in the experimental group (P < 0.05), and the modeling results were in line with expectations. A total of 1 249 differentially expressed microRNAs were identified in the two groups (P < 0.05; | log2 Fold Change | > 0.0). There were 14 microRNAs with significant differential expression, of which 2 were up-regulated and 12 were down-regulated. The results of biological function and pathway analysis indicated that miR-1247-3p, miR-132-5p, miR-21-3p, miR-363-3p, and 
    miR-451-5p with significant differential expression were significantly enriched in cell proliferation, differentiation, apoptosis and other biological processes as well as transforming growth factor-β signaling pathway. The results of real-time fluorescent quantitative PCR showed that the expression of miR-21-3p in the gastrocnemius muscle tissue of the experimental group was significantly lower (P < 0.001), which was consistent with the trend of the high-throughput sequencing results. Western blot results showed that the protein expression levels of transforming growth factor-β1 and p-Smad3 were significantly higher in the experimental group than the control group (P < 0.05). To conclude, microRNA plays a key role in the physiological and pathological processes of skeletal muscle atrophy. miR-21-3p may play a role in skeletal muscle cell metabolism by activating the biological activity of transforming growth factor-β1/Smad3 signaling pathway.
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    Mechanism of long non-coding RNA plasmacytoma variant translocation 1 involved in pelvic organ prolapse
    Hu Xinming, Qiao Yanhua, Wang Xiaofan, Li Linyu, Zhao Bing
    2023, 27 (5):  669-675.  doi: 10.12307/2023.092
    Abstract ( 409 )   PDF (1916KB) ( 61 )   Save
    BACKGROUND: Previous studies have shown that the expression of long non-coding RNA plasmacytoma variant translocation 1 (LncRNA PVT1) was significantly reduced in the uterine cardinal ligament of patients with pelvic organ prolapse through high-throughput sequencing. Transforming growth factor β1 signaling pathway is also one of the related pathways.
    OBJECTIVE: To investigate the effect of LncRNA PVT1 on pelvic floor ligament fibroblasts and its possible role in the pathogenesis of pelvic organ prolapse. 
    METHODS: Tissue samples of the pelvic floor ligament were extracted from rats and fibroblasts were isolated for primary culture. LncRNA PVT1 interference and overexpression vectors were constructed by lentivirus and added into fibroblasts. Morphology, proliferation, migration and apoptosis of fibroblasts were detected by immunofluorescence, MTT, cell scratch assay and flow cytometry test respectively. The mRNA and protein expressions of transforming growth factor-β1, Smad2/3 and c-Myc were detected by RT-PCR and western blot respectively. 
    RESULTS AND CONCLUSION: Overexpression of LncRNA PVT1 could significantly promote the proliferation of fibroblasts, improve the migration ability of cells, and inhibit the apoptosis of fibroblasts. The results of LncRNA PVT1 interference were opposite. In addition, the mRNA and protein relative expressions of c-Myc, transforming growth factor-β1 and Smad2/3 were significantly increased when LncRNA PVT1 gene was overexpressed, while the expressions of c-Myc and transforming growth factor-β1 were significantly inhibited when LncRNA PVT1 gene was interfered. To conclude, LncRNA PVT1 can affect the morphology and biological function of fibroblasts, which may be related to the transforming growth factor-β1/Smad2/3/c-Myc pathway. This study provides new ideas for the clinical diagnosis and treatment of pelvic organ prolapse.
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    Ephrin A receptor 2 DNA methylation increases in pancreatic beta cell apoptosis induced by homocysteine
    Zhang Qing, Gao Chunlan, Yu Feifei, Zhang Zhenghao, Ma Fang, Gao Yuan, Li Guizhong, Jiang Yideng, Ma Shengchao
    2023, 27 (5):  714-719.  doi: 10.12307/2023.062
    Abstract ( 458 )   PDF (1356KB) ( 59 )   Save
    BACKGROUND: Increased homocysteine levels lead to apoptosis of pancreatic β cells, but the exact mechanism remains unclear.
    OBJECTIVE: To explore the specific mechanism of DNA hypermethylation of Ephrin A receptor 2 (EphA2) and its promoter region in pancreatic β cells.
    METHODS: Mouse insulinoma cell lines (Min6) were cultured in vitro and divided into control group (0 µmol/L homocysteine) and homocysteine group (120 µmol/L homocysteine). After 48 hours of intervention in the cells, immunofluorescence and western blot were used to test the expression of apoptosis-related proteins Bax, Bcl-2, and Caspase-3 in pancreatic islet β cells of the two groups. The expression levels of DNA methylation-related proteins DNMT1 and DNMT3a were detected by western blot. Real-time fluorescent quantitative PCR (qRT- PCR) was used to detect the level of EphA2 mRNA. Western blot was used to detect the protein expression of EphA2. Nested methylation-specific PCR was used to detect the level of DNA methylation in the promoter region of EphA2.
