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10 September 2013, Volume 17 Issue 37 Previous Issue    Next Issue

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Gene transfection at different time affects the osteogenesis in traction area Hu Chun-bing, Wu Guo-ping, Liu Xi, Lan Yong-shu, He Xiao-chuan, Guo Li 2013, 17 (37):  6541-6547.  doi: 10.3969/j.issn.2095-4344.2013.37.001 Abstract ( 266 )   PDF (1903KB) ( 393 )   Save BACKGROUND: The previous studies have shown that electroporation-mediated recombinant plasmid pIRES-hBMP2-hVEGF165 can promote the early angiogenesis and new bone formation in the traction zone. OBJECTIVE: To investigate the effect of gene transfection at different time on the new bone formation in the traction zone during mandibular distraction in rabbits, so as to explore the optimal time for gene therapy and obtain a better treatment effect. METHODS: Forty-eight New-Zealand rabbits were employed. After accomplished the bilateral mandibular osteotomy and distractor implantation under general anaesthesia, the rabbits were randomly divided into four groups. The bilateral traction zones were transfected with the 2 μg (0.1g/L) recombinant plasmids pIRES-hBMP2-hVEGF165 at latency period, traction period and consolidation period, respectively, and the rabbits in these three groups received electroporation stimulation. The simple traction group only received traction without gene transfection. The rabbits in each group received traction at 3 days after modeling with the rate of 0.8 mm/d and once per day, and continuous traction for 10 days; three rabbits were sacrificed in each group at 1, 2, 4 and 8 week. The new tissues in the mandibular traction zone were cut for histological observation and histomorphometry analysis. RESULTS AND CONCLUSION: Histological examination and histomorphometry analysis showed that amounts of newly formed vessels, osteoblasts and mesenchymal cells of distraction period transfection group were greater than those of the latency period transfection group, consolidation period transfection group and simple traction group, and the new bone volume and the width of bone trabecula in the distraction period transfection group were higher than those in the other three groups (P < 0.05). It is better to transfect gene at the beginning of traction (distraction period) than at other stages of traction; in this way, we can obtain more remarkable effects on new bone formation. It suggests that the distraction stage is the optimal time for gene therapy. Figures and Tables | References | Related Articles | Metrics

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Bone remodeling of rabbit tibia diaphysis cortical bone under bending force Wang Yu, Li Cheng-mei, Li Yu-sheng, Zhang Zhi-gang 2013, 17 (37):  6548-6554.  doi: 10.3969/j.issn.2095-4344.2013.37.002 Abstract ( 350 )   PDF (1748KB) ( 444 )   Save BACKGROUND: The stress changes of bone are the main simulation sources for bone remodeling. At present, the research on the relationship between stress and bone remodeling focuses on the effect of axial stress on bone remodeling, while the research on bone remodeling under bending force is rare. OBJECTIVE: To observe the bone remodeling of rabbit tibia diaphysis cortical bone in growth phase under bending force. METHODS: A bone window with the diameter of 2.5 mm was made 0.5 cm from the medial distal epiphyseal line of rabbit proximal left tibia, and then ice saline was used to cool the tibia and memory alloy film. After cooling, the memory alloy stripe was straightened and implanted into the tibial bone marrow cavity through the bone window. The direction of memory alloy stripe implantation: two ends of the memory alloy stripe arrived to the lateral cortex after rewarming, and the curve apex arrived to the medial cortical bone, followed by the sutured wound. The continuous internal bending force was loaded on the rabbit tibia diaphysis through the shape memory alloy stripe implanted into the tibial bone marrow cavity in order to observe the bone remodeling mode of tibia diaphysis cortical bone under long-term bending force. RESULTS AND CONCLUSION: After 6 months continuous bending force, the thickness of cortical bone on the tension stress side of tibia diaphysis was decreased, while the thickness on the compressive stress side was increased, and no obvious bending deformation was observed. The thickness of the cortical bone on the compressive stress side was associated with the stress distribution on tibia. The tibia with integrity structure exhibited an inherent “resistance” to the bending stress, this indicating that the bone shaft of the long bone without fracture had no potential of force line correction basically. The results suggested that the bone remodeling model of tibia diaphysis cortical bone had the advantages of less trauma, controlled force, less influence on the growth and metabolism, no limitation on the animal activities and meet the physiological state, which was an ideal model of bone remodeling.   Figures and Tables | References | Related Articles | Metrics

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Short-time middle-load treadmill exercise affects the bone mineral density of hyperlipidemia male rats Wang Xiao-hong, Xu Chao, Zhang Zhuo, Guo Lian-ying, Zhou Bo 2013, 17 (37):  6555-6560.  doi: 10.3969/j.issn.2095-4344.2013.37.003 Abstract ( 283 )   PDF (1974KB) ( 329 )   Save BACKGROUND: Some studies have shown that hyperlipidemia can lead to osteoporosis in rats, and exercise can increase the bone mineral density of rats. But the effect of short-time exercise on the bone mineral density of hyperlipdemia induced osteoporosis male rats is unclear yet. OBJECTIVE: To investigate the effect of short-time middle-load treadmill exercise on the bone mineral density of hyperlipidemia male rats. METHODS: Twenty-six male Sprague Dawley rats were randomly divided into three groups: control group (n=8), hyperlipidemia group (n=9) and exercise intervention group (n=9). The rats in the control group were fed with normal diet, and the rats in the other two groups were fed with high-fat diet and lasted for 4 weeks to establish the hyperlipidemia models. The rats in the exercise intervention group received treadmill exercise 5 days per week for 4 weeks according to the following schedule: 15 m/min for 15 minutes in the 1st week, 15 m/min for 20 minutes in the 2nd week, and then 20 m/min for 20 minutes in the last 2 weeks. Slope grade of the treadmill was adjusted at 0°. At the end of experiment, the rats were sacrificed, and the bone mineral density of the right femur, the morphological change of tibia, the level of plasma alkaline phosphates and calcium content were examined. RESULTS AND CONCLUSION: Compared with the control group, the distal femur bone mineral density in hyperlipidemia group was significantly decreased (P < 0.05); histological analysis of the proximal tibia showed thinning and loss of bony trabeculae arrangement, the gap was widened, and a large amount of fat cells infiltration or integration into vacuoles in the marrow was observed, the plasma alkaline phosphates was significantly increased (P < 0.01). Compared with the hyperlipidemia group, distal femur bone mineral density in the exercise intervention group was increased (P < 0.05). After adjust body weight, the whole femur bone mineral density was significantly greater in exercise intervention group compared to hyperlipidemia group (P < 0.05). The histological analysis of the proximal tibia showed that the spaces of bone trabeculae decreased and the structure of bone trabeculae compacted, the alkaline phosphates activities were increased (P < 0.01). There were no significant differences in serum calcium and phosphates levels between groups. The results show that short-time middle-load treadmill exercise can increase the bone mineral density of hyperlipidemia male rats. Figures and Tables | References | Related Articles | Metrics

