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    18 June 2020, Volume 24 Issue 17 Previous Issue    Next Issue
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    Effects and mechanisms of sodium ibandronate in ovariectomy-induced osteoporosis rats
    Chen Chaoxiang, Zhou Shuping, Zhang Wei, Xiang Liang, Hou Wei, Wu Junxing
    2020, 24 (17):  2625-2629.  doi: 10.3969/j.issn.2095-4344.2576
    Abstract ( 307 )   PDF (20760KB) ( 110 )   Save

    BACKGROUND: Bisphosphonates are a novel inhibitor of bone resorption that can inhibit the activity and function of osteoclasts.

    OBJECTIVE: To observe the effects of sodium ibandronate on the expression of dentin matrix protein 1, Caspace3, Bcl-2 and Bax in condylar cartilage in osteoporosis rats.

    METHODS: Thirty-six female rats were randomly divided into sham group, osteoporosis group and sodium ibandronate group, twelve in each group. The sham group did not excise ovaries during surgery. Bilateral ovaries of rats were removed in the osteoporosis and sodium ibandronate groups. On the 7th day after operation, rats in the sodium ibandronate group were intraperitoneally given sodium ibandronate 10 μg/kg, once every 7 days. After 90 days, the rat ovaries were taken. Bone mineral density was measured in each group. The changes of condylar cartilage were observed by toluidine blue staining and TUNEL staining. The expression of dentin matrix protein 1 protein was detected by immunohistochemistry. The levels of Caspase3, Bcl-2 and Bax were detected by western blot assay. The study protocol was approved by the Animal Ethics Committee of Nanhua Hospital in China with the approval No. SLXD_201804010. 

    RESULTS AND CONCLUSION: Compared with the sham group or sodium ibandronate group, the bone mineral density in the osteoporosis group was significantly decreased (P < 0.05). The results of toluidine blue staining showed that the hypertrophic layer of condylar cartilage in the sodium ibandronate group was significantly thicker than that in the osteoporosis group. Compared with the sham group or sodium ibandronate group, the number of apoptotic cells in condylar cartilage and subchondral bone increased significantly in the osteoporosis group (P < 0.05). The expression of dentin matrix protein 1 protein was significantly lower in the osteoporosis group than the sham group, but it increased after treatment with sodium ibandronate (P < 0.05). Compared with the sham group, the expression of Caspase 3 and Bax in the osteoporosis group increased significantly, and the expression of Bcl-2 decreased. However, treatment with sodium ibandronate decreased the expression of Caspase 3 and Bax and increased the expression of Bcl-2 significantly. Overall, our findings reveal that sodium ibandronate can inhibit the apoptosis of condylar chondrocytes and the number of osteoclasts in osteoporotic state, which may be related to the regulation of Caspase 3, Bcl-2, Bax and dentin matrix protein 1 expression.

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    Relationship between degenerative lumbar disc disease and peripheral blood lymphocyte subsets
    Feng Yang, Yan Xu, Wang Yongkui, Yang Tengyue, Shang Lijie, Zhang Chunlin
    2020, 24 (17):  2630-2635.  doi: 10.3969/j.issn.2095-4344.2671
    Abstract ( 347 )   PDF (759KB) ( 47 )   Save

    BACKGROUND: Previous studies have suggested that the main causes of disc degeneration are heredity, aging, malnutrition and load history. The role of immune system in the process of intervertebral disc degeneration is not clear.

    OBJECTIVE: To observe the changes of peripheral blood lymphocyte subsets in patients with lumbar disc degeneration, and to study the relationship between the severity of lumbar disc degeneration and peripheral blood lymphocyte subsets.

    METHODS: Blood samples were collected from 76 patients with degenerative lumbar disc disease (experimental group) and 56 healthy volunteers (control group). The percentages of peripheral blood lymphocyte subsets including CD3+ T cells, CD4+ T cells, CD8+ T cells, CD19+ B cells, and CD3-CD16+CD56+ natural killer (NK) cells were measured by flow cytometry. The ratio of CD4+/CD8+ was calculated. The Pfirrmann grading standard was used to evaluate the grade of lumbar disc degeneration in the two groups, and the correlation between peripheral blood lymphocyte subsets and lumbar disc degeneration was further evaluated. The study protocol was approved by the Ethic Committee of the First Affiliated Hospital of Zhengzhou University in China with approval No. 2019-KY-285. Each participant signed informed consent prior to the study.

    RESULTS AND CONCLUSION: The degenerative grade of the experimental group was significantly higher than that in the control group (P < 0.05). The percentages of CD4+ T cells, NK cells and the ratio of CD4+/CD8+ in the experimental group were significantly higher than those in the control group (P < 0.05). The percentage of CD8+ T cells in the experimental group was significantly lower than that in the control group  (P < 0.05). There was no correlation between the grade of lumbar disc degeneration and lymphocyte subsets of the peripheral blood in the control group. In the experimental group, there was a linear positive correlation between the grade of lumbar disc degeneration and the percentage of CD4+ T cells, the ratio of CD4+/CD8+, and the percentage of NK cells (r=0.412, P=0.000; r=0.715, P=0.000; r=0.494, P=0.000), and there was a linear negative correlation between the grade of lumbar disc degeneration and the percentage of CD8+T cells (r=-0.737, P=0.000). Our results suggest that degenerative changes of the lumbar disc may be related to the changes of lymphocyte subsets in peripheral blood, and the increase of CD4+ T cells, NK cells and CD4+/CD8+ ratio may accelerate lumbar disc degeneration. Therefore, changes in the immune system may predict the occurrence of lumbar disc degeneration and may be a target for prevention and treatment of degenerative lumbar disc diseases.

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    Artesunate relieves subchondral bone osteoclast-mediated osteoarthritis pain
    Shalitanati•Wuermanbieke, Li Yicheng, Ren Jiangdong, Ma Hairong, Mu Wenbo, Cao Li
    2020, 24 (17):  2636-2641.  doi: 10.3969/j.issn.2095-4344.2672
    Abstract ( 479 )   PDF (25714KB) ( 51 )   Save

    BACKGROUND: Studies have shown that osteoclasts induce aberrant in growth of sensory nerves into the subchondral bone by secreting netrin-1, resulting in a reduced pain threshold in an osteoarthritis animal model. Therefore, we assume that inhibition of osteoclasts can alleviate sensory nerve-mediated pain symptoms.

    OBJECTIVE: To investigate whether artesunate inhibits subchondral bone osteoclasts and reduces sensory nerve-mediated pain, providing experimental data for the treatment of osteoarthritis pain using artesunate.

    METHODS: C57BL/6J male mice were randomly assigned to a sham operation group, a placebo group and an artesunate group, with 10 mice per group. The mice in the sham operation group were only subjected to right knee capsulotomy with no damage to the other structures. Moreover, there was no intervention after operation. In the other two groups, the mice received an anterior cruciate ligament transection of the right knee to establish the osteoarthritis model, followed by treatment with artesunate (artesunate group, 100 mg/kg per day) or equivalent volume of 5% NaHCO3 (placebo group) via intraperitoneal injection. Fourteen days after surgery, the footprint trial was performed, and the levels of tartrate-resistant acid phosphatase 5b (TRAcP5b), cathepsin K and carboxy-terminal telopeptide of type I collagen (CTX-I) in the peripheral blood were detected using ELISA. The knee joint specimens of each group were subjected to Safranin O-Fast Green staining, histological scoring, tartrate-resistant acid phosphatase (TRAP) staining and immunohistochemical staining with netrin-1 and calcitonin gene-related peptide (CGRP).