    RESULTS AND CONCLUSION: Compared with the control group, the expression of apoptosis-related proteins Bax and Caspase-3 in the pancreatic β cells was significantly increased in the homocysteine group, and the expression of Bcl-2 was significantly decreased; the mRNA and protein expression levels of EphA2 were significantly decreased (P < 0.05). Compared with the control group, the EphA2 DNA methylation level and the expression of DNMT1 protein in the pancreatic β cells were significantly higher in the homocysteine group (P < 0.05). To conclude, EphA2 DNA hypermethylation plays a significant role in homocysteine-induced pancreatic β cell apoptosis and DNMT1 may be involved in its hypermethylation process.
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    Risk factor analysis of metabolic syndrome and construction of a nomogram prediction model in middle-aged and elderly people
    Shen Lianwei, Zhu Hongliu, Wang Wei
    2023, 27 (5):  657-662.  doi: 10.12307/2022.1005
    Abstract ( 608 )   PDF (948KB) ( 70 )   Save
    BACKGROUND: With population aging and life-style variation in China, the prevalence of metabolic syndrome in the middle-aged and elderly is increasing. Therefore, it is necessary to screen the risk factors of the disease and construct a prediction model of the disease.
    OBJECTIVE: To investigate the risk factors of metabolic syndrome in middle-aged and elderly people in China and to construct the nomogram prediction model of metabolic syndrome, so as to provide reference for the prevention and control of metabolic syndrome in such populations. 
    METHODS: The data were from the 2015 follow-up data of the China Health and Retirement Longitudinal Study (CHARLS), with metabolic syndrome as a dependent variable. Nine variables including left hand muscle strength, right hand muscle strength, standing time, walking speed, time to get up from the chair, sex, age, smoking and drinking were involved to explore the related factors of metabolic syndrome in middle-aged and elderly people. Propensity score matching and descriptive analysis were performed on SPSS. The original data set was divided into training set and verification set with Rstudio. The training set was subject to univariate and multivariate regression analysis, nomogram model construction and internal verification, whereas the verification set was subject to external verification. 
    RESULTS AND CONCLUSION: (1) A total of 7 384 middle-aged and elderly people were screened, including 518 (7%) with metabolic syndrome and 6 865 (93%) without metabolic syndrome. (2) 6 865 subjects without metabolic syndrome was paired by 1:1 with propensity score matching to screen out 518 subjects as the control group and those with metabolic syndrome acted as the case group. The control group and the case group constituted the original data set. The original data set was randomly divided into the training set (728 subjects) and the validation set (308 subjects) at 7:3. (3) Construction of the model: according to the binary logistic regression analysis results of the training set (modeling cohort), five variables such as right hand muscle strength, walking speed, sex, drinking and standing time were selected to build the model. Evaluation of the model: the area under the receiver operator characteristic curve of the model was 0.877 (95% confidence interval: 0.851-0.902), indicating that the model has high discrimination and good fitting of the calibration curve, indicating that the model has a high calibration degree. (4) Internal verification of the model: the bootstrap method was used to generate new samples after 1000 repeated sampling with return, and the generated calibration curve fitted well. (5) External verification of the model: the model was tested by the verification set (verification queue), the area under the receiver operator characteristic curve was 0.89 (95% confidence interval: 0.854-0.926), indicating that the model has high discrimination and good fitting of the calibration curve, that is, the model has high calibration degree and good efficiency in the verification queue. (6) The prediction model of metabolic syndrome in the middle-aged and elderly constructed in this study has good reliability. The nomogram obtained from this can predict the probability of metabolic syndrome in the middle-aged and elderly according to the right hand muscle strength, walking speed, sex, drinking, age and standing time, which is conducive to the early detection, diagnosis and treatment of metabolic syndrome in the middle-aged and elderly and can be popularized in clinical practice. 
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    Screening key technical indicators for speed climbing based on machine learning
    You Guopeng, Wang Jianqing, Liu Fei, Yuan Qiang, Liu Haoyan, Wu Ying
    2023, 27 (5):  738-744.  doi: 10.12307/2023.096
    Abstract ( 440 )   PDF (1483KB) ( 117 )   Save
    BACKGROUND: Selecting key technical indicators to guide special training or competition is the key to improving competition performance. There are obvious differences in the techniques of male and female athletes and the key technical indicators of athletes of different sexes have not yet been known.
