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18 July 2023, Volume 27 Issue 20 Previous Issue    Next Issue

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Digital morphology of foramen magnum and occipital condyle: an observation based on three-dimensional reconstruction Wang Jian, Wang Xing, Li Kun, Gao Shang, Wang Chaoqun, Zhang Shaojie, Li Zhijun 2023, 27 (20):  3117-3122.  doi: 10.12307/2023.148 Abstract ( 461 )   PDF (1319KB) ( 78 )   Save BACKGROUND:  Lesions of structures around the ventral side of the brainstem are located in the deep surface of the craniocervical junction, surrounded by important structures. The far lateral approach is the basic surgical method, but sex and lateral differences should be considered in the approach. OBJECTIVE: To analyze the morphological characteristics of the foramen magnum and occipital condyle by three-dimensional reconstruction measurements, thereby providing anatomical parameters for the far lateral approach to the skull base in clinical practice.  METHODS: A total of 673 adult patients (448 males and 225 females) with head and neck CT scans were selected. The age ranged from 20 to 87. The length and width of the foramen magnum were measured. The area and index of the foramen magnum were then calculated by formulas. The length and width of the occipital condyle were measured, as well as the angle between the long and sagittal axis of the occipital condyle (O-S angle), the angle between the line connecting the midpoints of anterior and posterior foramen magnum and the line connecting the midpoint of anterior foramen magnum and the intersection of posterior occipital condyle and lateral foramen magnum (F-O angle), and the angle between the line connecting the midpoints of anterior and posterior foramen magnum and the line connecting the midpoint of anterior foramen magnum and the midpoint of the posterior wall of the hypoglossal canal (F-H angle). The sex and lateral differences of each index were analyzed. RESULTS AND CONCLUSION: There were statistical differences in foramen magnum indexes except between sexes (P > 0.05). The indexes of occipital condyle were significantly higher in males than in females (P < 0.05) except that the left occipital condyle width in males was lower than that in females (P > 0.05). The O-S angle in males was smaller than that in females, and there was a significant difference in the left O-S angle between sexes (P < 0.05). There was no statistical difference between right and left F-H angles. Significant differences were observed in occipital condyle length, occipital condyle width, O-S angle, and F-O angle (P < 0.05). These findings indicate that the parameters of the foramen magnum and occipital condyle were different in sex and on both sides. For the far lateral and transcondylar approaches, the right approach was relatively safe and exposed to a larger range. The length, width, and area of the foramen magnum, the length of the occipital condyle, and O-S angle in males were larger than those in females. Therefore, the sex difference should be considered in the far lateral approach. Figures and Tables | References | Related Articles | Metrics

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Effects of point-pressing of the spleen meridian on inflammatory factors and calcium homeostasis in the skeletal muscle of acute blunt contusion rats Liu Haichao, Wang Hao, Wang Shizhong, Lin Jianping, Jin Jing, Chen Shaoqing 2023, 27 (20):  3123-3128.  doi: 10.12307/2023.436 Abstract ( 376 )   PDF (1136KB) ( 308 )   Save BACKGROUND: Previous studies have shown that tuina can promote the repair of skeletal muscle injuries, but the specific mechanism is unclear. OBJECTIVE: To investigate the effect of point-pressing of the spleen meridian on inflammation and calcium homeostasis in skeletal muscle repair in rats with acute blunt contusion injuries.  METHODS: Thirty-two male Sprague-Dawley rats were randomly divided into blank group, model group, non-acupoint percussion group, and acupoint percussion group. Except for the blank group, animal models of acute gastrocnemius blunt contusion were established in the other three groups. In the non-acupoint pressing group, the intersection point of the same horizontal line of Yinlingquan acupoint (SP 9) and the midline of the gallbladder and bladder meridians was taken for percussion. In the acupoint pressing group, percussion was performed at the Yinlingquan acupoint. Point-pressing in the two groups was done once a day, for 7 consecutive days. Catwalk small animal gait analysis system was used to observe locomotor activity of rats. Hematoxylin-eosin staining was performed to observe the skeletal muscle morphology changes. ELISA method was used to detect the levels of inflammatory factors and the activity of sarcoplasmic/endoplasmic reticulum Ca2+ATPase (SERCA). Calcium concentration in the skeletal muscle was measured using the calcium assay. The level of apoptosis in the skeletal muscle was detected by TUNEL method. RESULTS AND CONCLUSION: Compared with the model and non-acupoint pressing groups, the average support phase was significantly increased in the acupoint pressing group (P < 0.01, P < 0.05). Aggregation of inflammatory cells and red blood cells was reduced in the acupoint percussion group, and skeletal muscle repair was also better in the acupoint percussion group than the non-acupoint pressing group. Compared with the model group, the levels of tumor necrosis factor α and interleukin-6 were significantly decreased in the acupoint pressing group (P < 0.01), and the SERCA activity was significantly increased (P < 0.01). Compared with the non-acupoint pressing group, the level of tumor necrosis factor α was significantly decreased (P < 0.01), the level of interleukin-6 changed insignificantly (P > 0.05), and the activity of SERCA was significantly increased in the acupoint pressing group (P < 0.01). Compared with the model and non-acupoint pressing groups, the concentration of Ca2+ was significantly decreased in the acupoint pressing group (P < 0.01, P < 0.05). Compared with the model group, the apoptotic rate was significantly reduced in the two point-pressing groups (P < 0.01). The apoptotic rate in the acupoint percussion group was significantly lower than that in the non-acupoint pressing group (P < 0.05). To conclude, percussion of the spleen meridian can effectively improve the motor function of rats with skeletal muscle injury and promote skeletal muscle repair. Its mechanism may be related to reducing inflammatory factors, promoting calcium pump function, maintaining calcium homeostasis, and thereby reducing cell apoptosis. Figures and Tables | References | Related Articles | Metrics

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Icariin promotes osteoblast proliferation and differentiation through a non-nuclear signaling pathway Mei Jie, He Qiang, Sun Xin, Yin Hong, Qian Weiqing 2023, 27 (20):  3129-3135.  doi: 10.12307/2023.495 Abstract ( 368 )   PDF (1830KB) ( 209 )   Save BACKGROUND: Estrogen deficiency can inhibit osteoclast proliferation and differentiation. Epimedium is a phytoestrogen that acts as a traditional kidney tonic herb, and its active ingredient, icariin, can produce some effects of estrogen. OBJECTIVE: To investigate the possible signaling pathways involved in the non-nuclear effects of icariin in promoting osteoblast proliferation and differentiation, as well as the possible mechanisms of phosphorylated protein cascade regulation in the signaling pathway by phosphorylated proteomics techniques.  METHODS: Primary osteoblasts were obtained from newborn Sprague-Dawley rats, cultured, and identified. The effect of icariin on osteoblast activity was measured by cell counting kit-8 method. The primary osteoblasts were passed to the third generation. After 24 hours, passaged cells were then divided into six groups: bovine serum albumin intervention group, PBS intervention group, icariin intervention group, icariin-bovine serum albumin coupling intervention group, ICI182780+icariin-bovine serum albumin coupling intervention group, and PD98059+icariin-bovine serum albumin coupling intervention group. The cells of each group were collected at 0, 5, 15 and 25 minutes and used for protein extraction, enzymatic digestion, peptide labeling and phosphorylated peptide enrichment. Phosphoproteomic analysis was performed using liquid chromatography-mass spectrometry technology RESULTS AND CONCLUSION: 10 μg/L Icariin significantly promoted the proliferation and differentiation of osteoblasts. Icariin-bovine serum albumin coupling was able to induce phosphorylation of ERK in osteoblasts without membrane permeation, confirming the non-nuclear effect. Compared with the icariin-bovine serum albumin coupling intervention group, there were 971 differentially expressed proteins (499 up-regulated and 472 down-regulated) in the icariin intervention group, 974 (509 up-regulated and 465 down-regulated) in the bovine serum albumin intervention group, 836 (377 up-regulated and 459 down-regulated) in the PBS intervention group, 231 (126 up-regulated and 105 down-regulated) in the ICI182780+icariin-bovine serum albumin coupling intervention group, and 150 (65 up-regulated and 85 down-regulated ) in the PD98059+icariin-bovine serum albumin coupling intervention group. Among them, 65 and 62 significantly expressed proteins were down-regulated (Fc value < 0.8) in the ICI182780+icariin-bovine serum albumin coupling intervention group and PD98059+icariin-bovine serum albumin coupling intervention group, respectively, most of which were involved in important biological processes, such as mRNA processing and stability regulation, RNA splicing, protein phosphorylation, and mitosis, as well as in signaling pathways analyzed by KEGG library, such as MAPK, protein ubiquitination, mTOR, and PI3K/Akt pathways. To conclude, icariin promotes osteoblast proliferation and differentiation through non-nuclear effects in relation to MAPK, protein ubiquitination, mTOR, and PI3K/Akt signaling pathways. Figures and Tables | References | Related Articles | Metrics