    RESULTS AND CONCLUSION: The percentage of ipsilateral contact area of the right hindpaw in the footprint trial was significantly higher in the sham operation group and artesunate group than the placebo group (P < 0.05), but there was no significant difference between artesunate group and sham operation group (P > 0.05). No significant differences were observed in the serum levels of TRAcP5b, cathepsin K and CTX-I between the groups (P > 0.05). Based on the Safranin O-Fast Green staining, the cartilage histology score was significantly lower in the sham operation group and the artesunate group than the placebo group (P < 0.05), but there was no significant difference between artesunate group and sham operation group (P > 0.05). TRAP staining indicated that compared with the placebo group, the Trap+ osteoclasts were significantly lower in the sham operation group and the artesunate group (P < 0.05), but no significant difference was found between artesunate group and sham operation group (P > 0.05). Compared with the placebo group, the netrin-1+ and CGRP+ sensory nerves in the subchondral bone were significantly decreased in the sham operation group and the artesunate group (P < 0.05), but the two indices had no significant differences between artesunate group and sham operation group (P > 0.05). Our findings from this study indicate that artesunate improves sensory nerve-mediated pain by inhibiting netrin-1 secreted by subchondral bone osteoclasts, and has therapeutic potential to alleviate osteoarthritis pain.

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    Expression and role of retinoblastoma RB1-inducible coiled-coil 1 in the subchondral bone of osteoarthritis
    Li Junyan, Fang Hang, Feng Xiaofeng, Cai Daozhang
    2020, 24 (17):  2642-2647.  doi: 10.3969/j.issn.2095-4344.2673
    Abstract ( 379 )   PDF (26025KB) ( 38 )   Save

    BACKGROUND: Osteoarthritis is mainly characterized by degeneration of the articular cartilage and reconstruction of the subchondral bone. The specific pathogenesis of osteoarthritis is still unclear. Most studies have used cartilage and subchondral bone as the main entry point to explore the molecular mechanism and signal pathway changes in the disease progression, providing new biological targets and research direction for the diagnosis and treatment of osteoarthritis.

    OBJECTIVE: To investigate the expression of RB1-inducible coiled-coil 1 (RB1CC1) in the subchondral bone during the development of osteoarthritis.

    METHODS: Eight-week-old C57 mice were randomly divided into experimental group and sham operation group. Experimental group was then randomly divided into two subgroups of 4 weeks and 8 weeks. In the experimental group, the tibia ligament of the right knee was cut off to dissociate the medial meniscus to induce osteoarthritis. In the sham operation group, only the joint capsule was cut without medial ligament resection and meniscus dissociation. The study was implemented with an experimental animal ethic approval from the Third Affiliated Hospital of Southern Medical University, China (approval No. 44007200038731) on December 13, 2017.

    RESULTS AND CONCLUSION: Compared with the sham operation group, the Osteoarthritis Research Society International scores were increased significantly in the experimental group. Compared with the sham operation group, the expression of collagen II was decreased, and RB1CC1 in the subchondral bone was gradually increased in the experimental group, which was consistent with the expression trend of BSP2. To conclude, with the development of osteoarthritis, the expression of RB1CC1 in the subchondral bone is gradually increased, which may be related to the increase of hyperplasia in the subchondral bone and remodeling.

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    Lycium barbarum polysaccharide intervenes with CD151 and matrix metalloproteinase 3 in the articular cartilage in a rabbit model of knee osteoarthritis 
    Zhao Fei, Ding Dong, Gong Fan, Huang Yonglu, Yao Zhanchuan, Li Xiaoliang, Luo Xiaojun, Yue Jianming
    2020, 24 (17):  2648-2653.  doi: 10.3969/j.issn.2095-4344.2562
    Abstract ( 395 )   PDF (22824KB) ( 29 )   Save

    BACKGROUND: Studies have shown that Lycium barbarum polysaccharide (LBP) has the functions of anti-aging, nerve protection, anti-fatigue, blood sugar control, anti-oxidation, and anti-tumor. It may have some protective effects against osteoarthritis of the knee, but have been rarely reported. CD151 and matrix metalloproteinase 3 (MMP-3) are two common cytokines for assessing knee osteoarthritis.

    OBJECTIVE: To observe the effect of LBP on the expression of CD151 and MMP-3 in rabbit osteoarthritis.

    METHODS: Sixty-four healthy 6-month-old white rabbits were randomly divided into four groups: blank group, model group, LBP group and normal saline group. Animal models of knee osteoarthritis were made using Hulth method in the rabbits except those in the blank group. The rats in the LBP and normal saline groups were fed with normal dose of LBP and normal saline for 4 weeks, and then the articular cartilage tissues were taken from the affected side at 12 weeks after modeling. The morphological changes of the articular cartilage were observed by hematoxylin-eosin staining. The expression levels and spatial distribution of CD151 and MMP-3 in articular cartilage was observed by immunohistochemical staining and western blot. Ethic approval was given by the People’s Hospital of Ningxia Hui Autonomous Region (approval No. 2014-30817).

    RESULTS AND CONCLUSION: immunohistochemistry staining and western blot results showed that the absorbance values and protein expression of MMP-3 and CD-151 were significantly lower in the LBP group than the normal saline and model groups (P < 0.05). Therefore, the expression of CD151 and MMP-3 in the articular cartilage of osteoarthritis was increased, and LBP could inhibit the expression of CD151 and MMP-3 in osteoarthritis, so as to slow down the occurrence of osteoarthritis.

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    Rhizoma drynariae flavonoids inhibits rat intervertebral disc degeneration by inhibiting nuclear factor-kappa B signaling pathway  

    Chen Min, Zhao Kai, Wang Juan, Hu Yong
    2020, 24 (17):  2654-2659.  doi: 10.3969/j.issn.2095-4344.2577
    Abstract ( 379 )   PDF (23338KB) ( 59 )   Save

    BACKGROUND: The combined action of mechanical compression, inflammatory factors, and matrix metalloproteinases leads to intervertebral disc degeneration. Rhizoma drynaria flavones have been shown to have anti-inflammatory effect. However, its protective effects against degeneration of the intervertebral discs remain unclear.

    OBJECTIVE: To investigate the potential molecular mechanisms underlying the protective effects of rhizoma drynaria flavones in a rat model of cervical intervertebral discs degeneration and analyze the clinical value.

    METHODS: Fifty Sprague-Dawley rats (half male and half female) were randomly divided into two groups: 10 in sham-operated group and 40 in degeneration group. Animal models of cervical intervertebral discs degeneration due to dynamic and static imbalance were made in the degeneration group. After successful modeling, 36 model rats were randomized into model group, high-dose group, middle-dose group and low-dose group (n=9 per group). Rats in the sham and model groups were intragastrically given 4 mL of normal saline per day, and those in the high-, middle- and low-dose groups were given 62.5, 125, and 250 mg/kg rhizoma drynaria solution, respectively. The C3-4, C4-5, C5-6, and C6-7 segments of the rat spine were taken. Immunohistochemistry and ELISA were used to detect expression of tumor necrosis factor α and matrix metalloproteinase 3 proteins. Quantitative RT-PCR was used to determine the mRNA expression of tumor necrosis factor α and matrix metalloproteinase 3. Western blot was used to detect the phosphorylation of IkBα.

    RESULTS AND CONCLUSION: Results from the immunohistochemistry, western blot, and RT-PCR, and ELISA showed that the phosphorylation of IkBα and expression of tumor necrosis factor α and matrix metalloproteinase 3 were significantly higher in the model group than the sham group (P < 0.01). However, interventions with rhizoma drynaria flavones, especially with low-dose rhizoma drynaria flavones, significantly lowered the phosphorylation of IkBα and expression of tumor necrosis factor α and matrix metalloproteinase 3. To conclude, high-, middle- and low-dose rhizoma drynaria flavones can exert protective effects against degeneration of the intervertebral discs by suppressing the nuclear factor-κB pathway and inhibiting the expressions of tumor necrosis factor α and matrix metalloproteinase 3. And low-dose rhizoma drynariae flavonoids have the optimal outcomes.