    OBJECTIVE: To select key technical indicators that affect the competition performance of elite speed climbing athletes, thereby providing direction and theoretical support for the scientific training of speed climbing.
    METHODS: The videos of speed climbing finals in the 2017-2021 World Championships, World Cup and China Climbing League were selected to obtain the competition samples of the top four male and female athletes (109 men and 117 women). Reaction time, contact time and segmental speed were collected and then random forest and XGBoost models of male and female speed climbing athletes were established using two-dimensional kinematic analysis. Accordingly, the influence of each technical indicator on the competition performance was calculated.
    RESULTS AND CONCLUSION: For male athletes, the root-mean-square error values of random forest and XGBoost models were 0.224 and 0.265, respectively, and r2 was 0.765 and 0.686, respectively. The key technical indicators were total contact time of the left hand, motion frequency of the right hand, total contact time of the right hand, speed of holds 3-6, motion frequency of the left foot, total contact time of the right foot, speed of holds 11-13, speed of holds 13-18, speed of holds 8-10, total contact time of the left foot, number of total holds of the left hand and motion frequency of the right foot. For female athletes, the root-mean-square error indicator of random forest and XGBoost models were 0.066 and 0.055, respectively, and r2 were 0.846 and 0.887, respectively. The key technical indicators were total contact time of the left hand, total contact time of the right hand, motion frequency of the right hand, motion frequency of the left foot, total contact time of the left foot, speed of holds 8-10, total contact time of the right foot, and speed of holds 10-11 and 13-18. To conclude, the technical indicators of the upper limbs (contact time sum and action frequency) on speed rock climbing competition are more influential than those of the lower limbs. The number of holds has no high influence on the competition results. The key segments for winning in male and female elite athletes are holds 3-6 and 8-11 respectively. Athletes should focus on optimizing the upper limb contact time of the key segments for winning and then optimize the lower limb contact time, so as to improve the competition performance.
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    The relationship between glucose transporter 1/4 gene polymorphisms and athletic ability
    Zhou Yinan, Yang Kun, Jiang Jiayi, Zhang Sizhuo, Zhu Yingwen
    2023, 27 (5):  745-750.  doi: 10.12307/2023.113
    Abstract ( 582 )   PDF (1067KB) ( 93 )   Save
    BACKGROUND: Scientific selection of athletes through genetic factors has become an important research hotspot in the field of competitive sports. Glucose transporter (GLUT) is an important carrier of glucose transmembrane transport. Recent studies have shown that GLUT1/4, which are the members of the GLUT family, have certain correlations between their gene polymorphisms and athletic abilities such as aerobic endurance and exercise sensitivity.
    OBJECTIVE: To review the association between GLUT1/4 gene polymorphisms and athletic ability and its mechanism and to provide evidence for the scientific selection of athletes.
    METHODS: The first author conducted a computer search in CNKI, WanFang, PubMed and Embase databases in January 2022, using “glucose transporter, gene polymorphism, exercise capacity, aerobic endurance, strength, athlete” as the Chinese search terms and “glucose transport protein, GLUT, GLUT1, GLUT4, SLC2A4, SLC2A1, single nucleotide polymorphism, SNP, athletic, sport performance, athlete, aerobic, endurance, power” as the English search terms. According to the inclusion criteria, 58 articles were finally included.
    RESULTS AND CONCLUSION: The main involved mutation sites include rs841853 in the GLUT1 gene and rs5418 in the GLUT4 gene. The research on the correlation between the two and athletic ability focuses on two aspects: aerobic endurance quality and exercise sensitivity, and both have significant correlations. In terms of the association mechanism of aerobic endurance quality, GLUT1 rs841853 single nucleotide polymorphism may affect endurance quality by affecting glucose uptake and lung ventilation capacity; and GLUT4 rs5418 single nucleotide polymorphism may regulate DNA methylation and transcription factors in the promoter region to yield excellent endurance quality. The polymorphism of this locus is thereby associated with athletic ability. In terms of exercise sensitivity, hypoxia-inducible factor-1α may play a central role in triggering the association between rs841853 polymorphism and hypoxic training sensitivity. Whereas, rs5418 polymorphism may lead to differences in insulin resistance in patients with impaired glucose regulation with different alleles undergoing aerobic training through the IRS-1/PI3K/AKT pathway. Therefore, selection of athletes based on GLUT1 and 4 gene polymorphisms may be a feasible method.
