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18 November 2018, Volume 22 Issue 32 Previous Issue    Next Issue

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Therapeutic targets of osteoporosis: correlation of age, body mass index, blood glucose and blood lipids with sclerostin and DKK-1 Zhang Shu-dong1, Zhao Si-cong2, Li Shi-fei1, Zhou Jian1, Yao Qi1, 3 2018, 22 (32):  5085-5090.  doi: 10.3969/j.issn.2095-4344.0395 Abstract ( 405 )   PDF (1056KB) ( 130 )   Save BACKGROUND: Chondrocytes have been shown to promote osteogenesis by inhibiting the expression of sclerostin. The key to regulating the balance between bone resorption and bone formation is sclerostin expression inhibition in the process of bone resorption. OBJECTIVE: To analyze the correlation of age, body mass index, fasting blood glucose (FBG) and serum lipid with sclerostin and DKK-1 in adult men. METHODS: Seventy healthy men aged 46-67 years who received physical examination were enrolled. Age, body height, body mass and medical history were collected. The FBG and blood lipids were detected. The serum levels of sclerostin and DKK-1 were detected by ELISA. RESULTS AND CONCLUSION: (1) The level of sclerostin was significantly different among participants with different ages, body mass indices and FBG levels (P < 0.05). The serum level of DKK-1 in 57-67-year-old participants was significantly higher than that in 46-56-year-old participants (P < 0.05). (2) There was no relationship between serum sclerostin and DKK-1 levels. (3) Multiple linear regression analysis showed that serum level of sclerostin was significantly associated with FBG and body mass index. Serum level of DKK-1 was significantly associated with age. Therefore, much attention should be paid to age, FBG and body mass index in studies on sclerostin and DKK-1. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Efficacy and safety of salmon calcitonin combined with calcium for viral cirrhosis with osteoporosis: study protocol for a randomized, parallel, controlled, 2-year follow-up clinical trial and preliminary results Wang Jing-feng1, Liu Tao2, Zhou Bai-sui3, Li Ai-qun4 2018, 22 (32):  5091-5096.  doi: 10.3969/j.issn.2095-4344.0544 Abstract ( 371 )   PDF (1179KB) ( 134 )   Save BACKGROUND: Chronic liver disease is one of risk factors for osteoporosis. At present, there are few studies on drug treatment for chronic liver disease combined with osteoporosis. For patients with osteoporosis, oral calcium therapy is often required, but the efficacy with calcium alone is not good. In patients with osteoporosis and cirrhosis, the relative deficiency of vitamin D in the body may result in osteomalacia and endocrine disorders. As an important calcium- and phosphorus-regulating hormone, calcitonin can diminish blood calcium and blood phosphorus, and further promote osteogenic differentiation. OBJECTIVE: To investigate the efficacy and safety of salmon calcitonin combined with calcium in the treatment of viral cirrhosis with osteoporosis. METHODS: This is a prospective, single-center, open-label, randomized, parallel, controlled, 2-year follow-up clinical trial. This study will include 102 outpatients and hospitalized patients with viral cirrhosis combined with osteoporosis in Yantai Affiliated Hospital of Binzhou Medical University, China. The 102 patients will be equally and randomly divided into two groups. The patients in the control group will receive oral calcium treatment, and the patients in the combination group will receive intramuscular injection of salmon calcitonin combined with oral calcium treatment, for 24 months. The primary outcome measure will be bone mineral density at the L2-4 at 24 months after treatment. The secondary outcome measures will include bone mineral density at the L2-4 before treatment, at 6 and 12 months after treatment; bone mineral density of the femoral neck and Ward's triangle at 6, 12 and 24 months after treatment; serum calcium, phosphorus, 1,25(OH)2D3, parathyroid hormone, osteocalcin, interleukin-10, interleukin-6, tumor necrosis factor-alpha and insulin-like growth factor 1 levels before treatment, at 6, 12 and 24 months after treatment; adverse reaction rate at 6, 12 and 24 months after treatment. This study protocol was approved by Medical Ethics Committee by Yantai Affiliated Hospital of Binzhou Medical University in July 2018 (2018071601). This study will be performed in strict accordance with the Declaration of Helsinki developed by the World Medical Association. Written informed consent will be obtained from each patient. This study was designed in March 2018. Patient recruitment and data collection will begin in October 2018. Participant recruitment will end in October 2019. Data analysis will be performed in January 2022. The whole study will be completed in February 2022. Dissemination plans include presentations at scientific conferences and peer-reviewed scientific publications. This study was registered with the Chinese Clinical Trial Registry (registration number: ChiCTR1800017732). Protocol version: 1.0. RESULTS AND CONCLUSION: Our team has published preliminary results for 64 patients. The 12-month follow-up results demonstrated that compared with the control group (n=32; oral calcium), serum calcium, 1,25(OH)2D3, interleukin-10, insulin-like growth factor 1 levels, and bone mineral density of L2-4, femoral neck and Ward's triangle were significantly increased, but serum phosphorus, parathyroid hormone and osteocalcin, interleukin-6, tumor necrosis factor alpha levels were significantly reduced in the treatment group (n=32; intramuscular injection of salmon calcitonin combined with oral calcium) (both P < 0.05). The trial will verify that salmon calcitonin combined with calcium is a suitable method for the treatment of viral cirrhosis with osteoporosis. Clinical data will confirm that their combination can remarkably improve bone mineral density of the lumbar spine and hip, improve bone biochemical indicators, and reduce the incidence of adverse reactions, with good long-term efficacy. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Can pravastatin reduce the risk of early stage of steroid-induced avascular necrosis of the femoral head? Wang Xiao-long1, Han Chao-qian1, Zhao Xiao-na2, Zhao Jian-min3, Liu Yu3 2018, 22 (32):  5097-5103.  doi: 10.3969/j.issn.2095-4344.0548 Abstract ( 341 )   PDF (5815KB) ( 107 )   Save BACKGROUND: Lipid metabolism and coagulation disorders as well as cell apoptosis have been shown to be closely related to the pathogenesis of steroid-induced avascular necrosis of the femoral head (SANFH). Meanwhile, pravastatin has been reported partially to exhibit the effects of lipid-lowering, anticoagulation and interfering the osteocyte apoptosis. OBJECTIVE: To investigate whether pravastatin can reduce the incidence of the early stage of SANFH in rabbits, and whether it can be used as an effective target for early intervention of SIFHN. METHODS: Sixty male New Zealand rabbits were randomly divided into three groups: model group (n=24, early stage of SANFH), intervention group (n=24, early stage of SANFH treated with pravastatin) and control group (n=12, treated with same volume of normal saline). The rabbit model of early stage of SANFH was established by injection of hormone combined with horse serum. All the rabbits were examined for serum lipids and coagulation indicators and underwent MRI examination at 2, 4 and 6 weeks after modeling. The rabbits were sacrificed for histopathological observation under light microscope. The ratio of empty lacuna was calculated, apoptosis of osteocytes was determined by TUNEL assay, and apoptotic index was calculated. RESULTS AND CONCLUSION: Compared with the model group, the necrotic degree in the intervention group was similar in the gross specimens and MRI performance, and there was no significant difference. The levels of cholesterol, triglyceride and low density lipoprotein in the intervention group were significantly higher than those in the control group at 2 and 4 weeks after modeling (P < 0.05). There was insignificant difference in all above levels between intervention and control