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    Mechanisms of Epimedium in the treatment of osteoarthritis based on network pharmacology and protein module
    Zhang Xiaoyun, Zhang Chi, Song Shilei, Han Jie, Chen Yueping, Yang Nan, Xu Canhong
    2020, 24 (17):  2660-2666.  doi: 10.3969/j.issn.2095-4344.2536
    Abstract ( 461 )   PDF (27814KB) ( 43 )   Save

    BACKGROUND: Pharmacological studies in modern Chinese medicine have shown that icariin has a very positive effect on osteoarthritis. Because of the complex chemical composition of Epimedium and its unclear mechanism underlying the treatment of osteoarthritis at the molecular level, network pharmacology is introduced to explain the potential chemical components and molecular mechanism of Epimedium in the treatment of osteoarthritis. This can provide a theoretical basis for future drug development and disease treatment.

    OBJECTIVE: To explore the molecular mechanism of Epimedium in the treatment of osteoarthritis based on network pharmacology.

    METHODS: TCMSP database was used to screen the active ingredients of Epimedium. TCMSP, Swiss Target Prediction and STITCH database were used to predict the regulatory targets of the active ingredients of Epimedium. OMIM, GeneCards and TTD database were used to predict the therapeutic targets of osteoporosis. The therapeutic target of Epimedium for osteoporosis was obtained by intersecting the therapeutic target of Epimedium and osteoarthritis. A drug-component-target-disease network was then constructed. The protein interaction was analyzed by STRING database, and the related signaling pathways and functions of protein modules were analyzed by DAVID database.

    RESULTS AND CONCLUSION: Twenty-three pharmacodynamic components of Epimedium were screened and 230 pharmacodynamic targets of Epimedium and 1 221 therapeutic targets of osteoarthritis were predicted. After crossing, 95 therapeutic targets of Epimedium for osteoporosis were obtained. Protein interaction analysis indicated that JUN, AKT1, RELA, MAPK1, IL6, CXCL8, MAPK8, MAPK14, FOS and IL1B were the core targets of protein interaction network. Key protein modules were mainly involved in interleukin receptor pathway, tumor necrosis factor signaling pathway, Toll-like receptor signaling pathway, T cell receptor signaling pathway, nuclear factor-kappa B signaling pathway and osteoclast differentiation pathway. They might play a role in the treatment of osteoarthritis by regulating many biological processes such as cell proliferation and apoptosis, immune cells and immune response, inflammatory factors and inflammatory response, and lipopolysaccharide cell response.

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    Changes of fibrotic factors in the recovery process of skeletal muscle strain
    Yang Ning, Huang Tao, Liu Xiaoran, Zhou Yue, Wang Ruiyuan, Li Junping
    2020, 24 (17):  2667-2674.  doi: 10.3969/j.issn.2095-4344.2573
    Abstract ( 402 )   PDF (29798KB) ( 26 )   Save

    BACKGROUND: Skeletal muscle injury is the most common sports injury, which can develop fibrosis and then impair muscle function. Varieties of fibrotic factors are involved in the repair of skeletal muscle and effectively regulate the process of tissue fibrosis.

    OBJECTIVE: To establish a rat model of acute gastrocnemius strain, observe the changes of fibrotic factors in the recovery of injury and explore the possible roles of fibrotic factors.

    METHODS: Totally 112 male Sprague-Dawley rats were randomly divided into control group (n=56) and injury group (n=56). The left gastrocnemius muscle was injured by a rat gastrocnemius pull-off device in the injury group. Each group was further subdivided into immediately group, 2 days group, 4 days group, 7 days group, 14 days group, 21 days group and 28 days group depending on the sampling time points. There were eight rats in each subgroup. The control group did not receive any intervention. The left gastrocnemius muscle of the injury group was strained by specialized equipment. Muscle fiber injury and inflammation were observed by hematoxylin-eosin staining. The expression of transforming growth factor β1 (TGF-β1), connective tissue growth factor (CTGF), matrix metalloproteinase-1 (MMP-1), and matrix metalloproteinase tissue inhibitor 1 (TIMP-1) were detected by western blot. The experiment was approved by the Sports Science Experimental Ethics Committee of Beijing Sport University.

    RESULTS AND CONCLUSION: Hematoxylin-eosin staining results showed that the muscle fibers of the gastrocnemius muscle arranged disorderedly and inflammatory cells were aggregated in the injury group. Compared with the control group, protein expression levels of TGF-β1 and CTGF significantly increased in the injury group from the 2nd day throughout the recovery (P < 0.05). MMP-1 expression significantly increased at the 2nd day after injury (P < 0.05) and gradually recovered until the 7th day (P > 0.05). TIMP-1 expression significantly increased from the 7th day until the 28th day (P < 0.05). These findings indicate that TGF-β1/CTGF signal pathway may be successively activated during the recovery of skeletal muscle strain, triggering the occurrence and development of skeletal muscle fibrosis. Meanwhile, MMP-1 and TIMP-1 protein may be activated to participate in the recovery of injury.

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    Autologous cortical bone block for repair of implant-site jaw bone defects  

    Wang Ning, Cui Tingting, Li Yongqi, Zhao Binbin, Zhong Weijian, Ma Guowu
    2020, 24 (17):  2675-2679.  doi: 10.3969/j.issn.2095-4344.2646
    Abstract ( 415 )   PDF (20394KB) ( 57 )   Save

    BACKGROUND: Autologous bone with good biocompatibility possesses the characteristics of bone conduction, osteoinduction and osteogenesis. However, the healing process after autologous bone grafting is still controversial. The focus of controversy is whether the bone graft is completely absorbed and replaced or whether it can retain cell viability for a long time.

    OBJECTIVE: To observe the histological changes in bone remodeling and regeneration after cortical bone grafting.

    METHODS: Six healthy beagle dogs were selected as the research animals. The maxillary premolars of experimental dogs were extracted and the bone plates with width of 10 mm, length of 15 mm and thickness of 2 mm were removed from the buccal side. A bone defect model was established. The corresponding size of cortical bone block was cut on the buccal side of the bilateral mandibular body. Cortical bone grafts were fixed in the maxillary defect area. On one side pure cortical bone block was used as grafting material; on the other side the implant was placed simultaneously with the bone graft. Samples were harvested at 3 and 6 months after bone grafting. The cell survival and bone graft resorption of autologous cortical bone graft were analyzed by gross observation and histological observation. The study protocol was approved by the Animal Ethics Committee of Dalian Medical University, China.

    RESULTS AND CONCLUSION: The size of the grafted bone was gradually reduced, and the margin was blunt and firmly bonded to the base bone. All implants lost after 6 months. A newly formed connection between the graft and the base bone was shown by hard tissue ground sections. The porosity of bone lacuna in the grafted bone block at 6 months was significantly lower than that at 3 months (P < 0.05). The grafted bone resorption rate at 6 months was significantly higher than that at 3 months (P < 0.05). These findings indicate that osseointegration exists between grafted cortical bone block and the base bone of the recipient area. Osteocytes in the graft bone partially remain alive. With the prolongation of healing period, the proportion of new bone cells increases, and the volume of the grafted bone gradually decreases. Osseointegration of the concurrent implant is disturbed due to the absorption of cortical bone graft.

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    Effects of dexamethasone combined with estrogen on the expression of interleukin-6, Caspase3 and Bcl-2 after spinal cord contusion in rats  

    Li Qinxuan, Wang Yizhi, Xu Yan, Zhang Huayang, Lü Qixuan, Zhang Cheng, He Zhengyun, Zhang Xiao, Yang Zheng
    2020, 24 (17):  2680-2685.  doi: 10.3969/j.issn.2095-4344.2591
    Abstract ( 282 )   PDF (27840KB) ( 31 )   Save

    BACKGROUND: The clinical use of drugs alone can well block specific pathological changes after spinal cord injury, but single medications often result in complicated adverse reactions. For example, estrogen can alleviate apoptosis caused by oxidative damage, but it can also activate inflammation. Therefore, the combined use of drugs has become a clinical treatment trend, and the mechanism of combined use is urgently needed to be clarified.

