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    18 February 2021, Volume 25 Issue 5 Previous Issue    Next Issue
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    Bone turnover characteristics in patients with alcohol-induced osteonecrosis of the femoral head
    Zheng Xiaolong, He Xiaoming, Gong Shuidi, Pang Fengxiang, Yang Fan, He Wei, Liu Shaojun, Wei Qiushi
    2021, 25 (5):  657-661.  doi: 10.3969/j.issn.2095-4344.2994
    Abstract ( 213 )   PDF (650KB) ( 34 )   Save
    BACKGROUND: The pathological changes of alcohol-induced osteonecrosis of the femoral head (ONFH) are the result of a combination of various factors, and its specific pathogenesis is inconclusive. Related studies have shown abnormal bone metabolism in patients with avascular necrosis of the femoral head.
    OBJECTIVE: To explore the bone turnover markers in patients with alcohol-induced ONFH and to explore the relationship between different stages of ONFH and bone metabolism.
    METHODS: This study retrospectively selected 193 male patients with alcohol-induced ONFH (necrosis group), including 35 cases of ARCO stage II, 131 cases of ARCO stage III and 27 cases of ARCO stage IV. Among them, there were 65 cases of a little drinking of alcohol 52 cases of moderate drinking, and 76 cases of heavy drinking. Another 182 healthy males undergoing physical examination with no history of drinking were taken as control group. The bone turnover markers [procollagen type 1 N-terminal propeptide (P1NP), C-terminal cross-linked telopeptides of type 1 collagen (β-CTX), molecular fragment of N-terminal osteocalcin (N-MID) and 25 hydroxyvitamin D (25OHD)] and biochemical indexes were tested and compared between two groups, followed by logistic regression analysis and correlation analysis. The study protocol was performed in line with the relevant ethic requirements of the First Affiliated Hospital of Guangzhou University of Chinese Medicine. All participants in the trial had been fully informed of the trial process.
    RESULTS AND CONCLUSION: In the necrosis group, P1NP, β-CTX, N-MID, 25OHD, serum calcium, uric acid, alkaline phosphatase, serum phosphorus levels were significantly higher than that of the control group (P < 0.05), and apolipoprotein A1 and high-density lipoprotein levels in the necrosis group were significantly lower than those in the control group (P < 0.05). Compared with patients with low level of alcohol, β-CTX and N-MID levels were significantly reduced in the patients with heavy drinking (P < 0.05). The P1NP level of patients with ARCO stage IV was significantly higher than that of patients with ARCO stage II and III (P < 0.05), and the β-CTX level of patients with ARCO stage IV was significantly higher than that of patients with ARCO stage III (P < 0.05). The correlation analysis results showed that alcohol intake levels were negatively correlated with β-CTX, alkaline phosphatase level was positively correlated with P1NP and β-CTX, and 25OHD was negatively correlated with low-density lipoprotein. Logistic regression analysis results showed that: P1NP (odds ratio=0.984, P=0.004), β-CTX (odds ratio=0.325, P=0.043), and high-density lipoprotein (odds ratio=2.622, P=0.014). To conclude, male patients with alcohol-induced ONFH have active bone formation and bone resorption, and obvious abnormalities in lipid metabolism. The progression of alcohol-induced ONFH can be predicted by these bone turnover markers.
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    Comparison of the Greulich-Pyle method, the CHN method and the China 05 method for assessing bone age in children and adolescents
    Pan Qile, Zhang Hong, Zhou Huikang, Cai Guang
    2021, 25 (5):  662-667.  doi: 10.3969/j.issn.2095-4344.2995
    Abstract ( 479 )   PDF (714KB) ( 45 )   Save
    BACKGROUND: In China, three bone age assessment methods have been widely used in the medical and sports fields, including the  Greulich-Pyle atlas method (GP method), CHN scoring method (CHN method), and China 05 method. A large-sample empirical study is required to determine which method is more suitable for assessing bone age of children and adolescents.
    OBJECTIVE: To provide a scientific evidence for appropriate bone age evaluation standards for children and adolescents in the eastern developed areas, by comparing the GGP method, CHN method and China 05 method based on samples of healthy children from Shanghai. 
    METHODS: A total of 4 152 healthy children and adolescents (2 185 boys and 1 967 girls) from the urban area of Shanghai were selected for the study. Their digital X-ray of the left hand and wrist were collected and evaluated by the GGP method, CHN method and China 05 method. The difference between the bone age and the chronological age was used to assess the applicability of different bone age standards. The study was approved by the Ethics Committee of Shanghai Research Institute of Sports Science, and informed consent was given by all parents of the enrolled students.
    RESULTS AND CONCLUSION: For the GP method, the difference between bone age and chronological age in both genders at the age of ≥ 8 years was -0.12 to -0.65 year with significant difference, except for 8-year-old girls. The significant age difference at the age of ≥ 9 years was 0.18 to 1.62 year, except for the 9-year-old age group. For the CHN method, the difference between bone age and chronological age among 6-17-year-old boys and 6-16-year-old girls was 0.42 to 1.56 years (P < 0.01). For the China 05 method, the difference between bone age and chronological age was 0.20 to 0.53 in 6-16-year-old boys (P < 0.01), 0.08 in 17-year-old boys (P > 0.05), and -0.60 in 18-year-old boys (P < 0.01); the age difference among 6-17-year-old girls was -0.01 to 0.56 year, and the difference was not significant in most age groups. Among the three methods, the result of China 05 method is relatively better, which is the best method that matches the current development of teenagers in Shanghai, suggesting that the China 05 method is more suitable for the eastern developed areas with economic level similar to Shanghai. All the three methods have some limitations. Considering the long-term growth trend of adolescents, it is necessary to revise the current bone age evaluation standards.
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    Delaying cartilage degeneration by regulating the expression of aquaporins in rats with knee osteoarthritis
    Liu Xin, Yan Feihua, Hong Kunhao
    2021, 25 (5):  668-673.  doi: 10.3969/j.issn.2095-4344.2996
    Abstract ( 226 )   PDF (758KB) ( 41 )   Save
    BACKGROUND: Down-regulation of aquaporins (AQPs) can delay cartilage degeneration in knee osteoarthritis, but its specific mechanism is undefined.
    OBJECTIVE: To investigate the effect of p38 MAPK signaling pathway regulating AQP expression on cartilage degeneration in a rat model of knee osteoarthritis. 
    METHODS: Sixty Sprague-Dawley rats were randomly divided into sham operation group (sham), model group, SB203580 low concentration group (SB203580-L,    7.5 mg/kg) and high concentration group (SB203580-H, 30 mg/kg), with 15 rats in each group. The modified Hulth method was used to construct the rat model of knee osteoarthritis. In the sham group, the joint cavity was only opened from the medial side of the right knee joint, without damage to the ligament and meniscus, and with preservation of the articular cartilage surface. Intraperitoneal injection of SB203580 with different concentrations was started at 1 week after surgery, once per week, for 8 weeks in total. The degree of knee joint swelling was measured. The pathological changes of cartilage tissue were observed by hematoxylin-eosin staining, and graded by the Mankin’s score. The expression levels of interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α) and matrix metalloproteinase-13 (MMP-13) in synovial tissue were detected by ELISA. The AQP1, AQP3 mRNA levels in cartilage tissue were detected by RT-PCR, and the protein levels of AQP1, AQP3, cleaved caspase-3, Bax, Bcl-2, p38MAPK and p-p38MAPK in cartilage tissue were detected by western blot. An approval for this study was obtained from the Animal Experimental Ethics Committee of Guangdong Second Traditional Chinese Medicine Hospital With an approval No. 20180125004
    RESULTS AND CONCLUSION: Compared with the sham group, the rats in the model group had severe cartilage injury, the Mankin’s score and the degree of joint swelling were increased significantly (P  < 0.05). Compared with the sham group, the levels of IL-1β, TNF-α and MMP-13 in synovial tissues, the cleaved caspase-3, Bax, and p-p38MAPK protein expression levels in chondrocytes, the mRNA and protein expression levels of AQP1 and AQP3 all increased significantly in the model group (P  < 0.05), whereas the expression level of Bcl-2 protein decreased significantly (P  < 0.05). Compared with the model group, the cartilage injury of rats in the SB203580-H group was improved significantly (P  < 0.05); the Mankin’s score and the degree of joint swelling were decreased significantly (P  < 0.05); the levels of IL-1β, TNF-α and MMP-13 in synovial tissues, the cleaved caspase-3, Bax, and p-p38MAPK protein expression levels in chondrocytes, and the mRNA and protein expression levels of AQP1 and AQP3 all decreased significantly (P  < 0.05), whereas the expression level of Bcl-2 protein increased significantly (P  < 0.05). There was no significant difference between the SB203580-L group and model group (P > 0.05). Therefore, blocking the p38MAPK signaling pathway is deduced to delay the degeneration of articular cartilage in knee osteoarthritis by inhibiting the expression of AQP1 and AQP3.