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Naringin intervention influences the formation and function of osteoclasts from mouse calvarial bone You Tao, Wang Lu, Ji Yan, Wu Xiao-hong 2013, 17 (37):  6561-6566.  doi: 10.3969/j.issn.2095-4344.2013.37.004 Abstract ( 253 )   PDF (1737KB) ( 344 )   Save BACKGROUND: Naringin can increase bonemorphogeneticprotein 2 gene expression and promote the proliferation and differentiation of multipotential stem cell line. In vitro cell experiment indicates that naringin can suppress osteoclast formation and anti-osteoporosis activity. OBJECTIVE: To prepare a mouse calvarial bone culture model containing naringin and to observe the effect of naringin with different concentrations on the formation and function of osteoclasts. METHODS: Calvarial bone was dissected out aseptically from the 4-day-old Sprague Dawley mice, and cultured in the culture medium containing 0, 1, 10 and 100 mg/L naringin. The effects of naringin on the number of tartrate-resistant acid phosphatase-positive osteoclast marker enzyme in calvarial bone and the concentration of calcium in the culture medium were detected after cultured for 1, 3, 7 and 10 days.  RESULTS AND CONCLUSION: After cultured for 1 day, the number of tartrate-resistant acid phosphatase-positive cells in calvarial bones and the calcium concentration in the culture medium had no difference between the groups. After cultured for 3 and 7 days, the number of tartrate-resistant acid phosphatase-positive cells was decreased and the calcium concentration was increased with the increasing concentration of naringin. At 10 days after culturing, this trend was most obvious. It showed that naringin could affect the number and function of tartrate-resistant acid phosphatase-positive cells in calvarial bones, and this effect showed a significant time- and dose-depend manner. The results show that naringin can not only promote the proliferation of osteroclasts, but also reduce the differentiation of osteoclasts or accelerate the apoptosis. Figures and Tables | Related Articles | Metrics

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Different frequencies of vibration strains affect the early proliferation and differentiation of osteoclasts Chen Guo-xian, Wang Guo-rong, Lin Zong-jin, Li Guo-shan, Lin Qun-xian, Huang Yi-ping, Guo Chun-xian, Luo Yuan-biao, Zeng Qing-dong, Chen Wei-yi 2013, 17 (37):  6567-6572.  doi: 10.3969/j.issn.2095-4344.2013.37.005 Abstract ( 352 )   PDF (1724KB) ( 443 )   Save BACKGROUND: Studies have shown that low-intensity and short-time vibration with a certain frequency can reduce the absorption of bone tissue and increase the quantity and quality of bone through promoting the proliferation and differentiation of osteoblasts.  OBJECTIVE: To investigate the effects of different frequencies of vibration strains on cycle, proliferation and differentiation potency of RAW264.7 cells cultured in vitro. METHODS: Passage 6 RAW264.7 cells in good conditions were randomly divided into six groups, and each group was induced cultured with Dulbecco’s modified Eagle’s medium containing receptor activator of nuclear factor kappa B ligand. The final concentration of receptor activator of nuclear factor kappa B ligand was adjusted to 50 μg/L, and then kept without changes. The non-loading group did not loaded with vibration strain, and the other five groups were loaded with 3-10 Hz, 15-35 Hz, 35-45 Hz, 50-70 Hz and 70-90 Hz vibration strains on the RAW264.7 cells respectively. The other vibration parameters were consistent; the vibration time was 15 min/time with the vibration intensity of 0.3 g, twice per day. The cell cycle and cell proliferation were detected at 3 and 6 days after loading of vibration strains. RESULTS AND CONCLUSION: After composite vibration loading for 6 days, the cell cycle phase in the vibration groups was changed to some extent when compared with the non-loading group. Compared with non-loading group, the cell number in the G1 phase of the vibration group was significantly increased (P< 0.01); the cell number in the S phase and G2+M phase of the vibration group was significantly decreased (P < 0.01); the vibration parameters in the vibration group were significantly decreased (P < 0.01). The results indicate that different frequencies of vibration strains can affect the cell cycle and proliferation, and can inhibit the proliferation and differentiation of osteoblasts.  Figures and Tables | Related Articles | Metrics

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Intermittent negative pressure affects tendon-bone healing after anterior cruciate ligament reconstruction Sun Zheng-ming, Ling Ming, Feng Wei-lou, Dong Xiang-hui, Liu Shi-zhang, Yi Zhi 2013, 17 (37):  6573-6579.  doi: 10.3969/j.issn.2095-4344.2013.37.006 Abstract ( 322 )   PDF (2222KB) ( 402 )   Save BACKGROUND: Intermittent negative pressure has been proven to promote the repairing of soft tissue and bone healing, but the effect of negative pressure on the tendon-bone healing after anterior cruciate ligament reconstruction has not been reported. OBJECTIVE: To research the effect of intermittent negative pressure on tendon-bone healing after anterior cruciate ligament reconstruction and on the biomechanics of tendon grafts. METHODS: A total of 24 New Zealand white rabbits were randomly selected to establish the models of anterior cruciate ligament reconstruction of autogenous semitendinosus. The hind leg of one side was selected randomly as the negative pressure group, and the joint of the negative pressure side was connected with the micro-negative pressure aspirator through drainage tube and maintained a low-intensity and intermittent negative pressure; the contralateral hind leg was as the control and connected with ordinary drainage tube. Drainage tubes of both sides were removed at the same time after 5 days. At 6 weeks after modeling, the joint fluid was drawn to detect the expression levels of interleukin-1β; femur-ligament-tibia complex was used for tension measurement of tendon graft, and histological observation of tendon-bone interface. RESULTS AND CONCLUSION: One rabbit had joint infection, and finally 23 rabbits were included in the study.Tension measurement results showed that maximum load for breakage in negative group pressure was significantly greater than that in the control group (P < 0.05). Histological observation found that the number of osteoblasts in the negative pressure group was significantly more than that in the control group, and the difference was statistically significant (P< 0.01). Detection of synovial fluid showed that iterleukin-1β content in synovial fluid of the negative pressure group was lower than that in the control group, and the difference was statistically significant (P < 0.01). Intermittent negative pressure may play an active role in tendon-bone healing and modeling of tendon graft after anterior cruciate ligament reconstruction. Figures and Tables | Related Articles | Metrics