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Mechanisms by which swimming exercise and diet control improve hypothalamic lesions in APOE-/- mice with high-fat diet Ding Linlin, Lu Taotao, Wei Wei, Li Yongxu, Lin Libin, Lin Zhicheng, Xue Xiehua 2023, 27 (20):  3136-3142.  doi: 10.12307/2023.471 Abstract ( 349 )   PDF (1511KB) ( 158 )   Save BACKGROUND: Swimming exercise and diet control are currently recognized as important interventions to improve metabolic abnormalities. Hypothalamic sirtuin 3 (SIRT3) and peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) are important factors regulating energy metabolism, but the correlation between the effects of swimming exercise and diet control on hypothalamic lesions in apolipoprotein E knockout (APOE-/-) mice with high-fat diet and the PGC-1α-SIRT3 pathway is still unclear. OBJECTIVE: To investigate the possible mechanism of swimming exercise and diet control in regulating hypothalamic lesions of APOE-/- mice with high-fat diet. METHODS: Fifty 20-week-old APOE-/- mice were randomly divided into normal diet group, high-fat diet group, diet control group, swimming exercise group and diet control+swimming exercise group, with 10 mice in each group. Except for the control group (normal diet), the other groups were fed with high-fat diet (21% fat, 0.15% cholesterol) for 8 weeks. After 8 weeks of feeding, the diet control group and the diet control+swimming exercise group were normally fed, and there were no changes in the other groups. The swimming exercise group and the diet control+swimming exercise group were given no weight-bearing swimming for 8 weeks in total. Mice were weighed at the same time each week, and changes in hypothalamic metabolites, including N-acetyl aspartate (NAA), choline complex (Cho), creatine (Cr), and myo inositol (MI) were detected by magnetic resonance spectrum of the hypothalamus. Morphological changes of mitochondria and myelin sheath in the hypothalamus were detected by electron microscopy. SIRT3 and PGC-1α protein expression was detected by western blot. RESULTS AND CONCLUSION: The body mass of the control group was significantly lower than that of the other four groups after high-fat diet feeding. After 8 weeks of intervention, the body mass of swimming exercise group, diet control group, diet control+swimming exercise group was significantly lower than that of high-fat diet group (P < 0.05), and the decreasing trend of diet control+swimming exercise group was the most obvious. Compared with the control group, the NAA/Cr and Cho/Cr of the high-fat diet group decreased significantly, and the MI/Cr increased significantly (P < 0.05). Compared with the high-fat diet group, the NAA/Cr and Cho/Cr of the diet control group, the swimming exercise group and the diet control+swimming exercise group increased significantly, while the MI/Cr decreased significantly (P < 0.05). The improvement effect on NAA/Cr and Cho/Cr of diet control+swimming exercise group was better than that of diet control group and swimming exercise group. The protein levels of SIRT3 and PGC-1α in the hypothalamus of diet control group, swimming exercise group and diet control+swimming exercise group were significantly higher than those of high-fat diet group (P < 0.05). The results of electron microscopy showed that in the high-fat diet group the mitochondria of mice were swollen, the crists were broken, disappeared or vacuolated, the myelin mechanism was blurred, most of the myelin was separated from the middle and edge, and the vacuolated degeneration was observed. The morphology and structure of mitochondria and myelin were improved after diet control and swimming exercise intervention. These findings indicate that both swimming exercise and diet control can effectively improve high-fat-induced hypothalamic lesions, and the combination of the two achieves better outcomes. The underlying mechanism is related to up-regulating the expression of mitochondria-related proteins PGC-1α and SIRT3, improving hypothalamic neurometabolism, and alleviating mitochondrial dysfunction. Figures and Tables | References | Related Articles | Metrics

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Screening and validation of neurogenic bladder miRNA-mRNA regulatory network Guo Shuhui, Yang Ye, Jiang Yangyang, Xu Jianwen 2023, 27 (20):  3143-3150.  doi: 10.12307/2023.492 Abstract ( 381 )   PDF (2365KB) ( 157 )   Save BACKGROUND: The miRNA-mRNA regulatory network plays an important role in various biological processes, but its specific mechanism in neurogenic bladder after spinal cord injury remains unclear. OBJECTIVE: To mine the related targets of neurogenic bladder based on bioinformatics and to construct the miRNA-mRNA regulatory network, verified by animal experiments. METHODS: The differential genes of neurogenic bladder were screened from GEO database to construct the miRNA-mRNA regulatory network. The differential genes were analyzed by gene ontology and Kyoto Encyclopedia of Genes and Genomes analyses using the David database. Protein-protein interaction analysis was performed using the STRING database. Key genes were identified by Cytoscape and ROC curves. Finally the key genes were validated by RT-PCR in a rat model of neurogenic bladder. RESULTS AND CONCLUSION: A total of 188 differential genes related to neurogenic bladder were identified. Gene ontology and Kyoto Encyclopedia of Genes and Genomes analyses revealed that they were enriched mainly in inflammatory response, cytokine-cytokine receptor interaction, and chemokine signaling pathway. Two key regulatory networks, miR-147-brain-derived neurotrophic factor and miR-362-5p-Scn9a, were identified based on the Cytoscape and ROC curve results. RT-PCR results showed that miR-147, miR-362-5p, and brain-derived neurotrophic factor were highly expressed in the neurogenic bladder rats, while Scn9a was lowly expressed compared with the control rats. Overall, these findings indicate that miR-147-brain-derived neurotrophic factor and miR-362-5p-Scn9a actas important regulators of pathophysiological processes such as inflammation and oxidative stress, which are expected to become new targets for the diagnosis and treatment of neurogenic bladder.   Figures and Tables | References | Related Articles | Metrics

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Neuroprotective effects of sodium butyrate and acetylation proteomics analysis in fluorosis rats Li Yangjie, Qi Rong, Zhang Xinyu, Cheng Jiaji, Zhou Ning, Cui Xue, Cheng Shuang, Wang Zhengdong, Yan Nan 2023, 27 (20):  3151-3157.  doi: 10.12307/2023.484 Abstract ( 460 )   PDF (1488KB) ( 237 )   Save BACKGROUND: Acetylation, one of the post-translational modifications of proteins, can induce changes in gene expression related to synaptic connection and memory storage by regulating chromatin structure. Sodium butyrate, a deacetylase inhibitor, has been confirmed to play a neuroprotective effect in various neurological disease models, but it has not been confirmed in animal experiments in the field of fluorine neurotoxicity, and its targets are still not comprehensive. OBJECTIVE: To study the effect of sodium butyrate on brain injury induced by fluorosis in rats and to preliminarily explore its possible mechanism. METHODS: Newly weaned male Sprague-Dawley rats were randomly divided into three groups. Rats in fluorosis group and sodium butyrate treatment group were given distilled water containing fluorine for 10 weeks. After 10 weeks of fluoride exposure, rats in the sodium butyrate treatment group were given 1 000 mg/kg sodium butyrate daily for 4 weeks. Fluorosis group and control group were intragastrically given the same amount of normal saline. Morris water maze was used to test the learning and memory ability of rats in each group. Pathological changes of the cerebral cortex were observed by hematoxylin-eosin staining. The acetylated modified proteins were identified by performance liquid chromatography-tandem mass spectrometry and their biological information was analyzed. RESULTS AND CONCLUSION: Compared with the control group, the body mass of rats decreased and the whole brain coefficient increased in the fluorosis group. Compared with the fluorosis group, the body mass of rats increased in the sodium butyrate treatment group, and the whole brain coefficient of rats decreased. Hematoxylin-eosin staining results showed cytoplasmic vacuolation, nuclear pyknosis, and perinuclear space widening in nerve cells of the fluorosis group. In the sodium butyrate treatment group, the number of normal nerve cells increased, cytoplasmic vacuolation decreased, and nuclear pyknosis decreased. Gene ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses showed that a large number of differential acetylated modified proteins were significantly enriched in the synaptic vesicle circulation, synaptic transmission, neurotransmitter transport, transmembrane material transport,and other pathways related to synaptic plasticity. The data of protein-protein interaction network were imported into Cytoscape software, and the top five key hub proteins were identified by different algorithms, which were Hsp8, Rab7a, Nsf, Ezr, and Cfl1. These proteins may be the pathogenic factors of fluorine neurotoxicity and the potential therapeutic targets of sodium butyrate. Figures and Tables | References | Related Articles | Metrics