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    MicroRNA-138-5p regulates chondrocyte proliferation and autophagy
    Ma Zetao, Zeng Hui, Wang Deli, Weng Jian, Feng Song
    2021, 25 (5):  674-678.  doi: 10.3969/j.issn.2095-4344.2997
    Abstract ( 247 )   PDF (664KB) ( 32 )   Save
    BACKGROUND: Abnormal autophagy in chondrocytes often leads to cartilage degeneration, thereby triggering osteoarthritis. Recent studies have found that microRNAs play an important role in chondrocyte autophagy; however, the molecular mechanism is yet unclear. 
    OBJECTIVE: To investigate the role of microRNA-138-5p (miR-138-5p) in the regulation of chondrocyte proliferation and autophagy activities, and to reveal its mechanisms. 
    METHODS: Chondrosarcoma cell line SW1353 were cultured in vitro and transfected with negative control miRNA or miR-138-5p mimic. Cell proliferation activity was measured by cell counting kit-8 assay, the expression of matrix metalloproteinases 1, 3, and 13 mRNA was measured by fluorogenic quantitative PCR. The endogenous LC3 subcellular location was detected by immunofluorescence staining. The miR-138-5p and SIRTI mRNA target sites were predicted using TargetScan 7.1 online tool. Autophagy-related proteins and AMPK signal proteins were detected by immunoblotting assay. 
    RESULTS AND CONCLUSION: Cells transfected with miR-138-5p mimic, compared with those transfected with negative control miRNA, showed lower proliferation activity, less LC3 puncta, and reduced expression of SIRT1, LC3-II, p-AMPK, but increased protein expression of p62 and increased mRNA expression of matrix metalloproteinases 1, 3, 13. There was a conserved miR-138-5p binding site in the 3’UTR region of SIRT1 mRNA. To conclude, miR-138-5p regulates SW1353 cell autophagy and proliferation through the SIRT1/AMPK signaling pathway. The up-regulated expression of miR-138-5p promotes the secretion of matrix metalloproteinases from chondrocytes, indicating that miR-138-5p plays an important role in the progression of osteoarthritis.
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    Expression of interleukin-24 in a mouse model of periapical periodontitis
    Zhang Mi, Wu Saixuan, Dong Ming, Lu Ying, Niu Weidong
    2021, 25 (5):  679-684.  doi: 10.3969/j.issn.2095-4344.2998
    Abstract ( 249 )   PDF (666KB) ( 30 )   Save
    BACKGROUND: Apical periodontitis, often accompanied by periapical bone destruction, is an inflammatory disease of the periapical tissue caused by mixed bacterial infections. Studies have shown that interleukin-24 plays an important role in inflammatory response and affects bone homeostasis as a pleiotropic immune regulator.
    OBJECTIVE: To explore the expression of interleukin-24 during periapical periodontitis in mice.
    METHODS: An induced animal model was constructed in mice to simulate the progression of human apical periodontitis. Twenty-five C57BL/6 female mice at 6 weeks of age were randomly divided into five groups. Mice in the experimental group were anesthetized and the pulps of both mandibular first molars were exposed to the oral cavity for 7, 14, 21, and 28 days, named 1-, 2-, 3-, and 4-week groups. The control group was given no treatments as 0-week group. Mandibular tissues were separated in each group after anesthesia, and then were fixed, decalcified and made into frozen slices of the molar region. Hematoxylin-eosin staining was used to observe the histopathological changes of periapical lesions. Immunohistochemistry staining and RT-PCR were used to detect the expression and changes of interleukin-24 in periapical lesions of mice. The study was approved by the Animal Ethics Committee of Dalian Medical University School of Stomatology. 
    RESULTS AND CONCLUSION: The results of hematoxylin-eosin staining showed that no obvious inflammatory response was observed in the normal periapical region of the 0-week group; the inflammatory cell infiltration area was continuously expanded and the periapical tissue was continuously destroyed in each experimental group, indicating that mouse model of periapical periodontitis was successfully established. The results of immunohistochemistry staining showed that there was almost no interleukin-24 expression in the 0-week group; the expression of interleukin-24 in the periapical region of each experimental group was significantly higher than that in the 0-week group (P < 0.05), and showed a peak expression in the 2-week group. The results of RT-PCR showed that the mRNA level of interleukin-24 in each experimental group was higher than that in the 0-week group (P < 0.05), and peaked at 2 weeks. Overall, our findings indicate that in the animal model of periapical periodontitis, the expression of interleukin-24 increases with a certain trend in the development of periapical lesions, suggesting that it may be involved in the progression of inflammation related to periapical periodontitis.
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    Resting-state functional magnetic resonance imaging evaluation of the brain’s default mode network in patients with sleep bruxism
    Jing Huimin, Yu Wenjuan, Wang Sijia, Chen Cong, Li Yifan, Wang Yonglan, Li Xin, Zhang Juan, Liang Meng
    2021, 25 (5):  685-689.  doi: 10.3969/j.issn.2095-4344.2999
    Abstract ( 224 )   PDF (698KB) ( 60 )   Save
    BACKGROUND: Sleep bruxism is a common oral accessory function, but the etiology is not yet clear. A psychological questionnaire survey on patients with nocturnal molars reveals that sleep bruxism is related to psychological factors, but the specific relationship and mechanism between them are not clear.
    OBJECTIVE: To analyze the changes of brain’s default mode network (DMN) in patients with sleep bruxism and to investigate the DMN status of patients with sleep bruxism. 
    METHODS: From November 2018 to May 2019, 20 patients with sleep bruxism (sleep bruxism group) diagnosed by polysomnography and 20 age-, gender-, and education-matched asymptomatic adults (control group) were recruited in the study. Brain resting-state functional MRI data were collected on a 3.0T scanner during 20:00 to 23:00. The MRI data were analyzed using independent component analysis method to extract the component of DMN. The one-sample t-test was used to make a network component template, and then the two-sample t-test used to compare the DMN components between the two groups.
    RESULTS AND CONCLUSION: Compared with asymptomatic controls, patients with sleep bruxism showed a significantly weaker functional connectivity of the precuneus within the DMN (t=-3.319, P < 0.05), indicating that patients with sleep bruxism show abnormal functional connectivity within the DMN at brain-resting state.
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    Expression and significance of interleukin-21 in intervertebral disc degeneration
    Yang Yang, Yao Yu, Shen Xiaotian, Liu Jiajia, Xue Jianhua
    2021, 25 (5):  690-694.  doi: 10.3969/j.issn.2095-4344.3000
    Abstract ( 253 )   PDF (679KB) ( 40 )   Save
    BACKGROUND: Interleukin-17, as the specific marker of Th17 cells, is highly expressed in degenerative lumbar disc tissue. Interleukin-21 plays an important role in proliferation and differentiation of Th17 cells. 
    OBJECTIVE: To test the expression of interleukin-21 in nucleus pulpous tissue and peripheral blood of patients who accepted the surgery of intervertebral disc degeneration and to analyze the relevant significance.
    METHODS: In total, 40 samples of nucleus pulpous tissues were obtained, 32 from intervertebral disc degeneration (IDD) and 8 from lumbar fracture (control group). Peripheral blood samples were collected from the same cohort. All patients were confirmed by physical examination and MRI and then admitted to the Spine Centre of Affiliated Hospital of Nantong University. We detected the positive signal of interleukin-21 in nucleus pulpous tissue through immunohistochemical method, and measured the levels of interleukin-21, interleukin-17 and interleukin-6 in the nucleus pulpous and peripheral blood by ELISA. Pearson’s correlation analysis was performed thereafter.  