    OBJECTIVE: To investigate the therapeutic effect of dexamethasone combined with estrogen treatment on the expression of interleukin-6, Caspase3 and Bcl-2 in rats post spinal cord contusion.

    METHODS: Forty-eight female Sprague-Dawley rats aged 4 weeks were randomly assigned to the following groups (n=12 per group): sham surgery group, normal saline group, dexamethasone group, combined treatment (dexamethasone and estrogen) group. The spinal cord contusion model was established by modified Allen’s method in all groups except for the sham surgery group. Rats in the sham surgery group and normal saline group were injected normal saline (0.2 mL/kg) at the lateral thigh muscle, those in the dexamethasone group were given dexamethasone (0.2 mg/kg), and those in the combined treatment group were treated with estrogen (0.04 mg/kg) and dexamethasone (0.2 mg/kg) at 30 minutes, 1, 2, 3, 4, and 5 days post-operation. Basso, Beattie and Bresnahan scoring was applied to evaluate the hindlimb motor functional recovery within 14 days post-operation. Expression of interleukin-6, Bcl-2 and Caspase3 was detected using immunohistochemistry at 3, 7, and 21 days post-operation.

    RESULTS AND CONCLUSION: The motor function of combined treatment group recovered earlier than that of dexamethasone group and normal saline group. Furthermore, during the whole recovery period, the Basso, Beattie and Bresnahan scores in the combined treatment group were significantly higher than those in the dexamethasone group (P < 0.05). The immunohistochemical results also indicated that the expression of interleukin-6 and Casepase3 in the combined treatment group was significantly lower than that in the dexamethasone group, and the expression of Bcl-2 was significantly increased in the recovery period (P < 0.05). To conclude, the combined treatment of dexamethasone and estrogen can effectively reduce the expression of proapoptotic factor Caspase3 and proinflammatory factor interleukin-6, and increase anti-apoptotic factor Bcl-2 to further promote functional recovery in rats after spinal cord contusion by impeding apoptosis and inflammation. 

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    Low-dose zoledronic acid regulates osteoclasts and osteoblasts in the extraction socket of ovariectomized rats  

    Cheng Yuting, Wu Chao, Huang Xiaolin, Li Fang, Shi Qianhui, Zhou Qian, Hong Wei, Wang Yong, Liao Jian
    2020, 24 (17):  2686-2693.  doi: 10.3969/j.issn.2095-4344.2597
    Abstract ( 277 )   PDF (88164KB) ( 30 )   Save

    BACKGROUND: Although zoledronic acid can effectively prevent bone loss in postmenopausal women, its effect and mechanism on the mandible are not clarified.

    OBJECTIVE: To study the morphological and pathological changes of mandibular tissue in ovariectomized rats treated with low-dose zoledronic acid and to investigate the regulatory effect and mechanism of RANKL/RANK/OPG signaling system in the inhibition of bone resorption by zoledronic acid.

    METHODS: Thirty-six adult female Sprague-Dawley rats were randomized into control, model and treatment groups. Animal models of osteoporosis were made by bilateral ovariectomy in the latter two groups, while the same amount of lipid tissues surrounding the ovary was removed in the control group. Three weeks after ovariectomy, the rats in the treatment group were given subcutaneous injection of 20 μg/kg zoledronic acid. The corresponding doses of saline were injected subcutaneously in the control and model groups. One week after treatment, the left mandibular molars were extracted from all the rats, and the rats’ jaws were separated and removed at 4 weeks after tooth extraction for detection. The residual alveolar bone of the extracted socket was observed by X-ray. The pathological changes of the mandibular cortex and cancellous bone were detected by hematoxylin-eosin staining. The number of apoptotic osteoblasts was detected by TUNEL apoptosis test. The expressions of receptor activator of nuclear factor-κ B ligand (RANKL), osteoprotegerin and nuclear transcription factor-κB (NF-κB) in the mandibular alveolar bone were detected by immunohistochemical technique. Western blot was finally used to detect the expression of RANKL and NF-κB at protein levels.

    RESULTS AND CONCLUSION: (1) At 4 week after tooth extract, subcutaneous injection of 20 μg/kg zoledronic acid effectively inhibited alveolar bone resorption and promoted new bone formation at the extraction socket. (2) Hematoxylin-eosin staining results showed that: in the model group, the cortical bone was thinned, and the trabecular bone was thinned and even ruptured. There were a large number of bone resorption lacunae but few osteoblasts in the model group. In the treatment group, the cortical bone was thickened and the trabecular bone had normal structure, with only a small amount of bone resorption lacunae and increased number of osteoblasts. (3) The number of apoptotic osteoblasts was significantly lower in the treatment group than the model group (P < 0.001). (4) Immunohistochemical staining results showed significantly decreased RANKL and NF-κB protein expressions (P < 0.001, P < 0.002) and significantly increased osteoprotegerin level (P < 0.001) in the treatment group than the model group. (5) Western blot results revealed that the protein expressions of RANKL and NF-κB in the model group were significantly higher than those in the control group, and treatment with zoledronic acid significantly reduced these protein levels (both P < 0.001). To conclude, zoledronic acid could inhibit the differentiation of osteoclasts by down-regulating the NF-κB signal pathway and meanwhile regulate the apoptosis of osteoblasts.

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    Effect of Xianling Gubao Capsule on alveolar bone mass in postmenopausal women with periodontitis: a cone-beam CT evaluation

    Zhang Yujing, Peng Yuzhi, Liu Baozhen, Jing Fang
    2020, 24 (17):  2694-2699.  doi: 10.3969/j.issn.2095-4344.2596
    Abstract ( 399 )   PDF (24421KB) ( 29 )   Save

    BACKGROUND: Xianling Gubao Capsule can reduce bone loss and promote the formation of osteoblasts. Its efficacy in osteoporosis and fracture has been preliminarily confirmed, but its efficacy in postmenopausal female periodontitis has not been clarified.

    OBJECTIVE: To assess the effect of Xianling Gubao Capsule on alveolar bone mass in postmenopausal women with periodontitis by cone-beam CT, providing an effective basis for its clinical use.

    METHODS: One hundred and thirty-five patients with periodontitis diagnosed and treated in the Department of Stomatology, First Affiliated Hospital of Guiyang College of Traditional Chinese Medicine from October 2015 to April 2017 were enrolled. All patents were randomly divided into blank group (basic treatment), control group (basic treatment+vitamin D) and experimental group (basic treatment+Xianling Gubao Capsule), with 45 patients in each group. All patients received oral hygiene and health education, and were guided to brush their teeth correctly and self-control plaques. Basic treatments for periodontal disease were performed to eliminate plaque and calculus by cleaning and curing the gums, and smoothing the root surface, once 3 months for 6 continuous months. The control group received oral Calci-D (600 mg of calcium and 125 U of vitamin D per tablet), one tablet per day, for 6 continuous months. The experimental group was orally given Xianling Gubao Capsule 1.0 g/time, 3 times per day, 3 months as a course of treatment, for two consecutive courses. Cone-beam CT was used to detect the changes of bone mineral density and height of alveolar bone defect in all patients before treatment, 3 and 6 months after treatment. Ethics approval was obtained from the First Affiliated Hospital of Guiyang College of Traditional Chinese Medicine on May 29, 2015 (approval No. K2015-186).