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Effects of Santong electroacupuncture on the activity of cytoplasmic phospholipase A2 in rats with spinal cord injury via the Rho/Rho kinase and MEK/ERK signaling pathways Yao Haihua, Min Youjiang, Hong Dongying, Wang Li, Lu Xiuyun, Yang Yihua 2023, 27 (20):  3158-3166.  doi: 10.12307/2023.447 Abstract ( 456 )   PDF (1521KB) ( 285 )   Save BACKGROUND: Cytoplasmic phospholipase A2 (cPLA2) is a novel strategy and target for the treatment of spinal cord injury. cPLA2 is regulated by RhoA/Rho kinase and mitogen-activated protein kinase (MEK)/extracellular regulated protein kinases (ERK) signaling pathways. Electroacupuncture can treat spinal cord injury by regulating the RhoA/Rho kinase and MEK/ERK signaling pathways. OBJECTIVE: To investigate the effects of electroacupuncture on the activity of cPLA2 after spinal cord injury by regulating Rho/ROCK (Rho kinase) and MEK/ERK signaling pathways. METHODS: After spinal cord injury modeling and Basso, Beattie, Bresnahan scoring, 72 female Sprague-Dawley rats were randomly divided into model group, electroacupuncture group, U0126 treatment group and Y27632 treatment group, with 18 rats in each group. Another 18 rats were used as sham operation group, in which only laminectomy was performed without spinal cord impingement. In the electroacupuncture group, Dazhui (GV 14), Yaoyangguan (GV 3), bilateral Ciliao (BL 32) and Zusanli (ST 36) were treated with electroacupuncture, once a day, 20 minutes each time, 14 times in total. Rats in the U0126 and Y27632 treatment groups were given U0126 and Y27362 subdural injection every other day, respectively. At the end of treatment, the rats were sacrificed after Basso, Beattie, Bresnahan scoring, and the spinal cord tissue was collected to detect the content of prostaglandin E2 and platelet activating factor by ELISA. TUNEL assay was used to detect the apoptosis rate of spinal cord nerve cells. Western blot was used to detect the protein expression of RhoA, ROCK II, MEK, ERK1/2, p-ERK1/2, cPLA2 and p-cPLA2 in the spinal cord. qRT-PCR was used to detect the mRNA expression of RhoA, ROCK II, MEK, ERK1/2 and cPLA2 in the spinal cord.  RESULTS AND CONCLUSION: Compared with the sham operation group, Basso, Beattie, Bresnahan score was significantly decreased in the model group (P < 0.01), while the number of apoptotic cells, the levels of prostaglandin E2 and platelet activating factor, and the expression of p-cPLA2 protein and cPLA2 mRNA in spinal cord tissue were significantly increased in the model group (P < 0.01). After treatment, the above indexes were significantly reversed in all the treatment groups except for the mRNA expression of cPLA2 in the electroacupuncture group (P < 0.01). Compared with the sham operation group, the expression of RhoA, ROCK II, and MEK protein and gene, the mRNA expression of ERK1/2, and the protein expression of p-ERK1/2 were significantly increased in the model group (P < 0.01). After treatment, the above indexes were significantly decreased in the three treatment groups (P < 0.01 or P < 0.05). To conclude, electroacupuncture can down-regulate the expression of factors related to Rho/ROCK and MEK/ERK signaling pathways, inhibit the activity of cPLA2, reduce the apoptosis of nerve cells, alleviate inflammatory responses, and finally achieve the therapeutic effect on spinal cord injury. Figures and Tables | References | Related Articles | Metrics

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Role of m6A RNA methylation modification in necrotic apoptosis in a rat model of brain injury Lin Shukai, Liu Zhonghai, Liu Zhen, Cai Jincheng, Wei Ruting 2023, 27 (20):  3167-3172.  doi: 10.12307/2023.187 Abstract ( 322 )   PDF (1179KB) ( 191 )   Save BACKGROUND: Mammalian central nervous system injury is dependent on the regulation of N6-methyladenosine (m6A) and is closely associated with necrotic apoptosis. At present, the relationship between m6A RNA methylation modification and necrotic apoptosis in rat models of traumatic brain injury has not been studied. OBJECTION: To study the relationship between m6A RNA methylation and necrotic apoptosis in a rabbit brain injury model and to provide experimental basis for the molecular mechanism of occurrence, development and prognosis of necrotic apoptosis following craniocerebral injury. METHODS: A total of 30 SPF male Sprague-Dawley rats were selected and randomized into sham operation group (n=10), traumatic brain injury group (n=10), and signal transducer and activator of transcription 1 (STAT1) inhibitor (NSC118218) group (n=10). The brain injury model was established using the modified Feeney method in the latter two groups, while the dura mater in the sham operation group was only exposed without striking. The levels of inflammatory factors, tumor necrosis factor ɑ, interleukin-6, interleukin-1β, interleukin-10, and high mobility group box 1 in the cerebral cortex of rats and brain water content were detected at 6 hours after brain injury. At the same time, the methylation level of total m6A RNA and the mRNA and protein expression levels of YTH domain family protein family 2 (YTHDF2), JAK1 and STAT1 were detected. RESULTS AND CONCLUSION: The levels of inflammatory factors, brain water content, and the expression of JAK1 and STAT1 in the traumatic brain injury group and NSC118218 group were significantly higher than those in sham operation group, while the proportion of m6A RNA methylation and the expression level of YTHDF2 were significantly lower than those in the sham operation group (P  <  0.05). The levels of inflammatory factors, brain water content, and the mRNA and protein expressions of JAK1 and STAT1 in the NSC118218 group were significantly lower than those in the traumatic brain injury group, while the proportion of m6A RNA methylation and the expression level of YTHDF2 were significantly increased (P  <  0.05). These findings indicate that the proportion of m6A RNA methylation and YTHDF2 expression decrease after brain injury, which activates the JAK/STAT signaling pathway, increases the expression of JAK1 and STAT1, and thus promotes necrotic apoptosis in cells. Figures and Tables | References | Related Articles | Metrics

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Role of LncRNA MALAT1 in myocardial autophagy reduction in aging rats after ischemic postconditioning Yang Huixia, Jie Yuzhen, Bai Zhigang, Jiao Yun, Yang Yong, Ma Tianlong, Ma Shengchao, Jiang Yideng 2023, 27 (20):  3173-3179.  doi: 10.12307/2023.478 Abstract ( 438 )   PDF (1474KB) ( 128 )   Save BACKGROUND: Ischemic postconditioning can alleviate myocardial ischemia-reperfusion injury, but the specific mechanism is not clear. OBJECTIVE: To investigate the mechanism of long non-coding RNA (lncRNA) MALAT1 in the reduction of autophagy levels in aging myocardium induced by ischemic postconditioning. METHODS: Twenty-seven Sprague-Dawley rats aged 22-24 months were randomly divided into three groups, with nine rats in each group: sham operation, ischemia-reperfusion, and ischemic postconditioning groups. Morphological changes of myocardial tissue were observed by hematoxylin-eosin staining and Masson staining. Rat myocardial cells (H9C2) were induced in vitro with 8 mg/mL D-galactose for 9 days and then divided into normoxia, hypoxia-reoxygenation, and hypoxia postconditioning groups. Western blot was used to detect the protein expression levels of LC3II/I and p62. Fluorescence quantitative PCR was used to detect the expression of lncRNA MALAT1 in aging myocardium and aging cardiomyocytes. Autophagy double-labeled adenovirus (RFP-GFP-LC3) was used to observe the changes of autophagic flux in aging cardiomyocytes. lncRNA MALAT1 interference fragment and overexpression plasmid were transfected into aging cardiomyocytes and the protein expression levels of LC3II/I and p62 were detected by western blot.  RESULTS AND CONCLUSION: Compared with the ischemia-reperfusion group, the myocardial tissue structure of the ischemic postconditioning group was basically clear, the nucleus was intact, and the deposition of blue collagen fibers in the myocardial tissue was reduced. Compared with the ischemia-reperfusion group, the expression of LC3II/I was decreased and the expression of p62 was increased in the ischemic postconditioning group (P < 0.05).  Compared with the hypoxia-reperfusion group, the expression of LC3II/I was decreased (P < 0.01) and the expression of p62 was increased (P < 0.05) in the hypoxia postconditioning group, and the number of intracellular autophagosomes and autophagolysosomes was decreased (P < 0.05). Compared with the ischemia-reperfusion group, the expression of MALAT1 in the aging myocardial tissue was decreased the ischemic postconditioning group (P < 0.01); compared with the hypoxia-reperfusion group, the expression of MALAT1 in aging cardiomyocytes was decreased in the hypoxic postconditioning group (P < 0.01). Compared with the hypoxia postconditioning+si-NC group, the expression of LC3II/I was decreased and the expression of p62 was increased in the hypoxia postconditioning +si-lncRNA MALAT1 (P < 0.01); compared with the hypoxia postconditioning+ad-NC group, the expression of LC3II/I was increased and the expression of p62 was decreased in the hypoxia postconditioning+ad-lncRNA MALAT1 group (P < 0.01). To conclude, the lncRNA MALAT1 mediated reduction of autophagy levels is an important mechanism underlying the protective effect of ischemic postconditioning in aging myocardium. Figures and Tables | References | Related Articles | Metrics