groups at 6 weeks after modeling (P > 0.05), but the levels in the intervention group were significantly lower than those in the model group (P < 0.05). There were no significant differences in the prothrombin time, and tissue plasminogen activator levels among groups at different time points after modeling (P > 0.05). Histopathological results revealed that the number of adipocytes and empty lacuna in the bone marrow cavity of the intervention group was less than that in the model group, and the sparse degree of trabecular bone was relatively mild. The ratio of empty lacuna in the intervention group was significantly higher than that in the control group at 2 and 4 weeks after modeling (P < 0.05). The ratio of empty lacuna in the intervention group was significantly lower than that in the model group at 6 weeks after modeling (P < 0.05), but had no significant difference from that in the control group (P > 0.05). The apoptotic index in the intervention group was significantly higher than that in the control group at different time points after modeling (P < 0.05), but there was no significant difference between intervention and model groups (P > 0.05). To conclude, pravastatin cannot reduce the risk of early SANFH, and cannot be used as an effective target for early intervention of SANFH. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Relationship between osteoporosis and incidence of knee osteoarthritis in Sprague-Dawley rats Zheng Zhao-kuan, Zhao Hong-pu, Lü Yu-ming, Zeng Jing, Gong Yi-xing, Huang Jian-wei 2018, 22 (32):  5104-5109.  doi: 10.3969/j.issn.2095-4344.0545 Abstract ( 262 )   PDF (4532KB) ( 96 )   Save BACKGROUND: Incidence of osteoporosis in patients with osteoarthritis is significantly higher than that of  healthy people, indicating that there are factors leading to osteoporosis and osteoarthritis. OBJECTIVE: To determine the effect of osteoporosis on the pathogenesis of osteoarthritis. METHODS: Twenty-eight rats were equivalently randomized into osteoarthritis and osteoporosis plus osteoarthritis (ovariectomy to induce osteoporosis) groups. One month after modeling, papain was injected into the left knee joint of each group to induce osteoarthritis, and the right knee joint in both groups served as the controls. RESULTS AND CONCLUSION: Compared with the right knee joint, osteophyte hyperplasia was observed at the left knee joint in both groups, and arthritis was more severe in the osteoporosis plus osteoarthritis group than in the osteoarthritis group. The Mankin scores in the osteoporosis plus osteoarthritis group were higher than those in the osteoarthritis group. The bone mineral density of L1-4 in the osteoporosis plus osteoarthritis group was lower than that in the osteoarthritis group. To conclude, osteoporosis can promote the development of osteoarthritis to some extent. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Differences in outcomes of different scraping therapy schemes for lumbar disc herniation based on metabonomics  Chen Li-hong1, Yue Rong-zhao1, Zhang Yong-yi1, Zhang Qin1, Wang Yan-jun1, Gui Qian1, Yang Min1, Xu Gui-hua1, Xie Tong2, Peng Lin-xiu2 2018, 22 (32):  5110-5116.  doi: 10.3969/j.issn.2095-4344.0556 Abstract ( 287 )   PDF (1586KB) ( 114 )   Save BACKGROUND: Scraping therapy has been widely used to treat lumbar disc herniation. The underlying mechanism can be studied through multiple links, pathways and targets using metabonomics. OBJECTIVE: To analyze the serum metabolic profile changes in lumbar disc herniation rats interfered with scraping therapy and differences in efficacy of different regimens by gas chromatography-mass spectrometry (GC-MS), and to explore the underlying mechanism to provide new ideas and basis for clinical treatment, and promote the normalization and standardization of scraping therapy. METHODS: Forty-eight Sprague-Dawley rats from Zhejiang Provincial Animal Center were randomly divided into Scraping therapy groups (corresponding channels with ecchymosis, corresponding channels without ecchymosis, acupoints with ecchymosis, acupoints without ecchymosis), model group, and blank control group (n=8 per group). A rat model of noncompressive lumbar disc herniation induced by autologous nucleus pulposus was established in the model and scraping therapy groups. Different scraping therapy programs were given to scraping therapy groups, once every other day, for 3 courses (3 days a course). The mechanical pain threshold before and after modeling was detected by electronic pain threshold detector. The serum endogenous differential metabolites were analyzed by GC-MS. RESULTS AND CONCLUSION: After modeling, the mechanical pain threshold in each group was significantly lower than before modeling (P < 0.01). According to the metabonomics, 14 potential biomarkers such as glycine, pyruvate acid and ornithine were screened out, and these potential biomarkers were involved in seven metabolic pathways, including glycine, serine and threonine metabolism, pyruvate metabolism and citric cycle. Scraping therapy played an anti-inflammatory effect on lumbar disc herniation rats by regulating the urea content, glycine, serine and threonine metabolism, pyruvate, and achieved analgesic effect through the adjustment of arginine and proline metabolism pathways. After comparing the contents of glycine, pyruvate acid, ornithine and urea in four groups of scraping therapy, the corresponding channels with ecchymosis group showed the best therapeutic effect on the abnormal metabolites. In the future, large sampled clinical trials are needed for further research. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Numerical analysis of fluid field changes in articular cartilage with micro-defects under compressive load  Li Xiao-ming1, 2, Men Yu-tao1, 2, Zhang Chun-qiu1, 2 2018, 22 (32):  5117-5122.  doi: 10.3969/j.issn.2095-4344.0387 Abstract ( 250 )   PDF (2212KB) ( 114 )   Save BACKGROUND: Mechanical study of articular cartilage has been focused on the deformation of solid phase, and there are few studies on interstitial flow. While interstitial flow is the core mechanism for maintaining the normal mechanical and physiological functions of cartilage. OBJECTIVE: To study the changes of cartilage fluid field of articular cartilage with micro-defects under compressive load, and to provide theoretical basis for better understanding of mechanical mechanism of cartilage injury. METHODS: The two-dimensional numerical model of fiber-reinforced and porous-viscoelasticity articular cartilage was established to simulate and parameterize the process of interstitial flow under compressive load. The variation of fluid field distribution of articular cartilage with micro-defects was obtained. RESULTS AND CONCLUSION: The increase in interstitial fluid pressure at the bilateral bottom corners of defect sites under compressive load increased the bearing capacity and prevented further damage. The reduction in the interstitial fluid pressure on the bilateral defects and the acceleration in the interstitial flow would promote the damage deteriorating to the surrounding area. In addition, due to stress concentration at the bilateral bottom corners of the defect and interstitial fluid pressure difference, it would make interstitial fluid flowed from the center stress of stress concentration into surrounding area. Our results show that injured cartilage has a self-regulating ability to delay damage. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Bushen Huoxue Decoction affects mRNA expression of various cytokines in cartilage of the rabbit models of osteoporotic osteoarthritis Chen Hai-xia1, Zhang Kai-wei2, Ma Wen-juan3, Shen Feng-jun2, Zhang Bo1 2018, 22 (32):  5123-5132.  