    RESULTS AND CONCLUSION: (1) Compared with the baseline, the alveolar bone defect height was significantly reduced in the experimental and control groups (P < 0.05), and bone mineral density was significantly increased in the two groups (P < 0.05). There were no changes in the alveolar bone defect height and bone mineral density in the blank group (P > 0.05). The improvement of alveolar bone defect height and bone density was better in the experimental group than the control group (P < 0.05). (2) With the increase of age and the aggravation of periodontitis, the alveolar bone defect height gradually increased and bone mineral density gradually decreased (P < 0.05). The alveolar bone defect height and bone mineral density of the patients aged from 46 to 50 were significantly improved after treatment (P < 0.05). The bone mineral density of the patients aged from 51 to 55 years increased significantly (P < 0.05), but the alveolar bone defect height had no significant changes (P > 0.05). The alveolar bone defect height and bone mineral density of the patients older than 55 years had no changes (P > 0.05). (3) Alveolar bone defect height and bone mineral density were significantly improved in patients with mild and moderate periodontitis (P < 0.05), while no significant change in alveolar bone defect height and bone mineral density was found in patients with severe periodontitis (P > 0.05). To conclude, the treatment of periodontitis by Xianling Gubao Capsule can significantly improve the alveolar bone defect height and bone mineral density, and cone-beam CT can be used to evaluate its clinical efficacy.

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    Establishing a model of femoral head necrosis by alcohol burning in Cyan-Shank Partridge Chicken: a feasibility analysis 
    Gu Jiangjiang, Zhao Fengchao, Cheng Qi, Tang Jinlong, Xu Shizhuang
    2020, 24 (17):  2700-2705.  doi: 10.3969/j.issn.2095-4344.2674
    Abstract ( 278 )   PDF (23341KB) ( 40 )   Save

    BACKGROUND: To date, the pathological mechanism of femoral head necrosis is still unclear, and the existing experimental animal models of femoral head necrosis all have shortcomings.

    OBJECTIVE: To establish a simple, reliable and stable animal model of femoral head necrosis in a Cyan-Shank Partridge Chicken, with similar biomechanical characteristics to human beings.

    METHODS: Twenty-four adult young Cyan-Shank Partridge Chickens were randomly divided into experimental group and control group (n=12 per group). One side of the femoral head was randomly selected for operation, and the other side was untreated (untreated group). In the experimental group, alcohol was used to burn the bone tissue in the drilled hole followed by implantation of the original bone. In the control group, the original bone was directly re-implanted after drilling. X-ray and CT scan examinations were performed 1 month after operation. The femoral head was then dissected for general observation, microCT observation and pathological staining. The study protocol was approved by the Animal Ethic Committee of Xuzhou Medical University in China (approval No. AEC2018-020).

    RESULTS AND CONCLUSION: All the Cyan-Shank Partridge Chickens were alive without wrongful death. They could walk upright with no obvious claudication at 1-3 days after operation. X-ray and CT results revealed collapse of the femoral head with unclear trabecular bone in the experimental group. General observation indicated that the femoral head was collapsed with no smooth surface in the experimental group. MicroCT results showed significantly reduced number of bone trabeculae and significantly increased trabecular thickness, trabecular spacing and bone volume fraction in the experimental group as compared with the control and untreated groups (P < 0.05). Pathological staining showed that there was no obvious necrosis but similar shape of the femoral head in the control and untreated groups, but the number of subchondral vessels was less in the control group than the untreated group. However, the cells arranged disorderly and obvious necrosis such as empty lacuna was observed in the experimental group. To conclude, the use of alcohol to burn the bone tissue of the femoral head of the Cyan-Shank Partridge Chicken can be used to establish a typical animal model of femoral head necrosis, which has similar pathological characteristics of human femoral head necrosis. 

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    Improvement for the construction of renal interstitial fibrosis model with unilateral ureteral obstruction in rats 
    Feng Hefei, Guo Tao, Huang Min, Su Wang, Xia Deyue, Peng Lan, Yu Yunbo, Sun Jie
    2020, 24 (17):  2706-2711.  doi: 10.3969/j.issn.2095-4344.2556
    Abstract ( 476 )   PDF (24382KB) ( 38 )   Save

    BACKGROUND: Unilateral ureteral obstruction (UUO) is a traditional surgical method that can cause renal interstitial fibrosis in rats in a short period of time (1-2 weeks), but it can develop many postoperative complications and result in a high mortality.

    OBJECTIVE: To optimize and improve the UUO operation in rats for reducing the incidence of postoperative complications and improving the survival rate of animals, and to detect the pathophysiological indicators of rats modeled by UUO, providing background data for the basic research on functional pharmacology.

    METHODS: Twenty male Sprague-Dawley rats were randomly divided into four groups, followed by traditional UUO ligation, improved UUO ligation, undissociated ureteral ligation, and opening the abdominal cavity with no ureter ligation (sham group), respectively. At 14 days postoperatively, the successful rate of renal interstitial fibrosis model, mortality rate, and the incidence of postoperative complications were compared between groups. Another 28 Sprague-Dawley rats, half male and half female, were randomly divided into improved UUO group and control group. Physiological indexes, including urine analysis, blood cells analysis, blood biochemical analysis, were tested as background information. Besides, the pathological changes of kidney tissues were compared between groups using hematoxylin-eosin staining and Masson staining at 14 and 21 days postoperatively.

    RESULTS AND CONCLUSION: Compared with the undissociated ureteral ligation group, the successful rate of modeling was higher in the traditional and improved UUO groups. The improved UUO surgery with double ligation of the middle ureter and no cutting of the middle ureter was characterized by easier operation, smaller surgical wound, faster modeling and higher successful rate of renal interstitial fibrosis model, lower postoperative mortality and lower incidence of complications in animals as compared with the traditional UUO surgery. The improved UUO group showed statistically significant differences from the sham group in blood biochemical indexes such as urea nitrogen, glutamate aminotransferase, and albumin (P < 0.05). Results from hematoxylin-eosin staining and Masson staining of the obstructed kidney revealed typical pathological features of renal interstitial fibrosis. 

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    Aerobic exercises alleviate renal oxidative stress injury in a mouse model of diabetic nephropathy 
    Liu Xiaochen, Wang Gaifeng, Zhang Shefeng
    2020, 24 (17):  2712-2717.  doi: 10.3969/j.issn.2095-4344.2675
    Abstract ( 340 )   PDF (24727KB) ( 35 )   Save

    BACKGROUND: Studies have confirmed that long-term regular aerobic exercise can effectively reduce blood glucose level and improve insulin resistance. It is an important measure for diabetic nephropathy, but the specific mechanism is unknown.

    OBJECTIVE: To investigate the effect of aerobic exercise on renal oxidative stress injury in mice with diabetic nephropathy, and explore its relationship with nuclear factor E2-related factor 2 (Nrf2)/heme oxygenase 1 (HO-1) signal pathway.

    METHODS: Male C57/BL6 mice were randomly divided into normal control group, model group, and aerobic exercise group, 8 mice in each group. Except for the control group, mice in the other two groups were intraperitoneally injected with 40 mg/kg streptozotocin for 5 consecutive days to induce diabetes. After 4 weeks, the 24-hour urine protein content > 30 mg suggested a successful modeling of diabetic nephropathy. After 8 weeks of modeling, mice in the aerobic exercise group underwent treadmill aerobic exercise for 8 weeks, 1 hour daily, 5 days per week. After 8 weeks of treatment, systolic blood pressure of the tail artery, 24-hour urinary microalbumin excretion rate, blood glucose, blood lipid, serum creatinine, and blood urea nitrogen levels were measured. The content of malondialdehyde and the activity of total superoxide dismutase of renal tissue homogenate were determined. The glomerular mesangial extracelluar matrix deposition, the level of reactive oxygen and the expression levels of Nrf2 and HO-1 in renal tissue were detected. Ethic approval was obtained from the Experimental Animal Ethic Committee of Henan Academy of Chinese Medicine in China (approval No. HPHT2015019).