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Jiajian Didang Tang inhibits glial scar formation in rats with acute spinal cord injury under the guidance of Tongfu Zhuyu therapy Wen Feng, Zhou Lei, Li Yang, Zheng Liming, Zhang Zhiwen, Fan Xiao, Wu Zijian 2023, 27 (20):  3180-3187.  doi: 10.12307/2023.529 Abstract ( 464 )   PDF (2398KB) ( 155 )   Save BACKGROUND: Acute spinal cord injury is a disease with high disability and mortality. An active and effective treatment in the acute phase is of great significance to prevent secondary injury. OBJECTIVE: To investigate the mechanism underlying Tongfu Zhuyu therapy in the treatment of acute spinal cord injury through observing the therapeutic effect of Jiajian Didang Tang on edema and recovery of limb function after acute spinal cord injury in rats. METHODS: A total of 60 Sprague-Dawley rats were randomly divided into sham group, model group, Didang Tang group, 2-(nicotinamide)-1,3,4-thiadiazole (TGN-020) group, and Didang Tang combined with TGN-020 group (combined group), with 12 rats in each group. Among them, the sham group only underwent laminectomy without spinal cord shock and the other groups used NYU percussion device to create an acute spinal cord injury model. After modeling, the sham and model groups did not take any treatment measures. The Didang Tang group was given 5.04 g/kg Didang Tang by intragastric administration, the TGN-020 group was given 5 mg/kg TGN-020 by intraperitoneal injection, and the combined group was given TGN-020 by intraperitoneal injection combined with Didang Tang by intragastric administration, once a day for 7 days. Basso, Beattie and Bresnahan score and Reuter score were evaluated on 1, 3, 5 and 7 days after operation. Spinal cord tissue was taken for general observation and water content measurement at 7 days after operation. The degree of tissue damage was detected by hematoxylin-eosin staining. The number of aquaporin 4/glial fibrillary acidic protein-positive cells in the spinal cord tissue was detected by immunofluorescence assay. The protein expression levels of Aquaporin 4, glial fibrillary acidic protein, chondroitin sulfate proteoglycan, and proliferating cell nuclear antigen protein in spinal cord tissue were detected by western blot. RESULTS AND CONCLUSION: The behavioral scores showed that compared with the model group, the Basso, Beattie and Bresnahan scores were significantly increased in the three administration groups (P < 0.05), while the Reuter scores were significantly decreased (P < 0.05). Moreover, there was no significant difference among the three administration groups (P > 0.05). Compared with the model group, the water content of spinal cord tissue was significantly decreased in the three administration groups (P < 0.05), while there was no significant difference among the three administration groups (P > 0.05). Hematoxylin-eosin staining results showed that the structure of the model group was disorganized, with a lot of scar tissue and syringomyelia. Compared with the model group, the tissue structure was relatively complete in the three administration group, with a small amount of scar tissue hyperplasia and syringomyelia, and the tissue morphology was effectively improved. Immunofluorescence staining results showed that compared with the model group, the number of aquaporin 4/glial fibrillary acidic protein-positive cells was less in the three administration groups, with lower brightness and reduced degree of cell swelling (P < 0.05). However, there was no significant difference among the three administration groups (P > 0.05). Western blot results showed that compared with the model group, the expressions of aquaporin 4, glial fibrillary acidic protein, chondroitin sulfate proteoglycan, and proliferating cell nuclear antigen protein in the spinal cord tissue of the three administration groups were significantly decreased (P < 0.05). While, there was no significant difference among the three administration groups (P > 0.05). To conclude, Jiajian Didang Tang can promote the recovery of limb function in rats with acute spinal cord injury under the guidance of Tongfu Zhuyu therapy. Its mechanism may reduce edema at the injury site by inhibiting the expression of aquaporin 4, so as to effectively inhibit the activation and proliferation of reactive astrocytes and reduce the formation of glial scar.    Figures and Tables | References | Related Articles | Metrics

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Effects of aerobic exercise on learning, memory, and hippocampal neuromorphology in mice with Alzheimer’s disease Zhang Yeting, Fu Yan, Li Xue, Wei Cuilan, Li Chuikun, Yuan Qiongjia 2023, 27 (20):  3188-3194.  doi: 10.12307/2022.673 Abstract ( 692 )   PDF (2117KB) ( 339 )   Save BACKGROUND: Exercise helps prevent and retard cognitive decline related to Alzheimer’s disease, dementia, and age. However, whether exercise prevents cognitive decline in patients with Alzheimer’s disease is related to the neuromorphological changes of the hippocampus is still unclear. OBJECTIVE: To investigate the effects of long-term aerobic exercise on learning and memory ability and neuromorphology of the hippocampus in mice with Alzheimer’s disease and its neuromechanism on Alzheimer’s disease. METHODS: Twelve wild-type mice aged 3 months were divided into two groups (n=6 per group): a wild exercise group and a wild control group. Mice in the wild exercise group were given an exercise intervention for 5 months, and mice in the wild control group had no intervention. Twelve APP/PS1 double transgenic mice with Alzheimer’s disease were divided into two groups (n=6 per group): a model exercise group and a model control group. Mice in the model exercise group were given an exercise intervention for 5 months, and mice in the model control group had no intervention. After the exercise intervention, the memory ability of mice was tested through the eight-arm maze test. Neuromorphological changes of the hippocampus in mice were observed using Nissl staining under transmission electron microscope.  RESULTS AND CONCLUSION: The results of the eight-arm maze test showed that the memory ability of mice in the wild exercise group was better than that in the wild control group and the model exercise group (P < 0.05), and the memory ability of mice in the model exercise group and the wild control group was better than that in the model control group (P < 0.05). The results of Nissl staining showed that the Nissl bodies with clear nuclei and nucleoli were clearly visible in the hippocampal dentate gyrus, CA3 and CA1 areas of mice in the wild exercise group and the wild control group. In the mice with Alzheimer’s disease, the Nissl bodies in the hippocampus, especially in the dentate gyrus and CA3 areas, were fuzzy, and the nuclei and nucleoli were difficult to distinguish. In particular, in the model control group, the structure of nerve cells was relatively fuzzy, some neurons were seriously damaged, nerve cells arranged loosely with large spacing interval. Under the transmission electron microscopy, the number of synapses in the hippocampal dentate gyrus was decreased in the model control group compared with the wild control group. Some synaptic clefts, presynaptic membrane, and postsynaptic membrane were blurred, and there were fewer vesicles in the presynaptic membrane. The density of postsynaptic dense zone was lower. Compared with the model control group, the number of synapses in the hippocampal dentate gyrus was increased in the model exercise group. The distribution of synaptic vesicles in the presynaptic membrane was dense and uniform, and the density of postsynaptic dense zone was increased. To conclude, exercise can improve the structure of hippocampal nerve cells in Alzheimer’s disease mice to a certain extent, which may be one of the neuromechanisms by which exercise improves the learning and memory ability of Alzheimer’s disease mice. Figures and Tables | References | Related Articles | Metrics