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Ultrasound effects on chondrocyte apoptosis and the expressions of caspase-8 and caspase-3 Guo Hua, Xia Lu, Zhou Jun, Chen Shi-ju, He Cheng-qi 2013, 17 (37):  6580-6586.  doi: 10.3969/j.issn.2095-4344.2013.37.007 Abstract ( 342 )   PDF (1824KB) ( 519 )   Save BACKGROUND: Ultrasound therapy can relieve pain and improve the movement function in patients with knee osteoarthritis, but there lacks of consistency in the literatures of ultrasound therapy. OBJECTIVE: To further identify the effectiveness of ultrasound therapy in the treatment of knee osteoarthritis. METHODS: Twenty-four rabbits were randomly divided into three groups: normal group, model group and ultrasound group. The rabbit in the normal group received no intervention; rabbits in the model group received anterior cruciate ligament transaction to establish the knee osteoarthritis model without any treatment; the rabbit in the ultrasound group received ultrasound therapy after modeling for 10 minutes once time, once per day,    0.3 W/cm2, 1 MHz and treated for 10 times. Hematoxylin-eosin staining was conducted for histological observation of rabbit articular cartilage; western blot and reverse transcription PCR assessment were used to assess the expressions of caspases-3 and caspases-8 in rabbit articular cartilage, while Terminal deoxynucleotidyl transferase dUTP nick end labeling was used to assess the ratio of chondrocytes apoptosis of rabbit knee articular cartilage. RESULTS AND CONCLUSION: The normal rabbit cartilage tissues and chondrocytes were neatly arranged in column; the middle cartilage layer of the model was thin; the chondrocytes were arranged disorderly and became less. After ultrasound therapy, the chondrocytes were rearranged neatly, and the number was increased. Compared with the normal group, the Mankin scores in the model group and ultrasound group were higher; the apoptotic rate of chondrocytes was higher in the model group and ultrasound group than in the normal group, and was also higher in the model group than in the ultrasound group. Compared with the normal group, the expressions of caspases-3 and caspases-8 were higher in the model group and the ultrasound group, while decreased after ultrasound therapy. The results indicate that ultrasound can improve the structure of cartilage tissues, decrease the expressions of caspases-3 and caspases-8 and reduce the apoptosis rate of chondrocytes. It is effective for the treatment of knee osteoarthritis with ultrasound therapy. Figures and Tables | Related Articles | Metrics

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Histological changes and safety of long-term acral nerve root stimulation electrode placement in rabbits Yan Peng, Zheng Wei-dong, Zhang Ji-kai, Tan Yun-bing, Li Gao-feng, Li Guang-chun, Song Cheng, Yang Xiao-yu 2013, 17 (37):  6587-6593.  doi: 10.3969/j.issn.2095-4344.2013.37.008 Abstract ( 351 )   PDF (908KB) ( 465 )   Save BACKGROUND: Studies have shown that sacral nerve-root stimulation based on anodes block technique can effectively reconstruct the bladder voiding function of the rabbits with spinal cord injury. But the corresponding technology of stimulating electrode has not been reported so far. OBJECTIVE: To design and develop the stimulating electrodes matching with both rabbit sacral nerve roots and anodal blocking technique, to observe the ultrastructure and morphological change of rabbit sacral nerve roots which implanted in electrode stimulation for a long-term and to assess the safety of stimulating electrodes. METHODS: Thirty New Zealand rabbits were included, 10 rabbits were randomly selected from them and sacrificed after anesthesia, and then cut the anterior roots of bilateral S2 and S3 immediately; after measuring the diameter under the light microscope, the sleeve type stimulation electrode matched with the diameter was made. The remaining 20 rabbits were randomly divided into control group and implantation group, with 10 rabbits in each group. In the implantation group, the stimulating electrodes were implanted into the forepart of S2 and S3 nerve roots after anesthesia, and then sacrificed after fed for half a year for collecting the samples. Then ultrastructure change of sacral nerve roots with the implantation was observed.  RESULTS AND CONCLUSION: Structure of nerve cells of sacral nerve roots remained in good condition under a light microscope after long-term implantation of the stimulating electrodes. No obvious degeneration of axons, no inflammatory infiltration and glial scar formation were observed. In the implantation group, myelins arranged closely without demyelination phenomenon, and there was no atrophy of neuronal nuclear, no nuclear sag, no increased nuclear decompression and heterochromatin in neurons under the light microscope. Immunohistochemical analysis showed, compared with the control group, there were no significant differences in the expressions of glial fibrillary acidic protein, Bax, Bcl-2 and Caspase-3 proteins of nerve roots in the implantation group. The stimulation electrode of rabbit sacral nerve root is developed successfully, that is, the implantation is simple and safe as it can be used for long-term implantation without histopathological changes and apoptosis. Figures and Tables | Related Articles | Metrics