    RESULTS AND CONCLUSION: Compared with the control group, the 24-hour urinary microalbumin excretion rate, levels of blood glucose, serum creatinine, blood urea nitrogen, total cholesterol, triglyceride, and low-density lipoprotein-cholesterol, and systolic blood pressure of the tail artery in the model group were significantly increased (P < 0.05), the glomerular mesangial extracelluar matrix was significantly accumulated (P < 0.05), and the levels of reactive oxygen species and malondialdehyde were significantly increased. Compared with the model group, the above-mentioned parameters were significantly decreased in the aerobic exercise group. Aerobic exercise significantly increased the activity of total superoxide dismutase, decreased the accumulation of glomerular mesangial extracelluar matrix, and up-regulated the expression of Nrf2 and HO-1 in the renal tissue. To conclude, aerobic exercise can alleviate renal oxidative stress injury in diabetic nephropathy mice, which may be related to the activation of Nrf2/HO-1 pathway.

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    Expression of cyclic RNA plasmacytoma variant translocation 1 in senescence of nucleus pulposus cells and its regulation mechanism
    Xie Mingzhong, Li Sen, Zhu Kai, Jiang Huacai, Qi Lisheng
    2020, 24 (17):  2718-2723.  doi: 10.3969/j.issn.2095-4344.2557
    Abstract ( 413 )   PDF (23231KB) ( 33 )   Save

    BACKGROUND: Cyclic RNA plasmacytoma variant translocation 1 (circPVT1) is involved in the senescence of fibroblasts, but the relationship of circPVT1 with nucleus pulposus senescence and its mechanism are still unclear.

    OBJECTIVE: To investigate the expression of circPVT1 in nucleus pulposus cell senescence and to explore its possible mechanism.

    METHODS: Human nucleus pulposus cells were cultured in vitro, and the senescence of nucleus pulposus cells was induced by ionizing radiation (5 Gy, 6 days). The expression of circPVT1 and let-7 mRNA was detected by real-time quantitative polymerase chain reaction (qRT-PCR). CircPVT1 siRNA and anti-let-7 were transfected into normal nucleus pulposus cells, which were divided into control group, si-NC+anti-NC group, si-circPVT1+anti-NC group, si-NC+anti-let-7 group, and si-circPVT1+anti-let-7 group. The expressions of circPVT1 and let-7 mRNA were detected by qRT-PCR. Cell counting kit-8 assay was used to detect the inhibition of cell proliferation. Plate cell clone formation assay was used to detect colony formation. Cell senescence was detected by SA-β-gal staining. The expressions of p21, p27, let-7 target high mobility group protein A2 (HMGA2) and KRAS were detected by western blot assay. Double luciferase activity assay was used to verify the relationship between let-7 and target regulation of HMGA2 and KRAS.

    RESULTS AND CONCLUSION: (1) Compared with normal nucleus pulposus cells, the expression of circPVT1 was decreased, while let-7 expression and the positive rate of SA-β-gal staining were increased in the irradiated cells (P < 0.05). (2) Compared with the control group and si-NC+anti-NC group, the si-circPVT1+anti-NC group appeared to have decreased expression of circPVT1 mRNA, HMGA2 and KRAS proteins and number of clones formed as well as increased let-7 mRNA expression, p21, p27 protein expression, cell inhibition rate and positive rate of SA-β-gal staining (P < 0.05). However, opposite changes were found in the si-NC+anti-let-7 group in relative to the control group (P < 0.05). (3) The expression of circPVT1 mRNA, clone formation, and expressions of HMGA2 and KRAS proteins in the si-circPVT1+anti-let-7 group were higher than those in the si-circPVT1+anti-NC group, and lower than those in the si-NC+anti-let-7 group. Let-7 mRNA expression, cell inhibition rate, positive rate of SA-β-gal staining, and expressions of p21 and p27 proteins in the si-circPVT1+anti-let-7 group were lower than those in the si-circPVT1+anti-NC group, and higher than those in the si-NC+anti-let-7 group (P < 0.05). Double luciferase activity assay showed that HMGA2 and KRAS were the targets of let-7. These findings indicate that inhibition of circPVT1 can inhibit the aging of nucleus pulposus cells. The mechanism may be through binding let-7 to inhibit the targeting of HMGA2 and KRAS proteins. 

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    Three-dimensional morphological characteristics of basal nuclei in children aged 3 or 4

    Xu Yangyang, Gao Shang, Su Baoke, Wang Yidan, He Yujie, Li Kun, Wang Haiyan, Li Xiaohe
    2020, 24 (17):  2724-2729.  doi: 10.3969/j.issn.2095-4344.2559
    Abstract ( 424 )   PDF (24983KB) ( 42 )   Save

    BACKGROUND: There are many studies on the morphology of basal nuclei of adult brain based on various imaging methods, but little is reported on the three-dimensional reconstruction measurement of basal nucleus morphology in children aged 3 or 4.

    OBJECTIVE: To establish a three-dimensional model of the basal nucleus in children aged 3 or 4 years relative to other age groups in order to explore the morphological basis for the neurodevelopment of the basal nucleus.

    METHODS: Fifteen cadavers of 3 or 4 years old children were selected and the basal nucleus was cut by thin layer. The image data set with accurate segmentation of its main structure was then completed. The cross sections of the main structures were analyzed, the diameter and angle of the related structures were measured in a specific section, and the differences between the sides were compared. The original data were re-sampled to establish a three-dimensional coordinate system of the basal nucleus. The morphological size of the main structure of the basal nucleus was measured, the anatomical identification was clearly identified, and the spatial information was quantified. Model and the public reference system were established. The base nucleus structures were presented three-dimensionally and dyed in different colors. The boundary between the structures was clearly displayed, and the internal structure was observed by rotating and cutting at any angle. The study protocol was implemented in line with the relevant ethical requirements of Inner Mongolia Medical University. Specimens were donated voluntarily, and children’s guardians signed informed consents.

    RESULTS AND CONCLUSION: (1) The reconstructed three-dimensional model could show the basal nucleus, and could be used to measure the volume of the basal nucleus and the related parameters of meridians. There was no significant difference between the sides of the reconstructed model. (2) The reconstructed three-dimensional structure could be rotated at any angle and at different speeds, and the corresponding scaling and transparent processing could be carried out to make the reconstructed structure image more realistic and realistic. In children aged 3 or 4, the reconstruction of the basal nucleus based on volume rendering can provide the morphological basis for stereotactic neuronavigation surgery as well as the diagnosis and treatment of epilepsy.

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    Treatment of deep second-degree burn wounds with the combination of biological dressing and epidermal cell implantation after controlled debridement 
    Shi Xueqin, Zhou Qin, Hu Dahai, Wang Yunchuan, Xie Songtao, He Ting, Liu Miaomiao, Qu Yanyan, Wang Jiani
    2020, 24 (17):  2730-2734.  doi: 10.3969/j.issn.2095-4344.2555
    Abstract ( 378 )   PDF (21695KB) ( 122 )   Save

    BACKGROUND: There are various methods for the treatment and repair of deep second-degree burn wounds, which have diverse effects and have no uniform standards. Therefore, new technologies need to be developed.

    OBJECTIVE: To observe the therapeutic effect of controlled debridement and traditional treatment on deep second-degree burn wounds.

    METHODS: A total of 80 patients with deep second-degree burns from June 2015 to June 2018 were enrolled and divided into a positive control group and a controlled debridement group, with 40 patients in each group. The positive control group was coated with Bangerkang burn bacteriostatic cream, and the oil gauze was pressure-wrapped. The dressing was changed at 3, 6, 9, and 12 days after the injury. In the controlled debridement group, epidermal cell suspension was prepared according to the requirements of wound preparation and was sprayed onto the wound surface. The wound was then covered with Recell Kit self-contained protective film. The film was removed to keep the wound dry on the 3rd day after operation, and then the wound was observed at 6, 9, and 12 days after operation. The study protocol was approved by the Ethics Committee of the Xijing Hospital of the Air Force Military Medical University. All patients volunteered to participate in the study and sign an informed consent. Patient information was registered online and appropriate treatment was performed according to a random (software online) assignment.