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    Circular RNA as a competing endogenous RNA is involved in the regulation of osteoarthritis
    Li Long, Li Guangdi, Shi Hao, Deng Keqi
    2023, 27 (5):  751-757.  doi: 10.12307/2023.068
    Abstract ( 402 )   PDF (923KB) ( 36 )   Save
    BACKGROUND: Osteoarthritis is one of the most common joint diseases and the leading cause of chronic joint pain and limited mobility in older adults. As an emerging important member of the competing endogenous RNA regulatory network, circular RNA plays an important role in the occurrence and development of osteoarthritis.
    OBJECTIVE: To overview circular RNAs, their biological functions, and competing endogenous RNAs and to emphatically review the research progress in the role of circular RNAs as competing endogenous RNAs in osteoarthritis.
    METHODS: CNKI, WanFang, VIP Chinese journal database, and PubMed databases were searched for relevant articles using “osteoarthritis, circular RNA, competing endogenous RNA” as Chinese and English search terms. We excluded outdated and repeated views, sorted out the retrieved literature, and selected 64 articles for review.
    RESULTS AND CONCLUSION: The mechanism of circular RNAs by which affect the occurrence of osteoarthritis is mainly to bind microRNAs as competing endogenous RNAs, thereby regulating the expression level of their target genes and regulating the related proliferation, apoptosis and differentiation of the cells to which they belong. Circular RNA as a competing endogenous RNA participates in regulating the proliferation, apoptosis, and autophagy of chondrocytes. Circular RNA as a competing endogenous RNA participates in regulating the formation and degradation of the extracellular matrix of chondrocytes. Circular RNA as a competing endogenous RNA is involved in regulating the production of inflammatory factors in chondrocytes. Circular RNA as a competing endogenous RNA is involved in regulating synovial cell apoptosis and autoimmune responses. Since circular RNAs are highly stable and conserved, the research on the competing endogenous RNA regulatory network mediated by circular RNAs will also open up new avenues for basic research on osteoarthritis, which help understand the pathogenesis of osteoarthritis more comprehensively. Circular RNA is also expected to be a high-quality early diagnostic marker and therapeutic target for osteoarthritis. To date, the competing endogenous RNA regulatory network is still in the verification stage. With the rapid development of bioinformatics, the competing endogenous RNA regulatory network needs to be further improved and supplemented.
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    Regulatory role of non-coding RNAs as potential therapeutic targets in spinal cord injury
    Li Zhichao, Tan Guoqing, Su Hui, Xu Zhanwang, Xue Haipeng
    2023, 27 (5):  758-764.  doi: 10.12307/2023.114
    Abstract ( 373 )   PDF (1229KB) ( 40 )   Save
    BACKGROUND: Although many studies have been carried out on the complex physiological and pathological mechanisms of spinal cord injury, there is still a lack of satisfactory treatment methods. Studies have shown that non-coding RNAs associated with secondary injuries, such as oxidative stress, inflammation, autophagy and apoptosis, are differentially expressed after spinal cord injury. Therefore, non-coding RNAs have potential as therapeutic targets after spinal cord injury.
    OBJECTIVE: Based on the latest research progress, to review the functional roles and regulatory mechanisms of non-coding RNAs in spinal cord injury, in order to provide new ideas for clinical treatment of spinal cord injury.
    METHODS: Using “spinal cord injury” AND “non-coding RNA” OR “microRNA” OR “long non-coding RNA” OR “circRNA” AND “oxidative stress” OR “inflammation” OR “autophagy” OR “apoptosis” as English and Chinese keywords, PubMed, Web of Science, CNKI and WanFang databases were searched. According to inclusion and exclusion criteria, 75 articles were reviewed.
    RESULTS AND CONCLUSION: After spinal cord injury, non-coding RNAs associated with secondary injury such as oxidative stress, inflammation, autophagy and apoptosis were significantly differentially expressed, resulting in differential expression of target genes and changes in cell functions. Regulating non-coding RNAs to antagonize oxidative stress, intervening microglia activation, polarization and inflammatory pathways to reduce inflammation, maintaining moderate levels of autophagy and keeping autophagy flux unobstructed, and inhibiting apoptosis may be the breakthrough points to promote the repair of spinal cord injury.