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Biological effects of peripheral nerve block on the spinal cord Li Xia, Zhang Yong, Zhang Jing 2013, 17 (37):  6594-6600.  doi: 10.3969/j.issn.2095-4344.2013.37.009 Abstract ( 475 )   PDF (930KB) ( 361 )   Save BACKGROUND: How does the peripheral nerve block work to inhibit the functions? What effects does it do to the spinal cord and the cerebral center? How does it regulate the peripheral nerve and how does it change anatomy of brain center? All above are still unknown. OBJECTIVE: To observe the effect of peripheral nerve block on the biology of spinal cord nerve cells. METHODS: Sixty New Zealand rabbits were selected and randomly divided into three groups: ischiadic nerve block group, subarachnoid block group and extradural nerve block group. Each group had 20 rabbits which were sub-divided into experimental group and control group with 10 rabbits in each subgroup. The rabbits in the three experimental groups were injected with lidocaine, bupivacaine and lidocaine at the middle point between femoral head and ischiadic tuberosity in ischiadic nerve block group, subarachnoid block group and extradural nerve block group, respectively. The rabbits in each control group were injected with normal saline at the same position. RESULTS AND CONCLUSION: After ischiadic nerve block, extradural nerve block or subarachnoid block for 30 minutes, the Nigeria’s bodies of the little round cells in the poliomyelia posterior horn laminae Ⅲ and Ⅳ and the polyhedral cells in the anterior horn laminae Ⅸ were less than those in the control groups. Nuclei leaned towards one side. c-Fos protein expression was decreased or showed no expression, suggesting that the cord nerve cell function of corresponding spinal segments was inhibited. The spinal pia mater of spinal cord had a hierarchical or fracture phenomenon after subarachnoid block. Figures and Tables | Related Articles | Metrics

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Simulation of intestinal content sensory nerves with ultrasonic detection system Li Jian-guo, Tang Cheng-xian, Yang Xue-feng, Shang Xian-hui, Cheng Jia-ping 2013, 17 (37):  6601-6606.  doi: 10.3969/j.issn.2095-4344.2013.37.010 Abstract ( 444 )   PDF (753KB) ( 447 )   Save BACKGROUND: The existed artificial anal sphincter cannot sense whether there are intestinal contents in the bowel and the state of intestinal contents, and the previous studies on the intestinal content signal did not included the different parts of colon and the signals of different intestinal content status. OBJECTIVE: To compare the differences of ultrasound signals of different intestinal content status in different parts of New Zealand rabbits detected with transmission-type ultrasonic signal detection system.  METHODS: Twenty healthy New Zealand rabbits were used; the rabbits were divided into groups according to distal part of proximal colon, distal colon and distal rectum. Five positions of solid stool particles were selected randomly as the detection point in each group. The voltage signals of stool were measured under solid, fluid, gaseous, and deflation status of the lumen. RESULTS AND CONCLUSION: In rectum, there were no significant differences in voltage signals between solid and gaseous conditions, while there were significant differences between fluid and deflation conditions (P=0.000). Multiple comparison showed there were significant differences between four conditions in distal part of proximal colon and distal colon (P=0). There were significant differences in voltage singles between rectum, distal colon and proximal colon under solid and fluid conditions (P=0). Multiple comparison showed there were significant differences in any part of large bowel for solid and fluid status (P=0). There were no significant differences in voltage singles between rectum, distal colon and proximal colon under gaseous and deflation conditions. There was interactive effect of intestinal contents between three parts of large bowel under the four conditions (P=0.000). Transmission-type ultrasonic detection system can distinguish artificial intestinal contents sensory nerves of various intestinal contents. And the application of ultrasonic detection system to detect the intestinal contents under different conditions can provide a new effective method for the research on artificial intestinal content sensory nerves. Figures and Tables | Related Articles | Metrics

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Deguelin treatment delays embryonic development of zebrafish: A gene chip analysis Li Wen-hao, Liu Rui-jin, Wu Xin-rong 2013, 17 (37):  6607-6612.  doi: 10.3969/j.issn.2095-4344.2013.37.011 Abstract ( 326 )   PDF (723KB) ( 531 )   Save BACKGROUND: Anticancer drug deguelin delays the embryonic development of zebrafish, but its exact mechanism is not yet clear. OBJECTIVE: To explore expression differences in genes and signaling pathways in deguelin-treated zebrafish embryos. METHODS: Zebrafish embryos were harvested from zebrafish fed according to the Zebrafish Book. Zebrafish embryos at 2-cell stage were selected and divided into two groups: control group treated with dimethyl sulfoxide incubator liquid, and experimental group treated with 0.6 μmol/L deguelin. Total RNA was extracted from the two groups at sphere stage. Then, gene chip technique was used to detect differentially expressed genes in the deguelin-treated zebrafish embryos. Real-time quantitative PCR was employed to validate microarray cluster analysis and pathway analysis to explore the mechanism of action of deguelin. RESULTS AND CONCLUSION: Chip results showed that 407 genes were upregulated more than three times and 461 genes were downregulated more than three times after deguelin treatment. PCR validation results were consistent with those of the chip. Fourteen pathways were identified by KEGG pathway analysis. Deguelin may play an important role by intervening cell metabolism growth and differentiation. Figures and Tables | Related Articles | Metrics

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Zebrafish embryonic brain cell apoptosis and c-fos gene expression after hypoxia reperfusion Chen Yan-chen, Zhao Dan, Qing Di, Cheng Dong-liang, Mao Jiao-yu, Wang Bin 2013, 17 (37):  6613-6619.  doi: 10.3969/j.issn.2095-4344.2013.37.012 Abstract ( 836 )   PDF (2760KB) ( 913 )   Save BACKGROUND: Foreign scholars have researched hypoxia reperfusion in zebrafish embryos, but there is no research on c-fos gene expression and the mechanism during zebrafish cerebral hypoxia reperfusion. OBJECTIVE: To observe the zebrafish embryonic brain cell apoptosis and expression of c-fos gene in brain tissues after hypoxia reperfusion. METHODS: Zebrafish embryos were selected at 48 hours post fertilization. Neonatal hypoxia reperfusion injury was simulated by gradually leading nitrogen (99.999%) into the device. After hypoxia treatment for 6, 12 and 24 hours, the embryos received reperfusion for 6 hours under normal oxygen concentration. The embryos in the control group received normoventilation (the dissolved oxygen concentration was about 7.0 mg/L). Acridine orange staining was performed to observe the effect of different hypoxia durations on the apoptosis of neurons in zebrafish, and then the c-fos gene expression was quantitative analyzed with real-time quantitative nucleic acid amplification detection system. And the expression level of c-fos gene was compared before and after hypoxia reperfusion. RESULTS AND CONCLUSION: A small amount of apoptotic brain cells could be detected in the control group, and the c-fos gene expression level was decreased; in the experimental group, the number of apoptotic cells was increased after hypoxia for 6, 12 and 24 hours, and the gene expression after hypoxia for 6 hours was increased distinctly. The results indicate that hypoxia can increase the c-fos gene expression in brain cells of zebrafish embryos, which may be one of the mechanisms of brain cell apoptosis increasing after hypoxia. Figures and Tables | References | Related Articles | Metrics