    RESULTS AND CONCLUSION: There were more positive signals of interleukin-21 in the IDD group than the control group. The expression levels of interleukin-21, interleukin-17 and interleukin-6 were significantly higher in the IDD group than the control group as detected by ELISA. The level of interleukin-21 was positively correlated with the expression of interleukin-17 and interleukin-6. These findings indicate that interleukin-21 is involved in inflammatory and autoimmune responses during the development of IDD.
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    Mechanism by which rapamycin protects spinal cord neurons in experimental autoimmune encephalomyelitis mice
    Xie Yang, Zhang Shujiang, Liu Menglan, Luo Ying, Yang Yang, Li Zuoxiao
    2021, 25 (5):  695-700.  doi: 10.3969/j.issn.2095-4344.3001
    Abstract ( 193 )   PDF (850KB) ( 33 )   Save
    BACKGROUND: There are increasing reports about autophagy, but the relationship between the level of autophagy in neurons and the neuroprotection mechanism is not clear.
    OBJECTIVE: To investigate whether rapamycin, an mammalian target of rapacmycin (mTOR) autophagy pathway inhibitor, could activate autophagy by mediating the P70s6k and mTOR protein levels to protect spinal cord neurons in experimental autoimmune encephalomyelitis mice. 
    METHODS: Fifty-four healthy female C57BL/6 mice were divided into three groups: control group, model group and treatment group, with 18 mice in each group. Mice in the model group and treatment group were injected with complete Freund’s adjuvant containing MOG35-55 and pertussis diluent for establishing models of experimental autoimmune encephalomyelitis. At the same time, the mice in the treatment group were given rapamycin (1 mg/kg per day), and those in the model and control groups were given the same amount of normal saline. The mice in the model and treatment were sacrificed at the peak of the onset, and the non-morbid mice, including those in the control group, were sacrificed after 4 weeks of feeding. The spinal cord tissue from each animal was taken to isolate the intumescentia lumbalis of the spinal cord. Nissl staining was used for pathological observation of the spinal cord tissue. Immunofluorescence double staining was used to observe the expression and co-localization of autophagy markers LC3 and NeuN in spinal cord tissue. Western blot was used to detect mTOR, P70S6K proteins and their phosphorylation levels in spinal cord tissue.
    RESULTS AND CONCLUSION: No mice in the control group had an attack, but those in the other groups developed experimental autoimmune encephalomyelitis to different extents. Compared with the model group, the treatment group had prolonged incubation time (P < 0.01), shortened progressive stage (P < 0.01), and decreased neurologic dysfunction score (P < 0.05). Compared with the control group, the model group had the significantly less number of Nissl bodies (P < 0.05), while the number of Nissl bodies in the treatment group was significantly higher than that in the model group, but still lower than that in the control group (P < 0.05). In the model group, LC3 was scattered in the spinal cord neurons and had no obvious dot-like aggregation, whereas in the treatment group, LC3 showed obvious dot-like aggregation, and its distribution was basically consistent with that of NeuN. The phosphorylation levels of mTOR and P70S6K proteins were highest in the model group, followed by the treatment group and control group in turn. To conclude, rapamycin might through inhibiting the phosphorylation levels of mTOR and P70S6K proteins activate the activity of autophagy to protect the spinal cord neurons in experimental autoimmune encephalomyelitis mice.
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    Triptolide improves motor dysfunction in rats following spinal cord injury
    Ma Binxiang, He Wanqing, Zhou Guangchao, Guan Yonglin
    2021, 25 (5):  701-706.  doi: 10.3969/j.issn.2095-4344.3002
    Abstract ( 193 )   PDF (897KB) ( 46 )   Save
    BACKGROUND: One of the main causes of high disability rate of secondary spinal cord injury is oxidative stress and inflammatory response. How to suppress secondary spinal cord injury is a hot topic of current research. 
    OBJECTIVE: To explore the improvement effect of triptolide on motor dysfunction after spinal cord injury and its possible mechanism.
    METHODS: Forty-eight healthy male Sprague-Dawley rats were randomly divided into sham operation group, spinal cord injury group and triptolide group, 16 rats in each group. The modified Allen method was used to establish the rat spinal cord injury model. Rats in the triptolide group received intraperitoneal injection of triptolide (0.1 mg/kg per day) 30 minutes after spinal cord injury. Both the sham operation group and spinal cord injury group were given the same amount of normal saline containing 1% dimethyl sulfoxide via the same route for 10 consecutive days. The sham operation group only underwent laminectomy without damaging spinal cord. Basso-Beattie-Bresnahan scoring method was used to evaluate hindlimb function of Sprague-Dawley rats at 7, 14, 21, 28, and 35 days after surgery. The thoracic spinal cord (T8-11) of the rats was collected on the 10th day after surgery for histological detection, western blot, and real-time quantitative PCR analysis.  
    RESULTS AND CONCLUSION: The behavioral scores in the spinal cord injury group and triptolide group increased with increasing days from injury, and the behavioral scores in the triptolide group were significantly higher than that in the spinal cord injury group at 14, 21, 28, and 35 days after surgery (P < 0.05). Hematoxylin-eosin staining results of the T8-11 sections at 10 days after surgery revealed severe edema, bleeding, and inflammatory cell infiltration in the longitudinal section of the thoracic spinal cord core area of the spinal cord injury group, and these abnormalities could be significantly reduced by triptolide treatment (P < 0.05). RT-PCR results showed that compared with the spinal cord injury group, the expression levels of tumor necrosis factor-α, p-STAT3 and p-JAK2 mRNAs in the spinal cord was significantly decreased (P < 0.05), and the levels of superoxide dismutase and catalase mRNAs were significantly increased in the triptolide group (P < 0.05). Findings from this study confirm that intraperitoneal injection of triptolide can moderately improve motor dysfunction after spinal cord injury. Its mechanism may be related to the abnormal activation of JAK/STAT signaling pathway regulated by triptolide.
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    A modified flap immediate implant is beneficial to soft tissue reconstruction in maxillary aesthetic area
    Zhang Bin, Sun Lihua, Zhang Junhua, Liu Yusan, Cui Caiyun
    2021, 25 (5):  707-712.  doi: 10.3969/j.issn.2095-4344.3003
    Abstract ( 254 )   PDF (791KB) ( 62 )   Save
    BACKGROUND: Studies have shown that conventional flap-guided bone tissue regeneration is a method commonly used  in the immediate implantation of the upper anterior teeth. It is an ideal method for repairing small bone defects, but it is difficult to achieve favorable osteogenesis effect in the tooth neck area.
    OBJECTIVE: To evaluate the clinical effects of modified flap immediate implant in maxillary aesthetic area. 
    METHODS: Sixty cases of immediate implant placement were investigated, and randomized into two groups: 30 received modified flap immediate implant in which a gingiva former with suitable size was screwed followed by buried healing (experiment group) and 30 received traditional flap immediate implant in which a cover screw was screwed followed by buried healing (control group). They were impressed digitally and restored with all-ceramic crown 6 months after the surgery. Implant retention rate, pink esthetic scores, gingival growth and labial bone absorption were detected and compared between two groups. 
    RESULTS AND CONCLUSION: The implant retention rates of both groups were 100%. Compared with the control group, the pink esthetic score and gingival growth were significantly increased in the experimental group (P < 0.05), and the labial bone thickness was significantly decreased (P < 0.05). The proximal, distal or mesial gingiva growth was increased a bit in the experimental group compared with the control group, but there was no significant difference between the two groups (P > 0.05). To conclude, the modified flap immediate implant is conducive to soft tissue reconstruction in maxillary aesthetic area.
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    Effects of Tongbi prescription hot compress combined with acupuncture on mRNA expressions of apoptosis-related genes,Caspase-3 and Bcl-2, in degenerative intervertebral discs
    Xu Yinqin, Shi Hongmei, Wang Guangyi
    2021, 25 (5):  713-718.  doi: 10.3969/j.issn.2095-4344.3004
    Abstract ( 173 )   PDF (757KB) ( 39 )   Save
    BACKGROUND: Acupuncture and hot compress of traditional Chinese medicine therapy have good curative effect on degenerative diseases of the intervertebral disc; however, the combination effect of these two methods on the apoptosis of intervertebral disc cells remains unclear.