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    MicroRNA, long non-coding RNA and circular RNA mediate the mechanism of decreasing uric acid, anti-inflammation and regulating bone metabolism in gout
    Shao Zichen, Li Huanan, Gu Bing, Zhang Xiaoyun, Sun Weikang, Liu Yongqian, Gan Bin
    2023, 27 (5):  765-771.  doi: 10.12307/2023.142
    Abstract ( 485 )   PDF (1147KB) ( 42 )   Save
    BACKGROUND: In the past decade, significant progress has been made in the understanding of the pathogenesis, effects, diagnostic methods and treatments of gout, but the pathogenesis of gout has not yet been clarified. A large number of experimental studies have shown that microRNAs (miRNAs), long non-coding RNAs (lncRNAs) and circular RNAs (circRNAs) are closely related to the occurrence and development of gout. 
    OBJECTIVE: To review the mechanism of miRNA, lncRNA and circRNA in different stages of gout development. 
    METHODS: The Chinese search terms were “Gout; Hyperuricemia; Gouty Arthritis; lncRNA; miRNA; circRNA; RNA” and the English search terms were “Gout; Arthritis, Gouty; Gouty Arthritis; Arthritides, Gouty; Gouty Arthritides; Hyperuricemia; RNA; Gene Products, RNA; RNA Gene Products; RNA, Non Polyadenylated; Non Polyadenylated RNA; Ribonucleic Acid.” PubMed, Embase, CNKI, WanFang, VIP and other databases were retrieved respectively. The search time range was 2013 to 2022. By reading the titles and abstracts of the literature, the low-quality, repetitive and irrelevant articles were excluded and 53 highly relevant articles were included in the review.
    RESULTS AND CONCLUSION: MiRNA, lncRNA, and circRNA are expected to become new targets and strategies for reducing uric acid levels, repairing immune balance, reducing inflammation and regulating osteoclast activation, which will become biomarkers for gout treatment. In the follow up study, we should strengthen the research on the mechanism of epigenetics in human body and also pay attention to the co-regulation of many kinds of non-coding RNAs.
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    Construction and application of myocardial ischemia model in miniature pigs
    Song Jian, Zhao Lei, Liu Aishi
    2023, 27 (5):  772-778.  doi: 10.12307/2023.098
    Abstract ( 506 )   PDF (1017KB) ( 50 )   Save
    BACKGROUND: With the increasing prevalence and mortality of ischemic heart disease, it is very important to study the pathophysiological mechanism, diagnosis and treatment of myocardial ischemia. By constructing animal models, researchers can deeply understand the pathophysiological mechanisms, therapeutic effects and drug safety of myocardial ischemia.
    OBJECTIVE: To summarize myocardial ischemia model methods so as to reveal the present situation and progress of myocardial ischemia animal models and to compare the advantages and disadvantages of various establishing methods, thereby providing reference for relevant clinical studies.
    METHODS: The databases of CNKI, WanFang Data Knowledge Service Platform, PubMed and GeenMedical were searched by computer from January 2000 to December 2021 using the keywords of “myocardial ischemia; animal models; miniature pigs.” Finally, 52 articles were included for review analysis according to inclusion and exclusion criteria.
    RESULTS AND CONCLUSION: At present, the clinical models of myocardial ischemia mainly include large animal species (dogs and pigs), rabbits, and rodents (rats and mice). Miniature pigs are the most ideal animal model donors for ischemic heart disease and have been widely used to prepare myocardial ischemia models. Among various methods for establishing acute myocardial ischemia models, open heart surgery and microcurrent stimulation have the characteristics of accurate positioning. Compared with the open thoracic surgery, closed thoracic surgery, drugs and microcurrent stimulation are simpler and more suitable for the clinical pathological process of acute myocardial ischemia. Moreover, the closed thoracic surgery that is minimally invasive and of high repeatability has become the preferred method. The main methods to construct the model of chronic myocardial ischemia include intima proliferation method, external chronic contraction method, microcirculation embolization method, high-fat diet method and subtotal ligation method. Microcirculation embolization can be used to explore the pathological mechanism of chronic myocardial ischemia caused by microcirculation disorders by blocking microvessels. In principle, both chronic extravascular contraction and subtotal ligation lead to chronic myocardial ischemia through extravascular mechanical compression, but subtotal ligation has a more controllable stenosis range and less damage to the natural structure of the vascular wall than chronic contraction. Intimal proliferation combined with high-fat diet is the most commonly used method to construct chronic myocardial ischemia model, as this combined method is closer to the clinical pathological mechanism of chronic coronary artery stenosis caused by atherosclerotic plaque. 