    RESULTS AND CONCLUSION: At 3, 6, 9, and 12 days after operation, the bacterial content, wound pain score, wound infection score and pro-inflammatory factor level in the controlled debridement group were significantly lower than those in the positive control group (P < 0.05). Moreover, there was no complication in both groups. These findings reveal that epidermal cell implantation combined with controlled debridement for deep second-degree burn wounds can achieve remarkable outcomes, which can significantly accelerate wound healing, reduce infection and alleviate the suffering of patients.

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    Ozone bath in the treatment of diabetic foot ulcer infection
    Qin Xinyuan, Wang Lei, Wang Jiangning
    2020, 24 (17):  2735-2741.  doi: 10.3969/j.issn.2095-4344.2676
    Abstract ( 776 )   PDF (27939KB) ( 73 )   Save

    BACKGROUND: Medical ozone is a strong oxidant, can kill bacteria, resist inflammation, improve circulation, relieve pain and promote regeneration. It has been reported that medical ozone has a positive effect on ulcer wounds such as caries.

    OBJECTIVE: To explore the clinical effect of ozone bath on the control of diabetic foot ulcer’s infection and the promotion of wound healing. 

    METHODS: According to the treatment methods, 100 patients with type 2 diabetic foot ulcer who met the inclusion criteria were divided into two groups randomly. In the experimental group, ozone bath was applied to the wound for 30 minutes after cleaning the wound with local normal saline; in the control group, air bath was applied to the wound for 30 minutes after cleaning the wound with local normal saline, once a day. Three weeks later, the bacterial control (quantitative analysis of bacteria, distribution of pathogenic bacteria), the degree of decline of inflammation index, the reduction of ulcer wound, the rate of wound healing and the visual analogue scale score of wound pain were counted in both groups. The expression levels of vascular endothelial growth factor, transforming growth factor-β and platelet-derived growth factor in wound tissues were measured before and after treatment. The protocols were approved by the Ethics Committee of Beijing Shijitan Hospital Affiliated to Capital Medical University. Patients and their family member signed the informed consent.

    RESULTS AND CONCLUSION: After 3 weeks of treatment, bacteria were still detected in 46 patients in the control group and 5 patients in the experimental group. The number of pathogenic bacteria was 73 before treatment and 66 after treatment in the control group, 74 before treatment and 7 after treatment in the experimental group. Bacterial quantitative analysis was significantly higher in the control group than in the experimental group after treatment. The indexes of ulcer healing, including the reduction of ulcer wound, the visual analogue scale score of wound pain, and the degree of decline of inflammation index, were better in the experimental group than those in the control group (P < 0.05). (4) The expression levels of vascular endothelial growth factor, transforming growth factor-β and platelet-derived growth factor in local wound tissues were significantly higher in the experimental group than in the control group (P < 0.05). The percentage of wound healing in the experimental group was significantly higher than that in the control group (P < 0.05). To conclude, ozone bath can significantly control the local infection of diabetic foot, reduce the distribution of pathogenic bacteria on the wound surface, relieve the inflammatory pain of the wound, stimulate the secretion of growth factor in wound, and improve the wound healing rate.

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    Current status of clinical trial registrations in tissue engineering based on ClinicalTrials.gov
    Lu Yan, Zhang Ting, Ouyang Zhaolian
    2020, 24 (17):  2742-2751.  doi: 10.3969/j.issn.2095-4344.2612
    Abstract ( 396 )   PDF (35155KB) ( 94 )   Save

    BACKGROUND: There is an imbalance between supply and demand for tissues and organs. Therefore, it is crucial to actively carry out basic research regarding tissue engineering and accelerate relevant clinical transformation.

    OBJECTIVE: To analyze the status of clinical trials registrations in tissue engineering and to provide reference and evidence support for clinical transformation research in tissue engineering.

    METHODS: Clinical trial data from the ClinicalTrials.gov were collected. Bibliometrics and mathematical statistics methods were adopted to analyze the status of clinical trial registration in tissue engineering from various aspects, including registration quantities, indications, sponsors, and stages.

    RESULTS AND CONCLUSION: There were 434 records of clinical trial registrations regarding tissue engineering worldwide, 65 of which were in China. Over half of the records were randomized controlled trials in China or in the world, and no Chinese institutions have participated in the 18 international multi-center clinical trials worldwide. These clinical trials covered a total of 57 indications all over the world and 37 indications in China, mainly concentrated in the fields of bone and cartilage, teeth, and skin. There were 275 sponsors all over the world, 33 of which were in China. Most of the clinical trials were in phase 1/phase 2. Through analyzing the status of clinical trial registrations regarding tissue engineering, there were less clinical transformation applications in tissue engineering. Although China had the second largest number of registrations in the world, there was still a certain gap with the United States. There were a certain number of clinical trials with high evidence levels, but no products with high potential value in the international market. More concerns were paid onto clinical transformation research in research institutions and medical institutions. Clinical applications mainly focused on the treatment of defects or functional defects in bone and cartilage, teeth and skin, which were mainly in the stage of safety or efficacy evaluation.

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    Metrology and visual analysis of English literature regarding osteoporosis in the past 5 years: based on the Web of Science 
    Zhang Qiang, Zhu Chan
    2020, 24 (17):  2752-2758.  doi: 10.3969/j.issn.2095-4344.2677
    Abstract ( 502 )   PDF (30940KB) ( 78 )   Save

    BACKGROUND: Osteoporosis has always been a hotspot in medical research. Visual analysis of English literature regarding osteoporosis can intuitively understand the research status, hotspots and frontiers in this field, and help researchers to hold on the research topic and direction.

    OBJECTIVE: To perform bibliometric, co-presence, co-citation, and visual analyses of English literature regarding osteoporosis, and to explore the relevant research status, research hotspots and dynamic frontiers in this field in the past 5 years.

    METHODS: The CiteSpace software was used for co-presence analysis of the authors, institutions, countries, and keywords and for co-citation analysis of cited authors, references, and journals in the 2015-2019 related literature included in the Science Citation Index Expanded (SCI-E) of Web of Science (WOS). 

    RESULTS AND CONCLUSION: A total of 2 980 articles were included. High-yielding countries include China, the United States, Japan, South Korea, and the United Kingdom. The main research institutions are Pittsburgh University, Harvard University, McMaster University, Oxford University, and other world-renowned universities. Shanghai Jiao Tong University wins a place in the orthopedics research. As high-yielding authors, Hong-Wen Deng focuses on genetic and osteoporosis mechanisms; Hiroyuki Kato, Yukio Nakamura, and Takako Suzuki jointly focus on the development of anti-osteoporosis drugs such as denosumab and ibandronate; Jonathan D Adachi focuses on vitamin D and osteoporosis and bone metabolism research; William D Leslie focuses on dual-energy bone mineral density assessment of osteoporosis; JA Kanis focuses on the appropriate population and timing for anti-osteoporosis. Osteoporosis International and Journal of Bone and Mineral Research published the most of research regarding osteoarthritis. New England Journal of Medicine, LanceT, JAMA-J and other top publications are also very concerned about osteoporosis research. The research discipline involves clinical and basic disciplines, with a focus on basic subjects. Advances in the development of anti-osteoporosis drugs are an important entry point for basic medical research. Lack of osteoporosis, zoledronic acid, glucocorticoid-induced osteoporosis, fat mass, oophorectomy, osteoporosis, mineral density, ovarian cancer, anti-osteoporosis, and bone metabolism is the frontier of osteoporosis research. In 2015-2016, animal research and senile osteoporosis were hotspots. In 2016-2017, the design of randomized trials, adult bone metabolism, and physical activity interventions were emerging areas of osteoporosis research. In 2017-2019, glucocorticoid-induced osteoporosis, mesenchymal stem cells and activation of cellular pathways were emerging areas of osteoporosis research. To conclude, CiteSpace can be used to bibliometrically analyze the English literature of osteoporosis and intuitively reveal the overall development trend of osteoporosis research in the past 5 years. Therefore, it can provide references for osteoporosis scholars in the reference content and development direction of the topic, and the results have certain significance.