doi: 10.3969/j.issn.2095-4344.0371 Abstract ( 252 )   PDF (1452KB) ( 130 )   Save BACKGROUND: Bushen Huoxue Decoction has been found to be effective for osteoarthritis and osteoporosis, but the underlying mechanisms are little reported. OBJECTIVE: To explore the effects of Bushen Huoxue Decoction on the mRNA expression levels of interleukin 1, interleukin 6, and transforming growth factor-β in the knee chondrocytes of female New Zealand female rabbit models of osteoporotic osteoarthritis. METHODS: Thirty female New Zealand rabbits were randomly divided into osteoporosis, osteoarthritis, and osteoporotic osteoarthritis groups. The right knee articular cartilages were removed, and the chondrocytes were isolated and cultured after modeling. Another seven rabbits were selected to prepare seven groups of serum containing drugs: normal group, low-, medium- and high-dose Bushen Huoxue Decoction groups, glucosamine hydrochloride group, alendronate sodium group and glucosamine hydrochloride plus alendronate sodium group. The chondrocyte proliferation in each group was detected by MTT assay. The mRNA expression levels of interleukin 1, interleukin 6, insulin-like growth factor-1 (IGF-1) and transforming growth factor-β (TGF-β) in the knee chondrocytes were detected by PCR. RESULTS AND CONCLUSION: MTT results showed that with the administration time increasing to day 7, the absorbance values in each group were significantly higher than those in the normal group (P < 0.05), except osteoarthritis and low-dose groups. Bushen Huoxue Decoction exerted effect in a concentration-dependent manner. PCR results: the order of mRNA expression level of interleukin 1 was as follows: osteoporotic osteoarthritis group > osteoarthritis group > osteoporosis group (P < 0.05). The mRNA expression level of interleukin 6 was the highest in the osteoporotic osteoarthritis group, followed by osteoarthritis group, the lowest in the osteoporosis group, and the level in the osteoporotic osteoarthritis and osteoarthritis groups was significantly higher than that in the osteoporosis group (P < 0.05). The order of mRNA expression levels of IGF-1 and TGF-β were the highest in the osteoporosis group, followed by osteoarthritis group, the lowest in the osteoporotic osteoarthritis group, and the difference was significant between osteoporosis and osteoporotic osteoarthritis groups (P < 0.05). Compared with the normal group, Bushen Huoxue Decoction significantly decreased the mRNA expression levels of interleukin 1 and interleukin 6, and significantly increased the mRNA expression levels of IGF-1 and TGF-β in a concentration-dependent manner. In summary, Bushen Huoxue Decoction can promote the proliferation of chondrocytes. The occurrence of osteoarthritis and osteoporosis is related to interleukin 1, interleukin 6, IGF-1 and TGF-β mRNA expression. There is a positive correlation between osteoarthritis and osteoporosis. Bushen Huoxue Decoction indirectly or directly protects and repairs injured chondrocytes. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Pilose antler regulates the expression of ADAMTS-4/TIMP-3 gene in the extracellular matrix in a rabbit model of osteoarthritis Sun Zhi-tao1, Feng Hua-long2, He Sheng-hua1, Lai Ju-yi1, Huang Fei-qiang2, Wang Ye-guang1 2018, 22 (32):  5133-5138.  doi: 10.3969/j.issn.2095-4344.0366 Abstract ( 322 )   PDF (1205KB) ( 142 )   Save BACKGROUND: Pilose antler has been shown to be effective for osteoarthritis, but its association with ADAMTS-4/TIMP-3 gene (main composition of extracellular matrix) is rarely reported. OBJECTIVE: To investigate the effect of pilose antler on the expression of ADAMTS-4/TIMP-3 gene in rabbit models of osteoarthritis, and to further elucidate the mechanism of pilose antler in the prevention and treatment of osteoarthritis. METHODS: Forty-two healthy female New Zealand white rabbits were selected and used for establishing the models of osteoarthritis by anterior cruciate ligament rupture. The rabbit models were then randomized into three groups: control group (normal saline), low-dose    (0.21 g/kg) and high-dose (0.84 g/kg) pilose antler groups. After intervention for 9 weeks, the content of hydroxyproline in both knees was detected. The expression levels of mRNA of cartilage target genes in each group were detected by RT-PCR. Protein expression levels of aggrecan and collagen type II were determined by western blot assay.  RESULTS AND CONCLUSION: Compared with the control group, the content of hydroxyproline in cartilage in the low-dose and high-dose pilose antler groups was significantly increased, and the content in the high-dose pilose antler group was significantly higher than that in the low-dose pilose antler group (P < 0.05). The order of mRNA expression level of ADAMTS-4 was as follows: high-dose pilose antler group < low-dose pilose antler group < control group (P < 0.05). The order of mRNA expression levels of metallopeptidase inhibitor 3, aggrecan and collagen type II was as follows: high-dose pilose antler group > low-dose pilose antler group > control group (P < 0.05). The expression levels of aggrecan and collagen type II were the highest in the high-dose pilose antler group, followed by the low-dose pilose antler group, and the lowest in the control group (P < 0.05). Our findings indicate that pilose antler can repair the injured cartilage by inhibiting the secretion of ADAMTS-4, increasing the secretion of metallopeptidase inhibitor 3 and preventing the degradation of aggrecan and collagen type II. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Interactions of Notch signaling pathway and miR-21 in peripheral nerve repair Yang Kai-yun, Sun Pei-pei, Wu Wen-liang, Liu Hai-chun 2018, 22 (32):  5139-5144.  doi: 10.3969/j.issn.2095-4344.0931 Abstract ( 242 )   PDF (1324KB) ( 92 )   Save BACKGROUND: Nerve injury repair is one of the major surgical hot spots in clinical and basic research. Small-gap sleeve method has been confirmed to have therapeutic effects on peripheral nerve injuries, but the detailed mechanism is unclear. The Notch signaling pathway and the microRNA system have gradually become potential hotspots in this field, but their interactions have not yet been clearly reported. OBJECTIVE: To observe the performance of bone marrow mesenchymal stem cells-packed small-gap sleeve in the treatment of peripheral nerve injuries and to explore the interactions of Notch signaling pathway and miR-21 during the repair process. METHODS: Models of 8 mm-recurrent laryngeal nerve defects were made in Sprague-Dawley rats and randomly divided into three groups: sham operation group, small-gap sleeve group and combined group (small-gap sleeve carrying bone marrow mesenchymal stem cells). After transplantation, neuroelectrophysiological results were measured, and the expression of Notch signal and miR-21 was determined by western blot assay. Bone marrow mesenchymal stem cells cultured in a small-gap sleeve undertook neuronal induction. By changing the expression of Notch/miR21, we detected the expression of neural markers and analyzed the activity of bone marrow mesenchymal stem cells. RESULTS AND CONCLUSION: Transplantation of bone marrow mesenchymal stem cells had therapeutic effects on peripheral nerve injuries as confirmed by neurostaining and electromyogram. During the neural differentiation of bone marrow mesenchymal stem cells, the expressions of Notch signal and miR-21 were significantly increased. Activation of Notch signal or miR-21 overexpression could strengthen the neural differentiation of bone marrow mesenchymal stem cells, while inhibition of Notch signal weakened the neural differentiation of bone marrow mesenchymal stem cells, and miR-21 lost its promotion effect. Overall, Notch signaling pathway and miR-21 are considered to promote the therapeutic effects of bone marrow mesenchymal stem cells combined with small-gap sleeve for peripheral nerve injuries, in which Notch signaling pathway has stronger promotion or inhibition roles. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effects of different test programs on the maximal fat oxidation in young females  Li Ying-ying, Peng Li, Bi Chen-chao 2018, 22 (32):  5145-5149.  