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    Research progress in promoting tendon to bone healing during anterior cruciate ligament reconstruction
    Xiong Bohan, Yu Yang, Lu Xiaojun, Wang Xu, Yang Tengyun, Zhang Yaozhang, Liao Xinyu, Zhou Xiaoxiang, He Lu, Li Yanlin
    2023, 27 (5):  779-786.  doi: 10.12307/2023.130
    Abstract ( 576 )   PDF (982KB) ( 46 )   Save
    BACKGROUND: Arthroscopic anterior cruciate ligament reconstruction is an important treatment for anterior cruciate ligament injury and fracture. Postoperative tendon to bone healing is closely related to the early postoperative rehabilitation and functional recovery of patients. In particular, the time and firmness of tendon to bone healing have become important indicators to evaluate the efficacy of anterior cruciate ligament reconstruction.
    OBJECTIVE: To summarize the research progress in promoting tendon to bone healing after anterior cruciate ligament reconstruction in recent years, hoping to inspire surgeons and related researchers and better serve basic research and clinical work.
    METHODS: Using “anterior cruciate ligament, tendon to bone Healing, knee joint, anterior cruciate ligament reconstruction” as English and Chinese search terms, related articles regarding tendon to bone healing were searched by computer in CNKI, WanFang Database, PubMed, and FMRS. The time period of literature retrieval was from January 2011 to December 2021, and systematic generalization, summary and analysis were conducted. The shortcomings and future prospects of the research are also discussed.
    RESULTS AND CONCLUSION: With the development of medical technology and research level, tendon to bone healing after anterior cruciate ligament reconstruction can be promoted from various aspects such as surgical method improvement, tissue engineering technology and postoperative rehabilitation optimization. However, there are also some limitations, such as a lot of research achievements to promote tendon to bone healing only stay on the animal experiment, especially the experiments of small animals (rat and rabbit). These animals are small in size and extremely different from humans in terms of biological mechanics and body mass; therefore, further research is required to explore whether these study results have the same effect in humans and whether there are long-term adverse reactions or not. There are still few studies on whether the combination of various methods to promote tendon to bone healing can produce a synergistic effect or an inhibitory effect, which may provide new ideas for further research on tendon to bone healing.
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    Application potential of naringin in bone tissue regeneration
    Zhang Min, Zhang Xiaoming, Liu Tongbin
    2023, 27 (5):  787-792.  doi: 10.12307/2023.056
    Abstract ( 506 )   PDF (786KB) ( 72 )   Save
    BACKGROUND: Bone tissue regeneration is currently an important research direction in the treatment of large-area bone defects. Naringin, as a natural flavonoid, has the potential to induce osteogenesis, which has attracted extensive attention in the field of bone tissue regeneration.
    OBJECTIVE: To review the main related effects and safety of naringin in promoting bone regeneration and its application in bone tissue engineering, thereby providing a theoretical basis for the clinical treatment of large-area bone defects.
    METHODS: The first author searched for articles regarding naringin-induced osteogenesis included in PubMed, WanFang, and CNKI databases from 2002 to 2021. The keywords were “naringin; bone defects; osteoinductive; osteoporosis; bone regeneration; osteoclasts; blood vessel; bone tissue engineering” in English and Chinese, respectively. Fifty-five eligible articles were finally reviewed.
    RESULTS AND CONCLUSION: Naringin has a good potential of inducing bone tissue production in anti-osteoporosis, promoting bone formation, inhibiting osteoclast formation and promoting angiogenesis by exerting estrogen-like effects or participating in the regulation of different signaling pathways such as bone morphogenetic protein, Wnt/β-catenin, PI3K/Akt and VEGF/VEGFR-2. In addition, naringin has the characteristics of safety, stability, easy extraction, and low cost. Therefore, naringin is a very promising natural bone induction small-molecule drug. In recent years, naringin combined with bone tissue engineering technology has improved the initial sudden release and bioavailability of drugs. However, in the future, we still need more experimental studies on the mechanism of induction osteogenesis in a more comprehensive and systematic way, and make full use of good bone tissue engineering technology to further verify the most suitable controlled-release carrier and local application effect of naringin. Thereby, naringin will be mature, stable and effective in the clinical treatment of bone defects as soon as possible.
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    Effect of hypoxic training on the oxygen sensing pathway
    Liu Yuan
    2023, 27 (5):  793-798.  doi: 10.12307/2023.111
    Abstract ( 468 )   PDF (824KB) ( 37 )   Save
    BACKGROUND: Hypoxic training is a training method that utilizes the dual stimulation of hypoxia and exercise to mobilize the potential of athletes and improve athletic performance. Oxygen sensing is the ability of cells to sense oxygen and adapt to oxygen, which is regulated by many key factors in cells, such as hypoxia-inducible factors, which in turn form an oxygen sensing pathway. This pathway plays an important role in hypoxic training.