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    Vascular endothelial growth factor intervention: a new approach for accelerating fracture healing
    Qian Zhouyao, Wang Yongping
    2020, 24 (17):  2759-2769.  doi: 10.3969/j.issn.2095-4344.2537
    Abstract ( 624 )   PDF (66091KB) ( 226 )   Save

    BACKGROUND: Vascular endothelial growth factor (VEGF) is a multifunctional cytokine that promotes angiogenesis, improves vascular permeability, maintains vascular stability, and has positive effects on fracture healing and bone regeneration. VEGF-based treatment is expected to become an emerging means for accelerating fracture healing.

    OBJECTIVE: To explore the effects of VEGF on fracture healing and to review its influencing factors and clinical applications.

    METHODS: The CNKI and PubMed databases were searched for articles related to VEGF effect on fracture healing published from January 2014 to March 2019 with the key words of “vascular endothelial growth factor, fracture healing” in Chinese and English, respectively. Initially 167 articles were retrieved, and only 85 eligible articles were included in result analysis after removal of unrelated or duplicate literature.

    RESULTS AND CONCLUSION: VEGF exists in different cells of the fracture end, and the expression in different cells has significant periodicity. VEGF promotes angiogenesis, enhances vascular permeability, and maintains the normal morphology and integrity of blood vessels, thereby promoting fracture healing and bone regeneration. Several physicochemical factors and cytokines can change the expression of VEGF, thus influencing the fracture healing process. VEGF-based tissue engineering therapies have significant advantages, and the main methods include: delivering VEGF into the affected area using tissue engineering materials; maintaining and improving VEGF expression using gene transfection technology; combining VEGF with other cytokines and exogenous stem cells; and applying bio-coatings based on the upregulation of VEGF. With the interdisciplinary development, VEGF-based tissue engineering treatment is expected to be a hot topic in medical research.

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    Biological function of circular RNA and its role in tissue repair

    Shi Lei, Sun Baolan, Zhang Suyao, Zhang Yuquan
    2020, 24 (17):  2770-2774.  doi: 10.3969/j.issn.2095-4344.2590
    Abstract ( 382 )   PDF (25550KB) ( 196 )   Save

    BACKGROUND: Circular RNA (circRNA) is a new endogenous non-coding RNA produced by the RNA shearing process. It has a closed loop structure, without 3' cap and 5' poly(A) tail. circRNA is widely distributed in eukaryotes, and has the characteristics of stable structure, evolutionary conservation, and abundant sources. Current research has shown that circRNA can function through diversified ways.

    OBJECTIVE: To review the classification and formation process of circRNA, its molecular properties, potential functions and its role in tissue repair.

    METHODS: A computer-based search for literature in CNKI and PubMed databases published from January 2005 to April 2019 was performed using the keywords of “circRNA, function, miRNA, injury” in English and Chinese, respectively. Finally, 39 articles were included for analysis.

    RESULTS AND CONCLUSION: Currently discovered circRNAs can be classified into four types according to their origins: exon-derived circRNA (exonic circRNA), exon-intron circRNA (EliRNA), intron-derived circRNA (ciRNA) and read-through circRNA (rt-circRNA). circRNA is rich, conservative and stable. Increasing studies have shown that circRNA has multiple functions, but mainly has the following four functions: sponge adsorption function, interaction with proteins, regulation of transcription, and participation in protein translation. Numerous studies have shown that circular RNA is involved in the regulation of tissue repair, and its expression profile is significantly related to tissue repair.

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    tau protein and Alzheimer’s disease
    Wang Jinchun, Liu Huiying, Cao Yunpeng
    2020, 24 (17):  2775-2781.  doi: 10.3969/j.issn.2095-4344.2598
    Abstract ( 576 )   PDF (926KB) ( 951 )   Save

    BACKGROUND: Deposition of neurofibrillary tangles is closely related to cognitive decline in Alzheimer’s disease, and tau protein is an important component.

    OBJECTIVE: To investigate the possible pathogenesis of Alzheimer’s disease and the role of tau protein hyperphosphorylation in the progression of Alzheimer’s disease.

    METHODS: The first author searched for relevant articles published from January 2001 to January 2019 in WanFang, CNKI, Vip, PubMed, and Embase with the key words of “Alzheimer’s disease; tau protein; β-amyloid cascade.”

    RESULTS AND CONCLUSION: Hyperphosphorylation of tau protein and formation of paired helix filaments are considered to be the basis of neuronal degradation in Alzheimer’s disease. tau protein may not depend on the cascade response triggered by β-amyloid deposition in the pathogenesis of Alzheimer’s disease, and tau protein-associated vaccine immunity has been verified to produce a curative effect in clinical trials. Further exploration on the relationship between tau protein, β-amyloid protein and Alzheimer’s disease is necessary to better understand the pathogenesis of Alzheimer’s disease, and provide a theoretical basis for the development of therapeutic drugs for Alzheimer’s disease.

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    Meta-analysis of bone metabolism indicators in patients with primary osteoporosis under different forms of exercises 
    Yuan Qiubao, Kong Haijun
    2020, 24 (17):  2782-2788.  doi: 10.3969/j.issn.2095-4344.2574
    Abstract ( 335 )   PDF (26946KB) ( 42 )   Save

    BACKGROUND: Physical exercise can significantly reduce bone mass loss, relieve pain and improve bone metabolism in osteoporosis patients, but there is no evidence-based evidence.

    OBJECTIVE: To investigate the effect of different physical exercises on the treatment of primary osteoporosis.

    METHODS: Randomized controlled clinical trials regarding the therapeutic effect of physical exercise on primary osteoporosis were screened. The physical exercise group was subjected to physical exercise, and the control group had no regular exercise during the test. The main outcome measures included bone mineral density of lumbar spine L2-L4, visual analog scale score, bone metabolism index (osteocalcin, total type 1 procollagen amino terminal peptide, urine pyridinium/creatinine, blood calcium, blood phosphorus). The included outcome indicators were meta-analyzed using the Review Manager 5.3 software.

    RESULTS AND CONCLUSION: A total of 25 randomized controlled trials were included. Meta-analysis results showed that physical exercise could effectively improve the bone mineral density of L2-L4 segments in primary osteoporosis patients (mean difference=0.06, 95% confidence interval [0.04-0.08], P < 0.000 01, I2=89%). Subgroup analysis results revealed significant differences in the control group and five-animal exercise & Yi-Jin-Jing group, setting-up exercise group, composite exercise group, and incremental exercise group compared with control group (P < 0.05), while there was no significant difference between other exercise groups and control group (P > 0.05). Physical exercise significantly reduced the pain as determined by the visual analog scale in osteoporosis patients (mean difference=-0.93, 95% confidence interval [-1.08 to -0.79], P < 0.000 01, I2=83%). Exercise intervention could improve serum osteocalcin, total type 1 procollagen amino terminal peptide and blood phosphorus levels, and reduce urine pyridinium/creatinine and serum calcium levels. However, there was no significant difference between exercise groups and control group (P > 0.05). The results of Egger’s and Begg’s tests indicated that publication bias of the included studies was at a low level. All these findings indicate that physical exercise has significant interventional effects on bone mineral density and pain in patients with primary osteoporosis.

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