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Constructing an artificial skin model of melanoma through seeding MV3 melanoma cells and HaCaT cells onto the de-epidermized dermis Ye Teng, Lu Bin, Lu Hong-guang 2013, 17 (37):  6620-6627.  doi: 10.3969/j.issn.2095-4344.2013.37.013 Abstract ( 323 )   PDF (545KB) ( 1005 )   Save BACKGROUND: We have built the three-dimensional human skin melanoma model with human epidermal keratinocytes and MV3 melanoma cells co-cultured on the de-epidermized dermis in vitro. OBJECTIVE: To establish a skin model of melanoma by mixed culture of MV3 melanoma cells and HaCaT cells on the de-epidermized dermis in vitro. METHODS: MV3 melanoma cells and HaCaT cells were mixed with different percentages and inoculated on the surface of de-epidermized dermis followed by a liquid culture and air-liquid culture, and then the tissue- engineered skin model was established in vitro. Routine biopsy immunohistochemical observation was performed on the constructed skin melanoma model. RESULTS AND CONCLUSION: Hematoxylin-eosin staining showed that MV3 melanoma cells were distributed on the surface layer of de-epidermized dermis and formed tumor masses, while the HaCaT cells were mixed growth with tumor cells and formed a typical epidermoid structure. Some tumor cells infiltrated into the surface or deep of de-epidermized dermis and showed a tumor foci distribution. The CK10, CK-pan and S-100 proteins were positive for immunohistochemical staining. With the increasing of MV3:HaCaT cell percentage, CK10 and CK-pan gradually down-moved from the surface, and changed from layer distribution to lumpy distribution, while the staining of S-100 protein was gradually distributed layer-by-layer, and some area showed tumor-like distribution. The results show that the skin model of melanoma can be in vitro constructed successfully by mixed culture of MV3 melanoma cells and HaCaT cells on de-epidermized dermis. Figures and Tables | Related Articles | Metrics

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Establishing a rat osteoarthritis model based on knee joint instability Zhang Rong-Kai, Yang Lu-kun, Ye Zhi-qiang, Wu Zhao-ya, Zhao Qing, Cai Dao-zhang 2013, 17 (37):  6628-6635.  doi: 10.3969/j.issn.2095-4344.2013.37.014 Abstract ( 540 )   PDF (2621KB) ( 689 )   Save BACKGROUND: Osteoarthritis is a kind of complex multi-cause degenerative joint diseases. It is important to prevent and treat osteoarthritis in the early stage. But it is difficult to obtain the sufficient amount of human bone specimens with early osteoarthritis which are suitable for the research. OBJECTIVE: To observe the pathological changes of articular cartilage in the rat osteoarthritis model established with joint instability. METHODS: The 10-week-old male Sprague Dawley rats were randomly divided into two groups: the experimental group and the control group. Rats in the experimental group underwent open surgery, involving both medial meniscectomy and medial collateral ligament transection. The rats in the control group only received capsular incision. The rats were sacrificed at 1, 2, 4, 6 and 8 weeks after operation to obtain the right knee joint specimens, and then histopathological examination was performed to analyze the changes of rat osteoarthritis disease. RESULTS AND CONCLUSION: The mild erosion, ulcers wear, severe wear, osteophyte formation and other pathological changes were observed during articular cartilage degeneration in the experimental group at 2, 4, 6 and 8 weeks after operation. The articular cartilage score at 2, 4, 6 and 8 weeks after operation in the experimental group was significantly higher than that in the control group (P < 0.05). The results showed that medial meniscectomy and medial collateral ligament transection can effectively establish the rat knee osteoarthritis model, and the pathological and morphological changes at 4 weeks after modeling are similar to the performance of early human knee osteoarthritis, so the model is the ideal model to research the early osteoarthritis. Figures and Tables | Related Articles | Metrics

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Eye acupuncture therapy affects intercellular adhesion molecule 1 expression in rat hippocampus of acute cerebral ischemia-reperfusion injury Pan Qian, Wang Zhe, Gao Yuan, Yu Dan, Wang Ying, Jing Huan, Wang Shou-yan, Wang Jian, Guan Hong-quan 2013, 17 (37):  6636-6640.  doi: 10.3969/j.issn.2095-4344.2013.37.015 Abstract ( 310 )   PDF (1551KB) ( 425 )   Save BACKGROUND: Significant increasing of intercellular adhesion molecule 1 expression can promote the occurrence of inflammatory response and increase brain tissue injury. OBJECTIVE: To investigate the effect of eye acupuncture point and non-point therapy on intercellular adhesion molecule 1 expression in rat hippocampus of acute cerebral ischemia-reperfusion injury model. METHODS: Forty Sprague Dawley rats were divided into normal group, sham-operation group, model group, eye acupuncture point group and eye acupuncture non-point group. Rats in the model group, eye acupuncture point group and eye acupuncture non-point group were used to establish the acute cerebral ischemia-reperfusion injury model with suture method, the suture was deep for 1.8-2.2 cm. In the eye acupuncture point group, the acupuncture points of hepatic region, upper energizer area, lower energizer area and kidney area were selected when cerebral ischemia-reperfusion occurred immediately and 30 minutes before drawing materials, then horizontal needling the acupuncture points around the orbit for 20 minutes. In the eye acupuncture non-point group, the puncture site was selected at 3 mm away from eye acupuncture non-point area, and then needled the puncture site with the same method in the eye acupuncture point group. RESULTS AND CONCLUSION: After eye acupuncture point therapy, the neurological deficit scores were decreased, and the intercellular adhesion molecule 1 expression in rat hippocampus was significantly decreased (P < 0.01). After eye acupuncture non-point therapy, there were no significant changes in neurological deficit scores and the expression of intercellular adhesion molecule 1 protein and mRNA in rat hippocampus. The results indicate that eye acupuncture can significantly improve the rat cerebral ischemia-reperfusion injury, and the mechanism may relate with reducing the intercellular adhesion molecule 1 expression in rat hippocampus. Figures and Tables | Related Articles | Metrics