    OBJECTIVE: From the perspective of how the body cells perceive oxygen and adapt to hypoxia during hypoxic training, to explore the physiological mechanism by which hypoxic training promotes exercise health and improves exercise performance.
    METHODS: A literature search of CNKI and PubMed was conducted using the keywords of “oxygen-sensing, hypoxic training, hypoxia inducible factor” in Chinese and English, respectively. After title, abstract and main text reading, 50 articles were finally included for review.
    RESULTS AND CONCLUSION: Hypoxia-inducible factors are important regulators in the oxygen sensing pathway. Hypoxic training can significantly upregulate the expression of hypoxia-inducible factor-1, enhance glucose metabolism, oxygen transport and angiogenesis in tissues, providing theoretical support for hypoxic training to prevent and alleviate hypoxia-induced cardiovascular diseases through the oxygen sensing pathway. The hypoxia-inducible factor signaling pathway may play its value as a new therapeutic target to improve human tissue function in the future and the elucidation of the mechanism by which hypoxia-inducible factors act on cellular sensing and response to oxygen levels opens up new fields for biology and medicine.
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    Effect of short-foot training on foot and ankle function in patients with flat feet: Meta-analysis and systematic review
    Feng Liang, Gong Shuhui, Huo Hongfeng
    2023, 27 (5):  799-804.  doi: 10.12307/2023.115
    Abstract ( 897 )   PDF (1834KB) ( 203 )   Save
    OBJECTIVE: To perform a Meta-analysis to comprehensively and quantitatively evaluate the effects of short foot training on foot and ankle function in patients with flat feet, and to provide a theoretical basis for more effective training in patients with flat feet.
    METHODS: Chinese and foreign literature databases such as Web of Science, The Cochrane Library, PubMed, Embase, CNKI database, VIP database, and China Biomedical Literature Database were searched. The search time was from database inception to January 31, 2022. Chinese database search terms and search formula: (flatfoot OR valgus foot OR stiff foot) AND (short foot training OR physical therapy). English database search terms and search formula: (flatfoot OR talipes valgus OR talipes calcaneovalgus) AND (short foot exercises OR physical therapy OR neurophysiotherapy). The Cochrane risk of bias assessment tool was selected to evaluate the quality of the included literature, and all included articles were analyzed for outcome indicators (navicular drop and foot posture index) using Revman 5.4 and Stata 12.0 software.
    RESULTS: A total of 12 papers with 443 subjects were included. Five were moderate risk of bias and seven were low risk of bias literature. Short-foot training could reduce navicular drop [standardized mean difference (SMD)=-0.59, P < 0.05). Subject age (regression coefficient: -1.563 9, P=0.004), body mass index (regression coefficient: -1.563 9, P=0.023) and intervention duration (regression coefficient: -1.445 6, P=0.042) influenced the overall effect of the included literature. Short foot training showed insignificant effects on foot posture index (SMD=-0.26, P > 0.05).
    CONCLUSION: Short-foot training can effectively reduce navicular drop in patients with flat foot but has no clear effects on foot posture index. Age, body mass index, and intervention time are the factors that influence the effectiveness of short-foot training interventions and are the factors that need to be considered when conducting relevant studies and organizing rehabilitation training for patients with flat feet. Short-foot training is recommended as a training tool for patients with flat feet. 
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    Effects of blood flow restriction training on muscle strength, muscle mass and physical performance in older adults: a Meta-analysis
    Pan Weimin, Wang Bing, Han Yabing, Li Ting, Song Jiaqi, Qin Huasheng, Liu Yang
    2023, 27 (5):  805-812.  doi: 10.12307/2023.097
    Abstract ( 631 )   PDF (1082KB) ( 121 )   Save
    OBJECTIVE: To systematically evaluate the effects of low-load blood flow restriction training and walking training with blood flow restriction on muscle strength, muscle mass and physical performance in older adults, thereby providing a theoretical basis for the scientific application of blood flow restriction to the prevention and treatment of senile sarcopenia. 
