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28 January 2019, Volume 23 Issue 3 Previous Issue    Next Issue

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Mechanism of XCT-790 reducing activity and osteogenesis of osteoblasts Liu Shaojin1, Wan Lei2, Qiao Rongqin2, Huang Hongxing2, Wang Jili1, Li Zhaozheng1 2019, 23 (3):  329-334.  doi: 10.3969/j.issn.2095-4344.0600 Abstract ( 521 )   PDF (655KB) ( 209 )   Save BACKGROUND: Wnt signaling pathway plays an important role in formation, differentiation and mature of osteocytes. As negative regulators of Wnt signaling pathway, DKK1 and Sost play negative roles in regulating bone mass and osteoblast differentiation. XCT-790, an inhibitor of estrogen receptor-related receptor α, can regulate the effect of estrogen signaling pathway on osteoclast function. OBJECTIVE: To observe the effects of XCT-790 on the proliferation of MG63 cells transfected with DKK 1 and Sost overexpression adenovirus vector and the expression of low-density lipoprotein receptor-related protein (LRP) 5, bone morphologic protein 2, osteopontin, osteoprotegerin, and connective tissue growth factor (CTGF) protein. METHODS: MG63 cells were divided into eight groups: blank control, overexpressed DKK1 (ad-DKK1), overexpressed Sost (ad-Sost), overexpressed DKK1+Sost (ad-DKK1+Sost), XCT-790-intervened blank adenovirus, XCT-790-intervened ad-Sost, XCT-790-intervened ad-DKK1-Sost groups. The cell viability was detected by MTT assay. The alkaline phosphatase activity was detected by alkaline phosphatase kit. Ca2+ concentration was detected by flow cytometer. Expression of LRP5, bone morphologic protein 2, osteopontin, osteoprotegerin and CTGF was detected by western blot assay. RESULTS AND CONCLUSION: Compared with the blank control group, ad-DKK1, ad-Sost, ad-DKK1+Sost and XCT-790-intervened blank adenovirus groups could significantly decrease the cell activity, alkaline phosphatase activity, increase calcium concentration and down-regulate the expression of LRP5, bone morphologic protein 2, osteopontin, osteoprotegerin and CTGF (P < 0.05). In the XCT-790-intervented ad-DKK1, ad-Sost, ad-DKK1+Sost groups, the cell activity, alkaline phosphatase activity, and the expression of LRP5, bone morphologic protein 2, osteopontin, osteoprotegerin and CTGF were significantly reduced, and the Ca2+ concentration was significantly increased (P < 0.05). In summary, XCT-790 can directly regulate Wnt signaling pathway, and can cooperate with DKK1 and Sost to inhibit Wnt signaling pathway, thereby decreasing the activity of osteoblasts, LRP5, bone morphologic protein 2, osteopontin, osteoprotegerin, and CTGF protein expression, and further decreasing differentiation, proliferation and osteogenesis of osteoblasts. 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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Gene expression profiles in osteoarthritis: a bioinformatic analysis  Dong Zhengquan1, Wei Lei2 2019, 23 (3):  335-340.  doi: 10.3969/j.issn.2095-4344.0797 Abstract ( 534 )   PDF (730KB) ( 598 )   Save BACKGROUND: Osteoarthritis is a threat for middle-aged and older adults. The development and occurrence of osteoarthritis is regulated by various genes, however, the signaling pathways remain unclear. OBJECTIVE: To investigate the gene expression profiles of osteoarthritis based on a bioinformatics analysis. METHODS: Totally 158 differentially expressed genes during the process of osteoarthritis were obtained from the analysis of synovial tissue data downloaded from the Gene Expression Omnibus database. The functional annotation of gene expression difference was analyzed by DAVID online database. The protein-protein interactions were evaluated by STRING online database. RESULTS AND CONCLUSION: Total 158 differentially expressed genes were identified, including 12 up-regulated genes and 146 down-regulated genes. The Gene Ontology annotation analysis and the KEGG pathway enrichment analysis were performed on DAVID online database, and the results showed that these differentially expressed genes were mainly involved in positive regulation of osteoclast differentiation and cell-cell adhesion, and these pathways were mainly involved in MAPK signaling pathway, P13K-Akt signaling pathway and ECM receptor. The protein-protein interactions of target genes were analyzed and the hub genes such as VEGFA, JUN, PRKACA, PXN and SPTAN1 were identified. In summary, the interaction of these genes may be involved in the process of osteoarthritis, and the network provides a potential target for the diagnosis and treatment of osteoarthritis. 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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Body composition and bone mineral density of juvenile basketball players versus ordinary middle school students: data from a middle school Zhang Yuantong 2019, 23 (3):  341-347.  doi: 10.3969/j.issn.2095-4344.0602 Abstract ( 540 )   PDF (640KB) ( 120 )   Save BACKGROUND: Studying the body composition and bone mineral density (BMD) of juvenile basketball players is not only related to the long-term development of basketball, but also related to the sustainability of national fitness. OBJECTIVE: To investigate the effect of basketball on the body composition and BMD in 14-year-old Han population, and to analyze the differences in BMD between sexes, and the causes. METHODS: Two hundred seventy 14-year-old middle school students of Han nationality in Zhengzhou were enrolled, 79 juvenile basketball players were included as athlete group (28 males, 51 females), 87 ordinary middle school students without consideration of height and body mass as control group 1 (32 males, 55 females), and 104 ordinary middle school students, whose height and body mass were similar to the juvenile basketball players as control group 2 (40 males, 64 females). The relevant data of daily life were obtained by questionnaire survey. The BMD of the middle segment of middle finger of handedness was measured by Aleris Metriscan. The height and body mass were detected, and the body mass index was calculated. The body composition was analyzed using DBA-550 Body Composition Analyzer, and the relative ratio of body mass was calculated. RESULTS AND CONCLUSION: The total water content of female juvenile basketball players was significantly higher than that of female ordinary middle school students (P < 0.05). The protein ratio of juvenile basketball players was significantly higher than that of ordinary middle school students with same sex (P < 0.05), especially in females (P < 0.01). There were significant differences in the fat and fat-free body mass between two groups with same sex (P < 0.01). The inorganic salt ratio of juvenile basketball players was significantly higher than that of ordinary middle school students of the same sex (P < 0.05). There was a significant difference in the BMD of middle finger between athlete group and control group 2 (P < 0.05), and the BMD in the control group 2 was significantly higher than that in the control group 1 of the same sex (P < 0.05). The daily physical exercise time of the juvenile basketball players was significantly higher than that of ordinary students, and the other indicators of life performance showed no obvious regularity. These results indicate that basketball plays a good role in improving the body composition, can reduce the fat content, improve the contents of protein, water, inorganic salts and fat-free body mass. Meanwhile, basketball can increase the BMD of middle finger, which can be used for preventing and treating obesity and osteoporosis. Additionally, the body composition and BMD indexes of basketball players can provide reference for the training and selection. 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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Low-intensity pulsed ultrasound can alleviate knee osteoarthritis pain and promote articular cartilage repair Luo Xianwen1, 2, Li Mingxing1 2019, 23 (3):  348-353.  doi: 10.3969/j.issn.2095-4344.0601 Abstract ( 787 )   PDF (672KB) ( 1797 )   Save BACKGROUND: Low-intensity pulse ultrasound (LIPUS) has been shown to effectively alleviate joint swelling and inflammation in patients with knee osteoarthritis. But its efficacy in the alleviation of osteoarthritis pain and in the repair of articular cartilage is rarely reported. OBJECTIVE: To evaluate the effect of LIPUS on alleviating knee osteoarthritis pain and treating articular cartilage injury. METHODS: Totally 120 patients with knee osteoarthritis were randomly divided into three groups, and treated with diclofenac sodium sustained release tablets (control group), LIPUS (LIPUS group) and their combinations (combination group), respectively. The visual analogue scale, Lequence index, Lysholm score, Noyes score, clinical efficacy and adverse reactions were observed at 1 and 3 months after treatment. RESULTS AND CONCLUSION: At 1 and 3 months after treatment, in the LIPUS and combination groups, the visual analogue scale score and the Lequence index were significantly lower than those in the control group, and the total effective rate was significantly higher than that in the control group (P < 0.01, P < 0.05). At 1 and 3 months after treatment, the Lysholm scores in the LIPUS and combination groups were significantly higher than those in the control group (P < 0.01). The Noyes scores in the LIPUS and combination groups were significantly lower than those in the control group at 1 and 3 months after treatment (P < 0.01). Additionally, there were two cases of drug-related headache and nausea in the control group, and no ultrasonic toxicity or drug-related adverse events occurred in the other two groups. These results imply that LIPUS is effective in the treatment of knee osteoarthritis, can effectively relieve pain and promote the repair of articular cartilage injury. It shows no effect on the effectiveness of drugs. 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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Expression of perilipin, comparative gene identification-58 and triglyceride in mouse skeletal muscle after 4 weeks of endurance training Liao Shuaixiong1, Zhang Guodong2, Song Gang1, 3 2019, 23 (3):  354-360.  doi: 10.3969/j.issn.2095-4344.0591 Abstract ( 484 )   PDF (730KB) ( 247 )   Save BACKGROUND: Comparative gene identification-58 and perilipins (2-5) are expressed in skeletal muscle. There are many studies addressing hormone sensitive lipase regulating steatolysis in skeletal muscle, but changes of triglyceride, perilipin family proteins and comparative gene identification-58 in skeletal muscle during exercise are little reported. OBJECTIVE: To investigate the effects of 4-week endurance training on the expression of perilipin family proteins and comparative gene identification-58 in C57BL/6J mice, and to explore their correlation with triglyceride level. METHODS: Eighty 6-week-old male healthy SPF C57BL/6J mice were randomly divided into control group and endurance training group according to the time-points pre-exercise, 1, 2, 3 and 4 weeks post-exercise. The first week of endurance exercise was adaptive training, and the formal training lasted for 4 weeks, 6 days weekly, the training time in the morning 8: 00-12: 00, with 16 m/minute, 5% degree slope for 40 minutes of treadmill exercise. Mice in the control group were allowed free access to food. The body mass and blood indexes were tested. The triglyceride level, expression of perilipin family proteins and comparative gene identification-58 in mouse gastrocnemius and soleus were detected. RESULTS AND CONCLUSION: Endurance training showed no effect on mouse body mass, blood glucose and free fatty acids, and significantly reduced blood total triglyceride, total cholesterol and insulin levels (P < 0.05), and increased the level of high density lipoprotein cholesterol (P < 0.05). The intramuscular triglyceride of the soleus muscle was higher than that of the gastrocnemius muscle. During the period of endurance training, intramuscular triglycerides firstly reduced and then rose (P < 0.05). In the working muscles (soleus and gastrocnemius) perilipin family proteins (except perlipin 2), comparative gene identification-58 mRNA and protein were significantly increased (P < 0.05). In summary, in the early stage of endurance training, changes in perilipin family proteins (except PLIN2), comparative gene identification-58 mRNA and protein expression participate in the regulation of hydrolysis, and their participation is weakened thereafter, indicating intramuscular triglyceride synthesis on fat is stronger than its hydrolysis. 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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Participation degree and control ability of human latissimus dorsi, trapezius, triceps, musculus and deltoid during cross support of hand ring: analysis on contribution rate of major muscle group Lan Qingshi 2019, 23 (3):  361-366.  doi: 10.3969/j.issn.2095-4344.0593 Abstract ( 631 )   PDF (732KB) ( 1169 )   Save BACKGROUND: There is a lack of large-sampled comparative study on the kinematics and surface electromyography (sEMG) between elite and general hand ring athletes undergoing cross bracing. OBJECTIVE: To explore the time and angle of cross support and the sEMG of main muscle groups in elite hand ring athletes, and to compare the differences between elite and general athletes. METHODS: Two high-speed cameras and MegawinT8 sEMG analyzer were used to collect the measurements in elite group (n=13, elite and first-level athletes) and general group (n=14, second-level athletes). The kinematics (time and angle) of positive/inverted/horizontal cross support and sEMG of trapezius, latissimus dorsi, teres major, deltoid, triceps brachii, pectoralis major, serratus anterior and biceps brachii were detected. RESULTS AND CONCLUSION: Compared with the general group, in the elite group, the positive cross support time increased by 58.9% (P < 0.01), left and right elbow angle was increased by 16.4% and 17.3% (P < 0.05), the angle between the left and right shoulder line and a horizontal surface was decreased by 46.9% and 49.2% (P < 0.01). The contribution rate of latissimus dorsi, teres major, and deltoid was increased by 28.8%, 108.1% (P < 0.01) and 5.8% (P < 0.05), respectively. The contribution rate of triceps brachii, pectoralis major, and biceps brachii was decreased by 47.8% (P < 0.01), 16.2% and 16.7% (P < 0.05). Compared with the general group, in the elite group, the inverted cross support time was increased by 81.6% (P < 0.01). The contribution rate of trapezius, teres major and triceps brachii was increased by 25.6%, 128.6% and 175.0% (P < 0.01), respectively. The contribution rate of serratus anterior and biceps brachii was reduced by 17.9% and 20.7% (P < 0.01). The horizontal cross support time, hip and knee angles were increased by 59.9% (P < 0.01), 9.0% and 6.9% (P < 0.05). The left and right wrist shoulder angles, and trunk angle was decreased by 20.4%, 22.1% and 31.1% (P < 0.01). The contribution rate of trapezius, latissimus dorsi and teres major was increased by 14.0% (P < 0.05), 508.3% and 18.2% (P < 0.01). The contribution rate of anterior serratus was decreased by 12.5% (P < 0.05). These findings indicate that control of the cross support time and the angle of the body position, and strengthening the participation degree and control ability of the latissimus dorsi, trapezius, triceps, musculus and deltoid contribute to completing the hand ring better. 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 kinesio taping on delayed onset muscle soreness in athletes Dong Qizheng 2019, 23 (3):  367-371.  doi: 10.3969/j.issn.2095-4344.0606 Abstract ( 611 )   PDF (597KB) ( 670 )   Save BACKGROUND: There are many studies on kinesio taping effects on delayed onset muscle soreness (DOMS) in general population, but its effects in athletes are little reported. Moreover, the assessment criterion is based on pain index, but not on biochemical factors. OBJECTIVE: To investigate the effect of kinesio taping on DOMS in athletes. METHODS: Forty-three athletes were recruited, and randomized into trial (n=21) and control (n=22) groups. All subjects underwent bilateral knee extension under 70% of maximum load for 5 courses (10 times a course, with 120 seconds in between) to create model of DOMS. Only the trial group received kinesio taping. The Visual Analogue Scale score, interleukin 6 and creatine kinase levels were determined at 0, 24, 48 and 72 hours after DOMS. RESULTS AND CONCLUSION: The Visual Analogue Scale scores at 0, 24, 48 and 72 hours after DOMS in the trial group were significantly lower than those in the control group (P < 0.01). The interleukin 6 level at 0 and 24 hours after DOMS in the trial group was significantly lower than that in the control group (P < 0.05 or P < 0.01). In the trial group, the Visual Analogue Scale scores increased firstly and then decreased (P < 0.01 or P < 0.05). The interleukin 6 level was on a decline, and the level was significantly decreased compared with 0 hour after DOMS (P < 0.05 or P < 0.01). The interleukin 6 level at 48 and 72 hours was significantly lower than that at 24 hours (P < 0.05). The creatine kinase level increased firstly and then decreased. Compared with 0 hour, there was a significant increase in creatine kinase level at 24 hours, and a significant decrease at 72 hours (P < 0.05). The creatine kinase level at 48 and 72 hours was significantly lower than that at 24 hours (P < 0.05). To conclude, kinesio taping significantly alleviates inflammatory reaction in athletes with DOMS, and shortens the recovery 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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A centrifuge device for high acceleration loading in rats: mechanobiological responses of animals in a high-acceleration environment Gao Zijian1, Yu Lu2, 3, Gao Lilan1, Zhang Chunqiu1, Zhang Xizheng2 2019, 23 (3):  372-377.  doi: 10.3969/j.issn.2095-4344.0588 Abstract ( 394 )   PDF (820KB) ( 301 )   Save BACKGROUND: Manned spacecraft launches and returns, and shipboard aircraft takes off and landing, will expose astronauts and pilots to high acceleration environment. High acceleration environment has an important impact on cells, organs, and bodies. The high acceleration centrifuge is a device that can provide high acceleration environment for cells and animals. It plays a role of basic platform in the study on mechanics biology and tissue engineering, especially biological laboratory. OBJECTIVE: To design a centrifuge for high acceleration loading in rats based on a centrifuge and to study the mechanobiological responses of rats in a high-acceleration environment. METHODS: Design process of centrifuge was as follows: determine parameters and select motor → design and validate of the centrifuge → select controller → manufacture and commission machine. Rats were loaded horizontally and vertically. Four groups including 4 G group, 8 G group, 10 G group and 20 G group and one control group were set up. The effects of different high acceleration environments on the physical condition and walking ability of rats were detected. RESULTS AND CONCLUSION: We designed and manufactured the centrifuge, and it included two parts: mechanical structure and control system. The machine run smoothly and had no noise, and could provide 0-40 G high acceleration environment and variable acceleration environment. After rats were subjected to different high acceleration loads, their bodies were inclined to one side and could not walk in a straight line, and accompanied by trembling, hindlimb congestion and other symptoms. The higher the high acceleration value, the greater the influence on rats. After a period of rest, the body could basically recover. Our test suggests that the centrifuge can provide a high acceleration environment for animal experiments, which can be used to study the mechanobiological responses of animals in high acceleration environment. 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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Vascular endothelial growth factor combined with mutant hypoxia-inducible factor 1alpha promotes angiogenesis Hu Liang1, Wang Junhai1, Wang Zhilie1, Xie Jinyuan1, Chen Deng1, Ding Fan2 2019, 23 (3):  378-383.  doi: 10.3969/j.issn.2095-4344.1024 Abstract ( 319 )   PDF (793KB) ( 342 )   Save BACKGROUND: Transcription and translation of vascular endothelial growth factor has been shown to be increased under ischemic and hypoxic conditions, which effectively improves angiogenesis and collateral microcirculation in vivo. Expression of mutant hypoxia-inducible factor-1α (HIF-1αmu) requires hypoxic condition, so its application is limited. OBJECTIVE: To study the effect of transfection of adenovirus-mediated- vascular endothelial growth factor combined with HIF-1αmu (Ad-VEGF-IRES-HIF-1αmu) into endothelial progenitor cells on the angiogenesis in the treatment of steroid-induced avascular necrosis of the femoral head. METHODS: Ad-VEGF-IRES-HIF-1αmu was transfected into endothelial progenitor cells, and the cell viability, morphology and cytopathic effect were observed. Endothelial progenitor cells transfected with Ad-VEGF-IRES-HIF-1αmu were implanted into the necrotic site of the animal model of steroid-induced avascular necrosis of the femoral head (experimental group: transfected with Ad-VEGF-IRES-HIF-1αmu; control group: suspension of endothelial progenitor cells; blank group: cell culture medium). After 10 weeks of transfection, expression of vascular endothelial growth factor, hypoxia-inducible factor1α and CD34 as well as microvessel density were detected. The vascular morphology was observed by ink perfusion. RESULTS AND CONCLUSION: The expression levels of vascular endothelial growth factor and hypoxia-inducible factor1α in the experimental group were higher than those in the control and blank groups (P < 0.05). The number of microvessels positive for CD34 was the argest in the experimental group. In the experimental group, the ink perfusion of femoral head showed new angiogenesis, the phenomenon of recanalization occurred in some vessels, blood vessels had a clear connectivity, and effective vascular veins evenly distributed into the defect area. The area of neovascularization in the experimental group was significantly larger than that in the control and blank groups (P < 0.05). Vascular endothelial growth factor combined with mutant hypoxia-inducible factor1α can enhance angiogenesis in the repair of steroid-induced avascular necrosis of the femoral head. 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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Ginsenoside Rg1 protects against hepatocyte apoptosis in a rat model of non-alcoholic fatty liver disease Xiao Yang1, Hou Yunhe2, Yin Xin3, Kang Feng3, Li Shude2, Yang Shikun4, Tao Jianping1 2019, 23 (3):  384-390.  doi: 10.3969/j.issn.2095-4344.0597 Abstract ( 454 )   PDF (1078KB) ( 494 )   Save BACKGROUND: Ginsenoside Rg1, the functional and active ingredient of Panax notoginseng and Panax ginseng, can alleviate oxidative stress and inhibit cell apoptosis. OBJECTIVE: To investigate the inhibitory effect of ginsenoside Rg1 on hepatocyte apoptosis in non-alcoholic fatty liver disease and the underlying mechanism. METHODS: Forty Sprague-Dawley rats were randomly assigned into four groups (n=10) after 2-week adaptive feeding, including control group (normal diet), model (22-week high-fat and high-sugar diet), low- and high-dose ginsenoside Rg1 groups. The rats in the ginsenoside Rg1 groups were fed with high-fat and high-sugar diet for 8 weeks, and then treated with low- and high-dose ginsenoside Rg1 for 8 weeks via gavage, respectively. The levels of oxidative stress indexes were tested by reagent color-developing method. The pathological changes of liver tissues were observed by heamtoxylin-eosin staining. The lipid droplet in the liver tissue was observed by oil red O staining. The apoptosis of hepatocytes was detected by TUNEL staining. The mRNA expression levels of Bax, Bcl-2, and Caspase-3 in the liver tissue were detected by Q-PCR. The protein expression levels of Bax, Bcl-2, Procaspase-3, Caspase-3 cleavage p17 in the liver tissue were determined by immunohistochemistry and western blot assay. RESULTS AND CONCLUSION: Hematoxylin-eosin staining results showed that compared with the control group, the hepatocytes in the model group arranged in disorder with numerous large fat lipid droplets, and inflammatory cell infiltration. After treatment with ginsenoside Rg1, the hepatocytes returned to normal arrangement and lipid droplets and inflammatory cell infiltration reduced. Oil red O staining results found that compared with the model and control groups, the lipid droplet in the hepatocytes were obviously reduced after treatment of ginsenoside Rg1 in a dose-dependent manner. TUNEL staining revealed the number of apoptotic hepatocytes in the ginsenoside Rg1s was significantly smaller than that in the model and control groups. Compared with the control and model groups, the contents of glutathione and superoxide dismutase in the ginsenoside Rg1 group were significantly increased, and the content of malondialdehyde was significantly decreased (P < 0.05). Compared with the control and model groups, there was a significant decrease in the mRNA expression levels of Bax and Caspase-3, and a significant increase in the mRNA expression of Bcl-2 (P < 0.05). Meanwhile, the protein expression levels of Bax and Caspase-3 cleavage p17 were significantly increased, and the protein expression levels of Bcl-2 and Procaspase-3 were significantly decreased (P < 0.05), which presented a dose-dependent manner. To conclude, ginsenoside Rg1 can relieve oxidative stress and suppress hepatocyte apoptosis in the rat model of non-alcoholic fatty liver disease, and further delays the disease development. 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 | Related Articles | Metrics

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Expression of RANKL in chronic periapical periodontitis in mice Zhang Hong, Ye Dandan, Dong Ming, Niu Weidong 2019, 23 (3):  391-395.  doi: 10.3969/j.issn.2095-4344.0398 Abstract ( 468 )   PDF (823KB) ( 940 )   Save BACKGROUND: Different degrees of bone resorption and granulation tissues appear in the apical region of chronic periapical periodontitis. RANK-RANKL-osteoprotegerin signaling pathway has been shown to be the key pathway for regulating bone resorption. OBJECTIVE: To detect the expression and role of RANKL in mouse apical periodontitis. METHODS: C57BL/6J mice were allocated into experimental and control groups. In the experimental group, were used to open the mandibular first molars exposed to the oral environment were open to create the model of periapical periodontitis. The mandibular tissue was collected at 1, 2, 3 and 4 weeks after modeling, and the periapical tissue changes were observed by hematoxylin-eosin staining. The expression level of RANKL at each time point was detected by immunohistochemical staining. RESULTS AND CONCLUSION: Hematoxylin-eosin staining results showed that the mouse model of chronic periapical periodontitis was successfully established. In the control group, only a few inflammatory cells were found in the apical area of mandibular first molars, and the periodontal tissue was intact. The extent of inflammatory infiltration was gradually increased from 1 week to 4 weeks after modeling, and the alveolar bone destruction gradually increased. The results of immunohistochemistry showed that the expression level of RANKL in the experimental group was significantly higher than that in the control group, the level increased significantly from 1 week to 2 weeks after modeling, continued to increase at 3 weeks, and reduced at 4 weeks (all P < 0.05). To conclude, the model of chronic periapical periodontitis is successfully established in mice. The expression level of RANKL in mouse chronic apical periodontitis is high and shows a clear trend, which may promote the progress of chronic apical periodontitis and bone destruction. 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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Protective effects of methane saturated saline on endotoxin-induced acute lung injury in rats Meng Qingxi1, Wang Weiheng2, Sun Aijun3, Xi Yanhai1, Ye Xiaojian1 2019, 23 (3):  396-403.  doi: 10.3969/j.issn.2095-4344.0599 Abstract ( 448 )   PDF (1129KB) ( 440 )   Save BACKGROUND: Methane has been shown to have significant antioxidant and anti-apoptotic effects in many diseases, such as intestine, liver, myocardium and other ischemia/reperfusion injury. The protective effect of methane on acute lung injury (ALI) has not been studied. OBJECTIVE: To investigate the protective effect of methane saturated saline on ALI induced by endotoxin in rats and its possible mechanism. METHODS: ALI in rats was induced by endotoxin (20 mg/kg). The experiment was divided into blank, control (methane saturated saline), ALI, ALI and methane (ALI treated by methane saturated saline) groups. After 24 hours, the wet/dry weight ratio of lung tissue was detected. Hematoxylin-eosin staining and electron microscopy were conducted. The contents of tumor necrosis factor α, interleukin 6, and interleukin 1β in alveolar lavage, and activity of malondialdehyde and superoxide dismutase in lung tissue were determined by ELISA. Cell apoptosis was measured by TUNEL. The content of caspase-3 in lung tissue was detected by western blot assay. RESULTS AND CONCLUSION: There was no significant difference in each index between blank and control groups. Compared with the blank group, the wet/dry weight ratio, inflammatory reaction in lung tissue, oxidative stress and apoptotic rate were significantly increased in the ALI and ALI and methane groups (P < 0.05). Compared with the ALI group, the wet/dry weight ratio, inflammatory cytokines, apoptotic rate, caspase-3 expression and malondialdehyde content were significantly decreased, and superoxide dismutase activity was significantly increased in the ALI and methane group (P < 0.05). In summary, methane saturated saline can treat ACI caused by endotoxin by reducing the level of oxidative stress in lung tissue, inhibiting the expression of inflammatory factors and reducing apoptosis. 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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Alteration and significance of E-selectin and P-selectin in the tissues surrounding fracture in mouse models Luo Wei1, Li Xianghe1, Yang Xianteng2, Li Senlei2, Zhang Jin2, Tian Xiaobin2, Sun Li2 2019, 23 (3):  404-408.  doi: 10.3969/j.issn.2095-4344.0603 Abstract ( 432 )   PDF (667KB) ( 135 )   Save BACKGROUND: E-selectin and P-selectin are important adhesion molecules involved in the inflammatory response in vivo, and inflammatory response plays a positive role in promoting early fracture healing. OBJECTIVE: To investigate the alterations and significance of E-selectin and P-selectin in the tissues surrounding fracture of mice. METHODS: Thirty-six BALB/c mice were selected for establishing stable tibia fracture model. The mouse models were randomly divided into 1-, 6-, 12-, 24-, and 48-hour and 3-day groups. The hematoma tissue surrounding the fracture was removed at corresponding time, and contralateral tibial tissues at the healthy side were used as controls. The concentrations of E-selectin and P-selectin in tissues were detected by ELISA. RESULTS AND CONCLUSION: The concentrations of E-selectin and P-selectin in the tissues surrounding the fracture were significantly higher than those in the healthy side (P < 0.01). E-selectin reached a peak at about 6 hours. P-selectin was the highest at about 24 hours. In summary, the concentrations of E-selectin and P-selectin in the tissues surrounding the fracture were significantly up-regulated, suggesting that E-selectin and P-selectin may mediate the migration and adhesion of inflammatory cells during the early stage of mouse fracture and play an important role in the pathological process of bone repair. 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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Platelet-rich fibrin repairs cartilage defect in a rabbit model of old traumatic knee arthritis in weight-bearing region Zou Chengda, Chen Xingguang, Su Guanghao, Zhang Ya, Wang Xiaodong 2019, 23 (3):  409-415.  doi: 10.3969/j.issn.2095-4344.0605 Abstract ( 430 )   PDF (998KB) ( 293 )   Save BACKGROUND: Platelet-rich fibrin not only exhibits anti-infective and anti-inflammatory properties, but also can promote tissue healing. It consists of various cytokines that can promote bone marrow mesenchymal stem cells (BMSCs) to differentiate into chondrocytes. OBJECTIVE: To explore the effect of platelet-rich fibrin on the repair of cartilage defect of old traumatic knee arthritis in weight-bearing region of rabbits. METHODS: Twenty-seven 6-month-old New Zealand rabbits provided by Experimental Animal Center, Soochow University, China were selected. Rabbit models of old traumatic knee arthritis cartilage defect in weight-bearing region (a depth of 3.0-4.0 mm, a diameter of 4 mm) were established by Hulth method. At 3 weeks, the granulation tissue was removed from the old defect via the original surgical incision, and the defect region was filled with the prepared filler. There were three groups, including platelet-rich fibrin, BMSCs and blank control groups. The joint was removed at 4, 8 and 12 weeks after repair for gross and histological observations. International Cartilage Repair Society scores and Wakitani’s scores were used to assess the repair outcomes. RESULTS AND CONCLUSION: Gross observation: at 4, 8, and 12 weeks after implantation, in the blank control group, abundant worn cartilage tissues, hyperplastic fibrous connective tissue and osteophyte, and articular cavity adhesion appeared. In the platelet-rich fibrin group, defect region was smooth, and the newly born tissue was similar to the surrounding normal tissue, which was better than that in the BMSCs group. Hematoxylin-eosin staining, Alysin blue staining, safranin-O staining, and type II collagen immunohistochemical staining showed that rabbit knee chondrocytes gradually increased, and chondrocytes showed type II collagen expression in the platelet-rich fibrin group. The degenerative chondrocytes at all layers were observed in the blank control group. There were few chondrocytes in the BMSCs group. International Cartilage Repair Society scores and Wakitani’s scores in the platelet-rich fibrinogen group were significantly superior to those in the blank control group (P < 0.01) and BMSCs group (P < 0.05). These results imply that platelet-rich fibrin can repair old cartilage defects in weight-bearing region caused by osteoarthritis, and its repairing ability is better than that of simple BMSCs. 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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Preserving ligamentum flavum for preventing dural adhesions after lumbar surgery in model rabbits Li Shuwen, Yang Zhe, Yin Heping, Wu Yimin, Bai Ming, Du Zhicai, Wang Yupeng, Meng Gedong 2019, 23 (3):  416-420.  doi: 10.3969/j.issn.2095-4344.0553 Abstract ( 433 )   PDF (702KB) ( 366 )   Save BACKGROUND: Formation of scar tissue and the consequent formation of adhesions with dura and nerve root is an important factor for postoperative outcomes of laminectomy. How to reduce or prevent the adhesion or scar formation has been paid much attention in the clinic.  OBJECTIVE: To explore the effect of preserving the ligamentum flavum on preventing dural adhesions after laminectomy in lumbar surgery. METHODS: Forty New Zealand white rabbits were randomly divided into ligamentum flavum resection group and ligamentum flavum preservation group (n=20 per group). All animals underwent L4-5 laminectomy, simulating nucleus pulpotomy. Postoperative dural adhesion was assessed according to Rydell-balazs and Nuss-baum criteria. RESULTS AND CONCLUSION: The operations were successfully completed, and there was visible scar in the operation area in the ligamentum flavum preservation group, the dura adhesion was easy to be separated bluntly from surrounding tissue. In the ligamentum flavum resection group, the scar tissue around the dura was very dense and the dural adhesions were serious, even osteoid was observed. The Rydell-balazs and Nuss-baum scores in the ligamentum flavum preservation group were significantly lower than those in the ligamentum flavum resection group (P < 0.05). To conclude, retaining the ligamentum flavum can reduce epidural scarring and adhesion of the dura to the surrounding tissue. 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 | Related Articles | Metrics

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Establishment and identification of a rabbit model of comorbid osteoarthritis and osteoporosis Ma Wenjuan1, Zhang Kaiwei2, Shen Fengjun2, Deng Zhiyuan3, Cheng Xiaojin4 2019, 23 (3):  421-426.  doi: 10.3969/j.issn.2095-4344.1025 Abstract ( 303 )   PDF (735KB) ( 140 )   Save BACKGROUND: The elderly often suffers from comorbid osteoarthritis and osteoporosis, but the relationship between osteoarthritis and osteoporosis is not yet clear. OBJECTIVE: To establish and identify an animal model of comorbid osteoarthritis and osteoporosis. METHODS: Thirty-seven female New Zealand white rabbits at 2 months of age were provided by the Animal Center of Guiyang Medical University, China. Of the 37 rabbits, 30 rabbits were selected and divided into osteoarthritis group, osteoporosis group, osteoarthritis and osteoporosis group. The remaining seven rabbits were used as normal control group. Different animal models of osteoporosis (excision of bilateral ovarian and intramuscular injection of methylprednisolone)/osteoarthritis (excision of anterior cruciate ligament)/osteoarthritis and osteoporosis (excision of bilateral ovaries and anterior cruciate ligament+intramuscular injection of methylprednisolone) were made in corresponding groups. After successful modeling, cartilage samples from each rabbit were isolated and cultured. Secretion of type II collagenase was compared among three groups. RESULTS AND CONCLUSION: (1) After excision of rabbit anterior cruciate ligament and bilateral ovaries and intramuscular injection of methylprednisolone, the bone density of osteporotic rabbits was reduced by more than 25%. Exfoliation of the articular cartilage could be observed from the joint appearance, and osteophyte formed markedly shown on the X-ray film of the knee joint obviously. Obvious articular cartilage damage was also observed on the MRI image. Therefore, the model of comorbid osteoarthritis and osteoporosis was successfully prepared by the above methods. (2) Chondrocytes could be successfully extracted from the rabbit knee joint cartilage in the three model groups. The ability of chondrocytes secreting type II collagenase was significantly lowered in the osteoporosis group compared to the osteoarthritis and osteoporosis group (P < 0.05). (3) To conclude, the animal model of comorbid osteoarthritis and osteoporosis was successfully established. 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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Different intensities of endurance training affect serum Irisin level and skeletal muscle protein expression of PGC-1alpha, FNDC5, and PPARdelta in mice with obesity induced by a high-fat diet Su Kunxia 2019, 23 (3):  427-434.  doi: 10.3969/j.issn.2095-4344.0613 Abstract ( 475 )   PDF (1567KB) ( 334 )   Save BACKGROUND: Peroxisome proliferators-activated receptor-γ coactivator-1 alpha (PGC-1α), fibronectin type III domain-containing protein 5 (FNDC5) and Irisin in mice have been shown to make a positive pathway of lipolysis, and PGC-1α can regulate the expression of peroxisome proliferator-activated receptor delta (PPARδ), which regulates lipolysis in skeletal muscle. Endurance training can effectively reduce the body mass of mice, but the effect of different intensities of endurance training on serum Irisin level, and PGC-1α, FNDC5, PPARδ expression in mice with obesity induced by a high fat diet is still unclear. OBJECTIVE: To study the effect of high- and low-intensity endurance training on serum Irisin level, and lipolysis-related proteins in mice with obesity induced by a high fat diet. METHODS: Four-week-old male C57BL/6J mice were fed normal diet for 1 week and then fed 8 weeks of high-fat diet to establish a mouse model of nutritional obesity. The control mice were fed a normal diet. After 1 week of adaptive treadmill training, the obese mice were divided into high- and low-intensity endurance training groups for treadmill training and the control group had no training. After 8 weeks, the body length, body mass and Lee’s index were measured and recorded. The level of Irisin in mouse serum was detected by ELISA. The expression levels of PGC-1α, FNDC5 and PPARδ in quadriceps femoris were detected by western blot assay. RESULTS AND CONCLUSION: High-fat diet could significantly increase body length, body mass and Lee’s index of mice, suggesting that high-fat-induced nutritional obesity mouse model was established. High-fat diet could significantly reduce the level of Irisin in serum and the expression of lipolysis-related proteins PGC-1α, FNDC5 and PPARδ in quadriceps femoris. Endurance training, especially high-intensity endurance training could effectively reduce the body length, body mass and Lee’s index of mice. Endurance training, especially high-intensity endurance training could effectively increase the level of Irisin in serum and the expression of PGC-1α, FNDC5 and PPARδ in quadriceps femoris. PGC-1α, FNDC5 and Irisin have consistent expression, suggesting that endurance training, especially high-intensity endurance training can up-regulate the expression of skeletal muscle PGC-1α, thereby increasing the expression of FNDC5 protein. FNDC5 is transformed into Irisin and released into the blood, thus increasing the level of lipolysis in mice, and achieving weight loss. 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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Ulinastatin improves rat liver metabolism after reduced-size liver transplantation  Gao Hongqiang, Liu Jing, Li Zhiqiang, Wang Hailei, Zhao Xiongqi, Zhang Shengning, Ran Jianghua, Li Li 2019, 23 (3):  435-440.  doi: 10.3969/j.issn.2095-4344.0594 Abstract ( 247 )   PDF (822KB) ( 161 )   Save BACKGROUND: Effects of ulinastatin in anti-inflammatory, viscera protection and improving microcirculation after liver resection or liver transplantation have been widely researched. However, the role of microRNA regulation mechanism of ulinastatin has not yet been reported. OBJECTIVE: To discover the variation of microRNA expression profile and the change of proteomics in rats after reduced-size liver transplantation using ulinastatin, and to investigate if there is a lien between differentially expressed miroRNAs and proteins, so as to provide an in-depth theoretical evidence for clinical application of ulinastatin. METHODS: Rat models of reduced-size liver transplantation were established using the Kamada double cuff method and then divided into two groups. Rats in the experimental and control groups were given intraperitoneal injection of ulinastatin (100 U/g) and normal saline (2 mL) respectively at 0, 12, 24, 36, and 48 hours after surgery. miroRNA chip and proteomic analysis were performed at 24 and 48 hours after transplantation. Then results were then imported into mirTarBase software for target prediction. RESULTS AND CONCLUSION: Compared with the control group, in the experimental group, there were 19 differentially expressed (> two-fold) proteins and 17 differentially expressed (> 1.5 fold) proteins in rats. A miRNA regulation channel: miR-181-a-5p and Gpx1 were found. To conclude, after reduced-size liver transplantation in rats, use of ulinastatin alters miR-181-a-5p expression, which improves liver metabolism through regulating Gpx1 expression. This may be one mechanism of action of ulinastatin. 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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Expression and functional identification of WT-/GOF-ADAMTS13 proteins Yu Shanshan, Lin Jiangguo, Fang Ying 2019, 23 (3):  441-446.  doi: 10.3969/j.issn.2095-4344.0604 Abstract ( 359 )   PDF (788KB) ( 1270 )   Save BACKGROUND: ADAMTS13 cleaves Von Willebrand factor (VWF) to regulate its size, thereby preventing aberrant platelet aggregation and thrombus. Eukaryotic cells are usually selected to express recombinant ADAMTS13. However, it still lacks a clear and efficient way for its expression and purification. OBJECTIVE: To find a suitable host cell and get stable cell lines to express WT-ADAMTS13 (wild-type) and GOF-ADAMTS13 (gain of function), so as to establish the purification method for high-purity ADAMTS13 proteins, thus studying their biological functions. METHODS: GOF-ADAMTS13 recombinant plasmid was obtained by site mutation kit with the WT-ADAMTS13 recombinant plasmid as a template. The expression effects of CHO-S, CHO-K1, and 293T cells on ADAMTS13 were compared to select appropriate host cells. The proteins were purified by Ni affinity chromatography and gel filtration. The purity and specificity of WT-ADAMTS13 and GOF-ADAMTS13 were identified by 7.5% SDS-PAGE and western blot assay. The interaction between WT-/GOF-ADAMTS13 and VWF-A2 was detected by atomic force microscopy to validate the activity of the two proteins. RESULTS AND CONCLUSION: Compared with CHO-S and CHO-K1 cell lines, 293T cell line was more suitable as expression host for WT-/GOF-ADAMTS13. After Ni column affinity chromatography and gel filtration purification, the concentration of purified WT-/GOF-ADAMTS13 was 103.7 and 149.7 mg/L, respectively with the purity of above 90%. Atomic force microscopy results showed that the adhesion frequencies of WT-/GOF-ADAMTS13 and VWF-A2 were 11.37% and 14.70%, respectively. These results suggest higher affinity of GOF-ADAMTS13 binding to VWF-A2 is consistent with recent ADAMTS13 conformational studies. 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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Protective effects and the mechanism of lipoic acid on cell model of Parkinson’s disease induced by lipopolysaccharide  Gao Hua1, 2, Li Yanxia1, Wang Dan1, Yang Xinling1 2019, 23 (3):  447-452.  doi: 10.3969/j.issn.2095-4344.0595 Abstract ( 357 )   PDF (1548KB) ( 555 )   Save BACKGROUND: Neuroimmune and inflammatory reaction have been shown to participate in the occurrence of Parkinson’s disease. Lipoic acid possesses anti-inflammatory and neuroprotective roles. OBJECTIVE: To investigate the protective effects of lipoic acid on PC12 cell models of Parkinson’s disease induced by lipopolysaccharide and the underlying mechanism.  METHODS: PC12 cells were divided into blank control, model (Parkinson’s disease model induced by lipopolysaccharide) and treatment (Parkinson’s disease model treated by lipoic acid) groups. PC12 cells were pretreated with lipoic acid for 2 hours, and then stimulated with lipopolysaccharide. The optimal concentration of lipopolysaccharide and lipoic acid was determined by cell counting-kit 8 assay. The expression of tyrosine hydroxylase was detected by immunohistochemistry. The levels of tumor necrosis factor α and interleukin 6 were detected by ELISA kit. The protein expression levels of nuclear factor-κBp65 and tyrosine hydroxylase were detected by western blot assay. RESULTS AND CONCLUSION: The optimal concentration of lipopolysaccharide was 600 μg/L for constructing cell model of Parkinson disease, and 25 μmol/L lipoic acid was the optimal intervention concentration. Lipoic acid could increase the number of tyrosine hydroxylase positive cells, reduce the release of inflammatory factors in cell model of Parkinson’s disease, inhibit the production of nuclear factor-κBp65 and increase tyrosine hydroxylase content. Therefore, lipoic acid exhibits its effects on cell model of Parkinson’s disease through inhibiting the nuclear factor-κB signaling pathway and reducing inflammation, thereby exerting neuroprotection. 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 microcirculation disturbance in model rat liver treated with Rougan Huaxian Granule  Wei Dongjue1, Ye Dongmei2, Zhang Fengying1, Zhou Ying1, Wang Zhenchang3 2019, 23 (3):  453-457.  doi: 10.3969/j.issn.2095-4344.0546 Abstract ( 507 )   PDF (733KB) ( 227 )   Save BACKGROUND: Chinese herbal compound Rougan Huaxian Granule has been shown to improve chronic hepatitis and liver fibrosis. However, whether it can improve liver microcirculation in multi-path, multi-target and multiple-link remains unclear. OBJECTIVE: To investigate the effects and differences of Rougan Huaxian Granule and its disassembled formulas on the regulation of microcirculation disturbance in model rat liver. METHODS: The rat model of liver fibrosis was established by CCl4 compound factors. Four weeks later, the model rats were randomized into six groups: model group, Dahuang Zhechong Pill group, Rougan Huaxian Granule 1, 2, 3 and 4 groups, treated with Rougan Huaxian Granule, Bushen Yanggan Formula, Yiqi Jianpi Foamula and Huatan Jiedu Formula, Huoxue Huayu Formula combined with Ruanjian Sanjie Formula, respectively. The remaining normal rats were used as control group. The drug (0.1 mL/kg) was given via gavage, once daily for 12 weeks. RESULTS AND CONCLUSION: Light microscopy showed that fibrous hyperplasia occurred in the liver mesenchyma, which divided the normal hepatic lobules into uneven cell masses (pseudolobule formation), suggesting the live fibrosis model was established successfully. Compared with the control group, the serum levels of NO and endothelin-1 were significantly increased in each group. With the progression of liver cirrhosis, the levels of NO and endothelin-1 were decreased significantly (P < 0.01), and the hepatic artery/vein activation time was shortened significantly (P < 0.05 or P < 0.01). Compared with the model group, the contents of NO and endothelin-1 in each treatment group were decreased in different degrees (P < 0.01), and the hepatic artery/vein activation time was prolonged (P < 0.01). Compared with Dahuang Zhechong Pill group, the contents of NO and endothelin-1 in Rougan Huaxian group 1 were significantly decreased (P < 0.05 or P < 0.01), and the hepatic vein activation time was prolonged (P < 0.01). In summary, the rat model of live fibrosis is successfully constructed. Rougan Huaxian Granule can promote the microcirculation of the liver and increase the hepatic microvascular flow rate, and delay the development of liver fibrosis. The mechanism may be related to the decrease in serum levels NO and endothelin-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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Effects of recombinant human bone morphogenetic protein-2 on the proliferation of breast cancer MCF-7 cells Liu Shuzhong1, Lao Lifeng2 2019, 23 (3):  458-463.  doi: 10.3969/j.issn.2095-4344.0578 Abstract ( 420 )   PDF (1684KB) ( 195 )   Save BACKGROUND: Clinical application of recombinant human bone morphogenetic protein-2 (rhBMP-2) has been found to increase the risk for malignant tumors, but the effect of rhBMP-2 on the proliferation of breast cancer MCF-7 cells and the underlying mechanism remain unclear. OBJECTIVE: To explore the effects of rhBMP-2 on the proliferation of breast cancer MCF-7 cells and the underlying mechanism in vivo and in vitro. METHODS: In vitro study: cell proliferation of MCF-7 under different concentrations of rhBMP-2 was determined by MTT assay and flow cytometry in serum-free condition. The cell cycle was detected by flow cytometry. The expression levels of p21 and cyclin E were detected by real-time PCR. The protein levels of p21, cyclin E and phosphorylation of PI3K/Akt were tested by western blot and real-time PCR. In vivo study: fourteen 6-week-old female nude mice were divided into two groups: experimental group (subcutaneous injection of rhBMP-2 plus MCF-7 cells), control group (subcutaneous injection of the same volume of MCF-7 cells). RESULTS AND CONCLUSION: In vitro study: rhBMP-2 markedly inhibited the proliferation of MCF-7 cells in serum-free condition. After 24-hour treatment, rhBMP-2 could increase the G1 cell ratio. rhBMP-2 could significantly increase the expression of p21 as well as significantly inhibit the expression of cyclin E and phosphorylation in the PI3K/Akt signaling pathway. In vivo study: the volume of subcutaneous tumor in the experimental group was smaller than that in the control group. Immunohistochemistry revealed that rhBMP-2 could significantly decrease the expression level of ki-67 in tumor tissues. Our study suggests that rhBMP-2 has significantly suppressive effect on the proliferation of breast cancer MCF-7 cells via PI3K/Akt pathway in vitro and in vivo. Therefore, we can provide the basic science data to avoid risks of breast cancer and to support the utilization of rhBMP-2 in the management of spinal fusion, treatment of bone ununion and other orthopedic disorders. 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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Resveratrol protects endothelial cells from oxidative damage and inhibits expression of pro-thrombosis molecules: the underlying mechanisms He Wei, Lou Zhenkai, Wang Bing, Li Xingguo, Zhao Chongyu, Zhao Xueling 2019, 23 (3):  464-469.  doi: 10.3969/j.issn.2095-4344.0598 Abstract ( 455 )   PDF (833KB) ( 233 )   Save BACKGROUND: Resveratrol has been shown to hold anti-oxidant and anti-arterial thrombosis effects. OBJECTIVE: To explore the mechanism of resveratrol in repairing endothelial cell injury and inhibiting deep vein thrombosis at molecular level. METHODS: Human umbilical vein endothelial cells were isolated and cultured. The cells were divided into three groups: control group (common medium), H2O2 group (cultured with 200 μmol/L H2O2 for 24 hours), and resveratrol plus resveratrol group (pretreated with 10, 20 and 30 μmol/L resveratrol for 2 hours, and then cultured with 200 μmol/L H2O2 for 24 hours). The cell viability was evaluated by MTT assay. The apoptotic cells were observed by Hoechst 33258 staining and fluorescence microscopy. The protein expression levels of P-selectin, P-selectin glycoprotein ligand-1, von Willebrand Factor and PI3-K/Akt/GSK3β and NF-κB signaling pathways were tested by western blot assay. RESULTS AND CONCLUTION: Compared with the control group, H2O2 could induce endothelial cell injury and resveratrol could repair endothelial cells injured by H2O2 (P < 0.05). The apoptotic rate in the H2O2 group was significantly higher than that in the control group (P < 0.05). Compared with the H2O2 group, the apoptosis rate in the H2O2 plus resveratrol group was significantly decreased (P < 0.05). Compared with the control group, the expression levels of P-selectin, P-selectin glycoprotein ligand-1, von Willebrand Factor, NF-κB and GSK3β in the H2O2 group were significantly increased (P < 0.05), the expression of Akt protein showed no significant change (P > 0.05), and the expression level of p-Akt protein was significantly down-regulated (P < 0.05). Meanwhile, resveratrol could reverse all above protein expression (P < 0.05). To conclude, resveratrol can repair human umbilical vein endothelial cells and inhibit the expression of P-selectin, P-selectin glycoprotein ligand-1 and von Willebrand Factor proteins, which may be related to PI3-K/Akt/GSK3β and NF-κB signaling pathways. 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 | Related Articles | Metrics

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Bushen Jianpi Recipe affects lipid metabolism and leptin expression in rats with osteoporosis receiving a high fat diet Li Ping1, Lin Yu1, Zhu Xi2, Zhang Yiyuan1, Xiao Lili1, Hua Peng1, Lin Xueyi1 2019, 23 (3):  470-475.  doi: 10.3969/j.issn.2095-4344.0592 Abstract ( 353 )   PDF (654KB) ( 431 )   Save BACKGROUND: Menopause can also lead to changes in lean tissue and fat composition. Osteoporosis is found to be more common in obese elderly than in non-obese people. Thereafter, leptin may be an important regulatory factor in the pathogenesis of osteoporosis. OBJECTIVE: To investigate the effect of Bushen Jianpi Recipe on lipid metabolism and leptin in high fat diet rats with osteoporosis. METHODS: Totally 220 SPF Sprague-Dawley rats aged 6 months were randomly divided into model group (n=170) and sham group (n=50). The model group underwent bilateral ovarian ligation and the sham group received the same intervention as the model group except that ovarian ligation was not performed. At 12 weeks after surgery, the rat models were randomized into normal, high-fat, high-fat plus Jiangu Granule (2 g/(kg•d)), and high-fat plus simvastatin (100 μg/(kg•d)) groups (n=40 per group). The administration began at 13 weeks after surgery via gavage. The normal and sham groups were given 2 mL of normal saline, once daily. After 2, 6, 12 and 24 weeks of treatment, the body mass was measured. The serum, lumbar vertebrae and proximal femurs were removed. The bone mineral density of the right tibia was measured by double energy X-ray. The concentration of serum osteocalcin, tartrate-resistant acid phosphatase isoform 5b (TRACP-5b), total cholesterol, triacylglycerol, leptin and leptin receptor were detected by ELISA. The mRNA expression of leptin and leptin receptor in bones was determined by qPCR. RESULTS AND CONCLUSION: The rat body mass in the high-fat plus Jiangu Granule group was higher than that in the normal and high-fat groups at the same time, but lower than that in the sham and high-fat plus simvastatin groups. The expression levels of leptin and leptin receptor, and osteocalcin level in the high-fat plus Jiangu Granule group were significantly higher than those in the normal and high-fat groups, and the contents of TRACP-5b, cholesterol, and triacylglycerol were significantly lower than those in the Leptin and leptin receptor, the BGP concentration were higher than those in the normal diet model group and the high fat diet model group. The concentrations of TRACT-5b, cholesterol, and triglycerides were lower than those in the normal and high-fat groups (P < 0.05). These results indicate that Bushen Jianpi Recipe can regulate lipid metabolism disturbance, reduce blood viscosity and improve the structure of blood vessel wall. It can also increase the expression of leptin and leptin receptor in serum and bones of rats, improve bone metabolism, and prevent and treat primary osteoporosis. 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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Screening differentially expressed proteins in the serum of patients with systemic lupus erythematosus combined with osteonecrosis of the femoral head by iTRAQ technology Fan Siqi1, Zeng Ping2, Zhou Yi3, Qin Gang2, Liao Xiaobo2, He Kaiyi2 2019, 23 (3):  476-481.  doi: 10.3969/j.issn.2095-4344.0583 Abstract ( 341 )   PDF (661KB) ( 298 )   Save BACKGROUND: Systemic lupus erythematosus (SLE) combined with osteonecrosis of the femoral head (ONFH) is a result of long-term application of steroids that induces death of active ingredients in femoral head. But there is a lack of specific biomarker for early diagnosis. OBJECTIVE: To screen the systemic markers of SLE with steroid-induced ONFH using isobaric tags for relative and absolute quantitation (iTRAQ) technology. METHODS: Serum of eight patients with SLE and steroid-induced ONFH was collected and used as the experimental group, and serum of 10 patients with SLE but without steroid-induced ONFH served as the control group. The differentially expressed proteins were screened and identified by iTRAQ technique and two-dimensional liquid chromatography-tandem mass spectrometry. RESULTS AND CONCLUSION: Totally 10 kinds of differentially expressed proteins were identified. There was significant difference between two groups. In the experimental group, the expression levels of copper basket protein, immunoglobulin IgG, immunoglobulin κ chain and complement C2 were significantly up-regulated. The expression levels of endothelial cell protein C receptor, human kallikrein-binding protein, pigment epithelium-derived factor, tetracycline, insulin-like growth factor 2, and thyroxine carrier protein were down-regulated. KEEG pathway analysis indicated that 10 kinds of proteins were closely related to Wnt signaling pathway, coagulation and complement cascades reaction signaling pathway and SLE signaling pathway. In summary, the differentially expressed proteins are obtained successfully in steroid-induced ONFH by proteomics, which provides theoretical basis for the diagnosis and pathogenesis of steroid-induced ONFH. 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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Distribution characteristics of polymorphism of human leukocyte antigen C of hepatitis B virus carriers in patients from Shenzhen Quan Zhanrou, He Liumei, Chen Hao, Hong Wenxu, Gao Suqing 2019, 23 (3):  482-486.  doi: 10.3969/j.issn.2095-4344.0586 Abstract ( 320 )   PDF (619KB) ( 313 )   Save BACKGROUND: Due to the development of human leukocyte antigen (HLA) typing techniques, there are a large number of studies reporting the relationship between hepatitis B virus (HBV) and HLA-A, -B, and -DR, but little is reported on the relationship between HBV and HLA-C. OBJECTIVE: To analyze the polymorphism of HLA-C of HBV carriers in Shenzhen. METHODS: PCR-based typing method was used to analyze the polymorphism of HLA-C locus alleles of 122 HBV carriers and 186 healthy volunteers (control group) in Shenzhen. RESULTS AND CONCLUSION: Twenty-four alleles for the locus HLA-C were detected in the control group, of which HLA-C *01:02, 07:02, 03:04 and 08:01 were the most common. In the HBV carrier group, 18 kinds of HLA-C alleles were detected, including the common alleles HLA-C*07:02, 01:02, 03:04, 08:01. HLA-C*12:02 was not detected in the HBV carries group. The detection rate of HLA-C*12:02 in the control group was significantly higher than that in the HBV carrier group. These results imply that the frequency distribution of HLA-C alleles of HBV carriers in Shenzhen also have a high degree of genetic polymorphism compared with the control group. There may be association between HBV carriers and HLA-C*12:02 alleles. HLA-C*12:02 allele possibly plays a negative role in HBV carriers in Shenzhen. 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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Platelet-rich plasma injection promotes repair of partial rotator cuff of tears: basic research and clinical trials Liu Yan1, Zhang Haoxu2, Liu Fulong3, Zhou Mei1, He Chengqi1, He Hongchen1 2019, 23 (3):  487-492.  doi: 10.3969/j.issn.2095-4344.0581 Abstract ( 597 )   PDF (674KB) ( 1280 )   Save BACKGROUND: There is no effective treatment for partial rotator cuff tears, and platelet-rich plasma (PRP) is a promising therapy. OBJECTIVE: To summarize the application status and progress of basic research and clinical trials addressing PRP injection in partial rotator cuff tear healing. METHODS: A computer-based search of PubMed, MedLine, Springerlink databases was performed for relevant articles published from 2008 to 2018 with “PRP, platelet-rich plasma, partial RCT, partial rotator cuff tear” as key words. Initially, 74 articles were retrieved, and finally 12 eligible articles were included for result analysis. RESULTS AND CONCLUSION: Few clinical trial or animal experiment addressing the present use of PRP injection in a partial rotator cuff tear are reported. There are more clinical trials about PRP applied in arthroscopic repair of full-thickness rotator cuff tear, but most of them do not show better outcomes than the control group. Some studies show that PRP injection applied in partial rotator cuff tear can obtain better outcomes than corticosteroid injection. The results of PRP injection in partial rotator cuff tear healing still need further investigation. The methods to prepare PRP are varied, whether PRP is injected to the precision site is also important. Further fundamental research and clinical studies are required to verify the method and the result of PRP injection in partial rotator cuff healing. 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 | Related Articles | Metrics