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Construction and expression of eukaryotic expression vector pcDNA3-ICOSIg Zhao Guo-hua, Zhang Rui, Xu Guo-yan, Wang Dong-mei 2013, 17 (37):  6641-6644.  doi: 10.3969/j.issn.2095-4344.2013.37.016 Abstract ( 364 )   PDF (1164KB) ( 414 )   Save BACKGROUND: So far, inducible co-stimulator is the important costimulatory molecule family member. Inducible co-stimulator can promote the activation of T cells proliferation and secretion, regulate Th1/Th2 cell polarization dependence, enhance B cell function which depend on the T cells. So, blocking the inducible co-stimulator may result the inactivation and no reaction of cloning in T cells, thus inducing the immune escape of tumor on the body. OBJECTIVE: To build a plasmid expression of inducible co-stimulator Ig, in order to observe the expression in rat body.  METHODS: cDNA encoding the extracellular domain of human inducible co-stimulator was prepared. The encode of the domain was fused with the gene of immunoglobulin IgG constant fragment (Ig) of encoding mouse, in order to build the inducible co-stimulator Ig fusion gene and the secreted eukaryotic expression vector pcDNA3-inducible co-stimulator Ig. Enzyme digestion of the recombinant and sequencing was performed, and then the positive liposome coated pcDNA3-inducible co-stimulator Ig was transferred into the muscle tissue of mouse right thigh. Western blot was used to detect the level of inducible co-stimulator Ig. RESULTS AND CONCLUSION: The sequencing confirmed that the size of target gene fragment pcDNA3- inducible co-stimulator Ig plasmid was exactly the same with the sequence of inducible co-stimulator published on Genebank, which indicated the successful of plasmid construction. After transferred into the mouse for 7 days, the liposome coated pcDNA3-inducible co-stimulator Ig was positively expressed in the mice serum, which showed that pcDNA3-inducible co-stimulator Ig could be expressed in the rat muscle cells. The results suggest that gene synthesis and recombinant technology can successfully construct the eukaryotic expression vector pcDNA3-inducible co-stimulator Ig. Figures and Tables | Related Articles | Metrics

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Construction and identification of eukaryotic co-expression vector carrying Myod1 and Myog Gao Hong-fei, Liang Bing-sheng, Shuang Wei-bing 2013, 17 (37):  6645-6651.  doi: 10.3969/j.issn.2095-4344.2013.37.017 Abstract ( 318 )   PDF (597KB) ( 477 )   Save BACKGROUND: Now it has cooperation and facilitative effete between myogenic regulatory factors through a long time study. So, gene therapy of double genes of Myod1 and Myog can obtain better effect, and can provide a new way for preventing denervated skeletal muscle atrophy. OBJECTIVE: To construct eukaryotic co-expression vector carrying Myod1 and Myog genes. METHODS: Full-length Myod1 gene and Myog gene cDNA were amplified by reverse transcription PCR, and then inserted into pVAX1 vector after digested to establish the recombined Myod1 and Myog eukaryotic co-expression vector pVAX1-Myod1-IRES2-Myog-IRES2-EGFP, and then identified with gene sequencing. The in vitro cultured 3T3 cells were transfected with pVAX1-Myod1-IRES2-Myog-IRES2-EGFP, and the expressions of Myod1 and Myog genes in the 3T3 cells were detected with western blot assay in order to identify whether the 3T3 cells could express the target protein correctly. RESULTS AND CONCLUSION: The sequencing results showed that the sequence length and base sequence of Myod1 and Myog cDNA in eukaryotic co-expression vector pVAX1-Myod1-IRES2-Myog-IRES2-EGFP were identical with the reported sequences. Myod1 and Myog protein band expressions were detected in 3T3 cells by western blot after transient transfection. The pVAX1-Myod1-IRES2-Myog-IRES2-EGFP, a eukaryotic co-expression vector of Myod1 gene and Myog gene is successfully constructed. Figures and Tables | Related Articles | Metrics

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Clinical applications of allogeneic osteochondral grafts from osteochondral tissue bank  Zhao Jian-li, Qi Jian-hong, Zhang Yan-ming, Yin Qi-pu 2013, 17 (37):  6652-6656.  doi: 10.3969/j.issn.2095-4344.2013.37.018 Abstract ( 350 )   PDF (614KB) ( 441 )   Save BACKGROUND: Full-thickness articular cartilage injury is notoriously difficult to be treated in the fields of orthopedics and sports medicine. Allogeneic bone and cartilage transplantation can offer a transparent cartilage with biological activity and biomechanical properties to repair full-thickness articular cartilage defects. Allogeneic osteochondral grafts from osteochondral tissue bank are adequate, and have a good prospect in the treatment of articular cartilage defects. OBJECTIVE: To summarize the drawn materials, preservation, quality control and clinical monitoring of allogeneic osteochondral grafts supported by the osteochondral tissue bank. METHODS: The first author searched PubMed and CNKI for the relevant articles published before 2013 using the key words of “tissue bank, knee, articular cartilage, preservation, transplantation” in English and Chinese, respectively. After retrieval, we summarized the drawn materials, preservation, quality control and clinical monitoring of allogeneic osteochondral grafts supported by the osteochondral tissue bank. RESULTS AND CONCLUSION: Nineteen of 194 retrieved articles were enrolled according to inclusive and exclusive criteria. The results show that allogeneic bone and cartilage transplantation is an effective method for the treatment of articular cartilage defects, and the establishment of the osteochondral tissue bank can provide safe and active tissues for the treatment of articular cartilage defects. Now, the osteochondral tissue bank is still in the initial stage. Figures and Tables | Related Articles | Metrics

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Exercise in the treatment of osteoporosis Wang Chun-yan, He Cheng-qi 2013, 17 (37):  6657-6663.  doi: 10.3969/j.issn.2095-4344.2013.37.019 Abstract ( 322 )   PDF (683KB) ( 580 )   Save BACKGROUND: Osteoporosis is a condition resulting in an increased risk of skeletal fractures due to a reduction in bone mass and degradation in bone microstructure. Treatment of osteoporosis usually involves the application of pharmacological agents as the first choice. Pharmacotherapy alone is not sufficient for the comprehensive management of osteoporosis, as pharmacotherapy has no effect on increasing muscle strength, improving balance ability, and preventing falls. Exercise, considered as an important non-pharmacologic therapy, plays a considerable role in the prevention of osteoporosis. The importance of therapeutic exercises in the treatment of osteoporosis has been recognized gradually. OBJECTIVE: To discuss the effect of exercise in the treatment of osteoporosis combined with some of the latest research literatures. METHODS: Clinical reports and mechanism researches about effects of different types of exercise on bone strength especially the treatment of osteoporosis were retrieved from CNKI database and Medline database with the key words of “osteoporosis, bone mineral density, bone strength, treatment, exercise, stress, tai chi, muscle, bone architecture” in both Chinese and English from January 2001 to February 2013. The repetitive researches and atypical reports were eliminated. RESULTS AND CONCLUSLON: Therapeutic exercises including aerobic exercise, resistance, impact movement, and vibration, which can safely increase bone strength and muscle strength, improve equilibrium function, and prevent falls and fractures. For spinal deformity patients, appropriate orthotics can improve the security and promote exercise therapy. As with drug therapy, therapeutic exercises are also individualized. Exercise program should be selected under the insurance of good compliance and security. Exercises that involve high strain rates seem to be more effective than others. However, all these benefits are comparatively small, and should be maintained by continuous exercises. Figures and Tables | Related Articles | Metrics

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Immune factors in spinal cord injury Xie Zhou-tong, Xu Hao, Chen Jian-mei, Liu Jian-jian 2013, 17 (37):  6664-6670.  doi: 10.3969/j.issn.2095-4344.2013.37.020 Abstract ( 466 )   PDF (611KB) ( 510 )   Save BACKGROUND: The glial scar is a major obstacle to the regeneration of the central nervous system, which plays a physical and chemical barrier role to central nervous system regeneration. How to effectively inhibit glial scar formation and promote axonal regeneration has great significance. OBJECTIVE: To explore the effect of the immune response in the formation of the glial scar after spinal cord injury. METHODS: A computer-based online search was performed in the PubMed database and the CNKI database for the articles on the mechanisms of spinal cord injury, immune cells and spinal cord injury as well as the glial scar formation after spinal cord injury from January 1990 to January 2013. The key words were “spinal cord injury, immunity, glia scar” in Chinese and English. The languages of the articles were limited in Chinese and English. Finally, 72 articles were included for the further analysis according the inclusion criteria. RESULTS AND CONCLUSION: After spinal cord injury, the effect of autoimmune response in which still been much debated. Studies showed that spinal cord injury can cause suppression of cellular immunity and non-specific immune function, causing immune function decreasing. Monitoring immune indicators and improving immune function have great significance in promoting the full rehabilitation of patients with spinal cord injury. Glial scar is the most important factor to prevent the repairing after spinal cord injury. Immune and inflammatory response is the main reason to secondary spinal cord injury. Degree of repair after spinal cord injury mainly depends on the protection of the residual neurons in order to keep them off secondary damage. However, autoimmune T cells can protect the residual neurons, the specific effect of T lymphocytes in glial scar formation after spinal cord injury is still unclear, and further exploration is needed. Figures and Tables | Related Articles | Metrics

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Expression of cytokines in the degenerative cartilages and synovial tissues of osteoarthritis patients Zhang Yan-hui, Gao Chun-yang, Li Shao-hua 2013, 17 (37):  6671-6675.  doi: 10.3969/j.issn.2095-4344.2013.37.021 Abstract ( 320 )   PDF (598KB) ( 595 )   Save BACKGROUND: The inflammation of osteoarthritis is mediated by chondrocytes and the synovial tissue-secreted cytokines. Articular cartilage and synovial tissue contains a variety of cytokines, which play important regulatory role in the repair of articular cartilage injury. OBJECTIVE: To analyze the relationship between chondrocytes, synovial tissue-secreted cytokines and osteoarthritis, as well as the effect of chondrocytes and synovial tissue-secreted cytokines on osteoarthritis. METHODS: A computer-based search was conducted in Wanfang database (www.wanfangdata.com.cn), PubMed database (www.ncbi.nlm.nih.guv/pubmed) by the first author from 2005 to 2010. The key words were “osteoarthritis, degeneration, cartilage tissue, cytokines”. A total of 146 articles were obtained through computer search, then reading the titles and abstracts for initial screen, 86 articles with the unrelated research purposes were eliminated, 40 articles with the repetitive content were excluded, and finally 21 articles related to the effect of various cytokines  in the degenerative cartilage and synovial tissue of the osteoarthritis patients were included for further analysis. RESULTS AND CONCLUSION: The cytokines mainly refers to the activated immune cells and some stromal cell-secreted non-specific regulate immune responses and inflammatory responses-mediated small molecule proteins, including lymphokines produced by the lymphocytes, as well as the single nuclear factor and other cytokines produced by monocytes macrophages. Synovial cell-secreted cytokines may partially explain the pathological process of osteoarthritis and play an important role in joint inflammation. Although, more and more scholars have pay attention to the effect of synovial cells and cartilage cell-secreted cytokines, but mainly studies the effect of exogenous cytokines on chondrocytes or synovial cells, and the effect of endogenous cytokines in the pathogenesis of osteoarthritis has not been extensively studied. Figures and Tables | Related Articles | Metrics