    METHODS: A computer-based search was conducted in PubMed, Embase, Cochrane Library, CNKI, WanFang, VIP, and CBM. Randomized controlled trials regarding the effects of blood flow restriction training on muscle mass, muscle strength and physical performance in older adults were included from inception to August, 2021. The literature quality was assessed using the bias assessment tool recommended by the Cochrane Collaboration by at least two evaluators and RevMan5.4 software was used to conduct meta-analysis of relevant data. Evaluation indicators contained three continuous variables including muscle strength, muscle mass and physical performance. Subgroup analysis was carried out for different test methods of outcome indicators included in the literature 
    RESULTS: A total of 16 randomized controlled trials, including 348 patients in total, were included according to the inclusion criteria. The quality of the literature was graded according to the Cochrane Collaboration’s risk of bias assessment tool, including 11 studies of grade B and 5 of grade C. The results of Meta-analysis showed that compared with high-load resistance training, low-load blood flow restriction training could improve muscle strength [standardized mean difference (SMD)=-0.14, 95% confidence interval (CI):-0.34-0.06, P=0.17] and muscle mass (SMD=0.08, 95% CI: -0.23-0.40, P=0.60) in older adults; compared with low-load resistance training, low-load blood flow restriction training and walking training with blood flow restriction could significantly improve muscle strength (SMD=0.57, 95% CI: 0.28-0.87, P=0.000 2); compared with walking training, walking blood flow restriction training significantly could improve muscle strength (SMD=0.62, 95% CI: 0.32-0.91, P < 0.000 1], but there was no significant change in muscle mass (SMD=0.28, 95% CI: -0.01-0.57, P=0.06) and physical performance (SMD=0.61, 95% CI: -0.04-1.26, P=0.07). The results of subgroup analysis showed that heterogeneity was reduced. The muscle strength via low-load blood flow restriction training and walking training with blood flow restriction was significantly better than that via low-load training by means of maximum voluntary contraction (SMD=0.95, 95% CI: 0.25-1.65, P=0.008) and isokinetic torque testing (SMD=0.80, 95% CI: 0.16-1.45, P=0.01], and the physical performance via walking training with blood flow restriction was significantly better than that via walking training by means of physical function evaluation (SMD=0.94, 95% CI: 0.35-1.52, P=0.002).
    CONCLUSION: Meta-analysis results of the existing 16 randomized controlled trials indicate that compared with the traditional resistance training and walking training, low-load blood flow restriction training and walking training with blood flow restriction can also increase muscle strength and muscle mass in older adults. Although the large-scale randomized controlled trials are still needed on the improvement of physical performance, low-load blood flow restriction training and walking training with blood flow restriction could be good ways to prevent elder population from sarcopenia. 
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    Effects of exercise on depression-like behaviors in chronic unpredictable mild stress rodent models: a systematic review and Meta-analysis
    Wang Xiaoge, Liu Jiwen, Yang Shuai, Bao Jinyu, Li Cui
    2023, 27 (5):  813-820.  doi: 10.12307/2023.069
    Abstract ( 637 )   PDF (1186KB) ( 148 )   Save
    OBJECTIVE: Depression is one of the five major causes of the global burden of diseases. Currently available antidepressants have low efficacy, slow onset, and serious side effects, while exercise is a better way to treat depression. This paper systematically evaluates the effects of exercise on depression-related behaviors in chronic unpredictable mild stress (CUMS) animal models. 
    METHODS: A systematic literature search from January 1, 2000 to February 28, 2022 was performed in WanFang, CNKI, PubMed, Embase, and Web of Science databases. Studies regarding the effects of treadmill running, swimming, and voluntary wheel running on forced swimming, tail suspension, and sugar water preference behavior of CUMS animal models were collected. Literature screening, data extraction and methodological quality assessments were completed by two researchers independently. The SYRCLE tool was used to assess the risk of bias in the included studies. RevMan 5.3 and Stata 13.0 analysis software were used for statistical analysis.
    RESULTS: A total of 23 controlled experimental studies were included, including 301 rodents in exercise group and 302 rodents in control group. Meta-analysis results showed that exercise significantly reduced the forced swimming latency [standardized mean difference (SMD)=-3.93, 95% confidence interval (CI): (-4.88, -2.98), P < 0.000 01] and tail suspension latency [SMD=-4.42, 95% CI: (-5.62, -3.23), P < 0.000 01] in CUMS modeling rats and mice. Exercise also increased the index of sucrose preference test [SMD=2.37, 95% CI: (1.62, 3.11), P < 0.000 01]. Subgroup analysis of exercise modes in the suspension test reduced heterogeneity [SMD=-3.68, 95% CI: (-4.16, -3.21), P < 0.000 01], but it did not affect the exercise effect.
    CONCLUSION: Exercise can effectively improve depression-like behaviors of forced swimming, tail suspension, and sucrose preference tests in CUMS model rodents. The mode of exercise may be the source of heterogeneity affecting the behavior of the suspension tail test. Modeling time, exercise stage, exercise mode, and exercise duration are not the main factors that affect the effect of exercise on depression.  
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