doi: 10.3969/j.issn.2095-4344.0393 Abstract ( 422 )   PDF (1057KB) ( 98 )   Save BACKGROUND: The exercise intensity corresponding to the maximal amount of fat oxidation per unit time contributes to losing weight. In recent years, the maximum fat oxidation intensity of the subjects is determined by the incremental load program. However, the anaerobic threshold is the turning point of aerobic metabolism to anaerobic metabolism, which has been shown to be related to the maximum fat oxidation. Whether individualized anaerobic threshold level can accurately detect the maximum fat oxidation is unknown. OBJECTIVE: To study the maximal fat oxidation under increasing load scheme and constant load scheme, and to explore the changes of maximal fat oxidation under the two schemes. METHODS: Sixty-six young females underwent treadmill exercise with increasing and constant load schemes, respectively. Gas metabolism indexes during exercise were collected to calculate the fat oxidation and the maximal fat oxidation under the two regimens. RESULTS AND CONCLUSION: The constant load scheme showed a slightly, but not significantly, higher maximum fat oxidation intensity compared with increasing load scheme (P > 0.05). Under increasing load, the maximal fat oxidation intensity was (54.60±9.02)%VO2max and (66.93±6.63)%HRmax, and under constant load, it was (56.71±6.65)%VO2max and (66.53±8.27)%HRmax. Moreover, under constant load, the respiratory exchange ratio and movement velocity corresponding to the maximum fat oxidative intensity were high (P < 0.01), the test time was short and the ratio of energy supplied by fats was low (P < 0.01). Considering the intensity of exercise and time consumption, it is recommended to detect the maximal fat oxidation intensity based on the individual anaerobic threshold level under constant load, especially the older adults who are weak, obese or cannot exercise for a long time. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effect of steady direct current electric field on the vascularization of chorioallantoic membrane Zhang Xin-yue1, Li Chen-jun1, 2, Li Yan2, Ren Kai-ge1, Pan Lian1, Chen Jun-chi1 2018, 22 (32):  5150-5156.  doi: 10.3969/j.issn.2095-4344.0377 Abstract ( 321 )   PDF (1817KB) ( 104 )   Save BACKGROUND: Tissue-engineered bone has become an important means of repairing maxillofacial bone defects due to its minimally invasive, safe and accurate characteristics. OBJECTIVE: The model of chick chorioallantoic membrane was loaded with different intensities of electric field, to detect the blood vessel growth-related parameters, and to explore the effect of electric field on force-vascularization of tissue-engineered bone, in order to search for a method that can promote force-vascularization of tissue-engineered bone, thus increasing the repair success rate. METHODS: The model of chick chorioallantoic membrane (Inshell model) was established using the chicken embryos hatched for 8 days. The models were randomized into A, B, C, D, and E groups, followed by loaded with 0, -0.75, -1.5, -3 and +1.5 V and steady direct current electric fields at different directions. The images were taken at 0, 12, 24, 36 and 48 hours. The average blood vessel length and growth angle were measured by Image J, and analyzed on SPSS 22.0 software.  RESULTS AND CONCLUSION: After loaded with different currents of electric fields, the average growth rate of vessel length in the groups A, B and C was 0.012, 0.023, and 0.047 5 mm/h, respectively. Independent sample t test results revealed that after loaded with -1.5 and 0 V for 48 hours, the average length of blood vessel was (4.22 ±0.436) and (3.21±0.31) mm/mm2, respectively, and the difference was significant (P < 0.01). After loaded with -1.5 and 0 V for 48 hours, the angle of blood vessels was (21.36±20.16)° and (82.17±50.44)°, respectively, and the difference was significant (P < 0.05). The chick chorioallantoic membrane around the electrode in the groups D and E degenerated and chick embryos died. These findings indicate that appropriate negative electricity promotes the growth of blood vessels, and changes the angle of blood vessel growth, suggesting that steady direct current electric field greatly affects chick chorioallantoic membrane. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effects of curcumin on apoptosis and proliferation of inflammatory chondrocytes induced by lipopolysaccharide Li Ya-nan, Ni Juan, Fang Yu-shun, Li Tao, Tan Hong-fei, Zhang Qing-song 2018, 22 (32):  5157-5162.  doi: 10.3969/j.issn.2095-4344.0539 Abstract ( 328 )   PDF (2411KB) ( 122 )   Save BACKGROUND: Curcumin has been shown to play antioxidative, anti-inflammatory and anti-apoptotic roles in osteoarthritis, but the underlying mechanisms remain unclear. OBJECTIVE: To investigate the effects of curcumin on apoptosis and proliferation of chondrocytes induced by lipopolysaccharide, and to explore the potential mechanisms, so as to provide new ideas for the treatment of osteoarthritis. METHODS: Chondrocytes were isolated from the 4-week-old Sprague-Dawley rats, cultured, and identified by toluidine blue staining. The chondrocyte inflammation model in vitro was established by lipopolysaccharide induction. The cells were divided into five groups: control group (common medium); model group (medium with 5 µg/L lipopolysaccharide); low-dose group (medium with 5 µg/L lipopolysaccharide +  5 µmol/L curcumin); medium-dose group (medium with 5 µg/L lipopolysaccharide + 10 µmol/L curcumin); high-dose group (medium with     5 µg/L lipopolysaccharide + 20 µmol/L curcumin). After 24 hours, the expression levels of interleukin 1β and transforming growth factor α in the cellular supernatant were detected by ELISA method. The apoptotic cells were detected by flow cytometry. Cell proliferation was tested by MTT assay. The expression levels of autophagy-related proteins and extracellular regulated protein kinases 1/2 were determined by western blot assay. RESULTS AND CONCLUSION: The levels of interleukin 1β and transforming growth factor α in the supernatant of chondrocytes induced by lipopolysaccharide were significantly higher than those in the control group (P < 0.01). The levels of interleukin 1β and transforming growth factor α in the supernatant treated with curcumin were lower than those in the model group, which was on a dose-dependent manner (P < 0.05, P < 0.01). After 24 hours of treatment, apoptotic rate of cells in the model group was significantly higher than that in the control group (P < 0.01). Compared with the model group, the cell apoptotic rate in each curcumin group was decreased, especially the high-dose group (P < 0.01). MTT assay showed that the cell proliferation ability in the model group was significantly lower than that in the control group (P < 0.01). Compared with the model group, the cell proliferation ability in each curcumin group was restored, and the effect was the best in the high-dose group. The results of western blot showed that the autophagic ability and the levels of autophagy marker (LC3-II) and autophagy related protein (Beclin-1) were significantly increased in each curcumin group compared with the model group(P < 0.05, P < 0.01), and the expression level of p-ERK1/2 protein was significantly increased (P < 0.05). Our findings suggest that 20 µmol/L curcumin cannot only inhibit apoptosis and relieve inflammatory of chondrocytes induced by lipopolysaccharide, but also has a certain antagonistic effect on lipopolysaccharide- induced inhibition of inflammatory chondrocyte proliferation, which may be via regulating the ERK1/2 pathway-activated autophagy. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Changes of microenvironment in a rat model of arthritis treated by the combination of methotrexate and diclofenac sodium Niu Zi-han1, Ran Xin-jian2, Mawlan•Mamut1, Pan Chuan-peng1, He Dong-hang3, Ren Cong-gai3, Xu Qi4 2018, 22 (32):  5163-5168.  