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Model of steroid-induced avascular necrosis of the femoral head: Analysis of different construction techniques Li Rui-qi, Zhang Guo-ping, Li Yi-jiong, Li Ya-li, Ren Li-zhong, Zhang Yu-chen, Wang Wei, Gao Hong-yang, Lü Ya-jun 2013, 17 (37):  6676-6681.  doi: 10.3969/j.issn.2095-4344.2013.37.022 Abstract ( 425 )   PDF (1004KB) ( 541 )   Save BACKGROUND: To construct a normal animal model of femoral head necrosis contributes to the research of the pathogenesis of femoral head necrosis, which can provide theoretical basis for the prevention and treatment of femoral head necrosis. OBJECTIVE: To research the experimental effect of lipopolysaccharide combined with dexamethasone injection in the induction of rabbit femoral head necrosis. METHODS: Thirty-six New Zealand white rabbits were randomly divided into model group (n=21) and control group (n=15). The rabbits in the model group were injected with 10 μg/kg lipopolysaccharide daily and continuous for 2 days, and then injected with 25 mg/kg dexamethasone daily for 3 days continuously. The rabbits in the control group were injected with the normal saline at the same volume. RESULTS AND CONCLUSION: After 4 weeks, the X-ray film of the rabbit in the model group showed the joint gaps were widened, the density was increased, the articular subchondral bone mineral density was increased, the femoral head was flat, trabecular bone was fuzzy, the boundaries between subchondral bone and cancellous bone was unclear, and the patchy high-density areas were observed in the femoral head with shortened femoral neck. The bone mineral density of partial femoral head was measured with dual-energy X-ray absorptiometry, and found that the bone mineral density of femoral head and the bone mineral content of the model group were significantly lower than those of the control group (P < 0.05). Histological section observation showed that the bone cell lacuna was empty and shallow, fat cells were increased and vascular thrombosis was observed, meanwhile, the osteonecrosis rate and lacunae rate of the survival animals were significantly higher than those in the control group. Dexamethasone combined with lipopolysaccharide can effectively construct the model of steroid-induced avascular necrosis of the femoral head. Figures and Tables | Related Articles | Metrics

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Expression of minichromosome maintenance 3 from the peripheral blood of fluorosis patients and the liver and renal function Zhang Ya-lou, Zhao Yang, Tang Li, Wu Qi-qing, Bai Sheng-bin, Zhong Jin-jie 2013, 17 (37):  6682-6688.  doi: 10.3969/j.issn.2095-4344.2013.37.023 Abstract ( 368 )   PDF (891KB) ( 429 )   Save BACKGROUND: Previous studies have shown that minichromosome maintenance 3 is related with fluorosis, but the expression of minichromosome maintenance 3 in fluorosis patients is not clear yet. OBJECTIVE: To analyze the mRNA expression level of minichromosome maintenance 3 in peripheral blood from patients exposed to fluoride and normal controls. METHODS: Eleven patients with mild fluorosis by drinking water (exposure group) and 11 cases of control (non-exposure group) were selected for research. SYBRGreen1 real-time quantitative PCR was used to determine the mRNA expression of minichromosome maintenance 3 in peripheral blood mononuclear cells, and the liver and renal function indicators were detected. RESULTS AND CONCLUSION: mRNA expressions of minichromosome maintenance 3 in the exposure group and non-exposure group were (0.573 60±0.102 59) and (0.550 0±0.171 81), respectively, and there was no significant difference between two groups (P > 0.05). There were no significant differences in the liver and renal function indicators between two groups. The results indicate that mild fluorosis has no significant effect on mRNA expression of minichromosome maintenance 3 in the peripheral blood mononuclear cells. More indicators are needed to compressively analyze the effect of fluoride on the liver and renal functions. Figures and Tables | Related Articles | Metrics

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Body shape and structure characteristics of national women’s handball team athletes Chen Hao, He Jiang-chuan 2013, 17 (37):  6689-6694.  doi: 10.3969/j.issn.2095-4344.2013.37.024 Abstract ( 538 )   PDF (652KB) ( 341 )   Save BACKGROUND: Studies have shown that the physical development potential is often associates with certain combinations of hand print features. OBJECTIVE: To analyze the body shape and structure characteristics of national women’s handball team athletes. METHODS: Fifty national women’s handball team athletes were collected from Shanghai, Beijing, Tianjin and Anhui provinces and PLA. The body shapes of the athletes were measured by the Research Institute of National Sports Council, and the hand prints were collected. The common factors of body shape and structure characteristics were analyzed with factor analysis method, including height, weight, forearm length, wrist circumference, hand length, hand width and index finger length and large-handedness index.  RESULTS AND CONCLUSION: The body shape and structure characteristics of national women’s handball team athletes could be divided into four types: long trunk lean type indicator, hand skeleton and muscle fiber development level index, flexible quality indicators as well as the alertness and mental level indicators. The long trunk lean type indicator was the main factor to affect body shape and structure characteristics of national women’s handball team athletes. Figures and Tables | Related Articles | Metrics

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Association between osteoprotegerin gene G1181C polymorphism and bone mineral density in postmenopausal women: A Meta-analysis Zhang Hao, Gan Lu, Guo Yan, Wang Rui-heng 2013, 17 (37):  6695-6700.  doi: 10.3969/j.issn.2095-4344.2013.37.025 Abstract ( 326 )   PDF (646KB) ( 311 )   Save BACKGROUND: Genetic studies have shown that there may be association between osteoprotegerin gene G1181C polymorphism and bone mineral density in postmenopausal women, but there are some differences in the conclusions. OBJECTIVE: To study the association between osteoprotegerin gene G1181C polymorphism and bone mineral density in postmenopausal women through Meta-analysis. METHODS: The PubMed database, Embase database, CNKI database and Wanfang database were searched for the observational studies about the association between osteoprotegerin gene G1181C polymorphism and bone mineral density in postmenopausal women. The standardized mean difference was considered as the effect indicator and calculated with random or fixed effect models according to the significance of heterogeneity. RESULTS AND CONCLUSION: A total of 9 articles were included with a total of 8 799 and 9 365 participants for which vertebral and spine bone mineral density had been measured, respectively. The analysis results showed that the bone mineral density of the postmenopausal women with the G allele of G1181C was significantly lower than that of the postmenopausal women with the C allele. Similar results were obtained in Caucasian population subgroup analysis. The Caucasian population subgroup analysis showed that the bone mineral density of the postmenopausal women with GG genotype of G1181C was lower than that of the postmenopausal women with GC and CC genotype. The results indicate that there is association between osteoprotegerin gene G1181C polymorphism and bone mineral density in postmenopausal women, and G allele is a risk factor for lower bone mineral density. Figures and Tables | Related Articles | Metrics