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Rongjin Niantong Fang for treating knee osteoarthritis by regulating cartilage matrix metabolism Zhao Zhongsheng, Zheng Ruoxi, Lin Jie, Chen Jun, Ye Jinxia, Fu Changlong, Wu Guangwen 2023, 27 (20):  3195-3201.  doi: 10.12307/2023.405 Abstract ( 434 )   PDF (1629KB) ( 136 )   Save BACKGROUND: Preliminary studies have found that Rongjin Niantong Fang can effectively inhibit the degradation of cartilage matrix. Long non-coding RNA GAS5 can act as a competitive endogenous RNA that adsorbs miR-21-5p to regulate the expression of matrix metalloproteins, thereby influencing the metabolism of articular cartilage matrix. However, whether Rongjin Niantong Fang regulates cartilage matrix metabolism through long non-coding RNA GAS5/miR-21 and plays a role in preventing and treating osteoarthritis needs to be further studied. OBJECTIVE: To explore the mechanism by which Rongjin Niantong Fang treats knee osteoarthritis by long non-coding RNA GAS5/miR-21 pathway regulating cartilage matrix synthesis and catabolism.  METHODS: Sixty male SPF-grade Sprague-Dawley rats aged 8 weeks were randomized into four groups (n=15 per group): blank group, model group, treatment group and control group. Knee osteoarthritis models were established using the modified Hulth method in the latter three groups. The blank group and the model group were intragastrically administered with normal saline, the treatment group was intragastrically administered with water extract of Rongjin Niantong Fang, and control group was intragastrically administered with glucosamine hydrochloride capsules, once a day for 12 weeks. After treatment, microstructure and morphological changes of cartilage were observed by hematoxylin-eosin staining and safranin O-fast green staining. The mRNA expression of long non-coding RNA GAS5, miR-21, and matrix metabolic factors in cartilage was detected by real-time PCR. The protein expression of matrix metabolic factors in cartilage was detected by western blot.  RESULTS AND CONCLUSION: In the model group, the four layers of the cartilage were disordered and arranged irregularly, the tide line and adhesion line were irregular or disappeared, chondrocytes proliferated and arranged disorderly, the subchondral bone proliferated, and a large amount of collagens and proteoglycans were lost. In the treatment and control groups, the four layers of the cartilage and tide line were recognizable, chondrocytes arranged regularly, the subchondral bone structure was relatively intact, and part of collagens and proteoglycans were lost. Compared with the blank group, the expression levels of long non-coding RNA GAS5, matrix metalloproteinases 3, 9, 13, and a disintegrin and metalloproteinase with thrombospondin motif-5 mRNAs and proteins in cartilage were significantly increased in the model group (P < 0.01 or P < 0.05), while the expression levels of miR-21, tissue inhibitors of metalloproteinase 3, type II collagen, and Aggrecan proteins and mRNAs in cartilage were significantly decreased in the model group (P < 0.01 or P < 0.05). Compared with the model group, the expression trends of these genes and proteins in cartilage were reversely changed in the treatment and control groups (P < 0.01 or P < 0.05). To conclude, Rongjin Niantong Fang can reduce the loss of collagen and proteoglycan in cartilage matrix, delay the degradation of cartilage matrix, and reduce the damage of cartilage morphology and structure by down-regulating the expression of long non-coding RNA GAS5, matrix metalloproteinases 3, 9, 13, and a disintegrin and metalloproteinase with thrombospondin motif-5, and up-regulating the expression of miR-21, tissue inhibitors of metalloproteinase 3, type II collagen, and Aggrecan in cartilage of knee osteoarthritis rats. Figures and Tables | References | Related Articles | Metrics

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Warm-needling moxibusion inhibits NLRP3 inflammasome activation and improves cartilage injury in a rabbit model of knee osteoarthritis Wu Yongli, Li Long, Liu Junwei, Liu Di, Wang Duo 2023, 27 (20):  3202-3208.  doi: 10.12307/2023.429 Abstract ( 566 )   PDF (1621KB) ( 179 )   Save BACKGROUND: Warm-needling moxibusion has a significant clinical effect on knee osteoarthritis, but its therapeutic mechanism remains unclear.  OBJECTIVE: To observe the effect of warm-needling moxibusion on the expression of NLRP3, matrix metalloproteinases 3, 13, and type I collagen in cartilage tissue of rabbit knee osteoarthritis. METHODS: Forty 6-month-old male New Zealand rabbits were randomly divided into blank group, model group, warm-needling moxibusion group, and NLRP3 inhibitor group, with 10 rabbits in each group. Except for the blank group, osteoarthritic models of the right hind limb were established in the other three groups using plaster cast fixation. After modeling, intervention with warm-needling moxibusion in the warm-needling moxibusion group was performed once a day and lasted for 4 weeks; in the NLRP3 inhibitor group, MCC950, an NLRP3 inhibitor, was injected into the knee joint cavity, once a week, for four times in total. Lequesne MG score was used for behavioral evaluation, hematoxylin-eosin staining was used to observe the morphological structure of cartilage tissue, and the expression of NLRP3, matrix metalloproteinases 3, 13, and type I collagen in chondrocytes was detected by immunohistochemistry and western blot. RESULTS AND CONCLUSION: Lequesne MG score: Compared with the blank group, Lequesne MG score increased in the other three groups after modeling (P < 0.05). After treatment, the Lequesne MG scores were significantly decreased in the warm-needling moxibusion group and NLRP3 inhibitor group compared with the model group (P < 0.05). Hematoxylin-eosin staining: compared with the blank group, the number of chondrocytes was decreased, the structural integrity was destroyed, and the Mankin’s score was significantly increased in the model group (P < 0.05). Compared with the model group, the distribution of chondrocytes was more uniform, the structure of cartilage layer was relatively clear, and the Mankin’s score was decreased in the warm-needling moxibusion group and NLRP3 inhibitor group (P < 0.05). Immunohistochemistry and western blot detection: compared with the blank group, the expression of NLRP3, matrix metalloproteinases 3 and 13 was significantly increased (all P < 0.05), and the expression of type I collagen was significantly down-regulated in the model group (P < 0.05). Compared with the model group, the expression of NLRP3, matrix metalloproteinases 3 and 13 were significantly decreased in the warm-needling moxibusion group and NLRP3inhibitor group (all P < 0.05), and the expression of type I collagen was significantly up-regulated (P < 0.05). To conclude, warm-needling moxibusion can inhibit the activation of NLRP3 inflammatory promoter, down-regulate the release of inflammatory factors, matrix metalloproteinases 3 and 13, reduce the inflammatory amplification reaction, and delay the degradation of type I collagen, thereby reducing cartilage injury and local damage of articular cartilage due to knee osteoarthritis. Figures and Tables | References | Related Articles | Metrics

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Effect of aerobic exercise on renal fibrosis in rats with chronic renal failure Liu Peng, Ma Gang, He Ruibo, Peng Peng 2023, 27 (20):  3209-3215.  doi: 10.12307/2023.421 Abstract ( 404 )   PDF (1519KB) ( 135 )   Save BACKGROUND: Renal fibrosis is the main characteristic and basic pathophysiological change of chronic renal failure, in which local imbalance of renin-angiotensin system homeostasis induced abnormal collagen metabolism may play a key role. Aerobic exercise is an important non-drug method for the prevention and treatment of various types of tissue fibrosis; however, its effect and mechanism on renal fibrosis have not been clarified.  OBJECTIVE: To elucidate the effect of regular aerobic exercise on renal fibrosis in a rat model of chronic renal failure and to explore the mechanism of collagen metabolic pathway and renin-angiotensin system.  METHODS: Forty-five 6-week-old male Sprague-Dawley rats were randomly divided into sham group, model sedentary group, and model exercise group, with 15 rats in each group. Animal models of chronic renal failure were established by 5/6 nephrectomy in the latter two groups. The sham and model sedentary groups were caged and fed quietly, while the model exercise group received 18 weeks of treadmill aerobic exercise training (60 minutes a day, 5 days per week). After exercise, caudal artery blood pressure was measured by non-invasive sphygmomanometer; urine was collected from metabolic cage for 24-hour proteinuria measurement; and blood sample was taken from the heart to detect serum urea nitrogen and serum creatinine. Renal histopathology was observed by periodic acid–Schiff and Masson staining, and glomerular sclerosis index and fibrosis index were obtained. Protein expression of type I collagen, transforming growth factor-β1, matrix metalloproteinase-2, matrix metalloproteinase-9, tissue inhibitor of metalloproteinase-1, angiotensinogen, renin, renin receptor, angiotensin converting enzyme, angiotensin converting enzyme 2, angiotensin II type 1 receptor, angiotensin II type 2 receptor, and Mas receptor in kidney tissue was determined by western blot. The activity of matrix metalloproteinase-2 and matrix metalloproteinase-9 in kidney tissue was detected by gelatin zymography.  RESULTS AND CONCLUSION: Compared with the model sedentary group, in model exercise group, blood pressure was decreased and renal function was improved (P < 0.05), glomerulosclerosis index and renal fibrosis index values were reduced (P < 0.05), the protein expressions of type I collagen, transforming growth factor-β1, tissue inhibitor of metalloproteinase-1, angiotensinogen, renin, angiotensin converting enzyme and angiotensin II type 1 receptor were downregulated (P < 0.05), the protein expression of matrix metalloproteinase-2, matrix metalloproteinase-9, angiotensin converting enzyme 2, angiotensin II type 2 receptor, and Mas receptor was upregulated (P < 0.05), the ratio of matrix metalloproteinase-2/tissue inhibitor of metalloproteinase-1, the ratio of matrix metalloproteinase-9/tissue inhibitor of metalloproteinase-1 and the activity of matrix metalloproteinase-2 in kidney tissue were increased (P < 0.05). To conclude, long-term aerobic exercise can alleviate renal fibrosis and delay the progression of renal failure induced by 5/6 nephrectomy in rats, and the mechanism may be related to the restoration of renin-angiotensin system homeostasis imbalance and improvement of collagen metabolism. Figures and Tables | References | Related Articles | Metrics