doi: 10.3969/j.issn.2095-4344.0368 Abstract ( 422 )   PDF (4701KB) ( 101 )   Save BACKGROUND: Rheumatoid arthritis is an autoimmune disorder that typically results in swollen and painful facet joints and morning stiffness. Its drug therapy is mainly the combination of non-steroidal anti-inflammatory drugs and anti-rheumatic drugs, however, the drug effects on the T-lymphocyte subset balance and the underlying mechanism remain unclear. OBJECTIVE: To explore the effect of the combination of methotrexate and diclofenac sodium on the balance of Th17/Treg cells in a rat model of collagen-induced arthritis. METHODS: After the male Wistar rat model of collagen-induced arthritis model was established, they were randomly divided into blank control, model, methotrexate, diclofenac sodium, methotrexate + diclofenac sodium, and positive control groups. They were assessed by measuring the arthritis index and toe swelling degree. After 4-week treatment, the expression levels of interleukin 10 and transforming growth factor β in the peripheral blood was detected by ELISA, and the change in Th17/Treg cells in the spleen was tested by flow cytometry; the mRNA expression levels of Foxp3 and ROR-γt in the spleen were examined by RT-PCR. RESULTS AND CONCLUSION: Compared with the model group, methotrexate, diclofenac sodium, and their combination could improve the general condition, reduce toes swelling degree, lower arthritis index (especially the methotrexate + diclofenac sodium group), reduce inflammation cell infiltration, and inhibit ROR-γt regulating Th17 cell differentiation and regulating the Foxp3-induced high differentiation of CD4+CD25+Foxp3+Tre, thereby restoring Treg/Th17 cell balance and improving the arthritis symptoms. The average gray value was significantly different among groups (P < 0.05). Except the down-regulation of interleukin 10 and interleukin 6 levels like diclofenac sodium, the combined therapy could down-regulate the expression level of transforming growth factor β, increase the Treg/Th17 cell proportion, and inhibit the expression of Foxp3 and ROR-γt. Additionally, methotrexate significantly reduced the expression level of transforming growth factor β. In summary, the combination of methotrexate and diclofenac sodium possesses a synergistic effect, which provides the theoretical foundation for the treatment of rheumatoid arthritis. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Establishment and comprehensive evaluation of animal models of steroid-induced avascular necrosis of the femoral head Tong Peng1, Wang Yang2, Liang Ying3 2018, 22 (32):  5169-5174.  doi: 10.3969/j.issn.2095-4344.0399 Abstract ( 302 )   PDF (5142KB) ( 156 )   Save BACKGROUND: Establishing a simple, feasible and high-efficient animal model of steroid-induced avascular necrosis of the femoral head (SANFH) is the main method to study SANFH. OBJECTIVE: To establish the animal models of SANFH, and to investigate the pathology, imaging, bone/serum biochemistry and hemorheology. METHODS: Twenty rabbits were randomly divided into two groups. The rabbits were subjected to injection of 10 mL/kg horse serum via the ear vein twice, with 14 days in between; at 14 days after the final venous injection, the gluteal muscle was injected with 8 mg/kg prednisolone acetate, once daily for 6 consecutive days, so as to establish the model of SANFH (experimental group). The remaining rabbits were injected with the same volume of normal saline (control group). At 8 weeks after modeling, X-ray and MRI examinations of the femoral head were performed. At 10 weeks after modeling, the bone/serum biochemistry, hemorheology, and histological observation of the femoral head were conducted. RESULTS AND CONCLUSION: X-ray: in the control group, the cortical bone was thick and the trabecular bone was clear. In the experimental group, the joint space became narrow, the cortical bone became thinner, the bone density decreased significantly, and the trabecular bone was severely in disorder. MRI examination showed that there was no necrotic femoral head in the control group, and the typical bilinear sign was observed in the experimental group. Bone biochemistry: the contents of hexosamine, calcium/hydroxyproline, hexose/hydroxyproline, and calcium in the experimental group were significantly lower than those in the control group (P < 0.05). Serum biochemistry: the calcium-phosphorus product, cholesterol, and calcium levels in the experimental group were significantly lower than those in the control group (P < 0.05), and the triacylglycerol level was significantly higher than that in the control group (P < 0.05). Hemorheology: the whole blood hyperviscosity, low shear viscosity, plasma viscosity and hematocrit in the experimental group were significantly higher than those in the control group (P < 0.05). Histological observation: in the experimental group, there were numerous necrotic osteocytes in the trabecular bone, and the non-necrotic bone cells were swollen; the nucleus was severely deformed and edge-polymerized due to the compression, and the phenomenon of nuclear fragmentation, dissolution, and emptying were observed. The number of multinucleated osteoclasts was significantly increased, and hematopoietic stem cells were significantly reduced. While, the control group exhibited no such phenomena. These results indicate that an ideal animal model of SANFH can be established successfully by the combination of horse serum and high doses of prednisolone acetate, and X-ray examination, MRI qualitative analysis, bone/serum biochemistry as well as hemorheology are ancillary diagnostic tools for SANFH. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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“Accordion” technology treats nonunion in model rabbits Feng Zhen1, Zhang Meng2, Wu Wei-na3 2018, 22 (32):  5175-5179.  doi: 10.3969/j.issn.2095-4344.0384 Abstract ( 305 )   PDF (4970KB) ( 127 )   Save BACKGROUND: “Accordion” technique achieves satisfactory outcomes in the treatment of nonunion, but there is a lack of basic research. OBJECTIVE: To verify that the “accordion” technique is effective for nonunion in New Zealand white rabbits. METHODS: The model of unilateral femoral fracture with noninfectious nonunion was created in the New Zealand white rabbits. The model rabbits were then divided into three groups: experimental group: treated with “accordion” technique (4 weeks of pressure with external fixator at a speed of 0.5 mm/d, then 2 weeks of retraction at the same speed, and finally 4 weeks of pressure); control group: external fixation and pressure; blank control group: only external fixation. The rabbits underwent X-ray examination at 2, 4, 6, 8 and 12 weeks. The fracture specimens were taken for hematoxylin-eosin staining and histological observation. RESULTS AND CONCLUSION: The X-ray examination of the experimental group suggested that the fracture  was healed, while the control group and the blank control group still did not heal (P < 0.01). Histological examination showed that the fracture stump of the experimental group changed from the nonunion state to the healing state, and the blank control and control groups showed no obvious changes. Thus, “accordion” technology is a simple, minimally invasive and efficient treatment method for nonunion. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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miR-21a-5p knockdown promotes the locomotor function recovery of mouse models of spinal cord injury Wang Wen-zhao, Su Yan-lin, Li Hong-fei, Shen Lin, Chen Jia-nan, Pan Xin-da, Jia Tang-hong 2018, 22 (32):  5180-5185.  doi: 10.3969/j.issn.2095-4344.0773 Abstract ( 315 )   PDF (5573KB) ( 103 )   Save BACKGROUND: The pathological changes following spinal cord injury (SCI) are complex, and there is a lack of effective treatment method. miRNAs are non-coding RNA molecules that post-transcriptionally regulate gene expression, which provides a novel treatment strategy for SCI. OBJECTIVE: To investigate the effect of miR-21a-5p on the locomotor function post-SCI in mice and explore the related mechanisms. METHODS: A total of 48 healthy C57BL/6 adult mice were randomly divided into SCI (SCI model was induced by Allen’s method) and sham 1 groups (n=24 per group). Microarray assay was used to find the differentially expressed miRNAs in the lesion tissues after SCI. An additional 72 mice were randomly divided into sham 2, antagomiR and control groups (n=24 per group). The SCI mice in the later two groups were given the intrathecal injection of antagomir-21a and antagomir, respectively. The locomotor function recovery of hind limbs was evaluated by Basso Mouse Scale score; the expression levels of SCI-related markers in the lesion tissues were detected. RESULTS AND CONCLUSION: The expression level of miR-21a-5p was significantly up-regulated in the lesion tissues after SCI (P < 0.05). AntagomiR-21a inhibited the expression of miR-21a-5p post-SCI. The Basso Mouse Scale scores showed a significant difference between sham and SCI groups (P < 0.05), the scores in the antagomiR-21a group was higher than those in the control group at 7 days postoperatively, and showed a significant difference on day 4 (P < 0.05). Compared with the control group, antagomiR-21a significantly suppressed the expression levels of fibronectin and CSPGs (P < 0.05), and significantly up-regulated the expression levels of neurotrophic factors (P < 0.05). Our results suggest that there is a up-regulation in the expression level of miR-21a-5p after SCI, and miR-21a-5p knockdown can suppress scar formation and promote secretion of neurotrophic factors, thereby improving motor functional recovery. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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A method of high purity primary neuron culture and high efficiency transfection Li Xing-tong1, Tang Hong-yan3, Ma Wei1, Yang Jin-wei1, 2, Wang Xian-bin1, Dai Yun-fei1, Li Jun-yan4, 2018, 22 (32):  5186-5190.  doi: 10.3969/j.issn.2095-4344.0383 Abstract ( 519 )   PDF (5504KB) ( 180 )   Save BACKGROUND: Neuron culture and cell transfection technologies are important means to study the development and pathophysiologic mechanism of neurons in vitro, but the purity and transfection efficiency of primary cultured cortical neurons are poor. OBJECTIVE: To establish a simple, efficient and stable method of culture and transfection method of primary neurons in vitro. METHODS: Primary cortical neurons were harvested from neonatal 1-day rat brains under aseptic condition, which was digested with 0.25% trypsin prior to centrifugation and made into cell suspensions, followed by being seeded into Neurobasal-A medium (5×109 L-1 per pore). The morphological characteristics of neurons were observed by inverted microscope; two neuron-specific markers (MAP2 and Tuj1) were used for immunolabeling to identify the cultured cells; the transfection efficiency of neurons was tested by Lipofectamine 2000 and Block-iT Transfection Kit. RESULTS AND CONCLUSION: The neurons cultured in vitro exhibited interconnected networks after culture for 7 days. All the cultured neurons displayed MAP2 and Tuj1 immunoreactivity. The highly effective transfection was observed under fluorescence microscope using Lipofectamine 2000 transfected neurons. In summary, the culture method of primary cerebral cortex neurons can be adopted to obtain highly purified neurons. Besides, high transfection efficiency of primary neurons can be realized by artificial liposome. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effect of acupuncture on the expression level of interleukin-6 in rats after a single eccentric exercise Bai Sheng-chao1, Chen Sheng-ju1, Wang Bo1, Wu Ying1, Shang Hua-yu2 2018, 22 (32):  5191-5196.  doi: 10.3969/j.issn.2095-4344.0540 Abstract ( 254 )   PDF (1294KB) ( 75 )   Save BACKGROUND: Interleukin-6, is a cytokine that has wide sources. Interleukin-6 increases in the skeletal muscle after exercise, which plays an important role in motor function recovery. Acupuncture at the skeletal muscle after exercise can improve the ultrastructure of skeletal mscle, alleviate injury and promote functional recovery. However, acupuncture effects on the interleukin-6 post exercise remains unknown. OBJECTIVE: To explore the influence of acupuncture intervention on interleukin-6 in rats after eccentric exercise. METHODS: Totally 114 male Sprague-Dawley rats were randomly divided into four groups: control group (n=6), exercise group (n=36), acupuncture group (n=36) and exercise plus acupuncture group (n=36). The rats in the exercise and exercise plus acupuncture groups were submitted to eccentric exercise on a downward treadmill at 16° decline. The rats in the acupuncture and exercise plus acupuncture groups were subjected to the oblique acupuncture at the triceps surae. Samples except the control group were collected at 0, 2, 24, 48, 72 and 120 hours after treatment (n=6 per group at each time point). Protein expression level of interleukin-6 in the soleus and rectus femoris was detected by western blot assay. The plasma level of interleukin-6 was determined by ELISA. RESULTS AND CONCLUSION: In the acupuncture group, the protein expression level of interleukin-6 in the soleus and rectus femoris and plasma level of interleukin-6 showed no significant changes (P > 0.05). After eccentric exercise, the interleukin-6 levels in the soleus, rectus femoris and plasma were significantly increased, and the protein expression level of interleukin-6 did not differ significantly between soleus and rectus femoris (P > 0.05). Acupuncture intervention after exercise, the protein expression level of interleukin-6 in the soleus was significantly reduced (P < 0.05), while interleukin-6 levels in the rectus femoris and plasma showed no significant change (P > 0.05). These findings imply that acupuncture cannot markedly change the interleukin-6 level in normal rats. Eccentric exercise can lead skeletal muscle to create more interleukin-6, and acupuncture intervention after eccentric exercise can reduce the amount of interleukin-6 at peak phase in the needling part. But its effect is local, not significant on the skeletal muscle of other parts and plasma. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effect of the combination of aerobic exercise and flavonoids from Lycium ruthenicum Murr on the inflammatory cytokine levels in a mouse model of alcoholic liver injury Li Di1, Chen Rui1, Chen Jia-qin1, Zheng He2, Zhou Bai-cun1, Mao Hai-feng1, 3, Chen Yi-lin1, Qu Hong-lin1, 3, Chen Wei1, Peng Qi1,Luo Chi-miao2 2018, 22 (32):  5197-5202.  doi: 10.3969/j.issn.2095-4344.0370 Abstract ( 251 )   PDF (1354KB) ( 99 )   Save BACKGROUND: There is still a lack of an effective treatment method for alcoholic liver injury. Extracts from Lycium ruthenicum Murr have been shown to play a role in immunoregulation, reducing blood lipid and glucose, antioxidation, prevention and treatment of cardiovascular disease and tumor. OBJECTIVE: To investigate the effect of aerobic exercise combined with flavonoids from Lycium ruthenicum Murr on alcoholic liver injury and its anti-inflammatory mechanism. METHODS: Forty healthy male Kunming mice were used to construct alcoholic liver injury models. The rat models were randomly divided into model, flavonoids, aerobic exercise and aerobic exercise + flavonoids from Lyceum ruthenicum Murr (combination) groups. Aerobic exercise and intragastrical administration of flavonoids from Lyceum ruthenicum Murr were given at 1 day after modeling for 6 weeks. Afterwards, the mice were fasted overnight and then the liver tissues were removed. Hematoxylin-eosin staining and histopathological scoring were conducted. The serum contents of glutamic pyruvic transaminase, glutamic oxaloacetic transaminase and total bilirubin were tested by biochemical analysis, and the mRNA expression levels of tumor necrosis factor α, nuclear factor κB, interleukin 6 and interleukin 10 in the liver tissues and blood were determined by real-time PCR. RESULTS AND CONCLUSION: Hematoxylin-eosin staining and histopathological changes: in the model group, the hepatic cord arranged in disorder, hepatic lobules were damaged severely, and there were abundant uneven fat vacuoles and inflammatory cells in the cytoplasm. The damage degree of liver tissues was significantly lowered in the flavonoids and aerobic exercise groups (P < 0.05). The necrotic hepatocytes, fat vacuoles and inflammatory cell infiltration were significantly decreased in the combination group (P < 0.01). Compared with the model group, the serum contents of glutamic pyruvic transaminase, glutamic oxaloacetic transaminase and total bilirubin in the combination (P < 0.01) and aerobic exercise (P < 0.05) groups were significantly reduced. Real-time PCR results: compared with the model group, the mRNA expression levels of tumor necrosis factor α, nuclear factor κB, interleukin 6 in the liver tissues and blood in the three treatment groups were significantly reduced (P < 0.05), especially in the combination group (P < 0.01), and the mRNA expression level of interleukin 10 was significantly increased. Our findings manifest that aerobic exercise and flavonoids from Lycium ruthenicum Murr can effectively reduce the expression levels of inflammatory cytokines. As a minimally invasive early detection method, circulating RNA detection of plasma inflammatory factors is an important marker in the screen, observation and prognosis of alcoholic liver injury. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Icariin affects the expression of fractalkine and its receptor CX3CR1 in cerebral cortex of natural aging rat models Zhang Xin1, Kang Zeng-jun2, Xue Juan1, Zhang Shun3, Liu Xiao-qing4, Liu Wei-hua1 2018, 22 (32):  5203-5208.  doi: 10.3969/j.issn.2095-4344.0385 Abstract ( 238 )   PDF (1464KB) ( 136 )   Save BACKGROUND: Chemokine fractalkine not only involves chemotaxis, adhesion, and inflammation, but also participates in the proliferation and apoptosis of nerve cells in brain tissue. OBJECTIVE: To observe the effect of icariin on the expression of fractalkine and its receptor CX3CR1 in cerebral cortex of natural aging rats. METHODS: Eighty 3-month-old male Sprague-Dawley rats were randomly divided into four groups: young group, natural aging group, low- and high-dose icariin groups. Rats in the young and natural aging groups were intragastrically administered equal amount of normal saline for 60 days. Starting from 16 months old, rats in the drug groups were given 30 and 60 mg/kg icraiin for 60 days. Pathological changes in brain morphology were observed by hematoxylin-eosin staining. Brain senescence was identified by β-galactosidase staining. The expression of fractalkine and CX3CR1 was detected by immunohistochemistry. The levels of interleukin 6 and tumor necrotic factor α were tested by ELISA. The expression and activity of caspase-3 was determined by western blot assay and colorimetry, respectively. RESULTS AND CONCLUSION: The expression level of fractalkine in the natural aging group was significantly lower than that in the young group. The expression of CX3CR1, the expression and activity of caspase-3, the number of positive particles of SA-β-gal, and the levels of interleukin 6 and tumor necrotic factor α in the natural aging group were significantly higher than those in the young group. After administration of icariin especially at high doses, the above phenomena were reversed. These results suggest that icariin can reduce the levels of inflammatory factors and inhibit apoptosis, which may be via regulating the expression of fractalkine and its receptor CX3CR1. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effect of Notch1/Hes1 signal regulating the expression of CCAAT/enhancer-binding protein alpha on the proliferation and differentiation of type II alveolar epithelial cells induced by hyperoxia Wan Feng-yun, Lu Hong-yan, Wan Xue-qing, Hao Xiao-bo, Zhu Yue 2018, 22 (32):  5209-5214.  doi: 10.3969/j.issn.2095-4344.0386 Abstract ( 327 )   PDF (1234KB) ( 131 )   Save BACKGROUND: Notch1/Hes1 signal and CCAAT/enhancer-binding protein alpha (C/EBPα) play important roles in lung development and injury. Preliminary studies have shown that hyperoxia can induce abnormal proliferation and differentiation of type II alveolar epithelial cells (AECII), which is related to C/EBPα expression. But, whether Notch1/Hes1 signal is involved remains unclear. OBJECTIVE: To investigate the effect of Notch1/Hes1 signal regulating C/EBPα on the proliferation and differentiation of AECII induced by hyperoxia. METHODS: Human AECIIs were cultured in vitro, and randomly divided into air group, air activator group (Notch pathway activator Jagged1 protein, 500 μg/L), hyperoxia group (95% O2 and 5% CO2 mixture gas administered at the speed of 3 L/minute for 10 minutes), hyperoxia activator group (500 μg/L Notch pathway activator Jagged1 protein added at 30 minutes before oxygen administration). AECIIs of each group were collected after 24 hours of intervention. The expression of Notch1, Hes1 and C/EBPα at mRNA and protein levels was determined by RT-PCR and western blot assay. The proliferation of AECII was measured by cell counting kit-8 assay. Flow cytometry labeled AQP5 was applied to detect the differentiation of AECII. RESULTS AND CONCLUSION: Compared with the air group, the mRNA and protein expression levels of Notch1, Hes1 and C/EBPα were increased significantly in the air activator group (P < 0.05), the proliferation of AECII was increased and the differentiation was reduced significantly (P < 0.05). In the hyperoxia and hyperoxia activator groups, the mRNA and protein expression levels of Notch1, Hes1 and C/EBPα were significantly decreased, the proliferation of AECII was decreased and the differentiation was increased significantly (P < 0.05). Compared with the hyperoxia group, the mRNA and protein expression levels of Notch1, Hes1 and C/EBPα were significantly increased in the hyperoxia activator group (P < 0.05), and the proliferation of AECII was increased and the differentiation was decreased significantly (P < 0.05). These results suggest that Notch1/Hes1 signal regulates the expression of C/EBPα and affects the proliferation and differentiation of AECII after exposure to hyperoxia. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Association of interleukin-6 polymorphism with serum level of interleukin-6 in endurance athletes Chen Jun-fei1, 2, Yan Yi1, Guo Jian-jun3, Sun Ming-xiao4, Tang Qiang2, Xie Min-hao5 2018, 22 (32):  5215-5221.  doi: 10.3969/j.issn.2095-4344.0853 Abstract ( 338 )   PDF (1442KB) ( 80 )   Save BACKGROUND: Aerobic capacity has a high heritability, and with the development of molecular biotechnology, molecular genetic marker screening is helpful for gene diagnosis and predicting aerobic capacity. OBJECTIVE: To observe the distribution characteristics of single nucleotide polymorphism (SNP) of interleukin-6 gene in endurance athletes, explore whether it can be used as a molecular genetic marker, and whether there is any association between interleukin-6 SNP and serum level of interleukin-6. METHODS: Two groups of subjects were selected. In the endurance group, 78 athletes of Han nationality and from northern plains who were engaged in endurance project (middle distance race, marathon race, or heel-and-toe walking race) for at least 2 years, and at least second-class sports grade were included. In the control group, 142 subjects who were Beijing city middle school students (nonobese or low body mass, 12-15 years old), and whose parents were of Han nationality from northern plains were included. Three SNP loci of interleukin-6 gene rs1800796 (-572C/G), rs2066992 (1430T/G) and rs13306436 (4477G/A) were typed by PCR-RFLP, and the serum level of interleukin-6 was tested by ELISA. RESULTS AND CONCLUSION: There was gender difference in -572C/G genotype distribution in the endurance group (P < 0.05), and the genotype distribution of the female endurance group was higher than that of the female control group (P=0.095). In the female endurance group CC genotype accounted for 61.1%, and CG+GG genotype accounted for 38.9%. In the female control group, CC genotype accounted for 43.0, and CG+GG genotype accounted for 57.0. The serum level of interleukin-6 of female athletes with -572C/G locus CC genotype in the female endurance group was lower than that in the female control group (P < 0.05). In addition, the serum level of interleukin-6 in athletes with -572C/G locus CC genotype in the endurance group was lower than that in the control group (P=0.070). The rs1800796 of interleukin-6 gene is a molecular marker of female endurance athletes. The serum level of interleukin-6 in female athletes with CC genotype at rs1800796 is lower than that in female controls. Rs1800796, rs2066992 and rs13306436 of interleukin-6 are not genetic markers for male athletes. Further study should focus on expanding the sample size. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Association of sarcopenia with osteoporosis: research and application Ma Jian-xiong1, 2, 3, Kuang Ming-jie1, 2, He Wei-wei1, 2, Chen Heng-ting1, 2, Wang Ying1, 2, Sun Lei1, 2, Lu Bin1, 2, Tian Ai-xian1, 2, Wang Yan1, 2, Dong Ben-chao1, 2, Bo Hao-hao1, 2, Cui Yu-hong3, Ma Xin-long1, 2 2018, 22 (32):  5222-5227.  doi: 10.3969/j.issn.2095-4344.0380 Abstract ( 310 )   PDF (1052KB) ( 107 )   Save BACKGROUND: As the increase in number of older adults, sarcopenia has drawn more and more attention. Importantly, high relevance exists in bone and muscle, osteoporosis and sarcopenia. OBJECTIVE: To review the relationship between sarcopenia and osteoporosis in order to remind when preventing and treating one of the diseases cannot neglect the important role of another disease. METHODS: Databases of WanFang, CNKI, VIP, PubMed, Embase and Medline were searched with the keywords of “sarcopenia, muscular atrophy, osteopenia and osteoporosis” in English and Chinese, respectively for the literature concerning sarcopenia and osteoporosis published before December 2017. The types of literature included journal article, conference literature, dissertation and review. Initially 1 479 articles were retrieved involving in 1 374 English and 105 Chinese articles, and finally 50 eligible articles were enrolled for summation based on the inclusion and exclusion criteria. RESULTS AND CONCLUSION: (1) Muscles and bones are co-regulated by various factors, and the muscle mechanical stress plays an important role in the development and maintenance of the bone. (2) Skeletal system possesses a regulatory effect on muscle, also includes osteoblasts and cytokines secreted by osteocytes. (3) Except for vitamin D, testosterone and corticosteroids, muscle and bone are collectively regulated by oestrogen and growth hormone, suggesting that abnormal changes in these hormones will lead to bone and muscle dysfunction. (4) The mechanism between osteoporosis and sarcopenia is very complex, including the effect of muscle contraction on bone, and endocrine regulation between bone and muscle. (5) Studying on the relevance between osteoporosis and sarcopenia is of instructive significance for the prevention of fractures following falls in older adults. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effect and mechanism of TRPC1 channel regulating function of vascular smooth muscle cells  Gao Chao1, Jia Xiao-ling1, Rong Li-jun2 2018, 22 (32):  5228-5233.  doi: 10.3969/j.issn.2095-4344.0557 Abstract ( 376 )   PDF (958KB) ( 92 )   Save BACKGROUND: The function of vascular smooth muscle cells is closely related to the occurrence of vascular diseases. TRPC1 channel can regulate the function of vascular smooth muscle cells. OBJECTIVE: To summarize the effect of TRPC1 channel on the fine function of vascular smooth muscle, and to discuss the underlying mechanisms. METHODS: A computer-based retrieval of Beijing University of Aeronautics and Astronautics Library, CNKI, Baidu Academic and PubMed Central Medline was conducted for the literature addressing vascular smooth muscle cells and TRPC1 channel from 1969 to 2017. The keywords were “vascular smooth muscle cells, TRPC1 channel, vascular smooth muscle cells function” in English and Chinese, respectively. Finally, 1 Chinese article and 44 English articles were enrolled. RESULTS AND CONCLUSION: TRPC1 channel is one of the non-selective cation channel superfamily TRP and widely expressed on vascular smooth muscle, which regulates vascular smooth muscle cell function. Since its discovery, the TRPC1 channel has been identified as one of the store-operated Ca2+ channels in cells due to its contribution to the store-operated Ca2+ entry. We have got a lot of research results on the TRPC1 channel effect on vascular smooth muscle cells by gene knockout, RNAi, mRNA blocking and specific blocking antibodies and other research methods. In this paper, the results of the current study on the effect of TRPC1 channel on vascular smooth muscle cells are summarized, and the possible mechanism is discussed. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Research advance, bottlenecks and challenges of microarray of micro organs Yang Peng-cheng, Qi Yan, Zhang Dong-hui 2018, 22 (32):  5234-5240.  doi: 10.3969/j.issn.2095-4344.0558 Abstract ( 531 )   PDF (1190KB) ( 211 )   Save BACKGROUND: Because the animal model of drug screening program has the shortcomings of high cost and low effectiveness, different types of tissue microarray of micro organs have been developed for early drug screening. OBJECTIVE: To review the advances in the field of tissue microarray of micro organs. METHODS: PubMed database was retrieved for the articles concerning chips of brain, heart, lung, live, kidney and skin published between 1997 and 2017. The keywords were “micro organs, tissue microarray, drug screen”. The advantages and disadvantages of different methods for preparing the same tissue microarray were summarized.  RESULTS AND CONCLUSION: Tissue microarray of micro organs has the similar function with native tissues, with low cost and high stability, so it can be used for high throughput drug screening of industrialization. Compared with the single cell tile cultured cells, tissue microarray of micro organs has the advantages of reflecting the survival state of the cells and metabolites, and the function parameters of tissues and organs. However, in the design and production of tissue microarray of micro organs, there are some challenges such as simulating environment in vivo, constructing three-dimensional cultivation system, how to promote or maintain cell maturation and perform easy detection. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effects of kinesio taping at different intervention time on the motor function of lower extremity after stroke: a Meta-analysis Wu Yan-hua, Li Zhuang-miao, Dong Xu 2018, 22 (32):  5241-5248.  doi: 10.3969/j.issn.2095-4344.0400 Abstract ( 416 )   PDF (1943KB) ( 174 )   Save BACKGROUND: Kinesio taping is a method of treating motor dysfunction of lower extremity after stroke, but there is still a lack of evidence-based medical evidence for its intervention time and efficacy. OBJECTIVE: To systematically review the effect of kinesio taping at different intervention time on the motor function of lower extremity after stroke. METHODS: PubMed, Web of science, Cochrane Library, EMbase, OVID (MEDLINE), CNKI, WanFang, VIP, CBM, FMRS and grey literature database (Opengrey) were retrieved, from inception to November 3, 2017. Two evaluators independently screened the literature, extracted the data and evaluated the quality according to the inclusion and exclusion criteria. Meta-analysis was conducted on RevMan 5.3. RESULTS AND CONCLUSION: A total of 15 studies involving 675 patients were included. The results of Meta-analysis showed that the trial group was better than the control group in the improvement of lower limb movement, walking and balance function in stroke patients (SMD=1.17, 95%CI (0.46, 1.87), Z=3.25, P=0.001; SMD=0.80, 95%CI (0.42, 1.18), Z=4.15, P < 0.000 1; SMD=1.18, 95%CI (0.59, 1.77), Z=3.94, P < 0.000 1). The results of subgroup analysis showed that the trial group was better than the control group in the improvement of lower limb movement, walking and balance function in patients with cerebral apoplexy after 4 weeks of treatment (SMD=1.73, 95%CI (0.71, 2.76), Z=3.33, P=0.000 9; SMD=0.85, 95%CI (0.54,1.16), Z=5.43, P < 0.000 01; SMD=0.96, 95%CI (0.54,1.39), Z=4.46, P < 0.000 01). The trial group was better than the control group in the improvement of lower limb movement and walking function in patients with cerebral apoplexy after 8 weeks of treatment (SMD=0.44, 95%CI (0.12, 0.75), Z=2.71, P=0.007; SMD=1.11, 95%CI (0.65, 1.56), Z=4.76, P < 0.000 01). These results indicate that 4-week kinesio taping adjuvant therapy is superior to the simple conventional treatment, rehabilitation therapy or other treatments, which can improve the lower limb movement, walking and balance function of stroke patients. After 8 weeks of treatment, it can improve the lower limb movement and walking function of stroke patients. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics