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08 June 2018, Volume 22 Issue 16 Previous Issue    Next Issue

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Bushen Jianpi Huoxue decoction increases the proliferation of Sost-overexpressed adenovirus transfected osteoblasts and alkaline phosphatase activity Wan Lei1, Huang Hong-xing1, Huang Hong2, Wang Fan3, Chai Shuang4, Wang Ji-li4, Liu Shao-jin4, Huang Jia-chun4 2018, 22 (16):  2461-2466.  doi: 10.3969/j.issn.2095-4344.0275 Abstract ( 354 )   PDF (1245KB) ( 136 )   Save BACKGROUND: Chinese herbs have been shown to affect osteoblasts through Wnt/β-catenin signaling pathway. Sclerostin (Sost) is an inhibitor of Wnt signaling pathway and an inhibitor of osteogenesis, and it is also an antagonistic factor that regulates the differentiation and proliferation of osteoblasts. Bushen Jianpi Huoxue decoction (BJHD) is the empirical formula in the prevention and treatment of osteoporosis at the Affiliated Orthopedics Hospital, Guangzhou University of Chinese Medicine. BJHD can promote osteogenesis and effectively treat osteoporosis. OBJECTIVE: To observe the effect of BJHD on the cell proliferation, alkaline phosphatase activity and expression levels of related proteins in Sost-overexpressed adenovirus transfected osteoblasts. METHODS: The osteoblasts were divided into two groups: blank control group (empty adenovirus transfection) and Sost overexpressed group (Sost overexpression and adenovirus transfection). Two groups were cultured with blank serum and medicine serum. The cell proliferation was tested by cell counting kit-8 assay, and alkaline phosphatase activity was tested by alkaline phosphatase kit. Expression levels of bone regulation-related proteins were tested by western blot assay. RESULTS AND CONCLUSION: Compared with the blank serum, medicine serum could increase the cell viability at 24, 48 and 72 hours in both groups (P < 0.01). Compared with the blank serum, medicine serum increased the alkaline phosphatase activity (P < 0.01). Compared with the blank serum, medicine serum in the blank control group decreased the expression of low-density lipoprotein receptor-related protein 5 and osteoprotegerin (P < 0.01), but showed no effect on the expression of osteopontin and tumor necrosis factor α. Medicine serum in the Sost-overexpressed group decreased the expression of low-density lipoprotein receptor-related protein 5, osteoprotegerin and osteopontin, increased the expression of tumor necrosis factor α (P < 0.01). These results indicate that BJHD can increase the proliferation of recombinant adenovirus transfected Sost-overexpressed osteoblasts and alkaline phosphatase activity, and regulate the expression of low-density lipoprotein receptor-related protein 5, osteoprotegerin, osteopontin, and tumor necrosis factor α. 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 low basal metabolic rate and mortality in older adults with hip fractures Sun Chun-sheng, Zheng Xin, Guo Kai-jin, Li Dong-ya, Li Cheng-yu, Wang Yi, Chang Bu-qing, Feng Shuo, 2018, 22 (16):  2467-2471.  doi: 10.3969/j.issn.2095-4344.0218 Abstract ( 358 )   PDF (1007KB) ( 137 )   Save BACKGROUND: Basal metabolic rate (BMR) is an important indicator of human energy metabolism, and low BMR leads to the dysfunction of liver and kidney. Low BMR is usually found in patients with hip fractures, but there is a lack of study on the relationship between mortality of hip fracture and low BMR. OBJECTIVE: To investigate the effect of low BMR on the 1-year mortality in older adults with hip fractures. METHODS: Totally 507 patients with hip fractures aged more than 60 years from January 2014 to March 2016 were included in this retrospective study. Age, sex, surgery or not, surgical pathway, duration from injury to surgery, hospitalized pulmonary infection, number and kind of comorbidities, and 1-year mortality were recorded. BMR on admission was recorded, and multiple Logistic regression analysis was applied.  RESULTS AND CONCLUSION: All patients were followed up for 13-15 months, and the 1-year mortality was 13.41% (68/507). The mortality in the low BMR group was significantly higher than that in the non-low BMR group (P < 0.05). Logistic regression analysis showed that older age, conventional treatment, number of combined medical diseases, hospitalized pulmonary infection, and low BMR are risk factors for 1-year mortality in older adults with hip fracture. These results imply that low BMR is strongly associated with 1-year mortality in older adults with hip fracture. BMR can reflect the nutritional status, neuroendocrine, cellular and energy metabolism. Thereafter, for older adults with hip fractures and low BMI, nutrition therapy, re-warming, and endocrine therapy may help reduce the trauma-induced mortality. 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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Clinical significance of beta-catenin and matrix metalloproteinase-13 in synovial tissue of osteoarthritis Li Yue-jun1, Zhu Wei-guo2, Fang Qin-zheng2, Gu Pei-lun1, Gao Peng1, Jian Yong-zhi1, Yang Peng-fei1, Dong Jin-bo2 2018, 22 (16):  2472-2477.  doi: 10.3969/j.issn.2095-4344.0294 Abstract ( 313 )   PDF (5342KB) ( 122 )   Save BACKGROUND: Role of Wnt/β-catenin signaling pathway in the pathogenesis of osteoarthritis has been proved by numerous animal experiments and human specimen trials. Wnt/β-catenin signaling pathway and matrix metalloproteinase 13 (MMP-13) play important roles in pathological process of osteoarthritis, and they may be related with each other. OBJECTIVE: To detect the expression levels of β-catenin and MMP-13 in the synovial tissue of osteoarthritis, and to investigate the relationship between β-catenin and MMP-1 in the pathogenesis of osteoarthritis. METHODS: Synovial tissues were obtained from the 54 patients undergoing total knee arthroplasty or knee arthroscopic diagnosis and treatment surgery and 12 healthy patients suffering from amputation caused by trauma. The samples were divided into non-, mild-, medium- and severe-osteoarthritis groups based on advanced Mankin scores. The expression levels of β-catenin and MMP-13 in the synovial tissues were detected by immunohistochemistry, and correlation analysis was conducted. RESULTS AND CONCLUSION: Immunohistochemistry results showed that MMP-13 was negative in the non-osteoarthritis group, positive in 5 (50%) samples in the mild-osteoarthritis group, positive in 11 (65%) samples in the medium-osteoarthritis group and positive in 23 (85%) samples in the severe-osteoarthritis group. β-Catenin was positive in 1 (8%) sample in the non-osteoarthritis group, positive in 7 (70%) samples in the mild-osteoarthritis group, positive in 14 (82%) samples in the medium-osteoarthritis group and positive in 25 (93%) samples in the severe-osteoarthritis group. The expression levels of β-catenin and MMP-13 in the synovial tissues presented a positive correlation (r=0.806, P < 0.001), and the levels increased with joint degeneration becoming severe. These results imply that there is a significant increase in the expression levels of β-catenin and MMP-13 in the synovial tissues of osteoarthritis compared with the non-osteoarthritis group, and β-catenin and MMP-13 exert effects in development of osteoarthritis. Beta-catenin and MMP-13 are closely associated with pathological changes such as cartilage degeneration and synovial inflammation, thus providing new theoretical direction and experimental basis for targeted gene therapy 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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Correlation of bone mineral density with skeletal muscle content and physical activity Liu Yun-ting1, Guo Hui2, Zhang Yi-min3 2018, 22 (16):  2478-2482.  doi: 10.3969/j.issn.2095-4344.0254 Abstract ( 552 )   PDF (1134KB) ( 172 )   Save BACKGROUND: Bone mineral density is the most important indicator for bone level. Osteoporosis is a worldwide problem, and there is no safe and effective radical treatment. It can only be prevented, so bone mineral density arise more attention. OBJECTIVE: To explore the relationship between skeletal muscle content, physical activity level and bone mineral density in older adults, so as to provide theoretical basis for preventing osteoporosis treatment. METHODS: Totally 111 subjects aged 40 to 84 years were recruited randomly. The bone mineral density was measured by double energy X-Ray bone density meter, and the human skeletal muscle content, fat content and percentage of skeletal muscle were measured by human body component analyzer. The variance analysis was used for multiple group comparisons in different age groups, LSD method for comparisons between groups, and Pearson correlation analysis for correlation analysis. RESULTS AND CONCLUSION: Bone mineral density was significantly related to the physical activity, body mass, and skeletal muscle content (r=0.30, 0.41, and 0.45, P < 0.01). For middle and older adults, skeletal muscle content, physical activity level and body mass were positively related to bone mineral density. Age and bone mineral density had a negative relationship. These results indicate that physical activity is an important factor affecting bone density, and higher level of physical activity is beneficial to maintain and improve bone mineral density. Increasing muscle mass and improving body composition can improve bone mineral density. 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-467g suppresses the osteogenic differentiation and mineralization of mouse preosteoblasts via down-regulation of Runx-2 expression Chang Zhi-qiang, Zhang Li-feng, Li Peng-fei, Ma Min, Guo Jun 2018, 22 (16):  2483-2488.  doi: 10.3969/j.issn.2095-4344.0827 Abstract ( 318 )   PDF (4468KB) ( 126 )   Save BACKGROUND: Preliminary studies have found miR-467g inhibits bone regeneration, however, there is little information about the underlying mechanism. OBJECTIVE: To explore the effect of miR-467g on osteogenic differentiation and mineralization of mouse preosteoblasts and the underlying mechanism. METHODS: C57 mouse preosteoblasts were isolated and cultured in vitro. The expression levels of Runx-2, Osterix, and Osteocalcin, as well as alkaline phosphatase and mineralization activities were determined by western blot, real-time PCR, alkaline phosphatase and Alizarin red staining, respectively. miR-467g-overexpressed preosteoblasts were constructed to investigate the effect of miR-467g on osteogenic differentiation and mineralization of preosteoblasts by lipofection transfection. Dual luciferase reporter assay was used to identify whether the 3’UTR of Runx-2 mRNA was a binding target of miR-467g. RESULTS AND CONCLUSION: The primary mouse preosteoblasts had a good osteogenic proliferation and differentiation ability in vitro. Expression level of miR-467g was decreased with the increase in osteogenic induction time. MiR-467g suppressed the osteogenic differentiation and mineralization of mouse preosteoblasts. Luciferase assay confirmed that miR-467g targeted Runx-2 directly. In summary, miR-467g can suppress the osteogenic differentiation and mineralization of mouse preosteoblasts via down-regulation of Runx-2 expression. 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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Geometric shapes of tissue-engineered cartilage exert effects on mechanical behaviors of a defected area Zhao Yong-zheng1, 2, Liu Hai-ying1, 2, Zhang Chun-qiu1, 2, Hu Ya-hui1, 2 2018, 22 (16):  2489-2495.  doi: 10.3969/j.issn.2095-4344.0819 Abstract ( 244 )   PDF (1854KB) ( 103 )   Save BACKGROUND: Uncertainty of repairing articular cartilage defects is highly associated with the mechanical behaviors of the defected area, and the mechanical environment varies with the defect shape, depth and load. OBJECTIVE: To study the mechanical behaviors of articular cartilage defects under physiological load by finite element analysis. METHODS: The axisymmetric model of articular cartilage injury and repair based on transversely isotropy was established using ABAQUS software. The mechanical behaviors of the defect zone repaired with different repair shapes (cylindrical, frustum of a cone, orthorhombic prism, elliptical column) and depths of tissue-engineered cartilage under compressive load were analyzed. RESULTS AND CONCLUSION: The simulation results showed that there were significant differences in the mechanical behaviors of the defect area repaired with tissue-engineered cartilage in different shapes and depths. The stress concentration was the most obvious at the middle-layer defect repair, and the stress distribution was more reasonable at the deep (whole) layer defect repair. Furthermore, the distribution of the stress field and the liquid flow field at the cylinder-shaped tissue-engineered cartilage repair was the closest to the normal cartilage. That is to say, the tissue-engineered cartilage in cylinder or frustum-cone shape is recommended to repair cartilage defect. Importantly, the middle-layer repair is inadvisable. 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 treadmill training on the fitness and activation of cerebral cortex in older adults and youth Li Jin-tian 2018, 22 (16):  2496-2501.  doi: 10.3969/j.issn.2095-4344.0245 Abstract ( 368 )   PDF (1120KB) ( 151 )   Save BACKGROUND: Aging process is associated with declines in physical fitness and cognitive function, and proper exercise can improve the body function. Treadmill training has advantages of non-full weight-bearing and adjustable trampling resistance, so it has been widely used in clinical rehabilitation training, but whether treadmill training can enhance the cognitive function of individual brain needs an in-depth study. OBJECTIVE: To explore whether the effect of treadmill training on older adults and youth is the same.  METHODS: Forty-three healthy volunteer college teachers in universities were recruited, and divided into older adult group (n=18, aged 60-75 years) and youth group (n=25, aged 25-45 years) according to the age characteristics. Each person was trained for 4 consecutive weeks of treadmill training. The 6-minute walking, 10-m walking, muscle strength of lower limbs and transcranial magnetic stimulation test were performed before training, at 4 weeks of training and 1 month after training. RESULTS AND CONCLUSION: Before training, the amount of physical activity in the older adult group was higher than that in the youth group, but the functional walking, muscle strength of lower limbs and degree of cerebral cortex activation were worse than those in the youth group. After 4 weeks of treadmill training, there was a significant improvement in the functional walking and muscle strength of lower limbs in both groups, the improvement rate of 6-minute walking and leg muscle strength showed no significant difference between groups, but the improvement rate of 10-m walking in the youth group was significantly higher than that in the older adult group. At 1 month after training, 4-week treadmill training exerted a continued effect on the 6-minute walking and leg muscle strength in youth, and 4-week treadmill training exerted a continued effect on the 10-m walking in older adults. The action threshold, motor evoked potentials and cerebral cortex movement area of either brains in older adults were poorer than those in youth. Four-week treadmill training showed no significant effect on the cerebral cortex activation in older adults and youth. In summary, 4-week treadmill training can effectively enhance the ability of functional walking and muscle strength of lower limbs in older adults and youth, and the treadmill training can achieve almost the same improvement and continuation effect between older adults and youth. However, whether treadmill exercise can improve the cerebral cortex activation in older adults and youth needs an in-depth study. 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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Mechanism by which dysferlin promotes repair of exercise-induced skeletal muscie injury in rats Huang Qiao-ting1, 2, Xu Jie1, Lin Jia-shi3 2018, 22 (16):  2502-2507.  doi: 10.3969/j.issn.2095-4344.0219 Abstract ( 433 )   PDF (1243KB) ( 117 )   Save BACKGROUND: Numerous studies concerning dysferlin focus on muscle diseases (such as muscular dystrophy), but the relationship between membrane repair after exercise-induced muscle damage and dysferlin is little reported. OBJECTIVE: To observe the cell membrane permeability and expression levels of dysferlin and calpain3 in the rat gastrocnemius after acute eccentric exercise, so as to provide an theoretical reference for exploring the molecular mechanism of muscle regeneration and the exercise therapy of muscular diseases. METHODS: Thirty-two Sprague-Dawley rats were randomly divided into four groups including control, 24, 48 and 72 hours post exercise groups. The membrane permeability and expression levels of dysferlin and calpain3 were determined by immunohistochemistry, western blot assay and qRT-PCR. RESULTS AND CONCLUSION: The activity of serum creatine kinase, and expression levels of calpain3 mRNA and dysferlin protein, as well as membrane permeability at 24 hours post exercise were significantly greater than those in the control group (P < 0.05 or 0.01). The expression level of dysferlin mRNA at 24 and 48 hours post exercise was significantly higher than that at 72 hours post exercise (P < 0.05). Therefore, the damage to the skeletal muscle cell membrane was the most severe at 24 hours after eccentric exercise. Due to Ca2+ influx, expression of calpain3 mRNA was activated, and then the damaged cell membrane was repaired by increasing the expression of dysferlin. 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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Correlation of four gene polymorphic loci with athlete endurance phenotype Wei Qi, Fan Jia-cheng, Du Ya-wen 2018, 22 (16):  2508-2513.  doi: 10.3969/j.issn.2095-4344.0863 Abstract ( 427 )   PDF (1073KB) ( 137 )   Save BACKGROUND: Increasing evidence shows that at least 155 genetic polymorphisms are associated with aerobic performance and elite endurance athlete status. OBJECTIVE: To study the association of α-actin 3 (ACTN3) gene, nuclear respiratory factor 2 (NRF2), β2 adrenergic receptor (ADRB2) gene and peroxisome proliferative activated receptor gamma coactivator 1 alpha(PPARGC1A) polymorphic loci with aerobic performance of rowing athletes and their interaction effect, thereby providing basis for understanding the mechanism of genetic polymorphisms acting on endurance athlete status. METHODS: A case-control experiment was designed to analyze the distribution characteristics of four gene polymorphism loci in 15 excellent rowing athletes and 50 common college freshmen. The association of four polymorphic loci with the aerobic performance-related indexes was analyzed using total genotype scores. RESULTS AND CONCLUSION: The preponderant polymorphism locus distributions of ACTN3, PPARGC1A and NRF2 in the athletes group were higher than those in the control group, and the NRF2 gene loci showed significant difference between two groups. In the athletes group, the mean values of VO2 max was significantly different among three genotypes. That is to say, these three genetic polymorphisms may be the biomarkers to predict the elite endurance athlete status, but the mechanism needs to be studied in depth. 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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Establishing a rat model of intervertebral disc degeneration using three methods Bai Rong-fei1, Zhang Zhen2, Lin Yi-feng3, Yuan Chao3, Wang Sheng-yu1, Fang Sheng1, Chi Li-ye1 2018, 22 (16):  2514-2519.  doi: 10.3969/j.issn.2095-4344.0221 Abstract ( 788 )   PDF (1296KB) ( 154 )   Save BACKGROUND: Numerous studies focus on animal models of intervertebral disc degeneration (IDD), but criteria for establishing the animal models of IDD have not been confirmed, and there is a lack of systematic comparison among models. OBJECTIVE: To compare the rat models of IDD established by puncturing at annulus, endplate injection and their combination, thus providing reference for IDD model selection. METHODS: Eighty Sprague-Dawley rats were equivalently randomized into four groups: puncturing group (puncturing at the annulus), endplate injection group (endplate injected with ethyl alcohol), combination group (puncturing at the L5-6 annulus and endplate injection at the same segment) and sham operation group. Three rats in each group were taken at postoperative 4, 8, and 12 weeks for X-ray examination to measure the disc height; and the discs were removed for histological observation and immunohistochemical staining. RESULTS AND CONCLUSION: The results of X-ray examination, hematoxylin-eosin staining and immunohistochemical staining all showed that the IDD degree was gradually aggravated in all groups except the sham operation group. At postoperative 4 weeks, compared with the sham operation group, in the endplate injection and combination groups, the percent disc height was significantly decreased, the pathological scores were significantly increased and the average gray value of collagen type I was significantly reduced (P < 0.05). At postoperative 8 and 12 weeks, compared with the sham operation group, the percent disc height in the other three groups were all significantly decreased, the pathological score was significantly increased, and the average gray value of collagen type I was significantly decreased (P < 0.05). Compared with the puncturing and endplate injection group, in the combination group, the percent disc height at postoperative 8 weeks was significantly decreased, and the average gray value of collagen type I at postoperative 12 weeks was significantly decreased (P < 0.05). These results suggest that the rat IDD model can be successfully constructed by above three methods. Puncturing at the annulus is easy to operate and control IDD progression, which can be used to study different stages of IDD. Endplate injection is suitable for the etiological study of IDD, and induces IDD earlier than puncturing, but the final results are similar. The combination method can significantly accelerate IDD aggravation, and thus is not time consuming. 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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Zuogui pill regulates DKK1 in the prevention and treatment of glucocorticoid-induced osteoporosis Zhang Zhi-da1, Ren Hui2, Shen Geng-yang1, Zhang Yu-zhuo1, Zhao Wen-hua1, Yu Xiang1, Shang Qi1, Yu Pei-yuan1, Liang De2, 2018, 22 (16):  2520-2525.  doi: 10.3969/j.issn.2095-4344.0230 Abstract ( 340 )   PDF (1436KB) ( 131 )   Save BACKGROUND: Chinese medicine is effective for preventing and treating glucocorticoid-induced osteoporosis, however, the underlying mechanism remains unclear. DKK1, an inhibitor of Wnt/β-catenin signaling pathway, can be up-regulated by glucocorticoid. Thereafter, DKK1 is an important target in the prevention and treatment of osteoporosis. OBJECTIVE: To explore the regulatory effect of Zuogui pill on DKK1 in the prevention and treatment of glucocorticoid-induced osteoporosis. METHODS: Eighteen three-month-old female Sprague-Dawley rats were randomly divided into three groups: control group, model group and Zuogui pill group. Rats in the model and Zuogui pill groups received the subcutaneous injection of dexamethasone to establish the model of glucocorticoid-induced osteoporosis. The Zuogui pill group rats were administrated Zuogui pill extracts, and the control rats were given the same volume of normal saline. At 1 month after modeling, the lumbar vertebrae were removed to test the bone mass and microstructures by micro-CT scanning. The biomechanical properties were detected by compression test. The mRNA expression levels of DKK1, Runx2 and CTSK were determined by Qpcr. The serum alkaline phosphatase activity was tested. RESULTS AND CONCLUSION: Compared with the control group, in the model group, the volumetric bone mineral density, trabecular bone volume fraction, trabecular number, and trabecular thickness were significantly decreased (P < 0.05), the trabecular separation and structure model index were significantly increased (P < 0.05). The serum alkaline phosphatase activity was on a decline. The mRNA expression level of DKK1 showed a significant up-regulation (P < 0.05). The mRNA expression level of Runx2 showed a down-regulated trend while mRNA expression level of CTSK showed an up-regulated trend. Compared with the model group, the Zuogui pill group showed significantly enhanced volumetric bone mineral density, trabecular bone volume fraction, and trabecular number (P < 0.05); the structure model index was significantly decreased (P < 0.05); the trabecular separation was reduced; the serum alkaline phosphatase activity was enhanced; the mRNA expression level of DKK1 showed a significant down-regulation (P < 0.05); the mRNA expression level of Runx2 showed an up-regulated trend while mRNA expression level of CTSK showed a down-regulated trend. The vertebral compressive strength in the Zuogui pill group was significantly higher than that in the model group (P < 0.05). In summary, Zuogui pill prevents and treats glucocorticoid-induced osteoporosis possibly through the down-regulation of mRNA expression of DKK1. 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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Roles of cytochrome C and Caspase-9 in steroid-induced avascular necrosis of the femoral head Gong Yu-lin1, Wang Yu-xin2, Zhao Zhen-qun3, Li Hong-bin1 2018, 22 (16):  2526-2531.  doi: 10.3969/j.issn.2095-4344.0761 Abstract ( 323 )   PDF (1855KB) ( 110 )   Save BACKGROUND: Steroid-induced avascular necrosis of the femoral head (SANFH) may be associated with apoptosis of osteocytes, but its regulatory mechanism remains unclear. Cytochrome C and Caspase-9 are key factors in the mitochondrial apoptosis signaling pathway, and their roles in SANFH still need further investigation. OBJECTIVE: To investigate the roles of cytochrome C and Caspase-9 in the SANFH. METHODS: Twenty New Zealand white rabbits were randomly divided into experimental and control groups (n=10 per group). Rabbits in the experimental group were injected with 7.5 mg/kg methylprednisolone, twice weekly, and those controls were subjected to the injection of same amount of normal saline. The treatment period was 8 weeks in both group.The rabbits were then sacrificed by air embolism to remove the bilateral femoral head specimens for histological and transmission electron microscope observations. The apoptotic cells were detected by TUNEL method and the expression levels of cytochrome C and Caspase 9 were detected by immunohistochemistry. RESULTS AND CONCLUSION: Eight weeks after intervention, compared with the control group, the experiment group had loose femoral head, thinned trabecular bone, increased adipocytes in the medullary cavity, and increased percentage of empty lacunae (P < 0.05); the apoptotic bodies were formed, and the apoptosis rate was increased (Р < 0.05); and the positive expression rates of cytochrome C and Caspase-9 were significantly increased (P < 0.01). Correlation analysis results showed that the apoptosis rate was positively correlated with the empty lacunae rate and the positive expression rates of cytochrome C and Caspase-9 (P < 0.01). These findings indicate that the abnormal release of cytochrome C and high expression of Caspase-9 are related to the apoptosis of osteocytes in 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 | Related Articles | Metrics

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Quantitative study on cytotoxicity of methylene blue to human nucleus pulposus cells by cell counting-kit 8 assay Feng Hua-long1, He Sheng-hua2, Huang Fei-qiang1, Lai Ju-yi1, Sun Zhi-tao2, Xu Fu-guang1, Lan Zhi-ming1 2018, 22 (16):  2532-2536.  doi: 10.3969/j.issn.2095-4344.0240 Abstract ( 528 )   PDF (1137KB) ( 266 )   Save BACKGROUND: Our preliminary study has shown that methylene blue exerts toxic effect on human nucleus pulposus cells in a concentration-dependent manner, but the toxic range remains unclear. OBJECTIVE: To determine the critical range of the cytotoxicity of methylene blue to nucleus pulposus cells by cell counting-kit 8 (CCK-8) assay. METHODS: The nucleus pulposus was from a patient with intervertebral disc herniation. The nucleus pulposus cells were extracted and cultured. Passage 1 cells mere used to make cell suspensions. The cells were divided into nine groups for culture: blank control (only the medium, CCK-8 solution), control (only medium, cells and CCK-8 solution), and methylene blue groups (1%, 0.5%, 0.1%, 0.05%, 0.01% and 0.005% of methylene blue). The absorbance values were measured by CCK-8 assay at 24, 48, 72 and 96 hours after incubation. The cell viability was calculated, and the color was observed. RESULTS AND CONCLUSION: The color in the control group was the darkest, and the color in the methylene blue groups became lighter with the concentration of methylene blue increasing, and the 0.05%, 0.01%, 0.005% and 0.001% methylene blue groups showed darker color similar to the control group. The absorbance values in the 0.1% methylene blue group were significantly less than those in the control group (P < 0.05). There were no significant differences in the absorbance values and cell viability between 1% and 0.5% methylene blue groups (P > 0.05). The absorbance values and cell viability in the 0.1% methylene blue group were significantly higher than those in the 0.5% methylene blue group, but were significantly less than those in the 0.05% methylene blue group (P < 0.05). Thus, methylene blue exerts cytotoxicity to human nucleus pulposus cells, and the critical value of toxicity is between 0.1% and 0.05% and close to 0.05%. However, the exact value needs a further investigation. 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 treadmill training on the locomotor function in a rat model of dorsal root ganglion resection An Pu-tian1, Zhu Wen-wen1, Li Mai-chao1, Cui Xiao-juan1, Zhou Yan1, Zhang Yi-meng1, Jin Li-xin2 2018, 22 (16):  2537-2541.  doi: 10.3969/j.issn.2095-4344.0220 Abstract ( 349 )   PDF (4328KB) ( 173 )   Save BACKGROUND: Peripheral nerve injury can lead to extensive changes in central nervous system, and exercise training can promote the recovery of locomotor function following central nervous system injury. OBJECTIVE: To observe the changes of locomotor function and the expression levels of vesicular glutamate transporter VGLUT1 in the spinal cord in a rat model of dorsal root ganglion resection after treadmill exercise and to explore the effect of treadmill training on the locomotor function after peripheral nerve injury. METHODS: Thirty-nine 10-week-old male Wistar rats were randomized into experimental (n=15), control (n=15) and sham operation (n=9) groups. The rats in the experimental and control groups received the dorsal root ganglion resection at L3 and L4 segments to establish the model of peripheral nerve injury under local anesthesia, while the rats in the sham operation group were only subjected to dorsal root ganglion exposure. The rats in the experimental group underwent 15 m/minute treadmill training at postoperative 7 days, while rats in the other two groups were in free movement. Gait analysis was performed at preoperative 3 days, postoperative 7, 14, 21, and 28 days, respectively, and the behavioral changes of rats were observed. The tissue sections were obtained from L3 segment at postoperative 7, 14, and 28 days to detect the expression levels of VGLUT1 in the spinal cord by immunohistochemistry. RESULTS AND CONCLUSION: The peroneal nerve function index in the experimental and control groups was lower than that before surgery and that in the sham operation group at postoperative different time points (all P < 0.05). The index in the experimental and control groups was the lowest on day 7 postoperatively (P < 0.05), then the index gradually increased, but was still lower than the preoperative level (P < 0.05). The index in the experimental group was significantly higher than that in the control group at postoperative 21 and 28 days (P < 0.05). The expression levels of VGLUT1 in the lamina IX in the experimental and control groups were significantly lower than those in the sham operation group at different time points after surgery (P < 0.01). The levels in the experimental group were significantly higher than those in the control group at postoperative 14 and 28 days (P < 0.05). The levels in both groups on a decline after surgery, especially the control group (P < 0.05). These results suggest that treadmill can promote the recovery of locomotor function post peripheral nerve 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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Baicalin increases connexin 43 expression in striatal astrocytes of rats with Parkinson’s disease Han Xue-jie1, 2, Halleda•Balikarthan2, Gao Hua3, Yang Xin-ling2 2018, 22 (16):  2542-2548.  doi: 10.3969/j.issn.2095-4344.0222 Abstract ( 332 )   PDF (2954KB) ( 125 )   Save BACKGROUND: In the pathogenesis of Parkinson’s disease, connexin 43 is an important regulatory protein, and the underlying mechanism of connexin 43 in reducing the damage of dopamine neurons is still unclear. Baicalin has been shown to inhibit the inflammation and oxidative stress of Parkinson’s disease, and can protect dopamine neurons from deformation in a rat model of Parkinson’s disease. OBJECTIVE: To investigate the effect of baicalin on the behavior and the expression of connexin 43 in astrocytes in a rat model of Parkinson’s disease. METHODS: Eighty Sprague-Dawley rats were enrolled. Ten randomly selected rats received no intervention (control group), and Parkinson’s disease was induced by unilateral bisite (median substantia nigra, striatum) lesions using 6-hydroxydopamine stereotaxic apparatus in the remaining rats. The model rats received the treatment of normal saline (model group), 125 mg/kg Madopar, 50 and 100 mg/kg baicalin, respectively, for 28 consecutive days. The changes in rotational behavior were observed by behavioral tests. The number of cells positive for tyrosine hydroxylase was determined by immunohistochemistry. The expression of connexin 43 was detected by immunohistochemistry and western blot assay. RESULTS AND CONCLUSION: Behavioral results: there was no rotational behavior in the control group before and after intervention. In the other four groups, the model group rats exhibited rotational behaviors at 5 minutes after apomorphine injection, all were at a speed of 13 r/min before treatment (P > 0.05). Compared with the model group, the number of rotations in the Madopar and high-dose baicalin groups was significantly decreased (P < 0.01). There was no significant difference in the number of cells positive for tyrosine hydroxylase in the nigra between model and low-dose baicalin groups (P > 0.05). The number of cells positive for tyrosine hydroxylase in the Madopar and high-dose baicalin groups was significantly greater than that in the model group (P < 0.01). There was a significant increase in the expression of connexin 43 in the model group compared with the control group (P < 0.01). The expression level did not differ significantly between model and low-dose baicalin groups (P > 0.05). The expression level in the Madopar and high-dose baicalin groups was significantly lower than that in the model group (P < 0.01). In conclusion, 100 mg/kg baicalin can significantly improve the rotational behavior of rats with Parkinson’s disease, significantly increase the number of tyrosine hydrogenase-positive cells, and alleviate the damage of Parkinson’s disease neurons caused by 6-hydroxydopamine. Additionally, baicalin can down-regulate connexin 43 expression in astrocytes of rats with Parkinson’s disease. 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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Variations of sodium chloride concentration affect conformational dynamics of human neuronal calcium sensor-1 protein: a molecular dynamics simulation study Zhu Yu-zhen1, Li Yun-xiang1, Cai Hao1, Zhang Qing-wen2 2018, 22 (16):  2549-2557.  doi: 10.3969/j.issn.2095-4344.0237 Abstract ( 397 )   PDF (6474KB) ( 131 )   Save BACKGROUND: Human neuronal calcium sensor-1 (NCS-1) protein with a high charge is extremely sensitive to solution temperature, but whether the change of ion concentration in the solution can change the protein structure and affect the physiological function has not been reported. OBJECTIVE: To investigate the molecular mechanism of variations of sodium chloride concentration altering conformational dynamics of human NCS-1. METHODS: Through molecular dynamics simulation, the first two minimum energy models (PDB id: 2LCP) were utilized as the starting states of each independent molecular dynamics simulation of the human NCS-1 protein. After energy minimization, two different initial structures were used to conduct three independent 500 ns molecular dynamics simulations with explicit solvent for human NCS-1 protein at 0.1, 0.3 and 1 mol/L sodium chloride concentration. RESULTS AND CONCLUSION: The slightly high concentration of sodium chloride increased the flexibility of loop L2, simultaneously expanded the global and local structure of NCS-1 protein. Increase in sodium chloride concentration induced loop L3 to adopt a collapsed state and reduced the connectivity between the starting and ending residues of loop L3. N-domain and C-domain interdomain correlation was weakened and the intradomain coupling strengthened. Formation number and the probability of the salt bridges were reduced dramatically, especially at 0.3 mol/L sodium chloride. These findings manifest that human NCS-1 protein is sensitive to the small variation of sodium chloride concentration, which may alter the key factors on protein conformation. Our study may provide the theoretical reference at the atomic structural insights for probing the conformational variations of human NCS-1 protein at the different aqueous solutions. 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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Morphological observation of broken ends of the mouse tail during wound healing and analysis of cell proliferation active regions Guo Jing-xu, Wang Hai-tao, Li Shu-wei 2018, 22 (16):  2558-2563.  doi: 10.3969/j.issn.2095-4344.0829 Abstract ( 548 )   PDF (5999KB) ( 186 )   Save BACKGROUND: Skin-derived precursor cells play a regenerative role in wound healing, which can restore the physiological and aesthetic features of the skin by mitosis, proliferation and migration to the injury region. OBJECTIVE: To observe the structure of the mouse tail and the healing of the broken end, and then to investigate the correlation of these characteristics with the repair of skin trauma at the tail. METHODS: The tails of 2-day-old Kunming mice were cut off using ophthalmic microsurgical scissors to establish the mouse model of hair follicle regeneration in the tail. Regions in which the cells proliferated actively were determined. Wound healing samples were regularly taken, and embedded using OCT. Wound healing of the broken end was routinely observed by pathological staining. Cell proliferation was observed by immunofluorescence. Expression of AP-1 in the healing site was identified by immunofluorescent staining. RESULTS AND CONCLUSION: The structure of the mouse tail was complex, with rich blood supply. Good healing was observed in the broken end of the mouse tail. The cells in the hair follicles and dermal papillary layer showed active proliferation and mitosis, and thereby participated in wound repair and regeneration. Fluorescence staining results found that the positive expression of AP-1 was mainly concentrated in the epidermis and dermal papilla. These findings indicate that the epidermis and dermal papilla possess a strong ability to regenerate. 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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Hydrolysis of Nanzhu fluid promotes proliferation and migration of human microvascular endothelial cells Cen Yan-hui1, 2, Lin Yong3, Lin Jiang1, Jia Wei1, Zhao Jing1 2018, 22 (16):  2564-2569.  doi: 10.3969/j.issn.2095-4344.0215 Abstract ( 395 )   PDF (6301KB) ( 123 )   Save BACKGROUND: Pearl has the tranquilizing effect, and it can be applied in the treatment of hypertension. However, there is little report on the prevention and cure effect and mechanism of hydrolyzed pearl liquid on hypertension. OBJECTIVE: To observe the effect of hydrolysis of Hepu pearl (hydrolyzed Nanzhu fluid) on the biological behavior and secretion of human microvascular endothelial cells. METHODS: Human microvascular endothelial cells were cultured and passaged. There were four groups, and the microvascular endothelial cells were incubated in the 200 μL culture medium containing nothing (control group), and 120, 60 and 30 mg/L hydrolysis of Nanzhu fluid. The cell proliferation and migration was detected by cell conuting kit-8 assay and Transwell assay respectively; the cell cycle distribution was tested by flow cytometry; the cell apoptosis was assayed by TUNEL method; the secretion of nitric oxide and reactive oxygen species was tested by nitrale reduetase and chemical fluorescence method, respectively.  RESULTS AND CONCLUSION: Compared with the control group, hydrolysis of Nanzhu fluid significantly promoted the proliferation of microvascular endothelial cells in a manner-dependent manner (P < 0.05), suggesting the optimal concentration was 120 mg/L. Compared with the control group, the percentage of cells in S and G2 phase was significantly increased, and the percentage of cells in the G1 phase was significantly reduced in the hydrolysis of Nanzhu fluid group (P < 0.05), indicating hydrolysis of Nanzhu fluid could promote cell cycle progression. Apoptotic cells with green-stained nucleus were invisible in both groups. The number of cell migration in the hydrolysis of Nanzhu fluid group was significantly more than that in the control group (P < 0.05). Compared with the control group, there was a significant increase in the nitric oxide secretion, and a significant decrease in the production of reactive oxygen species in the hydrolysis of Nanzhu fluid group (P < 0.05). To conclude, hydrolysis of Nanzhu fluid can promote the proliferation and migration of human microvascular endothelial cells in vitro, and has the function of promoting the secretion of nitric oxide and inhibiting reactive oxygen species secretion, implying its positive role in the protection of endothelial cell function. 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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Non-surgical treatment of lumbar intervertebral disc protrusion without blood stasis: a serum proteomic analysis Gui Yu-chang1, Xu Jian-wen2, Tai Zhi-hong1, Rao Yuan-sen1, Cao Yu-ju1, Yin Li-jun3 2018, 22 (16):  2570-2576.  doi: 10.3969/j.issn.2095-4344.0835 Abstract ( 359 )   PDF (1205KB) ( 129 )   Save BACKGROUND: Proteomics is a well studied research method, but its application in the non-surgical treatment of lumbar intervertebral disc protrusion (LIDP) is little reported. OBJECTIVE: To screen the differentially expressed proteins in patients with LIDP but without blood stasis before and after non-surgical treatment by proteomics. METHODS: Sixty patients with LIDP but without blood stasis were selected, and treated with non-surgical treatment for 4 weeks. The differentially expressed proteins were screened and identified by iTRAQ combined with LC-MS/MS. The bioinformatics analysis of the identified proteins was carried out, and the curative effectiveness was investigated. RESULTS AND CONCLUSION: Compared with those before treatment, the Visual Analogue Scale scores were significantly (P < 0.05), the Japanese Orthopedic Association scores were significantly increased decreased (P < 0.05), and the excellent and good rate reached 95.0% post-treatment. A total of 300 differentially expressed proteins were screened and 25 significantly expressed proteins were identified (P < 0.05). Bioinformatics analysis revealed that nine of the significantly expressed proteins were enriched to 15 KEGG signaling pathways. These results suggest that the use of Western medicine non-surgical treatment for the LIDP without blood stasis can achieve satisfactory results. Besides, complement C1qA, cDNA protein (FLJ60724), complement C4B frameshift mutation, cDNA protein (FLJ53025), mannose binding protein C, apolipoprotein B, hemoglobin α-1 globin chain variant, hemoglobin β subunit and cDNA protein (FLJ76254) may be the potential serum markers of the non-surgical treatment for the LIDP without non-blood stasis. 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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DEPTOR gene silencing promotes β-cell insulin secretion Qiu Hong, Lai Shu-chang, Pan Dao-yan, Wang Xiao, Shen Jie 2018, 22 (16):  2577-2582.  doi: 10.3969/j.issn.2095-4344.0278 Abstract ( 422 )   PDF (1267KB) ( 137 )   Save BACKGROUND: Mammalian target of rapamycin (mTOR) complexes are a key regulator of pancreatic beta cells mass and function. DEP-domain containing mTOR-interacting protein (DEPTOR) is a common part of mTOR complexes and whether DEPTOR loss in islet β cells affects insulin-secreting function has  never been identified. OBJECTIVE: To assess the alternation of insulin secretion by silencing DEPTOR gene in pancreatic β cells NIT-1 and to explore the underlying mechanism. METHODS: Three siRNA sequences for silencing DEPTOR gene were designed and constructed, which were transfected with lipofectamine into NIT-1 cells. There were six groups: blank transfection group (NIT-1 cells plus Lipofectamin), negative control group (NC-FAM), positive control group (GAPDH), siRNA deptor 1 group (siRNA deptor385), siRNA deptor 2 group (siRNA deptor766), and siRNA deptor 3 group (siRNA deptor1275). The transfection efficiency was determined by fluorescence microscope. The relative expression level of DEPTOR mRNA was detected by quantitative-PCR. Insulin secretion in the cell conditioned medium was determined by insulin ELISA kit. The expression level of DEPTOR downstream key protein was detected by western blot assay. RESULTS AND CONCLUSION: Specific green fluorescence accumulated in a punctated pattern under fluorescence microscope, indicating that the effectiveness of transfection was eligible. Quantitative-PCR results showed two (siDEPTOR385 and siDEPTOR766) of the three siRNA sequences could significantly disrupt the expression of DEPTOR mRNA, which had significant difference with negative control group (P < 0.05). The ELISA results showed that the total amount of insulin secretion in the effective transfected groups was significantly increased (P < 0.05). Western blot assay results showed the grey levels of p-s6 and p-4EBP-1 proteins were significantly elevated, while p-AKT of those former was slightly decreased. These findings suggest that siRNA technology can effectively silence the DEPTOR gene in NIT-1 cells, which improves β-cell insulin secretion in a manner of mTORC1 activation. 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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Combined intravitreal injection of triamcinolone acetonide and ranibizumab for age-related macular degeneration: study protocol for a randomized controlled trial Wei Fen, Xiao Xiu-lan 2018, 22 (16):  2583-2587.  doi: 10.3969/j.issn.2095-4344.0262 Abstract ( 400 )   PDF (987KB) ( 232 )   Save BACKGROUND: Age-related macular degeneration is a condition that leads to the deterioration of the macula in the elderly, and is characterized by the presence of drusen and degenerative changes of the retinal pigment epithelium and choroidal capillaries. It is a major cause of blindness worldwide. The commonly used strategies, including antioxidant therapy and dilation of blood vessels, have shown undesired outcomes in clinical practice. Ranibizumab, an anti-vascular endothelial growth factor fusion protein, is an angiogenesis inhibitor that has been used to treat wet (neovascular) age-related macular degeneration; however, patients treated with ranibizumab are prone to develop endophthalmitis, rhegmatogenous retinal detachment, retinal tears, and iatrogenic traumatic cataract. OJECTIVE: To determine the efficacy and safety of intravitreal injection of combination of triamcinolone acetonide and ranibizumab in the treatment of age-related macular degeneration in a randomized controlled trial. METHODS: Eighty patients (160 eyes) with age-related macular degeneration admitted at Jingzhou Hospital Affiliated to Tongji Medical School of Huazhong University of Science and Technology, China will be recruited. These patients will be randomly assigned to control or treatment group at a 1:1 ratio. The patients in the control group will be treated with intravitreal injection of triamcinolone acetonide (0.1 mL, once daily), while those in the treatment group will be treated with combined intravitreal injection of triamcinolone acetonide (0.025 mL, once daily) and ranibizumab (0.05 mL, once a month). All patients will undergo continuous treatment for 3 months, followed by a 6-month follow-up. The primary outcome measure is foveal retinal thickness before and 6 months after treatment. The secondary outcome measures include best corrected visual acuity, intraocular pressure, quality of life scores before and 6 months after treatment, and the incidence of adverse events at 6 months after treatment. This study was approved by the Ethics Committee of the Jingzhou Hospital affiliated to Tongji Medical College of Huazhong University of Science and Technology, China (approval No. 20170338), and will be performed in accordance with the Declaration of Helsinki. The participants will be informed of the study protocol and procedures and asked to sign an informed consent. Participant recruitment will be initiated in January 2018. Sample and data collection will begin in January 2018 and end in June 2018. The analysis of outcome measures and the completion of trial will be in September 2018. The results of this study will be disseminated through presentations at scientific meetings and/or by publication in peer-reviewed journals. This trial was registered with the Chinese Clinical Trial Registry (registration No. ChiCTR-IOR-17013865). DISCUSSION: We aim to confirm the safety and efficacy of the intravitreal injection of combination of triamcinolone acetonide and ranibizumab in the treatment of age-related macular degeneration. 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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Ranibizumab combined with photodynamic therapy for the treatment of advanced-stage exudative age-related macular degeneration: study protocol for a self-controlled trial and preliminary results Yu Ping, Wang Qing, Liu Ling-ling 2018, 22 (16):  2588-2592.  doi: 10.3969/j.issn.2095-4344.0263 Abstract ( 321 )   PDF (1113KB) ( 111 )   Save BACKGROUND: The lack of visual field in the visual center of older adults with aged-related macular degeneration (AMD) will have a serious impact on the activities of daily living. There are atrophic and exudative AMD according to the pathological manifestations. Patients with advanced stage of exudative AMD will develop aged-related degeneration in the macular area. Ranibizumab combined with photodynamic therapy is mainly used for current treatment of exudative AMD, but more attention is paid to patients with early exudative AMD. OBJECTIVE: To analyze vision recovery, retinal thickness, and leakage of choroidal neovascular lesions in patients with advanced stage of exudative AMD after treatment with ranibizumab plus photodynamic therapy, and further explore the therapeutic efficacy of the combined therapy. METHODS: The prospective self-controlled trial will be completed in the Department of Ophthalmology, Affiliated Hospital of Qinghai University in China. The study population will comprise 113 patients with advanced exudative AMD who are eligible for inclusion and exclusion criteria. Photodynamic therapy combined with intravitreal injection of ranibizumab will be given as the interventional therapy. RESULTS AND CONCLUSION: The primary outcome measure of the trial is to investigate the visual improvement in patients 12 months after treatment. Secondary outcome measures include logarithmic visual acuity chart scores, retinal thickness, and leakage of choroidal neovascular lesions assessed monthly at 1-12 months after treatment, and adverse events during follow-up. Preliminary results showed that at 12 months after treatment, visual acuity in 43 affected eyes was increased by ≥ 2 lines, and the average retinal thickness decreased by 111.21 μm as compared with before treatment. Leakage of choroidal neovascular lesions in most affected eyes stopped or the lesions were contracted. Findings from the trial are expected to provide reliable data on the efficacy of ranibizumab combined with photodynamic therapy in the treatment of advanced exudative AMD, thereby providing a basis for the rational use of this combined therapy in the treatment of advanced exudative AMD. This trial was registered with the Chinese Clinical Trial Registry (registration number: ChiCTR1800015410). Protocol version (1.0). 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 promotes bone and cartilage regeneration Zu Hai-yue1, Yi Xue-ting2, Zhao De-wei1 2018, 22 (16):  2593-2600.  doi: 10.3969/j.issn.2095-4344.0805 Abstract ( 395 )   PDF (1290KB) ( 154 )   Save BACKGROUND: Low-intensity pulsed ultrasound (LIPUS) is a non-thermal and non-invasive mechanical stimulation, which has achieved certain curative efficacy on bone and cartilage defects. OBJECTIVE: To retrospectively analyze the effects and mechanisms of LIPUS in bone and cartilage regeneration process, and to review the related cellular signals and tissue regeneration mechanism involved in the current achievement of basic research and clinical application, thus providing theoretical basis for clinics. METHODS: The first author retrieved Cochrane Library, PubMed, CBM, CNKI and WanFang databases using compute for the articles addressing LIPUS promoting bone and cartilage regeneration published from January 1990 to February 2017. The keywords were “LIPUS, calcium, integrin, nitric, oxide, prostaglandin, BMP” in English and Chinese, respectively. The articles published in authoritative magazines or recently published were preferred, and finally 80 articles were selected for result analysis. RESULTS AND CONCLUSION: The basic research concerning LIPUS involves cellular mechanics and tissue engineering. Especially with the support of molecular biology, there has been a major breakthrough in promoting bone and cartilage regeneration. LIPUS can stimulate cells and tissues to produce mechanical signals by mechanical wave, lead to changes in cytokines in the signaling pathways, further accelerate blood supply and metabolism, and finally promote the regeneration of bone and cartilage. Therefore, LIPUS is an effective treatment method. 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 post-activation potentiation induced by high-intensity dynamic and static kinetotherapies enhance muscle explosive power? Li Xin-rui1, 2, Luo Jiong1, 2, Song Gang1, 2 2018, 22 (16):  2601-2606.  doi: 10.3969/j.issn.2095-4344.0817 Abstract ( 320 )   PDF (997KB) ( 134 )   Save BACKGROUND: Increasing evidence has shown that high-intensity warming up, including dynamic and static kinetotherapies, can improve the performance of muscle explosive power. However, the training scheme to achieve the best effect remains unclear. OBJECTIVE: To discuss whether the high-intensity dynamic and static kinetotherapies can enhance post-activation potentiation of the upper and lower limbs, so as to fully understand the high-intensity warming up, and thus promote the training efficacy. METHODS: Ovid Medline, Elsevier SDOL, PubMed, and CBJ databases were retrieved for the articles addressing the post-activation potentiation of high-intensity dynamic and static kinetotherapies published before 2016. The articles were classified in accordance with Oxford Centre for Evidence-based Medicine Levels of Evidence. RESULTS AND CONCLUSION: The possible mechanisms of the post-activation potentiation include the phosphorylation of myosin regulation light chain, the increase in motor unit recruitment as well as the change of pinnate angle. The resistance exercise at an appropriate mode, including dynamic, static, and dynamic combined with static, significantly induces post-activation potentiation, and improves the explosive force. Post-activation potentiation is impacted by muscle contraction type, amount of exercise, exercise intensity, induction time, and history of the subjects (training experience, absolute muscle force, and muscle fiber composition). High-intensity stimulus of post-activation potentiation may be accompanied by the generation of fatigue, and the inappropriate recovery time may result in the imperfect performance of post-activation potentiation. 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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Characteristics and acute responses of eccentric exercise Xu Zhi-yong1, Yin Xin2, Huang Qiang-nian3, Xu Sheng-jia3 2018, 22 (16):  2607-2612.  doi: 10.3969/j.issn.2095-4344.0225 Abstract ( 381 )   PDF (1025KB) ( 136 )   Save BACKGROUND: Eccentric exercise displays many advantages over concentric and isometric contractions, and it has been applied in exercise training and rehabilitation. The existing researches focus on the underlying mechanisms at molecular and neural levels, and the specific adaptation after eccentric exercise may be related to the adaptive signaling pathway. OBJECTIVE: To review the unique physiological characteristics and mechanisms of eccentric exercise, thereby providing reference for in-depth understanding of eccentric exercise. METHODS: The relevant articles were searched in PubMed (1990-2017) and CNKI (2010-2017) with the keywords of “eccentric exercise, negative muscle work, eccentric exercise training, downhill run, concentric exercise, positive muscle work, concentric exercise training, and uphill running” in English and Chinese, respectively. Ultimately, the articles eligible for physiological characteristics, acute response and adaptive mechanism were enrolled. RESULTS AND CONCLUSION: Totally 98 articles were retrieved, and 54 pertinent papers were enrolled for analysis based on the inclusion and exclusion criteria. The mechanical, molecular and neural mechanisms of eccentric contractions differ from those of concentric and isometric contractions. Special metabolic and cardiorespiratory responses after eccentric exercise reflect the advantages in making programs of exercise training and rehabilitation. Particularly, compared with concentric exercise, fewer motor units are recruited during eccentric exercise and exert muscle a greater stimulation, which involves the process of exercise-induced muscle damage and the activation of satellite cells. The content of satellite cells after eccentric exercise is higher than that of concentric exercise, suggesting that satellite cells may play an important role in the muscle damage reconstruction caused by eccentric exercise. In conclusion, eccentric exercise is of great significance for exercise training and rehabilitation, while further study on the adaptive mechanism of eccentric exercise is necessary. 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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Metabolomics applied in Chinese medicine syndromes: a bibliometric analysis He Jing1, Sun Zhi-ling1, Xie Tong2, Jiao Wen-juan1, Zhang Yong-yi1 2018, 22 (16):  2613-2618.  doi: 10.3969/j.issn.2095-4344.0195 Abstract ( 348 )   PDF (1213KB) ( 133 )   Save BACKGROUND: Metabolomics is a branch of systems biology taking systematic study, high-throughput detection and data processing as means, information modeling and systematic integration as targets, which can be used for recognizing metabolic indexes, provide evidence for individualized diagnosis and treatment and guide syndrome differentiation in the clinic. OBJECTIVE: To analyze the literature features and research status of metabolomics applied in the field of Chinese medicine syndromes so as to provide reference for its application in Chinese medicine syndromes. METHODS: Databases of CNKI, WanFang, CBM, PubMed, Web of Science and Medline were retrieved for the articles addressing metabolomics applied in Chinese medicine syndromes published before June 2017. The literature database was established, and then the literature and research features were analyzed using bibliometrics and data mining. RESULTS AND CONCLUSION: Totally 499 articles were enrolled, including 371 journal articles from journals (74.35%), 30 conference papers (6.01), 98 dissertations (19.64), and the 371 journal papers were published in 124 journals (32 of Chinese core journals (45.28%), and 10 SCI cited journals (3.77%)). In the articles, 7 types of disease systems (mainly in digestive system and circulatory system) were classified according to the statistics, involving 23 diseases and 39 interventions. In summary, there is still a lack of standardized classification for metabolomics applied in Chinese medicine syndromes and the quality of literature is poor. We should conduct more animal experiments and explore the essence and intervention measurements of syndromes, thereby controlling the disease occurrence and 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 | References | Related Articles | Metrics

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Management of biosamples transferred from hospitals to laboratories: main problems and solutions Cheng Ming1, Jiang Feng2 2018, 22 (16):  2619-2624.  doi: 10.3969/j.issn.2095-4344.0217 Abstract ( 415 )   PDF (1112KB) ( 112 )   Save BACKGROUND: Translational medical research realization needs close collaboration between clinicians and basic scientists. Human disease-related biological samples play a key role in the translational medical research. OBJECTIVE: To discuss the necessity of the cooperation between clinicians and scientists, and to discuss the main problems in the management of biosamples that transferred from hospitals to laboratories. METHODS: The keywords were “translational medical research, biosample, biobank” in English and Chinese, respectively. CNKI, VIP, WanFang and PubMed databases were retrieved for the literature published from January 2010 to December 2016. Totally 124 eliglible articles were included, and some of them were selected to analyze the cooperation between clinicians and scientists RESULTS AND CONCLUSION: Problems in the management of bio-samples that transferred from hospitals to basic research laboratories include ethics, standardized operation, as well as collection, transport, preservation, management and intellectual property of bio-samples. Standard biosamples with high quality is the key to translational medical research. Establishing a perfect interests-coordination mechanism is of great significance for the management of biosamples that transferred from hospitals to laboratories and translational medical 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