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Depression-like behavior characteristics of rats with neuropathic pain and expression of mGluR5 and NMDAR2B in the left dorsal agranular insular area Hu Yue, Zhu Yuanliang, Wan Tenggang, Xu Fangyuan, Xu Zhangyu, Li Jiyang, Li Dan, Wang Jianxiong 2023, 27 (20):  3216-3223.  doi: 10.12307/2023.410 Abstract ( 438 )   PDF (2059KB) ( 333 )   Save BACKGROUND: The occurrence of neuropathic pain and the resulting depression may be related to the increased protein expression of metabotropic glutamate receptor 5 (mGluR5) and N-methyl-D-aspartate receptor type 2B (NMDAR2B) in the brain. OBJECTIVE: To explore the behavioral characteristics of pain and depression in rats with neuropathic pain caused by chronic compressive injury of the sciatic nerve and to observe the expression of mGluR5 and NMDAR2B proteins in the left dorsal agranular insular area of the rats with neuropathic pain. METHODS: Fifty Sprague-Dawley rats were divided into sham operation group (n=25; only the sciatic nerve trunk was exposed without ligation) and model group (n=25; the sciatic nerve was ligated to establish a chronic compressive injury model). Von Frey filaments were used to detect mechanical paw withdrawal threshold in rats. Depression-like behaviors were observed by the sugar water preference test and forced swimming test. The sciatic nerve function index and hematoxylin-eosin staining were used to observe the function and structure of the sciatic nerve. The metabolism of the sciatic nerve and brain was observed by [18]F-FDG PET-CT. The expression levels of mGluR5 and NMDAR2B in the left-brain dorsal agranular insular area were detected by western blot. RESULTS AND CONCLUSION: Compared with the sham operation group, the mechanical paw withdrawal threshold and sciatic nerve function index were significantly decreased in the model group at 1 week after operation (P < 0.05), and depression-like behaviors appeared and progressively worsened in 2-3 weeks (P < 0.05). Hematoxylin-eosin staining results showed that the pathological changes of the sciatic nerve in the model group were progressively aggravated. The average standardized intake value of the right sciatic nerve/liver in the model group was significantly higher than that in the sham operation group in 2-4 weeks after operation (P < 0.05). The expression levels of mGluR5 and NMDAR2B in the left dorsal agranular insular area were significantly higher in the model group than the sham operation group (P < 0.05). To conclude, the pathological manifestations, sciatic nerve function, neuropathic pain, and depression-like behaviors are not synchronized after sciatic nerve injury. mGluR5 and NMDAR2B in the left dorsal agranular insular area may be involved in the pathogenesis of neuropathic pain and depression-like behaviors caused by pain.  Figures and Tables | References | Related Articles | Metrics

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Screening and analysis of differentially expressed genes in synovial tissue of rheumatoid arthritis Wang Chenyu, Yao Jiawei, Xu Xiongfeng, Qiu Bo 2023, 27 (20):  3224-3229.  doi: 10.12307/2023.406 Abstract ( 498 )   PDF (1384KB) ( 411 )   Save BACKGROUND: Rheumatoid arthritis is an autoimmune disease. Research on rheumatoid arthritis mainly focuses on its pathogenesis and target therapy.  OBJECTIVE: To search for differential genes by bioinformatics analysis of rheumatoid arthritis synovitis gene chip datasets, and to establish and improve the gene regulation network of rheumatoid arthritis.  METHODS: GEO2R was used to analyze the differential expression of rheumatoid arthritis chips screened from the database. R language and DAVID were used for gene ontology enrichment analysis and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis of differentially expressed genes. String database was used for protein-protein interaction network analysis. Cystoscape software was used to obtain key genes. qPCR and western blot were used for experimental verification of some key target genes.  RESULTS AND CONCLUSION: There were 2 064 differentially expressed genes in the synovial tissue of patients with rheumatoid arthritis, including 625 up-regulated genes and 1 439 down-regulated genes. Differentially expressed genes are mainly involved in immune response-activated cell surface receptor signaling pathway, antigen receptor mediated signaling pathway and other biological responses. Ten key target genes, including epidermal growth factor receptor (EGFR), MYC, RELA, ITGB2, LCK, EPHB2, IRF4, NRAS, FN1, and MAPK1, were identified by the protein-protein interaction network analysis. The number of protein interaction lines based on the protein-protein interaction network and Cystoscape algorithm score were used to obtain five genes with high significance, including EGFR, MYC, RELA, ITGB2, and LCK, and the experimental verification of these genes was carried out by qPCR and western blot experiments. The mRNA and protein expression levels of EGFR, MYC, RELA, ITGB2, and LCK genes in the synovial tissue of patients with rheumatoid arthritis were significantly higher than those of the control group, and the expression of EGFR changed most significantly. To conclude, there are distinct gene expression characteristics in the synovial tissue of knee joint after rheumatoid arthritis, and EGFR is the most valuable differentially expressed gene in the synovium of rheumatoid arthritis. Figures and Tables | References | Related Articles | Metrics

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Effects of Sanhua Jiegu San on Wnt/beta-catenin signaling pathway in osteoblasts Duan Jiahao, Tan Xuyi, Lu Min, Kuang Gaoyan, Fang Chuanlong, Xiao Man 2023, 27 (20):  3230-3235.  doi: 10.12307/2023.488 Abstract ( 448 )   PDF (1052KB) ( 183 )   Save BACKGROUND: Sanhua Jiegu San is a common drug for clinical fracture and tendon injury, which have good effect. However, there is a lack of relevant experimental evidence, and its mechanism of action remains unclear.  OBJECTIVE: To observe the effect of Sanhua Jiegu San on Wnt/β-catenin signaling pathway in osteoblasts.  METHODS: Firstly, the optimal concentration of Sanhua Jiegu San for osteoblast intervention was screened by pre-experiment. Then, the proliferation of osteoblasts was tested by cell counting kit-8 method. The maturity of osteoblasts was detected by alkaline phosphatase assay. The mRNA expressions of alkaline phosphatase, Runt-related transcription factor 2, osteocalcin, osteopontin, collagenase I, Axin2, Dkk4, Lef-1, and Tcf-1 were detected by PCR. The protein expressions of β-Catenin and P-β-Catenin were detected by western blot method.  RESULTS AND CONCLUSION: After 5 days of osteoblast culture, the number of osteoblasts reached the maximum at the concentration of 10 mg/L. After 21 days of osteoblast culture, the proliferation of cells in different concentration groups of Sanhua Jiegu San increased, which was obvious in the 10 mg/L group, but tended to decrease at 20 mg/L. After 7 days of osteoblast culture, alkaline phosphatase expression was increased in the low, medium and high concentration groups, especially in the high concentration group (20 mg/L). After 7 days of osteoblast culture, mRNA expressions of alkaline phosphatase, Runt-related transcription factor 2, osteocalcin, osteopontin, collagenase I were increased in the low, medium and high concentration groups, especially in the high concentration group (20 mg/L). After 21 days of osteoblast culture, the mRNA expressions of Axin2 and Dkk4 were decreased in the low, medium and high concentration groups, while the mRNA expressions of Lef-1 and Tcf-1 were increased, especially in the high concentration group (20 mg/L). After 21 days of osteoblast culture, the expression of P-β-Catenin protein was decreased in the low, medium and high concentration groups, while the expression of β-Catenin protein was increased, especially in the high concentration group (20 mg/L). To conclude, Sanhua Jiegu San can regulate the Wnt/β-Catenin signaling pathway in osteoblasts, promote the mRNA expression of alkaline phosphatase, Runt-related transcription factor 2, osteocalcin, osteopontin, collagenase I, Lef-1, and Tcf-1 and the protein expression of β-Catenin, and decrease the mRNA expression of Axin2 and Dkk4 and the protein expression of P-β-Catenin, thereby promoting osteoblast proliferation. This may be the mechanism by which Sanhua Jiegu San promotes fracture healing. Figures and Tables | References | Related Articles | Metrics

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Effects of icariin on NRG1-ErbB signaling pathways in hippocampus of schizophrenia rats Liu Yunqin, Lin Li, Xiao Wenhao, Ji Qiuming, Liu Yanqin 2023, 27 (20):  3236-3241.  doi: 10.12307/2023.433 Abstract ( 571 )   PDF (1205KB) ( 189 )   Save BACKGROUND: Icariin is one of the effective active components of Epimedium, which is important for reinforcing kidney and strengthening yang. It can not only promote the function of the reproductive system, but also significantly improve schizophrenia. However, its specific mechanisms of action are still in study and exploration phase. OBJECTIVE: To explore the effects and mechanisms of icariin on cognitive function of schizophrenia rats.  METHODS: A total of 48 Sprague-Dawley rats were randomly divided into control group, model group, low-dose and high-dose icariin groups with 12 rats in each group. Except for the control group, schizophrenia models were established in the other groups by intraperitoneal injection of N-methyl-D-aspartate receptor antagonist, MK-801, once a day for continuous 14 days. After modelging, the rats in the low- and high-dose icariin groups were intragastrically given icariin, 25 and 50 mg/kg per day for 28 days, respectively, while those in the control and model groups were given the same amount of normal saline. After administration, behavioral detection, Nissl staining and FJB staining of the hippocampus were performed. Oxidative stress level, inflammatory factor level, and NRG1/ErbB4 signaling pathway protein expression were detected. RESULTS AND CONCLUSION: Compared with the model group, stereotyped behavior score, total distance of spontaneous activities and escape latency were significantly decreased, while times of crossing the platform were significantly increased in the two administration groups (P < 0.05). The stereotyped behavior score and escape latency (days 3-5) in the high-dose icariin group were significantly lower than those in the low-dose icariin group (P < 0.05). Compared with the model group, mitochondrial membrane potential and superoxide dismutase activity in the hippocampus were significantly increased, while the levels of malondialdehyde, interleukin-6, interleukin-1β, and tumor necrosis factor-α were significantly decreased in the two administration groups (P < 0.05). Compared with the low-dose icariin group, superoxide dismutase activity was significantly increased and the levels of malondialdehyde, interleukin-6, interleukin-1β, and tumor necrosis factor-α were significantly decreased in the high-dose icariin group (P < 0.05). Compared with the model group, expressions of NRG1 and ErbB4 proteins in the hippocampus were significantly decreased in the two administration groups (P < 0.05). Compared with the low-dose icariin group, the expression of ErbB4 protein was significantly decreased in the high-dose icariin group (P < 0.05). Nissl staining and FJB staining of the hippocampus showed that the number of Nissl bodies in the hippocampus was significantly reduced in the model group, while the number of FJB-positive cells in the CA1 and CA3 regions was significantly increased; compared with the model group, the number of Nissl bodies in the hippocampal was significantly increased in the low-dose and high-dose icariin groups, while the number of FJB positive cells in CA1 and CA3 regions was significantly decreased. To conclude, icariin can improve cognitive function in rats with schizophrenia. And its mechanisms may be related to regulating expressions of NRG1/ErbB4 signaling pathway proteins and reducing oxidative stress and inflammatory responses. Figures and Tables | References | Related Articles | Metrics

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Valve replacement with preservation of the mitral subvalvular apparatus promotes effective valve healing Yuan Ye, Liu Tao, Liu Hua 2023, 27 (20):  3242-3246.  doi: 10.12307/2023.409 Abstract ( 348 )   PDF (801KB) ( 146 )   Save BACKGROUND: Preservation of the subvalvular apparatus in valve replacement surgery remains controversial and there is no uniform and reliable standard for the selection of surgical procedures for valve diseases. OBJECTIVE: To explore and analyze the effect of valve replacement with preservation of the mitral valve subvalve apparatus to promote effective valve healing.   METHODS: This study enrolled 68 patients with mitral insufficiency as the main disease combined with or without aortic valve disease, who were admitted to Shiyan Taihe Hospital from January 2018 to December 2018 and underwent mitral valve replacement with preservation of the mitral subvalvular apparatus and mitral valve and aortic valve double valve replacement. There were 11 cases with preservation of the subvalvular apparatus of the full valves, 35 cases with preservation of the subvalvular structure of the posterior valve, and 22 cases with complete resection of the mitral valve and the subvalvular chordae tendineae. Echocardiography review was performed 6 months after operation, and cardiac function indexes such as left atrial end-diastolic diameter, left ventricular end-diastolic transverse diameter, left ventricular ejection fraction, left ventricular short-axis shortening rate, left ventricular inflow tract and outflow tract, valve function, and paravalvular healing were compared among the three groups. Complications within 6 months after surgery were recorded. RESULTS AND CONCLUSION: The cardiopulmonary bypass time, blocking time, ICU stay time, and postoperative hospital stay in the full-valve subvalvular apparatus preservation group and the posterior subvalvular apparatus preservation group were all lower than those in the mitral valve and subvalvular chordae tendineectomy group (P < 0.05). There were no significant differences in the levels of cardiac function indexes such as left atrial end-diastolic diameter, left ventricular end-diastolic transverse diameter, left ventricular ejection fraction, and left ventricular short-axis shortening rate among the three groups (P > 0.05). The left atrial anteroposterior diameter at the end of diastole and the left ventricular end-diastolic transverse diameter in the full-valve subvalvular apparatus group were obviously lower than those in the other two groups. The left ventricular ejection fraction and left ventricular short-axis shortening rate in the full-valve subvalvular apparatus group were obviously higher than those in the other two groups. The incidence of complications within 3 years postoperatively showed no significant difference among the three groups. To conclude, although there is no significant difference in the effects of mitral valve replacement with preservation of the mitral subvalvular apparatus and double valve replacement of the mitral valve and aortic valve on the cardiac function of patients with mitral insufficiency as the main disease combined with or without aortic valve disease. However, in terms of data structure, valve healing is more obvious in patients undergoing valve replacement surgery with the mitral subvalvular apparatus preserved, which has certain advantages. Moreover, there is no significant difference in the incidence of complications within 3 years after surgery. Therefore, valve replacement surgery that preserves the mitral subvalvular apparatus can promote valve healing, improve recovery time, promote early recovery and discharge, and reduce the economic burden of patients. Figures and Tables | References | Related Articles | Metrics

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Classification and morphological parameters of scapular foraminal defects and its clinical significance Chen Meixiong, Zheng Kai, Zheng Nansheng, Zhou Li, Xu Mingkui, Yuan Shiguo 2023, 27 (20):  3247-3252.  doi: 10.12307/2023.449 Abstract ( 700 )   PDF (1327KB) ( 110 )   Save BACKGROUND: Scapular foraminal defects can affect the clinical diagnoses and treatments of diseases around the scapula. However, the classification of scapular foraminal defects is lacking, and less is reported on the distribution and morphological parameters of scapular foraminal defects in Chinese people. OBJECTIVE: To investigate the classification, distribution, and morphometric parameters of scapular foraminal defects in dry scapulae in a Chinese population. METHODS: A novel classification system with three categories and six subtypes of scapular foraminal defects was defined. Category I corresponds to the foramina of the scapular body for nerves and blood vessels, Category II corresponds to the variation in non-neurovascular holes or defects, and Category III is a mixture of Category I and Category II. Category I comprises two subtypes: Type A, suprascapular foramina; Type B, nutrient foramina. The scapula is divided into three partitions that correspond to Types C to E in Category II. Category III includes only Type F, a mixture of Type A to Type E. The distribution, classifications and morphometric data for 336 unpaired dry scapulae from Chinese adults were analyzed. RESULTS AND CONCLUSION: The interobserver reliability was excellent. Of 336 scapulae, 92 scapulae had 118 scapular foraminal defects. The incidence rate of scapular foraminal defects was 27.38%, of which Category I to III accounted for 2.81%, 16.96%, and 7.61%, respectively. The proportion of Types A-F was 7.61%, 19.57%, 9.78%, 10.87%, 40.22%, and 11.96%, respectively. The proportion of Type E was the most. The total diameters of Types A-F were (8.09±1.29), (1.98±0.80), (6.60±3.10), (11.19±6.67), (16.80±11.04), and  (17.43±13.88) mm, respectively. The total diameter of Type F was the largest. The total areas of Types A-F were (37.62±19.48), (2.13±2.23), (32.52±24.85), (77.56±124.32), (150.69±181.34), and (109.98±193.50) mm2, respectively. The total area of Type E was the largest. To conclude, variations in scapular foraminal defects are common, which require the attention of clinicians to avoid interfering with the diagnosis due to the appearance of scapular foraminal defects. Attention should also be paid to the treatment at the same time to avoid complications caused by invasive operation.  Figures and Tables | References | Related Articles | Metrics