    OBJECTIVE: To explore the effects of Tongbi prescription hot compress combined with acupuncture on the mRNA expression of apoptosis-related genes, Caspase-3 and Bcl-2, in intervertebral disc cells in a rabbit model of cervical spondylosis.  
    METHODS: Twenty-four rabbits were randomly divided into normal group, model group, acupuncture group and combination treatment group, with six rabbits in each group. Except for the normal group, animal models of cervical spondylosis were established in the other three groups. After successful modeling, acupuncture treatment (30 minute once a day) and Tongbi Recipe hot compress combined with acupuncture therapy (30 minutes once a day) were done in the acupuncture group and combination treatment group, respectively. The cervical spine X-ray of each rabbit was taken and scored before modeling, at 30 days after molding as well as at the end of the treatment. The C3-4 and C4-5 intervertebral discs of each group were collected after treatment for 15 times. Hematoxylin-eosin staining was used to observe the morphological changes of C3-4 discs, and real-time quantitative PCR was used to detect the relative expression of Caspase-3 and Bcl-2 mRNA in C4-5 intervertebral discs. The study protocol was approved by the Animal Ethics Committee of Guizhou Medical University.
    RESULTS AND CONCLUSION: Compared with the model group, the cervical spine lesions in the acupuncture group and combination treatment group were significantly reduced, with a significant reduction in the X-ray scores (P < 0.05). However, there was no significant difference between these treatment groups. The injury of intervertebral disc tissue and scores in the acupuncture group and combination treatment group were lower than those in the model group, and the score of the combination treatment group was significantly lower than that of the acupuncture group (P < 0.05). Compared with the model group, the expression of Casepase-3 mRNA decreased and the expression of Bcl-2 mRNA increased in the acupuncture group and combination treatment group, and the combination treatment group had lower Casepase-3 mRNA expression and higher Bcl-2 mRNA expression than the acupuncture group (P < 0.05). To conclude, Tongbi prescription hot compress combined with acupuncture therapy can delay the degeneration of intervertebral disc, and its mechanism may be related to the up-regulation of Bcl-2 mRNA expression and down-regulation of Casepase-3 mRNA expression.
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    Effect of mecobalamine combined with mouse nerve growth factor on nerve function recovery after cervical spondylotic myelopathy surgery
    Luo Xuanxiang, Jing Li, Pan Bin, Feng Hu
    2021, 25 (5):  719-722.  doi: 10.3969/j.issn.2095-4344.3005
    Abstract ( 580 )   PDF (613KB) ( 41 )   Save
    BACKGROUND: Cervical spondylotic myelopathy is a common degenerative disease of the cervical spine in spinal surgery. Surgical decompression is the most effective method to prevent the further development of the disease. However, patients are often accompanied by residual neurological symptoms such as sensory and motor dysfunction after surgery. Currently, the drug treatment for postoperative neurological recovery of patients is still unclear clinically.
    OBJECTIVE: To investigate the effect of mecobalamine combined with mouse nerve growth factor on the recovery of nerve function after cervical spondylotic myelopathy surgery.
    METHODS: A total of 82 cases of cervical spondylotic myelopathy from June 2017 to September 2018 in the Affiliated Hospital of Xuzhou Medical University were enrolled in this study. All the patients were randomly divided into observation group and control group with 41 cases in each. Mouse nerve growth factor injection was applied to the control group, and mecobalamine injection was added to the research group. All the patients were treated for 2 weeks. The symptoms and signs in both groups were respectively recorded before and after the treatment, and postoperative residual neurological symptoms were evaluated by Neck Disability Index (NDI) scores and Japanese Orthopedic Association scores. 
    RESULTS AND CONCLUSION: At 1 month, 3 months, 6 months and 1 year after the operation, the NDI scores of the two groups were lower than those before the treatment, and the JOA scores were both getting higher than those before the treatment. The NDI scores in the observation group at 6 months and 1 year after the operation were significantly lower than those in the control group, and as well, the Japanese Orthopedic Association scores were significantly higher than those in the control group. These findings indicate that the combination of mecobalamine and mouse nerve growth factor is helpful to the recovery of nerve function in patients after cervical spondylotic myelopathy surgery, and the effect is better than that of mouse nerve growth factor alone.
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    Mechanism by which Wenban Decoction reduces homocysteine-induced apoptosis of myocardial microvascular endothelial cells in rats
    Zhang Wenwen, Jin Songfeng, Zhao Guoliang, Gong Lihong
    2021, 25 (5):  723-728.  doi: 10.3969/j.issn.2095-4344.3006
    Abstract ( 302 )   PDF (851KB) ( 54 )   Save
    BACKGROUND: Studies have shown that serum homocysteine concentration is an independent predictor of the prevalence and severity of coronary artery disease for patients with normal hypersensitivity C-reactive protein levels. 
    OBJECTIVE: To investigate the protective effect of Wenban Decoction on the apoptosis of rat cardiac microvascular endothelial cells (CMECs) induced by homocysteine by regulating PI3K/Akt signaling pathway. 
    METHODS: Rat CMECs were primarily cultured in vitro, and the cells were randomly divided into control group, model group and Wenban Decoction group (50 mg/L Wenban Decoction). The cells in the latter two groups were injured by 10 mmol/L homocysteine prior to the treatment. Cell counting kit-8 was used to detect the cell viability of each group. ELISA was used to determine serum lactate dehydrogenase, malondialdehyde, whole blood catalase, superoxide dismutase, glutathione peroxidase, interleukin 6, intercellular adhesion molecule 1, interleukin 1β, and tumor necrosis factor α. Flow cytometry was used to detect cell apoptosis after addition of LY294002 based on the treatment with Wenban Decoction. Western blot was used to detect the expression of PI3K, Akt, p-Akt, Bax, Bcl-2 and caspase3 protein in the cells. An ethic approval was given by the Animal Experiment Ethics Committee of Liaoning University of Traditional Chinese Medicine (approval No. 21000092018010).
    RESULTS AND CONCLUSION: Compared with the control group, the survival ability of CMECs in the model group was significantly reduced, the leakage of lactate dehydrogenase was significantly increased, which caused oxidative stress and the release of inflammatory factors, and finally led to a large number of apoptosis. Compared with the model group, Wenban Decoction improved the survival ability of CMECs, reduced the leakage of lactate dehydrogenase, significantly decreased the intracellular levels of interleukin 1β, intercellular adhesion molecule 1, interleukin 6 and tumor necrosis factor α (P < 0.05), as well as reduced the number of apoptotic cells. PI3K inhibitor reversed the inhibitory effect of Wenban Decoction on homocysteine-induced apoptosis of CMECs. To conclude, Wenban Decoction can significantly improve the survival ability of CMECs, reduce the leakage of lactate dehydrogenase, inhibit the level of oxidative stress and the release of inflammatory factors, and ultimately reduce the number of apoptotic cells, which is related to the inhibition of PI3K/Akt signaling pathway.
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    Effect of acupotomy therapy on the expression of Bcl-2/Bax in synovial tissue of collagen-induced arthritis rats
    Liu Qing, Wan Bijiang
    2021, 25 (5):  729-734.  doi: 10.3969/j.issn.2095-4344.3007
    Abstract ( 219 )   PDF (807KB) ( 35 )   Save
    BACKGROUND: It has been shown that acupotomy therapy can improve the symptoms of rheumatoid arthritis, reduce the swelling of the joints, and improve the function of the damaged joints in clinical practice. However, the specific mechanism has not been reported in animal experiments. 
    OBJECTIVE: To observe the effect of acupotomy therapy on the expression of Bcl-2 and Bax in synovial tissue of collagen-induced arthritis rats. 
    METHODS: Forty rats were prepared for the experiment. Ten of them were randomly selected as normal controls, and the other 30 rats were used to manufacture collagen-induced arthritis models via secondary immunization. After the modeling, 20 successful models were randomly selected for subsequent experiments and randomly divided into a model group and an acupotomy group, with 10 in each group. Normal group and model group remain untreated. The medial and lateral patellofemoral ligament, the tibial and fibular collateral ligament, the midpoint of the upper margin of the patella and the midpoint of the patellar ligament around the left knee joint were selected as points of acupotomy treatment, two of which were selected for each acupotomy session. Rats in the acupotomy group were treated once a week for 3 continuous weeks. The general condition of experimental rats was monitored daily, thickness of the left posterior toe was measured and arthritis index was evaluated. Hematoxylin-eosin staining was executed to observe the pathological morphology of the synovial tissue. Real-time quantitative PCR and immunofluorescence staining were implemented to measure gene expression and immunofluorescence intensity of Bcl-2 and Bax in synovial tissue, respectively. The study protocol was approved by the Animal Ethic Committee of Hubei University of Traditional Chinese Medicine with an approval No. 00127518.
    RESULTS AND CONCLUSION: Compared with the normal group, the thickness of the left posterior toe and arthritis index score of the model group were significantly increased (P < 0.01, P < 0.01). Compared with the model group, the left hind toe thickness and arthritis index score of the acupotomy group were decreased (P < 0.05, P < 0.05). Hematoxylin-eosin staining showed that: pathologically, the synovial layers of the model group were thicker, with infiltration of a large number of inflammatory cells. However, the synovial layers of the acupotomy group were fewer with less infiltration of inflammatory cells. RT-PCR showed that there was no significant difference in the expression of Bax mRNA in the synovial tissue between all the groups (P > 0.05). The expressions of Bcl-2 mRNA and Bcl-2/Bax mRNA were significantly reduced in the acupotomy group compared with the model group (P < 0.01, P < 0.01). Immunofluorescence detection indicated that the fluorescence intensity of Bcl-2/Bax in the acupotomy group was significantly reduced compared with the model group (P < 0.01). To conclude, the down-regulation of Bcl-2 expression and Bcl-2/Bax ratio in the synovial tissue may be one of its mechanisms for improving rheumatoid arthritis.
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    Comparison of knee degeneration after anterior cruciate ligament reconstruction with or without remnant preservation
    Xie Chongxin, Zhang Lei
    2021, 25 (5):  735-740.  doi: 10.3969/j.issn.2095-4344.3008
    Abstract ( 245 )   PDF (833KB) ( 33 )   Save
    BACKGROUND:At present, it is still unclear whether remnant-preserving anterior cruciate ligament (ACL) reconstruction can achieve better joint stability and reduce the degeneration of the articular cartilage compared with the traditional operation.
    OBJECTIVE: To compare knee joint degeneration after ACL reconstruction with and without remnant preservation. 
    METHODS: A total of 60 New Zealand rabbits were randomly assigned in equal numbers to a sham group (group A), a conventional ACL reconstruction group (group B), a remnant-preserving and tensioning ACL reconstruction group (group C), and a remnant-preserving and graft through sleeve ACL reconstruction group (group D). In the group A, the joint capsule was cut open without dissection of the ACL, and corresponding rabbit models were established in the other three groups. Healing conditions of the remnant and tendon graft between groups C and D were observed at 12 weeks post-surgery, and the biomechanical properties of the tendon graft were determined by the pull-off test. Cartilage degeneration was assessed by Mankin score, following hematoxylin-eosin and Masson staining of the medial tibial plateau, and chondrocyte apoptosis examined using TUNEL assay. Expression of matrix metalloproteinase 13, proteoglycan and Bax proteins was measured using western blot assay. 
    RESULTS AND CONCLUSION: Healing of remnant to the tendon grafts was not observed in groups C and D at 12 weeks post-surgery. In addition to the presence of articular cartilage degeneration, the Mankin score, cartilage apoptotic index, and expression of matrix metalloproteinase 13 and Bax proteins in groups B, C and D were all significantly higher than those of group A (P  < 0.05), and the expression of proteoglycan in the three groups was lower than that in group A (P < 0.05), with no significant difference among the B, C, and D groups (P  > 0.05). To conclude, there are no significant differences in articular cartilage degeneration following remnant-preserving and conventional ACL construction, and this may be associated with the absence of increased knee stability by remnant preservation. 
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    Expression of long chain non-coding RNA PGM5-AS1 in serum of renal transplant patients and its regulation of human glomerular endothelial cells
    Jiang Xin, Qiao Liangwei, Sun Dong, Li Ming, Fang Jun, Qu Qingshan
    2021, 25 (5):  741-745.  doi: 10.3969/j.issn.2095-4344.3009
    Abstract ( 221 )   PDF (720KB) ( 30 )   Save
    BACKGROUND: The expression of long chain non-coding RNA PGM5-AS1 (lnc RNA PGM5-AS1) is reduced in peripheral blood of patients with acute immune rejection after renal transplantation. The effect of its expression on human glomerular endothelial cell (HRGEC) survival and macrophage chemotaxis remains to be studied.
    OBJECTIVE: To investigate the expression of lnc RNA PGM5-AS1 in serum of patients with acute rejection of renal transplantation and non-acute rejection patients and its effect on proliferation, cell cycle, apoptosis and macrophages chemotropism of HRGECs.
    METHODS: Forty-six patients with renal transplantation were divided into acute rejection group (n=17) and non-acute rejection group (n=29) according to whether or not acute rejection occurred within 1 month after operation. Peripheral blood sample was drawn from each patient at 1 day before operation, 1, 2, 3, and 4 weeks after operation. qRT-PCR was used to detect the expression of PGM5-AS1 in serum of patients with or without acute rejection. si-control and si-PGM5-AS1 were transfected into glomerular endothelial cells, and the expression of PGM5-AS1 in the transfected cells was detected by qPCR. MTT was used to detect cell proliferation, flow cytometry was applied to detect apoptosis and cell cycle, ELISA was adopted to detect cellular inflammatory factor secretion, and Transwell was used to detect chemotaxis of macrophages. Approval for this study was obtained from the Ethics Committee of Zhengzhou People’s Hospital, and all patients signed informed consent prior to the participation in the trial.
    RESULTS AND CONCLUSION: Compared with non-acute rejection patients, patients with acute rejection to renal transplantation had significantly lower PGM5-AS1 expression in serum at 1, 2, 3, and 4 weeks after transplantation (P < 0.05). After PGM5-AS1 silencing, the expression of GM5-AS1 in HRGEC cells was significantly lower than that in the si-control group and normal control group (F=379.658, P < 0.05). MTT results showed that PGM5-AS1 silencing significantly inhibited HRGEC cell proliferation (P < 0.05). Flow cytometry results showed that PGM5-AS1 silencing induced HRGEC cell arrest in G0/G1 phase and increased apoptosis (P < 0.05). ELISA results showed that PGM5-AS1 silencing inhibited interleukin-13 expression and increased interleukin-6, interferon-γ and tumor necrosis factor-α expression (P < 0.05). Transwell results indicated that HRGEC cells silenced by PGM5-AS1 significantly increased the chemotaxis of macrophages (P < 0.05). All these findings indicate that PGM5-AS1 is lowly expressed in serum of patients with acute rejection of renal transplantation, and inhibition of PGM5-AS1 can promote HRGEC cell damage, which can be used as a peripheral blood diagnostic marker for early acute rejection, and may be a molecular target for the treatment of acute rejection.
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    Mechanism of “Ruxiang-Moyao” herbal pair in the treatment of knee osteoarthritis based on network pharmacology
    Cao Xuhan, Bai Zixing, Sun Chengyi, Yang Yanjun, Sun Weidong
    2021, 25 (5):  746-753.  doi: 10.3969/j.issn.2095-4344.3010
    Abstract ( 298 )   PDF (1347KB) ( 45 )   Save
    BACKGROUND: “Ruxiang-Moyao”  is a common combination of traditional Chinese medicine in the clinical treatment of knee osteoarthritis. However, the pharmacological mechanism is not yet clear. 
    OBJECTIVE: Using network pharmacology technology to explore the therapeutic target of “Ruxiang-Moyao” as a commonly used traditional Chinese medicine for clinical treatment of arthromyodynia in the treatment of knee osteoarthritis and the relevant mechanism. 
    METHODS: The chemical constituents of “Ruxiang-Moyao” were collected by TCMSP pharmacology database and analysis platform, and the possible bioactive constituents of frankincense and myrrh were screened according to the biological oral availability ≥ 30% and class drug properties ≥ 0.18. The possible targets of each active constituent were screened out using the protein database (Uniprot). GeneCards, OMIM, TTD and DrugBank databases were consulted to mine knee osteoarthritis-associated gene targets. The disease-drug protein target genes were obtained after the intersection of the above-mentioned screening data. The STRING database calculation and analysis algorithm was used to screen out important key genes to build a protein-protein interaction network, and the key target genes were uploaded to the PPI network graph. Cytoscape software was used to map the drug-target and disease-target visualization network, and DAVID online tool was further used for gene ontology (GO) analysis and Kyoto Encyclopedia of Genes and Gnomes (KEGG) pathway enrichment analysis. 
    RESULTS AND CONCLUSION: Eight bioactive constituents of frankincense and 45 bioactive constituents of myrrh were obtained. At the same time, 412 target proteins of “Ruxiang-Moyao,” 1889 genes related to knee osteoarthritis and 105 co-targets of drugs and diseases were detected. The protein-protein interaction network found that AKT1, TP53, IL6, TNF, JUN, and MAPK1 might be the key targets of  “Ruxiang-Moyao” in the treatment of knee osteoarthritis. The GO enrichment analysis identified 66 GO items, which are involved in cell response to hypoxia, negative regulation of epithelial cell proliferation, immune response, insulin-stimulated cell response, and positive regulation of fibroblast proliferation. The enrichment analysis of KEGG pathway identified 95 related signaling pathways, which are involved in inflammation, cell apoptosis and cell senescence. David enrichment analysis showed that the key target of “Ruxiang-Moyao” intervention for knee osteoarthritis was mainly related to several biological processes such as inflammatory response, cell apoptosis and immune system. Overall, “Ruxiang-Moyao” has the characteristics of multi-pathway and multi-target action in the treatment of knee osteoarthritis, and mainly has anti-inflammatory and analgesic effects. The key targets of its action and the involved biological process and signaling pathway have been preliminarily revealed, providing a new idea for the clinical prescription treatment of knee osteoarthritis in the future.
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    The role of Hedgehog signaling pathway in transforming growth factor beta1-induced myofibroblast transdifferentiation
    Nie Huijuan, Huang Zhichun
    2021, 25 (5):  754-760.  doi: 10.3969/j.issn.2095-4344.3011
    Abstract ( 269 )   PDF (876KB) ( 38 )   Save
    BACKGROUND: In recent years, increasing studies have shown that abnormally activated Hedghog signaling pathway is widely involved in the injury repair of systemic multi-tissue organ diseases, but its related role in myocardial fibrosis is still unclear.
    OBJECTIVE: To study the effect of blocking Hedgehog-Gli signaling pathway on epithelial-mesenchymal transition of myocardial fibroblasts induced by transforming growth factor-β1 (TGF-β1).
    METHODS: (1) Animal experiment: The model of myocardial infarction in mice was established by left coronary artery ligature. The pathological changes of myocardial tissue were observed by hematoxylin-eosin staining and Masson staining at 3, 7, and 14 days after infarction. The mRNA, protein and spatio-temporal expressions of Gli1 at different time points were detected by RT-PC, western blot and immunofluorescence. The mRNA and protein expression changes of Gli1 before and after adding inhibitor were detected using RT-PCR and immunofluorescence. (2) Cell experiment: The primary cultured myocardial fibroblasts of neonatal Sprague-Dawley rats were cultured by differential adherent method. The cultured myocardial fibroblasts were cultured with 0, 1, 5, and 10 μg/L TGF-β1 intervention. The expression of Gli1 mRNA and protein was detected by RT-PCR and western blot, respectively. Myocardial fibroblasts were induced by TGF-β1 of 10 μg/L for 24 hours, and the spatio-temporal expression of Gli1 protein and expression of epithelial-mesenchymal transition markers were detected by immunofluorescence. Gli1 and Smo in Hedgehog signaling pathway were specifically blocked by GANT61 and Cyclopamine for 24 hours, respectively, and 10 μg/L TGF-β1 was added. The mRNA expression level of Gli1 was detected by RT-PCR, and the spatio-temporal expression of Gli1 and expression of epithelial-mesenchymal transition markers were detected by immunofluorescence.
    RESULTS AND CONCLUSION: (1) Results of the animal experiment: As the time after infarction prolonged, the mRNA and protein expression levels of Gli1 in the mouse myocardium gradually increased, but the mRNA and protein expressions of Gli1 in the infarcted tissue decreased after addition of the Hedgehog pathway specific blocker. (2) Results of the cell experiment: After stimulation with TGF-β1, the epithelial mesenchyme of myocardial fibroblasts transformed into myofibroblasts (positive for a-SMA). As the intervention dose of TGF-β1 increased, the mRNA and protein expression of Gli1 gradually increased. After specifically blocking Gli1 and Smo in the Hedgehog signaling pathway for 24 hours, the mRNA and protein expression levels of Gli1 were inhibited, accompanied by the changes in the expression of E-Cadherin and Vimentin during epithelial-mesenchymal transition, indicating the existence of epithelial-mesenchymal transition. By the combination of in vivo and in vitro experiments, this study confirmed that the Hedgehog-Gli signal pathway is involved in the occurrence and development of myocardial fibrosis and myocardial fibroblast transdifferentiation.
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    Application of 3D-printed coplanar template combined with fixed needle technique in percutaneous accurate biopsy of small pulmonary nodules
    Xu Junma, Yu Yuechao, Liu Zhi, Liu Yu, Wang Feitong
    2021, 25 (5):  761-764.  doi: 10.3969/j.issn.2095-4344.3012
    Abstract ( 221 )   PDF (806KB) ( 43 )   Save
    BACKGROUND: Percutaneous lung biopsy is an important method to clarify the nature of lung nodules. However, the lungs are more active due to the presence of respiratory motion. Percutaneous lung biopsy, especially for small lung nodules, is difficult.
    OBJECTIVE: To introduce the application of 3D-printed coplanar template combined with fixed needle technique in percutaneous biopsy of small pulmonary nodules.
    METHODS: A total of 24 patients who had percutaneous lung biopsy in the Second Affiliated Hospital of Xuzhou Medical University from July 2018 to April 2019 were enrolled. Imaging examination indicated small pulmonary nodules with a nodule diameter of 8-30 mm in all the patients. According to the probability of malignancy, all tumors were of the middle and high risk grade, and there were indications for percutaneous lung biopsy. All the patients were randomized into two groups (n=12 per group): the control group underwent free hand biopsy, and the observation group underwent percutaneous lung biopsy guided by 3D printed coplanar template combined with fixed needle. The number of puncture needle adjustments, number of CT scans, positive rate of specimens, and incidence of complications were recorded and compared between the two groups. Approval for this trial was obtained from the Ethics Committee of the Second Affiliated Hospital of Xuzhou Medical University.
    RESULTS AND CONCLUSION: The number of puncture needle adjustments, the number of CT scans and the incidence of pneumothorax during the operation were significantly lower in the observation group than the control group (P < 0.05), whereas there were no significant differences in the positive rate of specimens and incidence of bleeding complications between the two groups (P > 0.05). These findings indicate that the 3D-printed coplanar template combined with fixed needle technique can relatively fix the target lesion, reduce the number of needle adjustments and number of CT scans, reduce iatrogenic radiation, and reduce the incidence of complications, especially pneumothorax.
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    Molecular mechanism of Eucommia ulmoides active ingredients treating synovitis of knee osteoarthritis: an analysis based on network pharmacology
    Li Yonghua, Feng Qiang, Tan Renting, Huang Shifu, Qiu Jinlong, Yin Heng
    2021, 25 (5):  765-771.  doi: 10.3969/j.issn.2095-4344.3013
    Abstract ( 252 )   PDF (1201KB) ( 66 )   Save
    BACKGROUND: Studies have shown that the active ingredients of Eucommia ulmoides can inhibit the growth of synovial fibroblasts of arthritis, but the molecular mechanism underlying  Eucommia ulmoides  treating synovial inflammation of knee osteoarthritis is not yet clear.
    OBJECTIVE: To investigate the possible molecular mechanism of  Eucommia ulmoides  ingredients treating synovitis of knee osteoarthritis based on the network pharmacology and bioinformatics. 
    METHODS: The active ingredients and targets of  Eucommia ulmoides  were screened by using the TCM systematic pharmacological analysis platform (TCMSP). The disease targets corresponding to synovitis of knee osteoarthritis were obtained from Gene Expression Omnibus (GEO). The protein-protein interaction network was constructed by STRING online database, analyzed and showed by the Cytoscape software. Gene ontology analysis and Kyoto encyclopedia of genes and genomes pathway enrichment analysis of the targets were conducted by R/Bioconductor. The enrichment results were obtained with a significant difference (P < 0.05). 
    RESULTS AND CONCLUSION: A total of 21 active ingredients of Eucommia ulmoides, including eugenol, eucommia glycoside, eucommia liposide A, quercetin, and 25 different expressed genes (7 up-regulated and 18 down-regulated differentially expressed genes) were obtained. The main biological processes of the differentially expressed genes were found different significantly in cytokine activity, cytokine receptor binding and activating transcription factor binding. The hub genes, such as interleukin-6, vascular endothelial growth factor A and chemotactic factor 8, may be regulated by the active ingredients to influence the signal of MAPK/NF-kappa B/Toll-like receptor signaling pathway. Based on the network pharmacology and bioinformatics approach, the study preliminarily validates the molecular mechanism of Eucommia ulmoides for treatment of synovitis of knee osteoarthritis, which may lay a foundation for further pharmacodynamics study and experiment.
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    An insight into the mechanism of Salvia miltiorrhiza intervention on osteoporosis based on network pharmacology
    Xiao Fangjun, Chen Shudong, Luan Jiyao, Hou Yu, He Kun, Lin Dingkun
    2021, 25 (5):  772-778.  doi: 10.3969/j.issn.2095-4344.3014
    Abstract ( 251 )   PDF (1540KB) ( 62 )   Save
    BACKGROUND: Salvia miltiorrhiza has been widely used in the treatment of osteoporosis in recent years, but its mechanism in human body is not clear.
    OBJECTIVE: To screen the main active components of Salvia miltiorrhiza and their corresponding targets by the method of network pharmacology, to construct the active components-action target-disease network of traditional Chinese medicine, and to explore the mechanism of Salvia miltiorrhiza in the treatment of osteoporosis. 
    METHODS: Firstly, the main active components and related targets of Salvia miltiorrhiza were selected based on the characteristics of pharmacokinetics in the Traditional Chinese Medicine System Pharmacology Database and Analysis Platform (TCMSP), and then the known therapeutic targets of osteoporosis were screened from the Online Mendelian Inheritance in Man (OMIM) and GeneCards database. The Salvia miltiorrhiza target and osteoporosis target were intersected to obtain the common target of traditional Chinese medicine and disease. The traditional Chinese medicine-active ingredient-target-disease regulation network was topologically constructed by using Cytoscape software, and the protein-protein interaction network and bar graph of traditional Chinese medicine-disease target were constructed by using String database to screen the core targets. Finally, the traditional Chinese medicine-disease target was analyzed by GO functional enrichment analysis and KEGG pathway enrichment analysis.
    RESULTS AND CONCLUSION: A total of 65 active ingredients of Salvia miltiorrhiza, 932 related targets, 3 264 disease targets, and 74 Chinese medicine-disease interaction targets were obtained through screening. The analysis based on network pharmacology indicated that the potential active ingredients of Salvia miltiorrhiza in treating osteoporosis mainly include luteolin, tanshinone IIa, cryptotanshinone, etc. These components can activate target proteins such as AKT1, interleukin-6, vascular endothelial growth factor A, and MAPK1 to activate PI3K-Akt, interleukin-17, hypoxia-inducible factor-1, AGE-RAGE signaling pathway in diabetic complications. These activated signaling pathways can directly or indirectly participate in cell differentiation and apoptosis, metabolism, oxidative stress, and inflammatory response in the treatment of osteoporosis. The role of Salvia miltiorrhiza can be further confirmed by its multi-component, multi-target, and multi-system effects on osteoporosis. However, the above conclusions and specific mechanism of action need to be further studied.
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    The current situation of knowledge and attitudes towards organ, eye tissue, body donation of residents in Shenyang
    Yang Xin, Jin Zhe, Feng Xu, Lu Bing
    2021, 25 (5):  779-784.  doi: 10.3969/j.issn.2095-4344.3015
    Abstract ( 229 )   PDF (648KB) ( 59 )   Save
    BACKGROUND: China has stopped using the organs from executed prisoners for transplantation since 2015, and organ donation has become the only source of organ transplantation. Therefore, it is very important to fully understand the current status of organ donation. 
    OBJECTIVE: To understand the current status of organ, eye tissue and body donation awareness and attitude of residents in Shenyang city, and to analyze the influencing factors. 
    METHODS: A total of 10 000 residents of Shenyang were sampled by multistage cluster sampling. There were 8 942 valid questionnaires, and the effective rate was 93.43%. 
    RESULTS AND CONCLUSION: During the survey, 8 209 people heard of organ donation accounting for 91.8% of all the respondents, and 6 251 people were willing to make a survey about organ/body donation, accounting for 69.9%. With cognition and intention as dependent variables and own situation as independent variable, we made a Logistic regression analysis and found that the cognitive level was related to gender, education level, family per capita monthly income, occupation, political affiliation and religion. The corresponding odds ratio (OR) (known/unknown) values were: for males, the OR=0.786; for the education level of primary education or below, OR=0.188; for the monthly income per capita of 2 000-5 000 yuan, OR=1.418; for farmer, OR=1.593; for communist youth league member, OR=1.313; for non-religious, OR=1.810. Respondents' attitudes were related to gender, age, education level, marital status, family per capita income, occupation, political affiliation and religion. The corresponding OR (willing/unwilling) values were: for male, OR=1.131; for age ≤ 30 years, OR=1.266; for primary education or below, OR=2.090; for married, OR=0.841; for the monthly income per capita < 2 000 yuan, OR=1.253; for public institution or government department, OR=0.740; for the member of Communist Party of China, OR=0.799; for no religious belief, OR=0.842 (P < 0.05). The above data confirm that Shenyang residents have a high awareness and willingness of organ, eye tissue and body donation in general; however, further publicity is still needed to establish an effective organ donation management system.
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    An insight into biomarkers of osteoarthritis synovium based on bioinformatics
    Song Shan, Hu Fangyuan, Qiao Jun, Wang Jia, Zhang Shengxiao, Li Xiaofeng
    2021, 25 (5):  785-790.  doi: 10.3969/j.issn.2095-4344.3016
    Abstract ( 269 )   PDF (1139KB) ( 38 )   Save
    BACKGROUND: Over 90% of patients with osteoarthritis suffer from synovial lesions, and the occurrence of synovitis may also promote cartilage degeneration. To explore the potential mechanisms of synovial lesions is beneficial to find precise treatment targets and achieve good long-term prognosis.
    OBJECTIVE: To identify candidate hub genes in the synovial tissues during the development of osteoarthritis by bioinformatics analysis, and to further provide new insights for osteoarthritis study.
    METHODS: The osteoarthritis synovial-associated chip data sets GSE82107, GSE12021, GSE55457, and GSE55235 were downloaded from the public database GEO, including 40 cases of osteoarthritic synovial tissue and 36 cases of normal synovial tissue. R software was used to screen differentially expressed genes  using Gene ontology function enrichment and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis, and hub genes are selected by STRING online analysis tool and Cytoscape software.
    RESULTS AND CONCLUSION: Total 447 differentially expressed genes were selected between osteoarthritis and healthy control, including 201 up-regulated and 246 down-regulated genes. These differentially expressed genes were mainly enriched in inflammatory response, regulations of extracellular matrix, collagen, blood vessel development, as well as MAPK signaling pathway, tumor necrosis factor signaling pathway, arachidonic acid metabolism. Eight hub genes were screened by analyzing the protein interactions, which included matrix metalloproteinase 9, interleukin 6, vascular endothelial growth factor A, JUN, prostaglandin peroxide enzyme 2, CXCL8, MYC, epidermal growth factor receptor. The hub genes selected by bioinformatics analysis such as vascular endothelial growth factor A, matrix metalloproteinase 9, JUN, and prostaglandin peroxidase 2 may be biomarkers for the diagnosis of osteoarthritis and potential targets for treatment.
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    Mechanism of DNA methylation in exercise intervention for osteoporosis
    Liu Bo, Chen Xianghe, Yang Kang, Yu Huilin, Lu Pengcheng
    2021, 25 (5):  791-797.  doi: 10.3969/j.issn.2095-4344.3017
    Abstract ( 331 )   PDF (736KB) ( 62 )   Save
    BACKGROUND: The dynamic balance between bone formation mediated by osteoblasts and bone resorption mediated by osteoclasts is the basis for maintaining the stability of the body’s bone tissue. The metabolic imbalance between them can cause bone loss and fine structure degeneration of the bone cells, leading to osteoporosis. 
    OBJECTIVE: To review the role of DNA methylation in osteoporosis and to explore the mechanism of exercise affecting DNA methylation and DNA methylation regulating bone metabolism.
    METHODS: A computer-based search of PubMed and CNKI databases was performed for relevant articles published from January 2002 to April 2020 with “DNA methylation; Osteoporosis; Exercise intervention; Mechanical stress; Osteogenic differentiation” as key words in English and Chinese, respectively. Initially, finally 52 eligible articles were included for result analysis. 
    RESULTS AND CONCLUSION: DNA methylation is a relatively conservative and stable apparent modification, which regulates gene expression, silencing and disease occurrence. Studies have shown that reduced methylation levels of genes such as β-catenin, Runx2, osteopontin (OPG) can promote their expression and activate Wnt Pathway, whereas the reduction of methylation level of Sclerosin, receptor activator of nuclear factor kappa B ligand (RANKL) and other genes can promote their expression, and inhibit Wnt pathway and reduce the ratio of OPG/RANKL, thereby affecting the proliferation, differentiation and function of osteoblasts and osteoclasts, and accordingly regulating dynamic equilibrium between bone formation and bone resorption. Osteoblasts and osteoclasts act as sensitive cells for mechanical stimulation. Bone can transform the mechanical load generated by exercise into biological stimulation that acts on the differentiation and function of related bone cells, thereby regulating bone metabolism. In vitro experiments have indicated that different forms of mechanical stress stimulations can change the methylation level of genes such as OPN and GNAS1 to regulate their expression, which has a positive effect on bone formation. Bone tissue is a mechanically sensitive tissue, and DNA methylation can regulate bone metabolism by regulating a variety of factors. 
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    Role and application of bone morphogenetic proteins in articular cartilage regeneration
    Deng Zhenhan, Huang Yong, Xiao Lulu, Chen Yulin, Zhu Weimin, Lu Wei, Wang Daping
    2021, 25 (5):  798-806.  doi: 10.3969/j.issn.2095-4344.3018
    Abstract ( 310 )   PDF (636KB) ( 47 )   Save
    BACKGROUND: Articular cartilage degeneration is the main cause of osteoarthritis. Bone morphogenetic proteins play an important role in cartilage regeneration and repair.
    OBJECTIVE: To review the research progress of bone morphogenetic protein in the process of articular cartilage regeneration.
    METHODS: A computer-based online search of PubMed and Elsevier databases was performed using the keywords “bone morphogenetic proteins, BMPs, arthritis, osteoarthritis, OA, cartilage, chondrocyte” in English. A total of 272 papers were retrieved, 96 of which were included in final analysis. Another 27 papers related to concepts were also included. Therefore, 123 papers are finally included.
    RESULTS AND CONCLUSION: Bone morphogenetic proteins participate in many biological processes including cell proliferation, differentiation, migration, and apoptosis, and play an important role in the formation of bone and cartilage. Bone morphogenetic proteins participate in a variety of signaling pathway cascades by binding to different receptors, which can protect articular cartilage from cartilage destruction caused by inflammation and trauma. Bone morphogenetic proteins alone or in combination with other cytokines can repair cartilage defects improve degenerative lesions, and promote the differentiation and regeneration of articular chondrocytes. However, there are still some practical problems that need to be solved for the widespread use of bone morphogenetic proteins in cartilage regeneration, such as the safety of drug transporters, the lack of effective biological scaffold materials, the optimal dosage and time point of use of biological agents, and their toxic and side effects. Future research will focus on how to solve the above problems. The widespread application of bone morphogenetic proteins will open a new era for targeted treatment of cartilage damage and cartilage degenerative diseases represented by osteoarthritis.
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    The global competitive situation of cardiac tissue engineering based on patent analysis
    Xu Dongzi, Zhang Ting, Ouyang Zhaolian
    2021, 25 (5):  807-812.  doi: 10.3969/j.issn.2095-4344.3019
    Abstract ( 231 )   PDF (688KB) ( 55 )   Save
    BACKGROUND: Tissue engineering is the best way to repair or replace organ failure or tissue defects. From the theoretical point of view to the research in many fields, tissue engineering has developed very rapidly. As cardiovascular disease has become one of the most dangerous factors that endanger human health, research on cardiac tissue engineering has made tremendous progress.
    OBJECTIVE: To provide advices and references for the development and innovation of cardiac tissue engineering based on patent analysis in the field of cardiac tissue engineering that reveals the global competitive situation of cardiac tissue engineering.
    METHODS: We qualitatively and quantitatively analyze the number and development tendency, technology birthplaces, target markets, applicants, inventors and technical fields of patents in the field of liver tissue engineering in nearly 20 years. The data visualization method is used to present the current status and tendency of global cardiac tissue engineering technology development and China's development status.
    RESULTS AND CONCLUSION: The technology for cardiac tissue engineering has been evolved rapidly and highly innovative, with invention patents accounting for 93.91%. The scale of technological development in the field of cardiac tissue engineering in China is not expanded as large as that in the United States, but the growth rate is far faster than that in the United States, and it has accumulated a certain number of high-quality technological achievements, becoming the second largest source of technology in the world. The United States is the most concerned target market in the field of cardiac tissue engineering, and the Chinese market currently ranks fourth. Chinese cardiac tissue engineering has developed rapidly in recent years, growing much faster than that in the United States. There are three institutions in China that has mounted to the front of the world in terms of technology development. Due to a lack of technological achievements with high potential market value, there is still a certain gap between China and the United States. Chinese inventors have achieved certain results in this field, mainly in enterprises. The main fields of technological innovation in this field are heart valve covering materials and cell scaffolds. Through an in-depth and comprehensive analysis of patents in the field of cardiac tissue engineering, it can be found that the global development of cardiac tissue engineering is active and innovative, but generally lacks high-quality results. The United States is the most concerned target market in this field and the most important technology birthplace, whereas China is at a stage of rapid development in this field. In order to improve technological competitiveness, enterprises and universities should strengthen cooperation and focus on research hot spots.
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    Neurogenesis and neuroinflammation under exercise: alteration and regulation
    Zhao Xiang, Wei Cuilan, Zhang Yeting
    2021, 25 (5):  813-820.  doi: 10.3969/j.issn.2095-4344.3020
    Abstract ( 246 )   PDF (703KB) ( 71 )   Save
    BACKGROUND: Studies have found that exercise can regulate neuroinflammation and adult hippocampal neurogenesis, and improve cognitive function. However, the interaction between exercise and neuroinflammation-induced changes in hippocampal neurogenesis and cognitive function is unclear.
    OBJECTIVE: To analyze and summarize the mechanism by which neuroinflammation-mediated exercise improves adult hippocampus neurogenesis and cognition.
    METHOD: Using “exercise; neuroinflammation; neurogenesis; Alzheimer’s disease; aging; depression; cerebral ischemia; traumatic brain injury; cognition” as keywords, we retrieved literature regarding the effects or mechanisms of exercise on neuroinflammation, adult hippocampus neurogenesis and cognition in CNKI and Web of Science, and logically analyzed and summarized the included studies.
    RESULTS AND CONCLUSION: Neuroinflammation can cause nerve damage, and may lead to cognitive impairment, while exercise can effectively produce anti-inflammatory effects in the brain, and promote adult hippocampal neurogenesis, thereby improving cognitive ability. Exercise cannot only directly increase the expression of neurotrophic factors in the hippocampus and promote hippocampal neurogenesis, but also affect the expression of neurotrophic factors in the hippocampus and the process of neurogenesis through its anti-inflammatory mechanism. However, there is still much work to be done to better understand how the neurogenic and inflammatory environment changes under exercise conditions, and how this altered process can be modulated to promote cognition.
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