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Effects of ligustrazine on inflammation and oxidative stress in rat cardiomyocytes induced by lipopolysaccharide Sun Fangyuan, Meng Jialei, Ma Yuhui, Geng Huan, Zhang Tao 2023, 27 (20):  3253-3258.  doi: 10.12307/2023.494 Abstract ( 335 )   PDF (1211KB) ( 142 )   Save BACKGROUND: Sepsis-induced myocardial injury is related to inflammatory response and oxidative stress injury. Previous studies have shown that ligustrazine has many functions such as anti-oxidation and maintaining calcium homeostasis. OBJECTIVE: To investigate the effects of ligustrazine on inflammation and oxidative stress in lipopolysaccharide-induced rat cardiomyocytes.  METHODS: Rat cardiomyocytes were induced by lipopolysaccharide (1 μg/mL) to establish septic cardiomyocyte inflammatory models. Twenty-four hours after intervention with different concentrations of ligustrazine (40, 80, 160 μmol/L), the samples were collected and detected. The effects of ligustrazine on the release of lactate dehydrogenase and reactive oxygen species from H9C2 cells induced by lipopolysaccharide were detected. Apoptosis was observed by fluorescence microscope. RT-qPCR was used to detect the changes of inflammatory factors such as interleukin-1 β, tumor necrosis factor-α, and interleukin-6 as well as the expression of Toll like receptor 4 and MyD88.  RESULTS AND CONCLUSION: Compared with the normal control group, the expression of lactate dehydrogenase and reactive oxygen species in H9C2 cells increased significantly in the model group (P < 0.05) and a large number of cells were stained with propidium iodide under fluorescence microscope. The expression levels of interleukin-1β, tumor necrosis factor-α, and interleukin-6 were significantly increased (P < 0.05) and the expression of Toll like receptor 4 and MyD88 genes were significantly increased in the model group compared with the normal control group (P < 0.05). Compared with the model group, ligustrazine treatment could effectively inhibit the release of lactate dehydrogenase and reactive oxygen species, reduce the number of H9C2 cells stained by propidium iodide, downregulate the expression of interleukin-1β, tumor necrosis factor α and interleukin-6, and suppress the expression of Toll like receptor 4 and MyD88 genes (P < 0.05). To conclude, ligustrazine may reduce intracellular inflammatory factors and oxidative stress by inhibiting the expression of Toll like receptor 4/MyD88. Figures and Tables | References | Related Articles | Metrics

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Role and application of bone morphogenetic protein 2 in the repair of osteochondral defects Wang Buyu, Zhang Yong, Li Feifei, Dong Xiaoyu, Deng Jiang, Ruan Shiqiang 2023, 27 (20):  3259-3265.  doi: 10.12307/2023.482 Abstract ( 399 )   PDF (1195KB) ( 288 )   Save BACKGROUND: Repair of osteochondral defects is a major challenge that orthopedic surgeons face today, because current treatments fail to achieve satisfactory results. With the development of tissue engineering in recent years, new hope has been brought for the repair of osteochondral defects. Bone morphogenetic protein 2 is an important factor involved in bone growth, development, and repair, with good osteoinductive effects. In recent years, it has been found that bone morphogenetic protein 2 also has an effect on cartilage development. Objective: To investigate the research progress of bone morphogenetic protein 2 in the repair of osteochondral defects in recent years and to provide new ideas and strategies for the treatment of osteochondral defects by summarizing the application of bone morphogenetic protein 2 in the repair of various types of bone defects. Methods: WanFang, CNKI, and PubMed databases were searched for literature regarding the mechanism of bone morphogenetic protein 2 in the repair of osteochondral defects and its application in osteochondral tissue engineering published between January 1, 2012 and July 1, 2022. The search terms were “Bone morphogenetic protein 2, Chondrocytes, Cartilage defects, Bone defects, Mesenchymal stem cells, MSC, Signaling pathways” in Chinese and English. Results and Conclusion: Bone morphogenetic protein 2 promotes chondrogenic differentiation of mesenchymal stem cells and can maintain the chondrocyte phenotype to maintain cartilage properties. Bone morphogenetic protein 2 controls chondrogenesis by regulating chondrocyte glucose metabolism. Bone morphogenetic protein 2 acts on osteogenesis through the classical Smad 1/5/8 signaling pathway, but the mechanism of chondrogenesis is not clear. The current research and application of bone morphogenetic protein 2 in tissue engineering mainly focus on the field of bone defect repair, and less is reported on cartilage defect repair. How to prevent bone morphogenetic protein 2 from promoting the transformation of chondrocytes to hypertrophic chondrocytes will be the focus of future research. Figures and Tables | References | Related Articles | Metrics

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hFOB1.19 application in bone tissue engineering Wang Yanyang, Xu Pu 2023, 27 (20):  3266-3272.  doi: 10.12307/2023.444 Abstract ( 1532 )   PDF (1381KB) ( 2508 )   Save BACKGROUND: The biological characteristics of animal osteoblast models widely used at this stage are quite different from those of human cells. The primary human osteoblasts isolated and cultured are difficultly purified, and the culture generations are limited. Osteosarcoma cells have the risk of abnormal growth, which cannot fully meet the research requirements. In 1995, Harris transfected human fetal limb cells with simian vacuolating virus 40, and screened a group of immortalized human osteoblast cell line hFOB1.19 that differentiated into earlier osteogenic progenitor cells. OBJECTIVE: To review the basic characteristics, culture methods and application of hFOB1.19 as a cell model of bone tissue engineering materials.  METHODS: Relevant literature published from 1995 to May 2022 was retrieved in CNKI, WanFang, SinoMed, PubMed, Web of Science, Medicine, and Cochrane Library databases. The keywords were “hFOB1. 19, human osteoblast cell line, immortalized cells, monkey vacuolating virus 40, bone tissue engineering, bone substitute materials, biomaterials research” in Chinese and English, respectively. Finally, 73 papers were screened for review.  RESULTS AND CONCLUSION: hFOB1.19 is highly similar to osteoblasts in vivo in terms of morphology, phenotype, karyotype, biological characteristics, and differentiation potential, which has the ability of rapid and stable growth. hFOB1.19 has been widely used in cell adhesion, proliferation, differentiation, mineralization, gene expression, and protein synthesis to test the biocompatibility and osteogenic properties of bone tissue engineering materials. hFOB1.19 can make up for the shortcomings of different animal cell sources, abnormal proliferation of osteosarcoma cells, difficult separation of primary osteoblasts, and few passages. It has stable performance and good proliferation in bone tissue engineering experiments. However, due to the special culture conditions, limitation of cell passage number and biosafety, it is not widely used in the field of bone tissue engineering materials. At present, the development of bone tissue engineering materials is fast, but there is a lack of a systematic and comprehensive research system and popularization of a unified research standard in the development process. This article summarizes the application of hFOB1.19 in different studies to provide researchers with guidance for method selection and improve relevant standards. Figures and Tables | References | Related Articles | Metrics

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Therapeutic effect of different-frequency repetitive transcranial magnetic stimulations on post-stroke cognitive impairment: a Meta-analysis Yin Yikun, Wang Jialin, Sun Junzhi 2023, 27 (20):  3274-3280.  doi: 10.12307/2023.150 Abstract ( 452 )   PDF (1086KB) ( 784 )   Save OBJECTIVE: Post-stroke cognitive impairment (PSCI) is mainly manifested as impairment of attention, orientation, memory and visuospatial functions. Repetitive transcranial magnetic stimulation can stimulate brain nerve cells, change the action potential of nerve cells, and improve brain cognitive functions such as speech, memory, attention, and executive ability. This study systematically evaluated the efficacy of repetitive transcranial magnetic stimulation in improving cognitive impairment in stroke patients. METHODS: Randomized controlled trials addressing repetitive transcranial magnetic stimulation for cognitive impairment in stroke patients were searched from CNKI, VIP database, PubMed, Cochrane Library, EBSCO, Web of Science and other electronic databases. The retrieval time was from the database inception to December 2021. Montreal Cognitive Assessment Scale, Mini-Mental State Examination, P300 event-related potential (latency, amplitude), and modified Barthel index were used as outcome indicators. The risk assessment of the screened literatures was carried out according to the Cochrane tool described in the Cochrane Handbook for Systematic Reviews of Interventions. RevMan5.4 software was used for Meta-analysis. RESULTS: A total of 16 randomized controlled trials with 847 patients were included. Meta-analysis results showed that Montreal Cognitive Assessment Scale [mean difference (MD)=3.40, 95% confidence interval (CI): 2.57-4.24, P < 0.000 1), Mini-Mental State Examination (MD=2.22, 95% CI: 0.78-3.65, P=0.002), auditory event-related potential P300 latency (MD=-27.11, 95% CI: -35.56 to -18.66, P < 0.000 1) and P300 amplitude (MD=1.91, 95% CI: 1.06-2.76, P < 0.000 1), and modified Barthel index score (MD=6.98, 95% CI: 3.41-10.56, P=0.000 1) were better in the experimental group than the control group. Subgroup analysis showed that low-frequency (≤ 1 Hz) repetitive transcranial magnetic stimulation had a slightly lower effect than high-frequency (> 1 Hz) repetitive transcranial magnetic stimulation in improving cognitive function of stroke patients, but there was no significant difference between them. CONCLUSION: Current evidence suggests that both high- and low-frequency repetitive transcranial magnetic stimulation therapies can effectively improve the cognitive function and daily living ability of stroke patients. Limited by the number and quality of included studies, the above conclusions need to be verified by more high-quality studies. Figures and Tables | References | Related Articles | Metrics